SDRC Members and Their Research Descriptions

Kevin D. Cooper, M.D.
Professor, Chairman of Dermatology
Principal Investigator
SDRC Director

Dr. Cooper is an accomplished investigator in the areas of immunodermatology, photobiology and pathophysiology of psoriasis. One focus of his laboratory is to identify and characterize cutaneous antigen presenting cell (APC) subsets in the epidermis and dermis of normal and inflamed skin. The emphasis is on mechanisms of UV injury and how UV creates an altered milieu in the skin which contributes to photodamage, microbial tolerance, and tumor promotion. In particular, these studies center on how different APC's, such as macrophages and Langerhans cells, achieve distinct states of activation and differentiation in the skin and whether these APC's differentially influence T lymphocyte function. These studies use a combination of human and nurine approaches with classical cellular immunological assays, immunostaining techniques, advanced multicolor flow cytometry utilizing up to six simultaneously acquired parameters, and cytokine gene expression. The large repertoire of cell biological and immunologic techniques both utilizes and enriches each of the cores. Results are revealing exciting new concpets regarding how innate immune mechanisms of the skin, such as Complement C3 proteolysis, can modify cutaneous APC precursors and the T cell mediated acquired immune response.

A second major area of investigation is psoriasis immunopathogenesis and the interactions between the immune system (T cells and macrophage), the extracellular basement membrane zone matrix (EDA fibronectin) and basal keratinocyte progession regulation. The creation of EDA+ and - vectors for inducible-expressing cell lines used the Cell and Molecular Technology Core, the use of
Laser Capture Microscopy to localize EDA Fn expression to DEJ-lining macrophages was the first skin application of this Morphology Core technology at CWRU, and the project is absolutely dependent upon tissue from the Translational Research Core. Dr. Cooper's expertise in lymphomas, dermatitis, and clinical immunology enriches many interactions in the SDRC.

Two recent representative publications:

1. Ting KM, Rothaupt D, McCormick TS, Hammerberg C, Chen G, Gilliam AC, Stevens S,
Culp L, Cooper KD. Overexpression of the oncofetal Fn variant containing the EDA splice-in segment in the dermal-epidermal junction of psoriatic uninvolved skin. J Invest Dermatol. 114:706-711, 2000.

2. Hammerberg C, Katiyar SK, Carroll MC, Cooper KD. Activated complement component 3 (C3) is required for ultraviolet inductio immunosuppression and antigenic tolerance. J Exp Med. 187:1138-1142, 1998.

Hasan Mukhtar, Ph.D.
Professor of Dermatology, Environmental Health Sciences, Physiology
and Biophysics, Radiation Oncology, and General Medical Sciences (Oncology)
Previous Pilot and Feasibility Study Recipient
SDRC Co-Director
Director, Animal Experimentation Core

Dr. Mukhtar is an accomplished investigator working in multidisciplinary areas of cutaneous biology related to skin pharmacology/carcinogenesis and photodynamic therapy. There are three major areas of investigation in his laboratory. The first area of research is related to defining the role of cutaneous cytochrome P450 and other enzymatic pathways in biotransformation of endogenous and exogenous chemicals to which skin comes in contact. He has shown, using molecular and biochemical approaches, that multiple inducible forms of cytochrome P450 exist in murine and human skin. Two members of this multi-gene family of enzymes known as CYP1A1 and CYP1B1 are predominantly expressed and induced in cutaneous tissue following exposure to chemical carcinogens and ultraviolet B radiation. His studies showing susceptibility to biological effects of skin carcinogenesis to genes of major histocompatibility complex suggest that cell-mediated immunity to chemical carcinogens serves to protect individuals by removing mutant cells before they can evolve into clinically apparent tumors.

Dr. Mukhtar's second area of research focus is on skin cancer prevention. His studies have shown that the polyphenolic fraction isolated from green tea acts as an anti-carcinogen and provides protection against all stages of multistage cutaneous carcinogenesis processes known as tumor initiation, tumor promotion, and tumor progression. These findings have received considerable national and international attention. Efforts in his laboratory are directed in defining cellular, biomechanical and molecular mechanisms of this broad anti-carcinogenic effect. His studies showed that epigallocatechin-3-gallate, the major polyphenol in green tea, results in induction of apoptosis and cell cycle arrest and inhibits activation of transcription factor NFKB in human carcinoma cells. His recent studies demonstrating that skin application of green tea polyphenol to human skin results in protection against solar erythema and the associated cellular and molecular effects raises the possibility that green tea may prove useful against the adverse effect of ultraviolet radiation in the human population.

The third area of research in Dr. Mukhtar's laboratory relates to Photodynamic Therapy (PDT) of cancer. PDT shows promise as a treatment modality for various dermatological disorders including psoriasis, alopecia areata, non-melanoma superficial skin cancer and patch and plague stage cutaneous T cell lymphoma. Dr. Mukhtar is studying the usefulness of phthalocyanine photosensitizers in the treatment of skin tumors. Using chemically and ultraviolet radiation-induced mouse skin tumors, he is evaluating the role of apoptosis in PDT-induced regression of skin tumors. His studies have shown that apoptosis occurs in vivo during PDT-induced shrinkage of mouse skin tumors. Cutaneous photosensitivity is an undesirable effect that accompanies photosensitizer based PDT. He is also comparing the mechanisms by which PDT causes tumor regression with those that cause cutaneous photosensitivity. If the metabolic pathways are distinctly different for tumor cell death vs. skin photosensitivity, then it may be possible to design PDT protocols which limit the major adverse reaction of PDT (i.e. cutaneous photosensitivity) without affecting the efficacy of PDT regression of tumors.

Two recent representative publications:

1. Ahmad N, Feyes DK, Agarwal R and Mukhtar H: Photodynamic therapy results in induction of WAF1/C1P1/P21 leading to cell cycle arrest and aptosis. Proc Natl Acad Sci USA 95:6977-6982, 1998.

2. Katiyar SK, Matsui MS and Mukhtar H: Ultraviolet-B exposure to human skin induces cytochromes P450 CYP 1A1 and CYP 1B1. J Invest Dermatol 114:328-333, 2000.

Nihal Ahmad, Ph.D.
Assistant Professor
Department of Dermatology
Principal Investigator, Pilot & Feasibility Project in this application

Dr. Ahmad is a recently recruited Assistant Professor in the Department of Dermatology. After Post-doctoral training in the laboratory of Dr. Mukhtar in 1998, Dr. Ahmad was appointed as an Instructor. His area of research focused on the molecular mechanism(s) involved in signaling pathways, cell cycle regulation and apoptosis in photocarcinogenesis and during photodynamic therapy of skin cancer. Accordingly, he is directing his efforts in elucidating molecular mechanism(s) of photocarcinogenesis. Effective July 1998, he was successfully able to obtain a Research Career Development Award (RCDA) from the Dermatology Foundation. This award was renewed for three consecutive years and is effective through June 2001. He has used this unique opportunity to define the role of signaling pathways in UV responses that may have relevance to skin cancer. As principal investigator, Dr. Ahmad obtained grants from the Ohio Cancer Research Associates (effective from July 1, 1999, for two years), where he has been investigating cancer preventive effects of a compound resveretrol found in grape skin and from Cancer Research Foundation of America (Jan 15, 2000-Jan 14, 2001; renewable) where he is defining the involvement of matrix metalloproteases, urokinase and angiogenesis as a mechanism of anti inflammatory effects of antioxidants. His applications to the NIH and the Department of Defense are pending.

Two recent representative publications:

1. Ahmad N, Gupta S and Mukhtar H: Involvement of retinoblastoma (Rb) and E2F transcription factors during photodynamic therapy of human epidermoid carcinoma cells A431. Oncogene 18:1891-1896, 1999.

2. Ahmad N, Gupta S and Mukhtar H: Green tea polyphenol epigallocatechin-3-gallate (EGCG) differentially modulates nuclear factor kappa B (NF-kB) in cancer cells vs. normal cells. Arch Biochem Biophys 376:338-346, 2000.

Melvin Berger, M.D., Ph.D.
Professor and Chief, Pediatric Immunology
Member, Immunology Research Area

The long-term interests of Dr. Berger's laboratory are on expression of complement receptors on neutrophils, and neutrophil activation. The function of complement degradation products and their receptors in regulating inflammation, in particular C3 and CR1, are areas of particular expertise.

In recent years his laboratory has turned attention to studying the role of epithelial cells in regulating inflammation. This work grew out of Dr. Berger's studies of the inflammatory response in the lung in cystic fibrosis, in which IL-10 was greatly reduced or absent from the lavage fluid of most CF patients. Further studies showed that the bronchial epithelial cells themselves are the major source of this immunoregulatory cytokine in the healthy lung. As bronchial epithelial IL-10 downregulated IL-8, and IL-8 is believed to be the major chemo attractant responsible for the excessive neutrophil influx that characterizes CF lung disease, these data thus suggest that the epithelium itself plays a major role in determining and regulating the local inflammatory and immunologic milieu. Studies in a mouse model of the chronic Pseudomonas aeruginosa infection in CF show that IL-10 knockout mice have a greater neutrophilic response, more severe inflammation and lung damage than their normal counterparts; while IL-10 treatment of normal mice ameliorates the local inflammation as well as the systemic effects of this infection, without exacerbating the burden of bacteria or causing other infectious complications. Dr. Berger is presently principal investigator of a multi-center clinical trial of IL-10 in CF patients, as well as pursuing investigations aimed at understanding the relationship between the CF basic defect and diminished IL-10 production. These studies are highly relevant to epidermal responses that occur during UV exposure and in many skin diseases, and Dr. Berger is an active member, along with a number of the SDRC members, of the epithelial inflammaation group at CWRU.

A new application for Dr. Berger's work became evident upon interactions with Drs. Kefei Kang and Kevin Cooper, who were also interested in Complement receptor-mediated regulation of inflammation and IL-10. This collaboration, supported by the SDRC cores, and which revealed that CR3, through iC3b, which is critical for both UV-induced immunosuppression and epidermal photodamage, triggers IL-10 production and inhibits IL-12 production. This work nicely integrates intersecting expertise within the SDRC and reveals fundamental processes relevant to both UV-exposed skin inflammation and bronchial epithelium.

Two recent representative publications:

1. Yoshida Y, Kang K, Berger M, Chen G, Gilliam AC, Moser A, Wu L, Hammerberg C, Cooper K: Monocyte induction of IL-10 and down-regulation of IL-12 by iC3b deposited in Ultraviolet-exposed human skin. J Immunol 161:5873-5879, 1998.

2. Jost C, Klickstein L, Wetzler E, Kumar A and Berger M: Intracellular storage and regulated plasma membrane expression of human complement receptor Type 1 in rat basophil leukemia cell transfectants. Blood 92:300-309, 1999.

W. Henry Boom, M.D.
Professor, Department of Medicine
Director, Tuberculosis Research Unit
Vice Chair for Research, Department of Medicine

Dr. Boom is Vice Chair for Research in the Department of Medicine and is the Director of the Tuberculosis Research Unit (TBU), which coordinates the research of a group interested in microbiology modification of macrophages. A great deal of intellectual and technical exchaange occurs between the TBU investigators, the SDRC Immunology/ Microgiology Thematic Group, and the Center for Aids Research.

Dr. Boom has examined the mechanisms by which M. Tb organisms modify macrophages to inhibit their class II MHC antigen processing for T cell recognition, as well as modify macrophage cytokine production, in particular IL-10, IL-12, and TGF beta. The resultant observations of modification of T cell activation and killing mechanisms are highly analogous to the UV systems under study by Drs. Cooper and Stevens, to the Candida-monocyte interactions under study by Drs. Ghannoum and Kang, and to the fibrosis model (TGF beta dependent).

Two recent reprsentative publications:

1. Fulton SA, Cross JV, Toossi ZT, Boom WH. Partial regulation of IL-12 by IL-10, TGF-b and IFN-g in monocytes infected with M. tuberculosis. J. Infectious Diseases, 178:105-14, 1998.

2. Noss EH, Harding CV, Boom WH. M. tuberculosis inhibits class II MHC antigen processing in murine bone marrow macrophages. Cellular Immunology, in press, 2000.

Susann Brady-Kalnay, Ph.D.
Assistant Professor of Molecular Biology & Microbiology
Previous Pilot and Feasibility Study Recipient

Dr. Brady-Kalnay is investigating the role of Receptor Protein Tyrosine Phosphatases (RPTPs) in signals transduced upon cell-cell contact. She is studying the role of PTPm in contact-mediated signaling as a model for how RPTPs function at a cellular level. Previously, she demonstrated that PTPm mediates aggregation by binding homophilically, i.e. PTPm on the surface of one cell binds to PTPm on an apposing cell. Dr. Brady-Kalnay then went on to determine that the homophilic binding site resides in the immunoglobulin. Interestingly, PTPm associates with another family of cell adhesion molecules called cadherins molecules, which are abundant in skin. Cadherins are cell adhesion molecules that play a critical role in cytoskeletal organization leading to cell junction formation and epithelial polarization. There is evidence to suggest that tyrosine phosphorylation of the cadherins, commonly observed in transformed cells, leads to loss of adhesive function and breakdown of cell junctions. Presumably these proteins are normally maintained in a dephosphorylated state, through the action of a phosphatase, which is crucial for the stabilization of adhesion. Her results suggest that PTPm may regulate the tyrosine phosphorylation state of cadherins and thus their adhesion. Dr. Brady-Kalnay is working to determine how cadherins and PTPm generate signals in response to cell adhesion and how this changes during epithelial transformation and epidermal differentiation, in collaboration with Dr. Eckert.

A second focus of the research in Dr. Brady-Kalnay's laboratory is to investigate the role of PTPs in regulation of desmosome structure and function. While significant progress has been made in the identification of desmosomal components, there is little known about junctional assembly and/or signal transduction at these sites. Tyrosine phosphorylation may be involved in the regulation of these junctions, and stimulation of tyrosine phosphorylation induces a redistribution and of desmosomal components. Additionally, there are indications that tyrosine phosphorylation of two desmosomal components, plakoglobin and plakophilin, may be important for their regulation. Preliminary data indicates that a tyrosine phosphatase is present in purified desmosome preps. Dr. Brady-Kalnay will clone this PTP and investigate its role in regulating the tyrosine phosphorylation or function of the proteins in the desmosome. Understanding desmosome regulation will have implications in skin disease research due to the fundamental role of desmosomes in the structure and integrity of skin. Interestingly, many skin disease are caused by disruption of the integrity of desmosomal cellular junctions either by autoimmune antibody production or aberrant expression of mutated proteins. Dr. Brady-Kalnay plans to analyze whether there are changes in expression of this desmosomal PTP in various skin diseases, and whether such changes are involved in autoantibody-mediated or other dys-adhesive blistering skin diseases.

Two recent representative publications:

1. Brady-Kalnay S, Mourton T, Nixon JP, Pietz G, Kinch M, Chen H, Brackenbury R, Rimm DL, Del Vecchio RL and Tonks NK: Dynamic interaction of PTPm with multiple cadherins in vivo. J Cell Biol;141:287-296, 1998.

2. Burden-Gulley SM, Brady-Kalnay. PTPmu regulates N-cadherin-dependent neurite outgrowth. J Cell Biol;144:1326, 1999.

Henri Brunengraber, M.D., Ph.D.
Chairman and Professor, Department of Nutrition
Member, Photocarcinogenesis/Photobiology/Photomedicine Research Area

Dr. Brunengraber is internationally recognized for his pioneering work in utilizing nutritional components of intermediary metabolism to diagnose and modify organ function and dysfunction. He is chairman of the Department of Nutrition, which is ranked number two nationally in NIH funding, and he has recently turned his expertise nutritional modification of skin function. An outgrowth of this interest led the Department of Nutrition and Dermatology to submit a center grant to the NIH. This Center grant application contains three R01 applications headed by Drs. Cooper (Dermatology), Brunengraber (Nutrition), and Mukhtar (Dermatology). For diseases such as diabetes and specific inborn errors of nutrient metabolism, Dr. Brunengraber developed artificial nutrients for parenteral and enteral nutrition, i.e. 1,3-diol ketone body esters. He also defined conditions of applicability of the non-invasive chemical biopsy of the liver, using stable isotopes and isotopomer analysis.

Over the last eight years, Dr. Brunengraber has designed and synthesized pyruvate esters and thioesters which are potential agents for preventing and treating organ reperfusion injury, reactive oxygen species, thiol depletion, and accumulation of reducing equivalents. The new compounds are dipyruvyl-acetyl-glycerol (DPG), pyruvyl-acetyl-dihydroxyacetone (PADA), and pyruvate N-acetylcysteine ethyl ester (PNACE). PNACE restored 90% of function in perfused working rabbit hearts after 25 min warm ischemia, while control hearts did not recover. In live swine, DPAG and PNACE decreased myocardial infarct size after 1 hr of coronary artery occlusion followed by reperfusion. He hypothesizes that the beneficial effects of pyruvate result from (i) anaplerosis of citric acid cycle intermediates, and (ii) inhibition of fatty acid oxidation and free radical production.

Currently Dr. Brunengraber is collaborating with Drs. Cooper, Ahmad, Mukhtar and Stevens to examine the skin UV-protective effects of the pyruvate esters and thioesters and by green tea polyphenols and silymarin by indices of oxidative stress, erythema, skin immune function and isotopic investigations of intermediary metabolism. Because DPAG and PADA can be given to deliver a therapeutic concentration of pyruvate without lactic acidosis and sodium overload, these may represent a novel nutritional UV-protective strategy. Similarly through this collaboration, an understanding of mechanism which antioxidants prevent UV responses in the skin will come. Exciting preliminary data reveals a marked inhibition of the UV-induced erythema response by pyruvate esters and green tea polyphenols in human skin. The Translational Research Core has been essential to these exciting new applications.

Two recent representative publications:

1. Previs, S.F., Hallowell, P.H., David, F., and Brunengraber, H. Limitations of the mass isotopomer distribution analysis of glucose to study gluconeogensis. Heterogeneity of glucose labeling in incubated hepatocytes. J. Biol. Chem. 273:16859, 1998.

2. Verhoeven, N.M., Schor, D.S.M., Previs, S.F., Brunengraber, H., Jakobs, C. Stable isotope studies of phytanic-oxidation: in vivo production of formic acid. Eur. J. Pediatrics 156: S87-83, (1997).

Arnold I. Caplan, Ph.D.
Professor of Biology
Previous Pilot and Feasibility Study Recipient
Member, Executive Committee

Dr. Arnold Caplan has a major interest in connective tissue research and specifically in characterizing changes associated with aging. The foundations of his research, which include formal collaborations with several other members of the SDRC, are in the premise that aging is a natural process and that many of the changes occur as a genetically programmed extension of normal development. Aging involves changes in cellular and extracellular components which decrease the organism's ability to function optimally. It is clear that such changes are reflective of a continuation of programmatic events coded in the organism's genome. Dr. Caplan's experimental approach to these issues is to provide detailed analyses of selected extracellular components of skin from early human development through adulthood into senescence. He and his collaborator are investigating the chemistry and biosynthesis of skin proteoglycans as multivalent organizational components in the extracellular matrix and at the cell surface with emphasis on dermal molecules. By focusing his efforts on the chemistry and biosynthesis of this specific family of complex macromolecules as a function of development and aging, he has provided new insights into aging process in skin.

Dr. Caplan's connective tissue research program in aging of both skin and skeletal tissues has brought into the SDRC a number of investigators who have provided expertise in physical chemistry, biochemistry, cellular and developmental biology and human genetics. Their investigative approach is focused on basic aspects of extracellular matrix components of human dermal fibroblasts (both cell lines and primary isolates) and whole skin from specimens which range in age from first trimester of embryology to 90 year old. Dr. Caplan is a major organizing force in pulling together an interest group in skin equivalents.

In addition, Dr. Caplan has served on the Executive Committee from the initiation of the SDRC. In that capacity, he has been instrumental in advising the SDRC Director.

Two recent representative publications:

1. Carrino DA, Sorrell JM, Caplan AI. Age-related changes in the proteoglycans of human skin. Arch Biochem Biophys; 373:91-101, 2000.

2. Sorrell JM, Carrino DA, Baber MA, Asselineau D, Caplan AI. A monoclonal antibody which recognizes a glycosaminoglycan epitope in b dermatan sulfate and chondroitin sulfate proteoglycans of human skin. Histochem J. 31:549-558, 1999.

Shukti Chakravarti, Ph. D.
Assistant Professor
Departments of Medicine and Genetics
Previous Pilot and Feasibility Study Recipient

Dr. Shukti Chakravarti's laboratory is interested in extracellular matrix (ECM) biology within the context of normal development and disease. The amorphous interstitial ECM hydrates tissues and maintains tissue architecture for optimal functioning. It also plays a key role in presenting nutrients and growth factors to cells and possibly in regulating immune responses. Her work uses a combination of genetic and biochemical approaches to understand the role of specific proteoglycans in these matrices.

Lumican, a prototypic member of the interstitial extracellular matrix, is present in the cornea, dermis, articular cartilage, cardiac and muscle connective tissues. It is believed to regulate collagen fibril structure needed for corneal transparency, as well as elastic and tensile strength of dermal and muscle connective tissues. She has recently cloned the gene for lumican and completed its chromosomal mapping in human and mouse. Lumican-deficient mice have been generated in her laboratory by targeted disruption of the gene to understand its functions in vivo. This work was supported by an SDRC Pilot and Feasibility project. The mice have a functionally compromised ECM that affects the cornea, skin and bone. The skin manifestations include extremely loose and fragile skin, dramatically reduced biomechanical properties and abnormally thick collagen fibrils viewed by transmission electron microscopy. These features resemble Ehlers-Danlos syndrome and the potential for using the mutant mouse as a model for human Ehlers-Danlos syndrome is currently being explored.

Two recent representative publications:

1. Chakravarti, S, Magnuson, T., Lass, J.H., LaMantia, C., Jepsen, K.J. and Carroll, H. Collagen fibril defects affecting skin and cornea in lumuican-deficient mice, J. Cell Biol. 141:1277-1286, 1998.

2. Dunlevy JR, Chakravarti S, Gyalzen P, Vergnes JP and Hassell JR.: Cloning of mouse keratocan cDNA and localization of the gene to distal chromosome 10. Mammalian Genome 9:316-319, 1998.

Lloyd A. Culp, Ph.D.
Professor of Molecular Biology and Microbiology
Professor of Oncology/General Medical Science
Previous Pilot and Feasibility Study Recipient

Dr. Culp's research interests interface nicely with those of the SDRC. His laboratory analyzes the mechanisms of extracellular matrix adhesion of human dermal fibroblasts (separate populations of papillary and reticular) and how these mechanisms are altered in vitro and in vivo during aging of human skin. In addition to "normal" populations of papillary and reticular dermal fibroblasts from individuals of different ages, full-thickness fibroblasts are analyzed from the NIA Repository, as well as several populations of dermal fibroblasts from Down's syndrome patients (with Down's as a possible model of premature aging).

Fibronectin is an adhesion-promoting glycoprotein abundant in dermal matrices for which these cell types have integrin receptors that transmit appropriate signals to initiate a wide variety of physiological responses. Fibronectin contains two alternatively-spliced (from pre-mRNA) type III homology repeats--EDa and EDb-- that can be enriched in the dermis under specific situations. In order to study the potential adhesion-promoting/modulating roles of these spliced homology repeats (90 amino acids each), his laboratory has subcloned each type III homology repeat from III7 up to III12, including EDa and EDb, in a wide variety of combinations. His was the first laboratory to do so. These minigenes have been expressed in bacteria, the recombinant proteins purified, and these proteins tested for adhesive and signaling activity. These spliced sequences do indeed promote adhesion of specific cell types, including human dermal fibroblasts. Furthermore, they induce signaling via tyrosine phosphorylation of FAK (Focal Adhesion Kinase) and pp130cas. The kinetics of these phosphorylation reactions are different from those on the type III repeat III10 containing the RGDS sequence recognized by several integrins. Therefore, the significance of these signaling mechanisms is now available for detailed analyses in human papillary and reticular populations, possible perturbation in Down's cells and during normal aging processes. These applications are actively being studied in psoriatic uninvolved skin basal stem cells in collaboration with Drs. McCormick and Cooper, where he is a collaborator on a funded NIH project, and in collaboration with Drs. Eckert and Caplan on a next-generation skin equivalent project.

Two recent representative publications:

1. Culp LA, Lin WC, Kleinman NR. Tagged tumor cells reveal regulatory steps during earliest stages of tumor progression and micrometastatis. Histol Histopathol;14:879-886, 1999.

2. Culp LA, Lin WC, Kleinman NR, Campero NM, Miller CJ, Holleran JL. Tumor progression, micrometastasis, and genetic instability tracked with histochemical marker genes. Prog Histochem Cytochem; 33: 329-348, 1998.

Richard L. Eckert, Ph.D.
Professor of Dermatology and Physiology and Biophysics, Biochemistry, Oncology
and Reproductive Biology
Director, Cell Culture and Molecular Technology Core
Member, Executive Committee
Previous Pilot and Feasibility Study Recipient

Dr. Richard Eckert is an internationally recognized leader in the area of gene regulation and structural protein function in keratinocytes. His laboratory studies (i) involucrin gene regulation, (ii) involucrin structure/function, (iii) MAPK signaling pathways in keratinocytes, (iv) structure and role of S100 proteins in signal transduction, (v) the role of human papillomavirus in the genesis of cancer, and (vi) the role of retinoids as regulators of keratinocyte differentiation. His laboratory has extensive experience with all forms of molecular technology (promoter function analysis, DNA/protein interaction, protein purification, production of recombinant proteins, transgenic mouse technology, gene transfer, antisense-knockout, etc.) and methods of culture for skin cells. He also has extensive experience in protein biochemistry methods (biochemical isolation, peptide sequencing, peptide synthesis, etc.) and has an active program in the biology of epidermal wound healing.

Notable recent achievements of the Eckert laboratory include (i) determination of the sequence of the human keratinocyte envelope precursor protein, involucrin, (ii) identification of a role for C/EBP, AP1, POU domain and Spl transcription factors as regulators of involucrin promoter activity, (iii) identification, using transgenic mice, of a single AP1 site as being required for expression of involucrin gene in epidermis, (iv) development and characterization of a transgenic mouse model for HPV-dependent skin cancer, (v) elucidation of a MAPK signaling cascade that is responsible for TPA-dependent activation of hINV promoter activity, (vi) identification of S100 proteins as transglutaminase substrates, signaling proteins and precursors of the keratinocyte cornified envelope, (vii) characterization of a new tumor suppressor protein that suppresses keratinocyte proliferation, and (viii) characterization of an efficient retinoid antagonist.

In the previous application, Dr. Eckert directed the Cell Culture Core and Co-Directed the Molecular Biology Core. He was supported by a P&F to generate transgenic models of skin cell function. This project was successful and is now supported as a five-year RO1 to study involucrin transgene expression. This project has recently been renewed for a second time. These studies have (i) characterized the human involucrin promoter and (ii) resulted in the application of this promoter tool to develop transgenic models of human papillomavirus-dependent human cancer. Dr. Eckert's cancer studies interface in a very direct way with those of Dr. Mukhtar, who studies chemical and photo-induced skin carcinogenesis. Some of the animal models developed in his laboratory are now being used in collaborative projects with Dr. Ellen Rorke and Dr. Mukhtar.

Two recent representative publications:

1. Agarwal C, Efimova T, Welter JF, Crish JF, Eckert RL. CCAAT/enhancer-binding proteins. A role in regulation of human involucrin promoter response to phorbol ester.
J Biol Chem 274:6190-6194, 1999.

2. Efimova T, Eckert RL. Regulation of human involucrin promoter activity by novel protein kinase C isoforms. J Biol Chem. 275:1601-1607, 2000.

Robert Fairchild, Ph.D.
Adjunct Assistant Professor, Department of Pathology, CWRU
Department of Immunology and Urology, Lerner Research Institute
Cleveland Clinic Foundation

Dr. Fairchild has had a long-term interest in transplant immunology and contact hypersensitivity of the skin. He has characterized the critical role of effector CD8+ T cells and regulating CD4+ T cells in contact dermatitis and the role of distinct antigen presenting cell populations in independently activating thse T cells. He has made a number of novel observations regarding the role of chemokines in T cell recruitment and tissue inflammation in both contact dermatitis and transplant rejection models of skin and solid organs. Dr. Fairchild regularly interacts with the SDRC Immunology Thematic Group in the SDRC speaaker serices, in the Pathology Department-sponsored Cleveland T Cell Awareness Club, in periodic joint lab meetings with SDRC members in the Department of Dermatology, and in the utilization of the Morphology Core. Dr. Fairchild's interactions with the SDRC represent a critical venue for dialogue in skin immunology.

Two recent representative publications:

1. Kondo T, Morita K, Watarai Y, Auerbach MB, Taub DD, Novick AC, Toma H, Fairchild RL. Early incareased chemokine expression and production in murine allogeneic skin grafts is mediated by natural killer cells. Transplantation; 69: 969-977, 2000.

2. Koga S, Auerbach MB, Engeman TM, Novick AC, Toma H, Fairchild RL. T cell infiltration into class II MHC-disparate allografts and acute rejection is dependent on the IFN-gamma-induced chemokine Mig. J Immunol; 163:4878-4885, 1999.

Stanton L. Gerson, M.D.
Professor of Medicine; Chief, Hematology/Oncology
Co-Investigator, Pilot & Feasibility Project in this application

Dr. Gerson's interest is in DNA repair 06-alkylguanine DNA alkyltransferase and its inhibitor, benzylguanine which is known to sensitize cells to BCNU has led to pre-clinical and clinical studies of biochemical modulation by benzylguanine of AGT in cutaneous malignancies. This group has identified high AGT in T-cell lymphomas, and, biochemical monitoring of AGT depletion following benzylguanine treatment. This led to the development of a CTEP approved clinical trial using the combination of benzylguanine and locally administered BCNU for patients with advanced T-cell cutaneous lymphomas, in collaboration with Dr. Wood. This project, originally supported by SDRC cores and enrichment programs, is now funded by NIH as an R21.

In addition, Dr. Gerson's laboratory group will be involved in a Phase II clinical trial using the systemic combination of intravenous benzylguanine and BCNU in the management of advanced malignant melanoma. Dr. Gerson's laboratory group provide the pre-clinical modeling for appropriate drug administration and his laboratory was essential to the design of a Phase I clinical trial which established the biochemical modulatory dose of benzylguanine which would completely deplete AGT as 12mg/m2. This is the dose which will be used in Phase II clinical trials. Dr. Gerson continues to develop the pre-clinical models of benzylguanine with BCNU and temozolomide and is keenly interested in translational research studies using lymphocytes as a pharmacodynamic marker and tumor biopsies to specifically determine how effective benzylguanine inhibition of AGT as in cutaneous malignancies.

Dr. Gerson also has extensive experience in long-term transfection of bone marrow cells for gene therapy applications. As Director of the Bone Marrow Stem Cell Procurement Core for the Cancer Center, he provides a key resource for human immunocyte precursors. He works actively with Dr. Gilliam in the development of monocyte-transformed cells with secretory potential in GVH and scleroderma, and is a mentor on Dr. Gilliam's K08 proposal. He is also a key member of the SDRC-sponsored GVH multidisciplinary research group, involving also Drs Gilliam, Stevens, Levine, and Laughlin, and which has designed and is executing collaborative studies on patients with cutaneous and GI GVH.

Two recent representative publications:

1. Davis BM, Roth JC, Liu L, Xu-Welliver M, Pegg AE, Gerson SL. Characterization of the P140K, PVVP (138-140) MLK, and G156A O6-methylguanine-DNA methyltransferase mutants: implications for drug resistance gene therapy. Hum Gene Ther; 10:2769-2778, 1999.

2. Qin X, Liu L, Gerson SL. Mice defective in the DMA mismatch gene PMS2 are hypersensitive to MN induced thymic lymphoma and are partially protected by transgenic expression of human MGMT. Oncogene; 18:4394-4400, 1999.

Mahmoud A. Ghannoum, M.Sc, Ph.D.
Professor of Dermatology
Director, Mycology Reference Laboratory
Scientific Director, Center for Medical Mycology
Principal Investigator, Pilot & Feasibility Project in this application

Dr. Ghannoum's laboratory has been devoted to two main research areas: a basic science area focusing on determination of virulence factors responsible for the pathogenesis of Candida albicans and Cryptococcus neoformans infections, and a clinical laboratory area concentrating on development of reference methods for anti-fungal susceptibility testing.

Dr. Ghannoum's basic studies focus on the roles of fungal lipids and sterols, encapsulation, virulence and endothelial cell damage. Recently, Dr. Ghannoum obtained evidence that secretion of phospholipase by Candida is involved in the development of hematogenously disseminated candidiasis and tissue invasion in murine models. In an effort to prove the role of phospholipase in the virulence of C. albicans, Dr. Ghannoum has purified and characterized the dominant and cloned the gene encoding it.

Of particular relevance in the clinical laboratory area is the microdilution method Dr. Ghannoum developed for determining fungal drug susceptibility. This method is reproducible, easy to perform and has both intra- and inter-laboratory agreement. Importantly, it has utility as a predictor of treatment success for patients with acute AIDS-associated cryptococcal meningitis, and is applied to Candida and dermatophytes.

In an effort to delineate the pathogenicity factors of fungi and yeasts causing skin infections, Dr. Ghannoum is utilizing the methods and approaches developed in his previous investigations of fungal virulence. To date, skin fungal virulence has received little or no attention even though infections with these agents are increasing and may cause devastating systemic infections in the immunocompromised host, especially in those whose neutrophil and macrophage function is deficient. Dr. Ghannoum is developing a new direction in collaboration with Dr. Kang on a pilot project to study candidal-skin monocyte/macrophage bidirectional signaling and cytokine induction resulting in novel findings that were recenty published (1). Preliminary data from that institutional support study now forms the basis of a P&F application for a second year of pilot funding.

Dr. Ghannoum is also an important link of the SDRC to the Center for AIDS Research (CFAR) and the TB research unit. Collaborations involve studies of thrush in clinical trials at University Hospitals of Cleveland as well as studies from the Uganda CFAR international effort, for which Dr. Ghannoum is a member of the CFAR Clinical International Core Facility.

Two recent representative publications:

1. Xiong J, Kang K, Liu L, Yoshida Y, Cooper KD, Ghannoum MA. Candida albicans and candida krusei differentially induce human blood mononuclear cell interleukin-12 and gamma interferon production. Infect Immun; 68:2464-2469, 2000.

2. Leidich SD, Ibrahim AS, Fu Y, Koul A, Jessup C, Vitullo J, Fonzi W. Mirbod F, Nakashima S, Nozawa Y, Ghannoum MA. Cloning and disruption of caPLB1, a phospholipase B gene invovled in the pathogenicity of Candida albicans. J Biol Chem; 273:26078-26086, 1998.

Anita C. Gilliam, M.D., Ph.D.
Assistant Professor of Dermatology, Pathology and Oncology
Past Pilot and Feasibility Study Recipient
Director, Cellular and Molecular Morphology Core
Co-Director, Animal Experimentation Core
Principal Investigator, Pilot and Feasibility Project in this Application

Dr. Anita Gilliam's research is in the area of cutaneous immunobiology, with an emphasis on the molecular basis of autoimmune disease and graft versus host disease (GVHD). She is using murine models for scleroderma, a fibrosing autoimmune disease of unknown etiology, to dissect out the early immunologic events in skin fibrosis, in which increased collagen production by fibroblasts is thought to be driven by monocyte production of TGFb1. The ultimate goal is to develop interventions which can be tested in the mice as potential therapies for patients with scleroderma. Collaborative studies are in progress with Dr. Stanton Gerson, Director of Hematology/Oncology, to manipulate bone marrow stem cells by transduction with inhibitors of monocytes or TGFb1, promote their differentiation into monocytes in vitro, then transplant these genetically altered cells into mice with sclerodermatous graft versus host disease in order to inhibit fibrosis. These studies have direct relevance to graft versus host disease in humans, a common consequence of allogeneic bone marrow transplantation for hematopoietic malignancies.

A separate but related project involves the characterization of GVHD in patients who have received cord blood transplantation. Their immune systems remain immature for a prolonged period of time, and it is not clear how GVHD occurs in these individuals. This project is in collaboration with Drs. Stanton Gerson, Mary Laughlin, Alan Levine, and Seth Stevens.

As a dermatologist/dermatopathologist of the Cellular and Molecular Morphology Core, Dr. Gilliam regularly interacts with SDRC investigators on various projects involving immunopathology and histopathology of human and experimental animal skin. In one such collaborative study, Dr. Gilliam is working with Dr. Peter Heeger (CWRU Nephrology) on a model using skin graft rejection to study the complex immunologic events of allograft recognition surrounding rejection or tolerance induction. She also works with Dr. Kevin Cooper's (CWRU Dermatology) Immunobiology group as a consultant for projects related to photoimmunobiology of the skin (effects of UV light on human and mouse skin with respect to cutaneous cytokine profiles, and immunohistochemistry of monocytes, Langerhans cells and lymphocytes). Her clinical role as dermatopathologist for the CWRU Melanoma and Lymphoma Multidisciplinary Conference groups has led to collaborations with Dr. Gary Wood (CWRU Dermatology), who studies the immunologic phenotypes of cutaneous lymphomas, and with Dr. Timothy Spiro (CWRU Hematology/Oncology) and with Drs. Ahmad and Mukhtar (Dermatology).

Two recent representative publications:

1. McCormick LL, Yan Zhang, Tootell E, and Gilliam AC. Anti-TGFb treatment prevents skin and lung fibrosis in murine sclerodermatous graft versus host disease: a model for human scleroderma. J Immunol 163:5693-5699, 1999.

2. Gilliam AC, Kremer IB, Yoshida Y, Stevens SR, Tootell E, Teunissen MB,
Hammerberg C, Cooper KD. The human hair follicle: a reservoid of CD40+ B7-deficient Langrhans cells repopulate epidermis after UVB exposure. J Invest Dermatol; 110:422-427, 1998.

Tariq Haqqi, Ph.D.
Associate Professor of Medicine /Rheumatology and Dermatology
Previous Pilot and Feasibility Recipient

Tariq Haqqi, Ph.D. is a molecular immunologist and immunogeneticist who was recruited to CWRU in 1989. Dr. Haqqi is an excellent example of a talented scientist with extensive expertise whose interests have broadened to encompass fundamental problems in immuno and molecular dermatology. Dr. Haqqi's major interest is in collagen-induced arthritis. In particular, he is studying molecular mechanisms of induced protective immunity against collagen-induced arthritis. He is also interested in gene expression analysis in human cartilage/chondrocytes in response to inflammatory cytokines and has recently reported the identification of several novel TNF-a responsive genes in human chondrocytes. His collaborative studies with Dr. Mukhtar's laboratory also identified and characterized new genes expressed in chemically induced tumors in the skin of SENCAR mice. Dr. Haqqi has also developed a unique method of transcription profiling and this method is now being used in a collaborative study with Dr. Mukhtar to identify novel transcripts associated with mammalian UV-response. Most recently Dr. Haqqi is collaborating with Dr. Mukhtar to define the fate of naturally occurring substances in preventing arthritis. Their recent study published in PNAS is a beginning in this direction.

Two recent representative publications:

1. Valujskikh, A., D. Matesic, A. Gilliam, D. Anthony, T. M. Haqqi, and P. S. Heeger. T cells reactive to a single immunodominant self-restricted allopeptide induce chronic skin graft rejection in mice. J. Clin. Invest. 101:1398-1407, 1998.

2. Haqqi TM, Anthony DD, Gupta S, Ahmad N, Lee MS, Kumar GK, Mukhtar H. Prevention of collagen-induced arthritis in mice by a polyphenolic fraction f green tea. Proc Natl Acad Sci USA; 96:4525-4529, 1999.

Vincent C. Hascall, Ph.D.
Staff and Head of Connective Tissue Biology Section
Department of Biomedical Engineering
Lerner Research Institute, Cleveland Clinic Foundation

Major Research Interests of Dr. Hascall are: structure, function and metabolism of hyaluronan and proteoglycans in connective tissues, including the epidermis. In his labortory he established a research program focusing on cartilage matrix biochemistry.

Dr. Hascall studies the structure, function and metabolism of macromolecules of the extracellular matrix of connective tissues, particularly hyaluronan and proteoglycans. A major focus is on the role of hyaluronan in the epidermis. Epidermal keratinocytes are surrounded by an intercellular matrix enriched in hyaluronan. These cells actively synthesize and catabolize this macromolecule, which has a half life of less than a day in this tissue. With colleagues at the University of Kuopio in Finland and at the University of Michigan, Dr. Hascall has developed a rat keratinocyte cell line that undergoes complete epidermal stratification and cornification when cultured on collagen rafts at an air interface and in the absence of a feeder layer. This model system is proving versatile and exceptionally useful for various studies focusing on epidermal biology. He is actively engaged in interactions with the skin equivalent interest group.

Two recent representative publications:

1. de La Motte CA, Hascall VC, Calabro A, Yen-Lieberman B, Strong SA. Mononuclear leukocytes preferentially bind via CD44 to haluronan on hman intestinal mucosal smooth muscle cells after virus infection or treatment with poly (I.C). J Biol Chem; 274:
30747-30755, 1999.

1. Goodstone NJ, Hascall VC, Calabro A. Differential effects of Staphylococcus aureus alpha-hemolysin on the synthesis of hyaluronan and chonroitin sulfate by rat chondrosarcoma chondrocytes. Arch Biochem Biophys; 350: 26-35, 1998.

Steven Haynesworth, Ph.D.
Associate Professor, Biology

Dr. Haynesworth studies bone marrow-deerived mesenchynal stem cells and interacts with Drs. Caplan and Gerson to understand their differentiation into dermal fibroblastic cells vs. osteogenic cells. He has developed a monoclonal antibody which detects a newly discovered epitope on stem cells, endoglin. His work also interacts with the GVH group and the skin equivalent group, and he participates in each.

Two recent representative publications:

1. Koc ON, Gerson SL, Cooper BW, Dyhouse SM, Haynesworth SE, Caplan AI, Lazarus, HM. Rapid hematopoietic recovery after cooinfusion of autologous-blood stem cells and culture-expanded marrow mesenchymal stem cells in advanced breast cancer patietns receiving high-dose chemotherapy. J Clin Oncol;18:307-316, 2000.

2. Fleming JE Jr, Haynesworth SE, Casiede P, Baber MA, Caplan AI. Monoclonal antibody against adult marrow-derived mesenchymal stem cells recognizes developing vasculature in embryonic human skin. Dev Dyn; 212:119-132, 2000.

Peter S. Heeger, M.D.
Assistant Professor of Medicine and Pathology
Previous Pilot and Feasibility Study Recipient

Dr. Heeger's program centers around understanding the mechanisms of allogeneic graft rejection. The research is comprised of several components. One component uses mouse models of skin and cardiac transplantation to study the mechanisms of T cell mediated rejection (with specific emphasis on indirect allorecognition) and induction of immunologic tolerance to allografts. A second arm of the research program focuses on the use of a highly sensitive ELISPOT assay for immunologic monitoring of renal allograft recipients in humans. The funds for skin allograft rejection provided by the original Pilot and Feasibility program through the SDRC provided the foundation for these studies, which are now funded through the NIH and the Department of Veterans Affairs.

Two recent representative publications:

1. Valujskikh A, Heeger PS. T cells responsive through the indirect pathway can mediate skin graft rejection in the absence of interferon-gamma. Transplantation; 68:1016-1019, 2000.

2. Valujskikh A, Matesic D, Heeger PS. Characterization and manipulation of T cell immunity to skin grafts expressing transgenic minor antigen. Transplantation; 68:1029-1036, 1999.

Frederick P. Heinzel, M.D.
Associate Professor of Medicine
Principal Investigator, Pilot & Feasibility Project in this application

Dr. Fred Heinzel's research addresses the regulation and function of pro-inflammatory cytokines in two murine models of infectious disease pathology: leishmaniasis and endotoxemia. In the mouse model of leishmaniasis, Dr. Heinzel has studied the immunoprotective, Th2 T cell inhibiting effects of recombinant interleukin-12 in susceptible strains of mice and the importance of endogenous IL-12 in directing curative Th1 T cell responses in normally resistant strains of mice. Dendritic cells were identified as the source of IL-12 in vivo and an essential role for CD40 engagement in IL-12 production and cure of disease was determined. Current NIH-funded projects are aimed at developing new forms of immunotherapy of established, progressive leishmaniasis, which fails to respond to IL-12 treatments alone. Successful approaches include "resetting" of the CD4 phenotype by transient depletion with anti-CD4 antibodies, followed by cytokine-directed treatment (recombinant IL-12 and anti-IL-4 antibody). Alternatively, pretreatment of susceptible mice with activating anti-CD40 antibodies or soluble CD40 ligand protects against infection and he is now studying whether CD40 activation cures established disease as well. The mechanisms underlying these therapeutic interventions, such as induction of IL-12 receptors on CD4 cells, the role of CD8 Thl cells, synergistic pro-inflammatory cytokines and cytokine-independent activation of inducible nitric oxide synthesis are currently under study.

Through the efforts of the SDRC, Dr. Heinzel has recently entered into a focus on the skin immunobiology of leishmaniasis. As a mentor for Dr. Inger Kremer, a postdoctoral fellow in the Department of Dermatology working under a Dermatology Foundation fellowship, he has developed an interest in how initial skin infection conditions may effect the above IL-12/CD40 relationships. This has high relevance for UV effects and skin-based vaccine efforts, and he has formulated the data and hypothesis into a P&F proposal.
In addition, Dr. Heinzel has collaborated on another skin application examining the role of FLT-3 Ligand in expanding dendritic antigen presenting cells in the dermis. This was presented as a platform talk at the recent SID91). The recruitment of a scientist of Dr. Heinzel's expertise into the cutaneous biology aspects of his science is the direct result of the proactive SDRC's enrichment and connectedness activities.

Two recent representative publications:

1. Heinzel, F.P. and R.A. Maier, Jr. IL-4 independent acceleration of cutaneous leishmaniasis in susceptible BALB/c mice following treatment with anti-CTLA4 antibody. Infect Immun. 67:6454-6460, 1999.

1. Heinzel FP, Rerko RM. Cure of progressive murine leishmaniasis: interleukin 4 dominance is abolis had by transient CD4(+) T cell depletion and T helper cell type 1-selective cytokine therapy. J Exp Med;189:1895-1906, 1999.

Sudha Iyengar, Ph.D.
Assistant Professor of Epidemiology and Biostatistics
Director, Genotyping Facility, MetroHealth Campus

Dr. Iyengar's is involved in two funded genetics projects for dermatologic diseases. She is the PI of a project to recruit patients and families with alopecia areata (AA) from the general population. The cloning of the recessive gene for alopecia universalis (AU) on human 8p12 from a large inbred kindred lead her to hypothesize that genes for AA also segregate in smaller families. Using an affected relative pair strategy, she is examining a broader AA phenotype by collecting such families and genotyping candidate genes. This project, now in its second year of funding, has both an epidemiologic and a molecular genetic component. In collaboration with Dr. Animesh Sinha at the Weill Medical College of Cornell University, epidemiologic surveys were conducted in two clinics in the New York Metropolitan area to recruit patients with a family history of AA. Of 120 AA index cases that contacted, 22 probands with a living affected family member have been recruited. Additional recruitment and genotyping for polymorphisms at the HLA-D locus is ongoing in cases with and without a family history. These interim data from this study demonstrate that AA aggregates in families. Additional candidate gene analysis (eg. AU locus) and genome scan of AA multiplex families from this cohort should facilitate identification of genes controlling pathogenesis.

Dr. Iyengar is also a collaborator on a multi-center project to map genes for sarcoidosis, a systemic granulomatous disease of unknown etiology that likely involves exposure to some environmental agent in a genetically susceptible host. The goal of this project is to identify sarcoidosis susceptibility genes and determine how these genes and environmental risk factors interact to cause sarcoidosis. The affected sibling pair linkage analysis will be utilized to scan the genome for linked chromosomal regions. Dr. Iyengar's laboratory will be performing the genome scan for the consortium. Fine mapping of newly identified candidate regions and genes will be performed in conjunction with other members of the consortium.

As a new member, Dr. Iyengar anticipates vigorous interaction with other SDRC members. The nature of this interaction will be through the SDRC cores and through collaborative projects with other active members. At this juncture, two cores, the translational research core and the animal experimentation core are of key interest for the alopecia areata and the sarcoidosis projects. The translational research core will aid in recruitment of patients and families for both projects. If novel candidate genes are identified using either the candidate gene approach or the genome scan approach, the animal experimentation core will be utilized to generate knockout and transgenic mice to further comprehend the biologic function of the genes.

Two recent representative publications:

1. El-Meanawy M.A., Schelling J.R., Pozuela F., Churpek M.M., Ficker E., Iyengar S., Sedor J.R: Use of serial analysis of gene expression (SAGE) to generate kidney expression libraries. Am J. Physiol (in press).

2. Sirugo G., Iyengar S., Castiglione C.M., Kidd J.R., Pakstis A.J., Kidd K.K.: Analysis of extended haplotypes at DRD2 with new STRP and SNP sites. Am J Hum Genet 63: A221, 1998.

James W. Jacobberger, Ph.D.
Associate Professor
Division of General Medical Sciences, Cancer Research Center
Member, Cell Biology Research Area
Director, Flow Cytometry Core within the Cell Culture and Molecular Technology Core

Dr. Jacobberger received his Ph.D. in Microbiology at the University of Rochester (1983). His post-doc was in Experimental Pathology at the University of Colorado Health Sciences Center (1983-85). He joined CWRU in 1985 as Assistant Professor of Developmental Genetics and Anatomy and currently is Associate Professor of Oncology with tenure in the Division of General Medical Sciences, Cancer Center with joint appointment in the Department of Genetics. His Ph.D., post-doc, and early professorship emphasized Analytical Cytology applied in the areas of malaria, virology, and tumorigenic transformation. This interest is reflected in his establishment of a successful flow cytometry core facility (1985-present), partially supported by an NIH Cancer Center grant, that currently provides analysis and cell sorting for more than 30 different laboratories per year at CWRU. This facility, which has seen a marked increase in usage by SDRC members over the past few years currently offers a volume discount to SDRC PI's, and will now be an official service of the SDRC Cell Culture and Molecular Technology Core. He holds elected office for the International Society for Analytical Cytology; is an associate editor of the journal, Cytometry; and, reviews major instrumentation grants for NIH and NSF. He has received his own major instrumentation grant for a state-of-the-art flow cytometry from NIH.

Dr. Jacobberger's interests are cell cycle regulation and tumorigenesis. In the past, his laboratory has (1) shown that SV40 large T antigen is rate limiting for G1 transit and functions in a concentration dependent manner, (2) developed an in vitro/vivo model of astroglioma, (3) developed multiparametric cell cycle analytical technology that includes quantification of intracellular proteins and DNA content by flow cytometry. Current projects in the laboratory are (1) further development of multiparametric cell cycle analysis, and (2) dissection of the defects in TGF-b signaling in epithelial cancer cell lines. Dr. Jacobberger's expertise has been critical for a number of SDRC members' studies using flow cytometry of cell cycle proteins and intracellular cytokines.

Two recent representative publications:

1. Frisa PS, Lanford RE, Jacobberger JW. Molecular quantification of cell cycle-related gene expression at the protein level. Cytometry;39:79-89, 2000.

2. Jacobberger JW, Sramkoski RM, Zhang D, Zumstein LA, Doerksen LD, Merritt JA, Wright SA, Shults KE. Bivariate analysis of the p53 pathway to evaluate Ad-p53 gene therapy efficaacy. Cytometry;38:201-213, 1999.

Kefei Kang, M.D.
Associate Professor of Dermatology
Co-Director, Cell Culture and Molecular Technology Core

Dr. Kefei Kang has a long-standing research interest in immunodermatology. He has been engaged in studies of atopic dermatitis, lupus erythematosus, cutaneous T cell lymphoma and medical mycology. In particular, investigations have focused on the relationship of IgE-dependent and T cell mediated mechanisms in atopic dermatitis, as well as clinical studies

Currently Dr. Kang's active projects focus on antigen presenting cells of the skin and their immunoregulatory cytokines. Specifically, he has been examining: 1) IL- 10 and IL- 12 expression of human Langerhans cells (LC), dendritic antigen presenting cells of the dermis, and monocytes/macrophages; 2) the role of UV radiation in modifying these cells and cytokines; and 3) the mechanism of monocyte/macrophage activation in human skin via integrin ligation and cytokines; 4) cutaneous immunologic effects on antigen-presenting cells of candidal invasion.

He has found that CD 11b+ macrophages infiltrating human epidermis after in vivo UV exposure potently produce IL-10 and LC in human epidermis can synthesize IL-12. Moreover, in human skin, UVB can initiate early induction of IL-10 over IL-12 preferentially in expanding dermal monocytic/macrophagic population. Furthermore, he has found that adhesion molecules involve in the induction of cytokines. His recent studies have shown that ultraviolet exposure of human skin causes cellular fibronectin (ligand for b1 integrin) and iC3b (ligand for b2 integrin) deposition in contiguity with CD1lb+ monocytes, both cellular fibronectin and iC3b interaction with monocytes induce IL-10, but iC3b downregulates IL-12. In the iC3b study, he has established a collaboration with Dr. Melvin Berger, Professor of Pediatrics, an expert in complement interactions with leukocytes and infectious diseases.

Dr. Kang, in collaboration with Dr. Mahmoud Ghannoum, Director of Mycology Research Center, is examining microbial induction of monocyte cytokine profiles in cutaneous fungal infections. They have found that clinical Candida isolates that differ in their virulence differentially induce immunoregulatory cytokine IL-12 production. The molecular mechanisms involved in Candida albicans suppression of IL-12 expression is currently under investigation.

Two recent representative publications:

1. Xiong J, Kang K, Liu L, Yoshida Y, Cooper KD, Ghannoum MA: Candida albicans and Candida krusei differentially induce human blood mononuclear cell IL-12 and gamma interferon production. Infect Immun;68:2464-2469, 2000.

2. Kang K Gilliam AC, Chen G, Tootel E, Cooper KD. In human skin , UVB initiates early induction of IL-10 over IL-12 preferentially in the expanding dermal monocytic macrophagic population. J Invest Dermatol;111:31-38, 1998.

James W. Kazura, M.D.
Professor of Medicine; Chief, Division of Geographic Medicine
Previous Pilot and Feasibility Recipient
Member, Executive Committee

The focus of Dr. James Kazura's laboratory is on the mechanisms which regulate differentiation and expansion of T helper cell subsets in filariasis. Using mice as models, he has determined that bias towards development of the Th2 subset (i.e., increased production of IL-4 and IL-5 relative to IFN-g and IL-2) is in part regulated by the activity of the counter-regulatory endogenous IL-4. The skewing towards Th2 development can be reversed by IL-12, a product of B cells and macrophages which increase NK activity and expansion. Current experiments are directed at defining the structural constraints on parasite allergens that induce Th2 differentiation and/or diminish IL-12 productive in vivo. Both diseases of is interest filariasis and malaria, involve percutaneous introduction of infection and antigen and thereby skin antigen presenting cells involvement in the skewing are a source of dialogue with SDRC immunologists. As a new member of the Executive Committee, Dr. Kazura's international experience and excellence as an immunologist will help shape the international opportunities available through SDRC interactions.

Two recent representative publications:

1. Bockarie MJ, Alexander NDE, Kazura JW, Bockarie F, Griffin L, Alpers MP. Treatment with ivermectin reduces the high prevalence of scabies in a village in Papua New Guinea. Acta. Tropica. (In press).

2. Bucci K, Kastens W, Connelly M, Hollingdale MR, Shandar A, Alpers MP, King CL, Kazura, JW. HLA supertypes and T-cell IFN-g and TNF-a responses to polymorphic and non-polymorphic epitopes of Plasmodium falciparum liver stage antigen-1 in Papua New Guinea. Clin Exp Immunol (in press).

Timothy J. Kinsella, M.D.
Professor and Chair, Department of Radiation Oncology
Member, Photcarcinogenesis/Photobiology/Photomedicine Research Area

Dr. Kinsella's laboratory studies mechanisms of ionizing radiation (IR) damage and repair in human tumors with emphasis on altering IR damage/repair processes using modulators of nucleotide metabolism. His laboratory uses both in vitro and in vivo human tumors systems to characterize the regulation of three key enzymes (thymidine kinase, thymidylate synthase, ribonucleotide reductase) involved in DNA repair synthesis following IR damage. These research initiatives have a clear translational component involving the clinical development and testing of both nucleoside analogs and nucleotide inhibitors as radiosensitizers in radioresistant human tumors such as high grade brain tumors and melanomas. He has completed the pre-clinical evaluation of a new oral prodrug for IUdR-mediated radiosensitization and is submitting an investigator-initiated IND to the FDA to begin the initial Phase I study.

Since his recent relocation to Case Western Reserve University School of Medicine in 1998, in collaboration with the SDRC, he is also involved in the clinical development of a new photosensitizer, silicon phthalocyanine (Pc4) for photodynamic therapy of recurrent or locally advanced primary skin cancers and skin metastases. The pre-clinical toxicology and pharmacology of Pc4 was recently completed by the NCI drug decision network in collaboration with CWRU. Initial Phase I clinical testing for primary and metastatic skin cancers is scheduled to begin at the Skin Diseases Research Center and University Hospitals Ireland Cancer Center in late 2000. Dr. Kinsella has gained a wealth of experience in Phase I and II drug testing of radiosensitizers while at the National Cancer Institute (1980-1987) and University of Wisconsin (1987-1998) and will play a major role in the clinical testing of Pc4.

Two recent representative publications:

1. Hwang HS, Davis TW, Houghton JA, Kinsella TJ. Radiosensitivity of thymidylate synthase-deficient human tumor cells is affected by progression through the G1 restriction point into S-phase: implications of fluoropyrimidine radiosensitization. Cancer Res;60:92-100, 2000.

2. Berry SE, Garces C, Hwang HS, Kunugi K, Meyers M, Davis TW, Boothman DA,
Kinsella TJ. The mismatch repair protein, hMLH1, mediates 5-substituted halogenated thymidine analogue cytotoxicity, DNA incorporation, and radiosensitization of human colon cancer cells. Cancer Res;59:1840-1845, 1999.

Nanette R. Kleinman, D.V.M.
Assistant Director, Animal Resource Center
Consultant, Animal Experimentation Core

Nanette R. Kleinman, D.V.M. is Associate Director for Veterinary Services, Animal Resource Center at Case Western Reserve University. In 1995 she became Diplomate, American College of Laboratory Animal Medicine. To SDRC members through the Animal Experimentation Core, she provides advice on animal protocols, animal models and animal health.

1. Sell DR, Kleinman NR, Monnier VM. Longitudinal determination of skin collagen glycation and glycoxidation rate predicts early death in C57BL/6NNIA mice. FASEB J. 14:145-156, 2000.

2. Culp LA, Lin WC, Kleinman NR. Tagged tumor cells reveal regulatory steps during earliest stages of tumor progression and micrometastasis. Histol Histopathol;14:879-886, 2000.

Neil J. Korman, Ph.D., M.D.
Associate Professor of Dermatology
Co-Director, Translational Research Core

Dr. Korman heads the Clinical Trials Unit of the Department of Dermatology and the Immunofluorescence laboratory. The major focus of Dr. Neil Korman's research has been the characterization of the antigens involved in autoimmune blistering diseases of the skin. Specifically, he has concentrated his efforts on the bullous pemphigoid antigens which are hemidesmosome associated structures which are an integral part of the epidermal basement membrane zone. The principal long term objective of his research laboratory has been to develop an understanding of the biological significance in health and disease of the epidermal bullous pemphigoid antigens. The lab has focused its efforts towards isolating bullous pemphigoid antigens from human skin and determining their interactions with other components of the epidermal basement membrane zone. Hemidesmosome maturation will be studied by investigating the site of synthesis of the bullous pemphigoid antigens. The antigenic specificity of skin bound antibodies in patients with bullous pemphigoid has been compared to the antigenic specificity of the circulating antibodies. This research will enable the characterization of the types of molecular interactions that may occur in hemidesmosomes and may lead to a better understanding of the autoimmune response in bullous pemphigoid along with the development of new knowledge relevant to improved diagnosis and treatment of this important skin disease.

Another area of active investigation involves studying the role that various epidermal basement membrane components may play in the invasion of basal cell carcinomas. Basal cell carcinomas have been shown to have significant abnormalities in numerous defined molecules of the hemidesmosome anchoring fibril complex at both the protein and the mRNA level. These findings support the hypothesis that BCC tumor cells may possess an impaired ability to synthesize the normal complement of epidermal BM components and that the absence of a normal BM facilitates tumor invasion. This purpose of this work is to characterize the mechanisms responsible for the abnormalities found in the epidermal basement membrane in basal cell carcinoma with the goal of developing a better understanding of the invasive behavior of this most common of all human cancers.

A new area of investigation involves a collaboration, coordinated by the SDRC with Drs. Eckert, Hascall, Haynesworth, McCormick and Caplan on an interdisciplinary effort aimed at developing an advanced in vitro skin equivalent that will be useful not only for basic science mechanistic studies but also for applied studies in the development of new dermatologic agents. Dr. Korman's involvement in this project will draw upon his expertise in the epidermal basement membrane, and complements the expertise of the group in epidermal biology, immunocytes of the skin, mesenchymal fibroblastic stem cells, and dermal matrix, respectively.

Two recent representative publications:

1. Korman, NJ. New and emerging therapies in the treatment of blistering diseases. Dermatol Clin;18:127-137, 2000.

2. Chopra A, Maitra B, Korman NJ. Decreased mRNA expression of several basement membrane components in basal cell carcinoma. J Invest Dermatol;110:52-56, 1998.

Kent C. Kwoh, M.D.
Associate Professor, Medicine
Division of Rheumatology

Dr. Kwoh brings top-level expertise in clinical outcomes and international study design to the SDRC and its Translational Research initiatives. He is studying the effects of educational interventions and genetic factorson osteoporosis in teenage girls and the development of progrms to benefit the health of aging veterans. His funded studies on VA computer-assisted access to specialists, includes a component for skin disease, is critical to determining how best to bring quality and effective dermatology care to veterans at the VA outpatient sites in the Northeastern Ohio region (17 sites in all) which are too remote to be physically staffed by the VA Dermatology staff. He is also a critical collaborator in organizing the Dermatology component of the Twins Day Festival evaluations.

Two recent representative publications:

1. Hoffman GS, Druker Y, Cotch MF, Locker GA, Easley K, Kwoh CK. Wegener's Granulomatosis: patient-reported effects of disease on health, function and income. Arthritis Rheum 41:2257-2262, 1998.

2. Ibrahim SA, Snow JS, Harper DL, Norris D, Kwoh CK, Baker DW. Racial variation in the use of anticoagulant therapy in heart failure: a study of elderly, hospitalized patients. J Gen Int Med 15:134-137, 2000.

Michael E. Lamm, M.D.
Professor and Chairman, Department of Pathology
Previous Member, Executive Committee

Dr. Lamm has been a pioneer in mucosal immunity and IgA. In that regard, his findings with respect to the mucosal immune system were a forerunner to the concept of the skin immune system. His recent work has suggested that mucosal IgA acts, not only as an immunological barrier in secretions of mucous membranes, but also can (1) neutralize viruses inside epithelial cells that express the polymeric Ig receptor (secretary component) and (2) complex antigens in the mucosal lamina propria and transport them through the mucosal epithelium by the same route taken by free IgA. The first newly-proposed function, intracellular neutralization of viruses, envisions that under certain circumstances antibodies can act as defenses against intracellular pathogens in addition to the traditional immunological mode of cell-mediated immunity. The second new function suggests that mucosal IgA can function as an excretory immune system to rid the body of local antigens and, thus, protect the systemic circulation from an overload of immune complexes. In addition to the general relevance of the immune system to skin diseases, his research is potentially pertinent to diseases in which IgA and/or IgA immune complexes participate, e.g., dermatitis herptiformis and Henoch-Schonlein purpura. As a member of the Executive Committee, Dr. Lamm has provided valuable insight to research efforts in immunodermatological diseases within the SDRC, and coordination of resources with the Department of Pathology and Immunology Training Grant.

Two recent representative publications:

1. Huant YT, Miller CJ, Wong V, Fujioka H, Nedrud JG, Lamm ME. Replication and budding of simian immunodeficiency virus in polarized epithelial cells. Virology;257:24-34, 1999.

2. Lamm ME. Current concepts in mucosal immunity. IV. How epithelial transport of IgA antibodies relates to host defense. Am J Physiol;274:G614-617, 1998.

Mary Laughlin, M.D.
Assistant Professor, Medicine,
Division of Hematology and Oncology

Dr. Laughlin is the Director of the allogeneic Bone Marrow Transplant Program at University Hospitals and the CWRU Cancer Center. She specializes in cord blood transplantation and the characterization of T cell mechanisms present in cord blood T cells that may be responsible for the reduced severity fo GVH with this protocol. She is a highly active and contributory member of the SDRC-sponsored GVH multidisciplinary research group and is responsible for initiating the clinical protocol that generates the coordinated skin and gut biopsies.

Two recent representative publications:

1. Laughlin MJ, Rizzieri DA, Smith CA, Moore JO, Lilly S, Martin P, Carrier C, Stevens CE, Rubinstein P, Kurtzberg J. Engraftment and reconstitution of immune function post-unrelated cord blood transplant in an adult with Philadelphia chromosome positive acute lymphocytic leukemia. Leuk Res 22(3):215-219, 1998.

2. Kadereit S, Mohammad SF, Miller R, Woods KD, Listrom C, Alali A, McKinnon K, Iacobucci ML, Jacobberger JW, Sramkoski MR, Laughlin MJ. Reduced NFAT1 protein expression in human umbilical cord blood T lymphocytes. Blood 94:3101-3107, 1999.

Paul V. Lehmann, M.D., Ph.D.
Associate Professor of Pathology
Previous Pilot and Feasibility Study Recipient

Dr. Lehmann's primary field of interest is T cell biology and T cell mediated pathology with emphasis on autoimmune diseases. Being the barrier between the external and internal

antigenic universe, the skin plays a critical role in immune defense, hence also in T cell biology and T cell mediated pathology.

As far as T cell biology is concerned, one focus of studies of Dr. Lehmann's laboratory is to understand under what conditions the encounter with antigen initiates a T cell response and when does it lead to the inactivation of these T cells, resulting in tolerance. This question underlies cutaneous allergic responses, possibly including psoriasis, where environmental antigens cause unwanted immune pathology in the skin. The second aspect of this question pertains to cutaneous tumors, like melanoma, where the T cell system fails to reject the malignant cells. Dr. Lehmann had major surprises addressing these basic issues , including the discovery that protocols that were thought to induce T cell inactivation actually induce immunity of a certain (Th2) type. Similarly, in recent studies in collaboration with Dr. M-Tary Lehmann, tumors that have been thought to induce T cell tolerance, also induce Th2 type immunity. Because immunity has different manifestations, Dr. Lehmann studied what it takes to activate one or the other. He learned that the pro-inflammatory T cell response is induced when the antigen is encountered in a micro environment in which the cells of the innate immune system have been activated, including by the presence of IL-12 inducing mycobacteria or CpG-rich microbial DNA. In the absence of such irritation, Th2 immunity develops, by default. A major discovery, supported by SDRC P&F grant, was that these different types of T cell responses are not mediated by Th1/Th2 cells, that were thought to secrete blocks of mutually exclusive sets of cytokines, but that T cells differentiate into memory cells that can produce only one cytokine at a time.

As far as the basic mechanisms that underlie autoimmune diseases are concerned, this pertains to conditions such as pemphigus vulgaris, scleroderma and systemic lupus erythematosus, we learned that different types of autoimmune responses can lead to manifestation of the disease, or its suppression. We also completed studies which suggest that autoimmune responses are of low avidity, because negative selection purges the high affinity end of the repertoire, which provides a new rational for therapeutic.

The above breakthroughs have been possible through the development of ELISpot technology by Dr. Lehmann, whose SDRC P&F funding led to his development of a rapid, fully-automated image analysis system which significantly improves the throughput and sensitivity of ELISpot technology in immunologic detection of lymphokine-producing cells. Dr. Lehmann's collaboration is also an important part of the NIH-awarded work of Dr. Heeger on skin rejection and Dr. Stevens on T cell activation.

Two recent representative publications:

1. Lehmann PV. Selective peptidomimetic blockers of autoantigen presentation: a novel therapeutic approach to autoimmune disease. Trends Pharmacol Sci;21:79-80, 2000.

3. Karulin AY, Hesse MD, Tary-Lehmann M, Lehmann PV. Single-cytokine-producing CD4 memory cells predominate in type 1 and type immunity. J Immunol; 164:
1862-1872, 2000.

Alan D. Levine, Ph.D.
Associate Professor of Medicine, Pathology, Pharmacology, General Medical Sciences (Oncology)

The major research activity in Dr. Levine's laboratory is in immune tolerance, immune protection, and apoptosis during host defense systems. The intestinal mucosa is the largest lymphoid organ, as measured by both the number of resident leukocytes and the size of its surface area exposure to the environment (greater than a tennis court). In addition, the wall of the gut is continuously bathed by bacteria, parasites, fungi, amoebae, viruses, mitogens, toxins, and immunogenic food proteins. Therefore, a complex multi-tiered host defense system has evolved in the gut and can be divided into three areas: (1) Barrier exclusion by an actively regenerating epithelial cell monolayer. (2) Innate inflammatory responses mediated by local synthesis of pro- and anti-inflammatory cytokines. (3) Acquired immune responses regulated by T lymphocytes.

Dr. Levine's laboratory focuses on the mechanisms by which each of these systems is regulated. In particular, (1) they study the biochemical signaling pathways that maintain the balance between epithelial cell proliferation, differentiation, and apoptosis as these cells migrate along their basement membrane from the base of the crypt to the luminal surface. In animal models of acute and chronic intestinal inflammation, (2) they investigate the expression and function of pro-inflammatory and anti-inflammatory cytokines in response to gut injury induced by ischemia, peritonitis, and chronic intestinal inflammation, a model for human inflammatory bowel disease (IBD). Finally, using animal models and purified human mucosal T cells in vitro, (3) they explore the effects of co-stimulatory and accessory molecules, antigen presenting cells, accessory cells, immune cytokines, plasma membrane microdomains (lipid rafts), and the interstitial extracellular matrix on T cell activation, regulating immune protection, immune tolerance, and immunopathology in the normal and inflamed intestines from patients with IBD and graft versus host disease (GVHD).

Dr. Levine is involved in the SDRC through his involvement in the GVH multidisciplinary research grou, his generous co-mentorship of a number of SDRC faculty, including Drs. Stevens and Singer, and his technical interactions.

Two recent representative publications:

1. Grossmann, J., Mohr, S., Lapetina, E.G., Fiocchi, C. and Levine, A.D. "Sequential activation of select caspases in intestinal epithelial cells, primed for detachment-induced apoptosis." Amer. J. Physiol. 274:G1117-G1124, 1998.

3. Hurst, S.D., Cooper, C.J., Sitterding, S.M., Choi, J.-H., Jump, R.L., Levine, A.D., Barrett, T.A. "The differentiated state of intestinal lamina propria CD4+ T cells results in altered cytokine production, activation threshold and costimulatory requirements." J. Immunol., 163; 5937-5945, 1999.

Henry Lim, M.D.
Professor and Chairman, Department of Dermatology
Henry Ford Hospital

Dr. Lim is an active member of the Department of Dermatology, despite being located in Detroit's Henry Ford Hospital, i.e., 2 ½ hour drive and a 20 minute flight. Because Henry Ford Hospital is a CWRU-affiliated hospital for student education and research collaborations, a direct video-conferencing between Ford and CWRU is set up, and it is used regularly to enact meetings between SDRC members and Dr. Lim. In addition, Dr. Lim physically visits CWRU on a regular basis. He is taking an increasingly active role in advisory research faculty on photobiology and photomedicine applications, as well as in complement biology and general faculty development.

Two recent representative publications:

1. Nomura, N, Lim, HW, Levin, JL, and Sassa, S. Effect of soluble complement receptor type I on porphyrin-induced phototoxicity in guinea pigs. J Photochem Photobiol B: Biol., 42:28-31, 1998.

2. Nair, RP, Stuart P, Henseler T, Jenisch S, Chia NVC, Westphal E, Schork NJ, Kim J, Lim HW, Christopher E, Voorhees JJ, and Elders JT. Localization of psoriasis susceptibility locus PSORS1 to a 60 kilobase interval telomeric to HLA-C. Am J Hum Genet, 66:1833-1844, 2000.

Edward V. Maytin, M.D., Ph.D.
Full Staff, Dermatology
Assistant Staff, Biomedical Engineering
Lerner Research Institute
Cleveland Clinic Foundation

Dr. Maytin's primary research area involves gene regulation in the epidermis. Dr. Maytin is interested in regulatory proteins (transcription factors) that control epidermal gene expression, and has focused on two particular families of transcription factors: the C/EBPs (CCAAT/Enhancer Binding Proteins), and the AP-2 family (Activator Protein 2). Both are important in the differentiation program of the epidermis, a tightly-regulated process which results in the transition of immature basal cells into mature squames. In one project, Dr. Maytin is testing the hypothesis that C/EBPs may represent master regulators for epidermal differentiation. The prediction is that overexpression of various C/EBPs through a variety of gene-delivery techniques will alter the epidermal differentiation program, either promoting or preventing it. The research promises to improve our understanding of important gene-regulatory mechanisms in the skin during health, after injury, and in various disease states. In a second related project, Dr. Maytin is studying how cellular differentiation and transcription factors can enhance the treatment outcome in a therapeutic technique called photodynamic therapy (PDT).

Dr. Maytin arrived at the Cleveland Clinic Foundation in January 2000, noting that prior communications with faculty members at the SDRC served as a positive influence for his recruitment here. The potential to utilize the expertise in Core A (Cellular and Molecular Morphology) and to access their extensive bank of epidermal differentiation markers (antibodies, probes) should prove useful for both of Dr. Maytin's projects. Core B (Cell Culture and Molecular Technology) is anticipated to provide expertise in gene-delivery techniques; in particular, Dr. Maytin plans to use the core to create adenoviral vectors designed to deliver C/EBP factors into keratinocytes for studies on differentiation. Dr. Maytin also would like to utilize the power of the Functional Genomics Program within Core D (Animal Experimentation ).

Two recent representative publications:

1. Maytin EV, Habener JF, Transcription factors C/EBPa, C/EBPb, and CHOP (gadd153)
expressed during the differentiation program of keratinocytes in vitro and in vivo. J Invest Dermatol 110: 238-246, 1998.

2. Maytin EV, Lin J, Krishnamurthy R, Batchvarova N, Ron D, Mitchell PJ, Habener JF, A temporally- and spatially-defined network of transcription factors C/EBPa, C/EBPb, and AP-2 determines differentiation-specific expression of keratin K10. Dev Biol 216:
164-181, 1999.

Thomas S. McCormick, Ph.D.
Assistant Professor of Dermatology
Member, Immunology/Microbiology Research Area

Dr. McCormick's research is focused on macrophages and the participation of these cells in the cutaneous immune response. Macrophages are important participants in the inflammatory milieu, as they produce many of the cytokines essential for the inflammatory response to ensue. Macrophages have also been implicated in mediating inflammatory responses involved with graft rejection and in initiating signaling for hyperproliferation.

Attachment and migration through the vasculature is an integral part of the macrophage response to psoriasis. Integrins are essential cell surface molecules which anchor cells to their substrata via specific adhesion molecules. The expression of integrins associated with changes in cell cycle proliferation has been previously described. In psoriasis, macrophages that are recruited to the site of inflammation and keratinocyte hyperproliferation may be responsible for the deposition of a component of the extracellular matrix fibronectin, that has been indicated as participating in the upregulation of keratinocyte hyperproliferation. The interaction of keratinocyte integrin receptors with the fibronectin splice variant EDA fibronectin may provide part of the signaling mechanism that results in keratinocyte hyperproliferation, a hallmark of the psoriasis phenotype.

Our work is focused on the signaling pathways associated with keratinocytes that encounter altered fibronectin products. The role of the macrophage as either an active participant in this stimulation, or as a facilitator of fibronectin assembly is currently being investigated. Using current molecular tools, we are constructing inducible expression vectors that overexpress the fibronectin splice variants. Our long-range plan is to use these overexpression variants, which we have transfected into fibroblast cell lines, in skin equivalent models in combination with macrophages as well as other cells of the immune lineage to test the keratinocyte response to both macrophage and immune cell cytokines in the presence of extracellular matrices that are present in psoriatic skin.

Dr. McCormick's broad expertise is applied to collaborations with Dr. Heinzel on Leishmaniasis and FLT3 Ligand APC's, Drs. Cooper and Kang on psoriasis, and Dr. Mukhtar on UV immuno-carcinogenesis.

Two recent representative publications:

1. Katiyar, S.K., Challa, A., McCormick, T.S., Cooper, K.D., and Mukhtar, H. Prevention of Ultraviolet B-induced immunosupression in mice by green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG): Alterations in IL-10 and IL-12 production by EGCG play a critical role. Carcinogenesis;20:2117-2124, 1999.

2. Ting, K.M., Rothaupt, D., McCormick, T.S., Hammerberg, C., Chen, G., Gilliam, A.C., Stevens, S.R., Culp, L., and Cooper, K.D. 1999. Oncofetal fibronectin containing EDA splice variant is overexpressed in the dermal epidermal junction and penetrates to the proliferative basal keratinocyte compartment in psoriasis. J Invest Dermatol.
114:706-711, 2000.

William C. Merrick, Ph.D.
Professor, Department of Biochemistry
Professor, Division of General Medical Sciences (Oncology)
Director, Molecular Biology Core Laboratory

Dr. Merrick's laboratory has been interested in the mechanism and regulation of eukaryotic protein synthesis, which they have studied through the use of unique in vitro assays using purified components. Much of the purification and characterization of the translation factors has been performed in my laboratory while elements relating to regulation have been done in collaboration with others, most notably Dr. Jolinda Traugh. In the broad area of translation mechanism, current areas of focus are: structure/function of eEF1A as analyzed by site-directed mutagenesis in yeast; sequential function of initiation factors, especially eIF4A, eIF4B and eIF4F which are required for binding mRNA to 40S subunits; analysis of the mechanism of RNA helix unwinding by eIF4A and eIF4B; analysis of the mechanism of mRNA "scanning" as associated with internal initiation and re-initiation.

The laboratory will continue to examine the pathway of initiation complex formation, but it is anticipated that the use of physical means will be more a part of these studies to allow quantitation of the affinities of the various components of translation for each other. These efforts will also be enhanced by the availability of proteins expressed in either E. coli or yeast. Beyond this there are two specific projects which will be pursued. Our second area of research is to evaluate the binding of aminoacyl-tRNA to translation factors. Currently, a crystal structure exists for the prokaryotic ternary complex, EF1AoGDPNPoPhe-tRNA, but there is no equivalent structure for the related eukaryotic proteins, eEF1A, eIF2 or eIF2A. Efforts to obtain such information in the eukaryotic system will use a variety of approaches including chemical cross-linking, site-directed mutagenesis and attempts at crystallization.
Dr. Merrick is Director of the Molecular Biology Core Laboratory, which is co-funded by the SDRC, the Cancer Center and the CFAR. He has extensive experience in protein sequencing and peptide separation, and in advising SDRC members on structural biology applications available in this core and the Structural Biology Center.

Two recent representative publications:

1. Rogers, Jr., G. W., Richter, N. J. and Merrick, W. C. Biochemical and kinetic characterization of the RNA helicase activity of eukaryotic initiation factor 4A. J. Biol. Chem. 274, 12236-12244, 1999.

2. Richter, N. J., Rogers, Jr., G. W., Hensold, J. O. and Merrick, W. C. Further
biochemical and kinetic charaterization of human eukaryotic initiation factor 4H. J. Biol. Chem. 274, 35415-35424, 1999.

Vincent M. Monnier, M.D.
Professor of Pathology and Biochemistry
Previous Pilot and Feasibility Study Recipient

Dr. Vincent Monnier's research focuses on the role of the Maillard reaction in the aging of long-lived proteins, in particular skin collagen. His laboratory has demonstrated that reducing sugars form protein crosslinks in aging collagen involve oxidant stress. One of those crosslinks, pentosidine, was found to form in skin collagen at a rate inversely proportional to maximal longevity in eight vertebrate animal species. Pentosidine was also found to increase with age of the donor in peripheral T-cells and to correlate with the severity of diabetic complications.

In ongoing studies, Dr. Monnier and associates found that carboxymethyl-lysine (CML), the major advanced glycation end product in aging skin and skin undergoing actinic elastosis, can bind redox active transition metals such as copper. The copper bound to protein leads to protein degradation in presence of H202. Using CML-rich protein from serum of uremic patient that was immunoprecipitated with a CML antibody, they found that such protein generated free radicals in presence of the spin trap DMPO and H202.

With the help of SDRC funding, Drs. Monnier and Sell tested the hypothesis that UVB is a major mechanism for CML formation in the sun exposed skin. The mechanism appears to involve UVB induced oxidation of polyunsaturated free fatty acids (PUFAs) to form glyoxal, i.e. the immediate precursor of CML. The significance of their research lies in the fact that both normal and photooxidation induced skin collagen aging appears to proceed, surprisingly, by very similar mechanism. Excessive binding of redox active copper in sun exposed CML-rich lesions may lead to site specific generation of oxidative stress and contribute to actinic carcinogenesis.

Dr. Monnier's work has involved every core of the SDRC cores, and he has been an enthusiastic participant in the enrichment programs, the P&F review process, and general advisement.

Two recent representative publications:

1. Saxena, AK, Saxena, P, Wu, X. Obrenovich, M, Weiss, MF, Monnier, VM. Carboxymethylation of protein lysines generates sites for redoxactive and divalent metal binding. Biochem. Biophys. Res. Comm. 260: 332-8, 1999

2. Sell DR, Kleinman N, Monnier VM. Glycation and glycoxidation of skin collagen predicts longevity in C57NNia/BL mouse. FASEB J. 14:145-56, 2000.

Joseph H. Nadeau, Ph.D.
Professor of Genetics
Co-Director, Animal Experimentation Core

Dr. Nadeau's research focuses on the genetic and phenotypic dissection of selected mouse models of human multifactorial disease traits. The goal of this work is to develop general methods for analyzing complex traits in experimental systems and learn about the particular disease models. These studies use a combination of expression assays for large numbers of genes, measurement of metabolite levels and enzyme activities in normal and variant mice with emphasis on folate metabolism and inositol phospholipid metabolism, and linkage analysis of genes controlling these traits as well as disease susceptibility genes. With these various assays, we have identified new mouse models for hyperhomocysteinemia or reduced methylenetetrahydrofolate reductase (MTHFR) activity. In addition, we have identified anomalies in gene expression profiles in these mice and we have begun to define new networks of gene expression control. Other current work focuses on positional cloning and characterization of the Disorganization mutation, a single gene that causes an extra-ordinary variety of birth defects, the genetic control of susceptibility to testicular germ cell tumors, and the genetic and developmental control of drug-induced birth defects. Finally, we have pioneered the mathematical analysis of comparative genetic maps as a means for studying genome organization and evolution. Current work focuses on the evolution of gene families and physiological pathways.

Finally, we are developing an interdisciplinary program to identify and characterize many new mouse models of human genetic disease. These mutants are being created with chemical mutagenesis and are identified and characterized in collaboration with research programs in the School of Medicine. In collaboration with the SDRC (Drs. Cooper, Gilliam, Eckert and Mukhtar), the emphasis is on new models of skin cancer, skin inflammation and cutaneous structure. It is anticipated that SDRC's support of Dr. Nadeau's program, through the Animal Experimentation Core, will lead to new models of skin disease.

Two recent representative publications:

1. Nadeau JH, Singer JB, Matin A, Lander ES. Analyzing complex genetic traits
with chromosome substitution strains. Nat Genet:221-5, 2000.

2. Matin A, Collin GB, Asada Y, Varnum D, Nadeau JH. Susceptibility to testicular germ
cell tumours in a 129.MOLF-Chr 19 chromosome substitution strain. Nat Genet. :237-240, 1999.

Anna-Liisa Nieminen, Ph.D.
Assistant Professor of Anatomy
Confocal Microscopy Facility
Consultant, Cellular and Molecular Morphology Core

Dr. Anna-Liisa Nieminen's research interests concern the cellular and molecular mechanisms underlying necrotic and apoptotic cell death. In particular, her laboratory is applying new techniques of laser scanning confocal microscopy to characterize ion homeostasis, mitochondrial function, protease and phospholipase activation, membrane permeability and other biochemical parameters during the pathogenesis of lethal cell injury. Dr. Nieminen has over ten years of experience in digitized video microscopy and over five years of experience in the techniques of laser scanning confocal microscopy.

A phenomenon called the mitochondrial permeability transition plays an important role in cell injury. Increases of mitochondrial free Ca2+, formation of reactive oxygen species, and oxidation of mitochondrial pyridine nucleotides and glutathione promote the mitochondrial permeability transition that, in turn, leads to mitochondrial depolarization and uncoupling of oxidative phosphorylation. Recently, she showed the occurrence of the mitochondrial permeability transition in models of oxidative stress and excitotoxicity. Inhibitors of the permeability transition, like cyclosporin A and trifluoperazine, reduce lethal cellular injury in these models. These findings offer new strategies to rescue cells and tissues from irreversible toxic injury.

The second focus of her research is to define cellular mechanisms underlying apoptotic death in different cell types. Specifically, she is studying how mitochondria are involved in apoptotic cell death. Along this line of investigation, she is collaborating with Dr. Mukhtar to examine the possible anti-apoptotic effect of green tea on human carcinoma cells using confocal microscopy. She is also collaborating with Drs. Mukhtar and Oleinick to define the role of mitochondria in apoptotic death following photodynamic therapy.

Dr. Nieminen's lab is on the same floor as the Dermatology labs in the BRB, which greatly facilitates her core service functions.

Two recent representative publications:

1. Byrne AM, Lemasters JJ, Nieminen AL. Contribution of increased mitochondrial free Ca2+ to the mitochondrial permeability transition induced by tert-butylhydroperoxide in rat hepatocyte. Hepatology;29:1523-1531, 1999.

2. Heiskanen KM, Bhat MT, Wang HW, Ma J, Nieminen AL. Mitochondrial depolarization accompanies cytochrome c release during apoptosis in PC6 cells. J Biol Chem; 274:
5654-5658, 1999.

Nancy L. Oleinick, Ph.D.
Professor of Radiation Oncology
Member, Executive Committee

Dr. Oleinick's laboratory is investigating the mechanisms of cell killing by Photodynamic Therapy (PDT), focusing on a variety of tumor types that originate in or metastasize to the skin. The research is part of a multidisciplinary Program Project Grant funded by the National Cancer Institute (N. L. Oleinick, Principal Investigator) that is directed to the preparation and photophysical evaluation of new photosensitizers of the phthalocyanine type (Project 1); elucidation of the molecular and cellular mechanisms of PDT with phthalocyanines (Project 2), and investigation of the responses of normal and tumor tissue to PDT (Project 3). The aims of the project Dr. Oleinick directs are to elucidate the pathway for signal transduction triggered by PDT which leads to apoptosis, to sensitize cells to PDT, to develop transgenic tumor cell systems for studying apoptosis in vivo, and to evaluate the mutagenicity of PDT. She has found that PDT induces a series of molecular signals, including release of the membrane lipid second messengers, inositol trisphosphate and ceramide, mobilization of calcium ion, and activation of stress protein kinase cascades and proteases. The interaction of pro- and anti-apoptotic signals is being deciphered in PDT-treated cells. Dr. Oleinick's laboratory collaborates with Dr. Mukhtar's in studies on the cytotoxic mechanisms of PDT in vivo and in vitro, and the preclinical work of Pc 4 has been supported by SDRC cores and the enrichment program. Our lead photosensitizer, the phthalocyanine Pc 4, is now in the final stages of pre-clinical testing by the NCI's Drug Decision Network, in preparation for clinical trials. It is anticipated that, once an Investigational New Drug number is obtained, the first trial will be for cancers in the skin and will be a joint effort of the Skin Diseases Research Center and the CWRU/Ireland Comprehensive Cancer Center. The Translational Studies Core of the SDRC will play a major role, as will the Cell Morphology and Molecular Biology Core. Dr. Oleinick has been a key member and supporter of the SDRC.

Two recent representative publications:

1. Xue Ly, He J, Oleinick NL. Promotion of photodynamic therapy-induced apoptosis by stress kinases. Cell Death Differ;6:855-864, 1999.

2. Xue LY Qiu Y, He J, Kung HJ, Oleinick NL. Etk/Bmx, a PH-domain containing tyrosine kinase, protects prostate cancer from apoptosis induced by photodynamic therapy or thapsigargin. Oncogene;18:3391-3398, 1999.

Eric Pearlman, Ph.D.
Assistant Professor of Medicine (Geographic Medicine)
Previous Pilot and Feasibility Study Recipient

Dr. Pearlman's laboratory focuses on mechanisms of pathogenesis induced by the parasitic helminth that causes River Blindness (onchocerciaisis) which is prevalent throughout West and Central Africa and in Central America. Two projects are currently underway to investigate corneal and skin disease induced by this parasite. The project on corneal pathology involves the use of a murine model in which animals are injected intracorneally with parasite antigens, and regulation of inflammatory cell recruitment is determined by specific depletion of suspected mediators. The second project is on human skin disease and takes the following approaches: a) characterization of skin lesions in acute, early stage of onchocercal skin disease; b) temporal analysis of expression of chemokines in skin of infected individuals after topical application of antihelminthics. (This treatment, which is used routinely to determine levels of infectivity, induces a papular response that is very similar clinically and histologically to early stage onchodermatitis); c) a murine model of skin disease has been established to determine the requirements for specific mediators in recruitment of inflammatory cells to the skin. Studies on infected individuals are conducted in collaboration with the World Health Organization in Ivory Coast, West Africa. Dr. Pearlman's work is highly relevant to all skin diseases which involve eosinophils, and was supported by a P&FS award.

Two recent representative publications:

1. Pearlman, E., C.A. Garhart, D.J. Grand, E. Diaconu, E.R. Strine, and L.R. Hall. 1999.
Temporal recruitment of neutrophils and eosinophils to the skin in a murine model for
onchocercal dermatitis. Amer J. Trop Med Hyg. 61:14-18.

2. Pearlman E, Toe L, Boatin BA, Gilles AA, Higgins AW, Unnasch TR. Eotaxin expression in Onchocerca volvulus-induced dermatitis after topical application of diethylcarbamazine. J Infect Dis;180:1394-1397.

Thomas G. Pretlow, M.D.
Professor of Pathology, Oncology, Urology, and Environmental Health Sciences Consultant, Cellular and Molecular Morphology Core

Dr. Tom Pretlow's work and long-term interests are concentrated in two areas: the investigation of pre-neoplastic lesions and the development of prostate cancer models that will be useful for the investigation of clinical problems in humans.

His interest in pre-neoplastic lesions began in the late 1970s when he investigated phenotypic changes described by others in pre-neoplastic hepatic lesions in mice, called "enzyme-altered foci" by the several earlier investigators who had discovered them, and developed several methods for the purification of the cells that composed these lesions from other liver cells. More recently, Dr. Pretlow has found that alterations of multiple phenotypic markers occurs in morphologically normal portions human prostates from patients with prostate cancer. These alterations have included both enzyme-histochemical changes and antigenic changes. His work with pre-neoplastic lesions in liver, colon, and prostate led him to believe that enzyme-histochemical and antigenic changes in pre-neoplastic lesions precede morphological changes and, in many organ systems, are the first indications of neoplastic transformation. The approaches used with these organs are likely to be highly applicable to the abnormalities in skin cancer.

Dr. Pretlow's laboratory will provide expertise in laser capture microscopy for this purpose, a device which was purchased in part by the Department of Dermatology for utilization by SDRC membership through the core mechanism.

Two recent representative publications:

1. Hao XP, Willis JE, Pretlow TG, Rao JS, MacLennan GT, Talbot IC, Pretlow TP. Loss of fragile histidine triad expression in colorectal carcinomas and premalignant lesions. Cancer Res;60:18-21, 2000.

2. Qiu Y, Robinson D, Pretlow TG, Kung HJ. Etk/Bmx, a tyrosine kinase with a pleckstrin-homology domain, is an effecto phosphatidylinositol 3'kinase and is involved in interleukin 6-induced neuroendocrine differentiation of prostate cancer cells. Proc Natl Acad Sci USA;95:3644-3649, 1998.

Scot C. Remick, M.D.
Associate Professor of Medicine, Division of Hematology/Oncology
Program Leader, Developmental Therapeutics
Director, International Clinical Coordination Center Core Facility, CFAR
Member, Cutaneous Oncology Research Area
Current Pilot and Feasibility Study applicant

Dr. Remick joined the faculty of CWRU in October 1996 and is Program Leader of the Developmental Therapeutics core program of the Cancer Center and serves as Director of the newly established International Clinical Coordinating Center (ICCC) core facility of CWRU Center for AIDS Research (CFAR). His clinical research interests focus on early mechanism- based phase I studies of novel antineoplastic agents and phase II/III clinical trials in cancer and HIV infection with a focus on AIDS-related neoplasms, and quality-of-life outcome measurements. As Program Leader of the Developmental Therapeutics Program, Dr. Remick will have primary responsibility for coordinating and directing the phase I clinical trial of silicon phthalocyanine (Pc4)-photodynamic therapy (PDT), which is a novel second generation photosensitizer that was synthesized at CWRU. The Skin Diseases Research Center will closely collaborate on the development of Pc4-PDT for treatment of cutaneous malignancies at our institution. An exciting P&F on Pc4-PDT mechanisms of action in human in vivo skin cancer was submitted for this application by Dr. Remick, but was felt to be more appropriately funded in the next year, when the anticipated IND will be awarded and the clinical trial would actually begin.

Dr. Remick has clinical expertise in the management of AIDS-related malignancies and has published extensively in this area. He has developed an oral combination chemotherapy regimen for HIV-related lymphoproliferative disease and continuous infusion chemotherapy schedules for AIDS-related Kaposi's sarcoma in an effort to improve the therapeutic index of existing cytotoxic agents. He is interested and has published on the expanding spectrum of neoplastic disease in HIV infection and collaborated on the prevalence of HHV-8 in non-AIDS-defining neoplasms. Dr. Remick's interests in AIDS-related Kaposi's sarcoma clearly complements an area of research focus within the Skin Diseases Research Center. His expertise and participation is a key element in a developing multidisciplinary program in Kaposi's sarcoma.

Two recent representative publications:

1. Remick, S.C., Sedransk, N., Haase, R., Craffey, M., Subramanian, N., Dowlati, A., Nazeer, T., Ramnes, C., Blanchard, C., Mastrianni, D., Balducci, L., Horton. J., and Ruckdeschel, J.C. Oral combination chemotherapy in the management of AIDS-related lymphoproliferative malignancies. Drugs 58:99-107, 1999.

3. Levitan N, Dowlati A, Shina D, Craffey M, Mackay W, DeVore R, Jett J, Remick, SC, Chang A, Johnson D. Multi-institutioal phase I/II trial of paclitaxel, cisplatin, and etoposide with concurrent radiation for limited-stage small-cell lung carcinoma. J Clin Oncol;18:1102-1109, 2000.

Ellen A. Rorke, Ph.D.
Assistant Professor of Environmental Health Sciences
Previous Pilot and Feasibility Study Recipient
Current Pilot and Feasibility Study Applicant

Dr. Rorke studies retinoids effects on keratin gene expression and retinoid control of cell proliferation. Following HPV immortalization, cervical and epidermal keratinocytes are 10-100 fold more sensitive to retinoids including trans-retinoic acid. The major area being addressed is the elucidation of the mechanism underlying this altered cell responsiveness to retinoids following cell immortalization with the Human Papillomavirus (HPV). She has shown this change to occur independently of HPV oncogene expression and without changes in retinoid receptor expression. The second research area is the role of cell cycle proteins during growth inhibition by t-RA, TGFb, and interferon g in normal and HPV immortalized cervical cells. HPV oncogenes bind to and target two tumor suppresser proteins (p53 and RB) for proteolytic degradation. It is of interest to determine if their expression as well as the expression of other cell cycle regulators (such as cyclins and cyclin dependent kinases) are necessary for growth inhibition. The third research area focuses on the effects of environmental carcinogens (i.e. benzo[a]pyrene, TCDD, etc.) on epithelial cell transformation. Her work demonstrated that normal keratinocytes but not HPV immortalized ones undergo cell death following carcinogen insult. Furthermore, only the HPV immortalized cells can be transformed in vitro by these chemicals. Studies are being conducted on DNA damage, repair and mutagenesis to understand why immortalized cells are at increased risk of transformation.

Dr. Rorke actively collaborates with Drs. Mukhtar and Eckert, and provides an excellent resource for SDRC members who contemplate using retinoids in their work. she utilizes mainly the Animal Core, the Cell and Molecular Technology Core, and the Morphology Core.

Two recent representative publications:

1. Crish JF, Bone F, Balasubramanian S, Saim TM, Wagner T, Yun J., Rorke EA, Eckert RL. Suprabasal expression of the human papillomavirus type 16 oncoproteins in mouse epidermis alters expression of cell cycle regulatory proteins. Carcinogenesis; 21:
1031-1037, 2000.

2. Eckert RL, Crish JF, Balasubramanian S, Rorke EA. Transgenic animal models of human papillomavirus-dependent disease. Int J Oncol;16:853-870, 2000.

David R. Sell, Ph.D.
Assistant Professor of Pathology
Previous Pilot and Feasibility Study Recipient

The overall research focus is on the study of amino-carbonyl reactions of sugars with long-lived proteins as the basis for post-translational modifications of proteins during aging. Since oxidation is involved in many of these reactions, they have been referred to as glycoxidation. Dr. Sell has found skin collagen extremely useful to study these reactions. Because of its slow turnover, these products accumulate in this tissue during aging and certain disease processes such as diabetes and uremia. Furthermore, skin samples which are readily available through biopsies from human volunteers (Translational Study Core) and animals (Animal Experimentation Core) are being examined for such studies.

The most striking observation from this research is that levels of these products in skin are influenced by both genetic and environmental factors. Recently, we have become interested in the study of these products in relation to photooxidation and aging. The exact biochemical events of photoaging are not fully understood. However, the deleterious effects of sunlight have often been attributed to oxidation based upon a free radical mechanism, similar to that proposed for chronological aging. A significant finding is that glycoxidation independent of sunlight exposure increases proportionally with age in skin inversely related to species longevity and can predict individual longevities in mice when studied longitudinally. One biochemical marker for the assessment of glycoxidation is pentosidine, a difunctional protein crosslink consisting of a pentose sugar crosslinked to lysine and arginine residues. A further marker is N?-(carboxymethyl)lysine (CML), an oxidized form of lysine produced by the reaction of reducing sugars with lysine in the presence of oxygen. However, recent evidence suggests a mechanism of CML formation through polyunsaturated fatty acids (PUFA), most likely through the reactive intermediate glyoxal. Levels of CML progressively increase with age in unexposed human skin and are accelerated by actinic solar elastosis of photoaged skin. Our current focus is on mechanisms and processes that influence the formation of such products in skin and their interventions.

Two representative papers:

1. Sell DR, Lane MA, Johnson WA, Masoro EJ, Mock OB, Reiser KM, Fogarty JF, Cutler RG, Ingram DK, Roth GS, Monnier VM. Longevity and the genetic determination of collagen glycoxidation kinetics in mammalian senescence. Proc. Natl. Acad. Sci. USA 93:485-490, 1996.

2. Sell DR, Kleinman NR, and Monnier VM. Longitudinal determination of skin collagen glycation and glycoxidation rates predicts early death in C57BL/6NNia mice. FASEB J. 14:145-156, 2000.

Nora G. Singer, M.D.
Assistant Professor of Pediatrics and Internal Medicine
Previous Pilot and Feasibility Study Recipient

Dr. Nora G. Singer's research interest is in T-cell development and autoimmunity. Her primary interest has been in the role of CD6 as a T-cell co-stimulatory molecule in autoimmunity. Prior to leaving her former institution she generated a monoclonal antibody that appears to recognize a novel CD6 ligand expressed in skin. Her current interests are in cloning the cDNA for this novel CD6 ligand, and in understanding its function in normal and diseased skin, and in thymic development.

Dr. Singer is currently developing murine models in which to test the function of T-cell CD6 and its epithelial ligands using CD6-overexpressing transgenic mice, TCR transgenic mice and autoimmune-prone mice. She hopes that elucidation of the CD6-dependent path of co-stimulation will lead to manipulation therapeutically of CD6 and its ligands to treat tissue specific manifestations of autoimmune disease.

Two recent representative publications:

1. Singer NG, McCune WJ. Prevention of infectious complications in rheumatic disease patients: immunization, Pneumocystis carinii prophylaxis, and screening for latent infections. Curr Opin Rheumatol;11:173-178, 1999.

3. Singer NG, McCune WJ. Update on immunosuppressive therapy. Curr Opin Rheumatol;10:169-173, 1998.

J. Michael Sorrell, Ph.D.
Senior Research Associate, Biology
Member of Cell Biology Thematic Area

Dr. Sorrell is engaged in the study of extracullular matrix molecules of the dermis and their modification by age and production by papillary vs. reticular dermal fibroblasts. He has developed a set of monoclonal antibody reagents for detecting proteoglycans on the surfaces of cells. He is extremely active in collaborative and technical exchanges between Dr. McCormick's Dr. Cooper's, Dr. Eckert's and Dr. Hascall's labs in regard to skin equivalents.

Two recent representative publications:

1. Carrino, D.A., Sorrell, J.M., and Caplan, A.I. "Age-Related Changes in the Proteoglycans of Human Skin", Arch. Biochem. Biophys, 373:91-101, 2000.

2. Sorrell, J.M., Seavolt, M.B., Baber, M.A., Carrino, D.A., Asselineau, D.A., and Caplan, A.I. "Development and Microdomain Distribution of Cell Surface Markers for Human Dermal Fibroblasts". Submitted, Exp. Cell Res, 2000.

Timothy P. Spiro, M.D.
Assistant Professor of Medicine
Hematology/Oncology Division
Member, Oncology Research Area

Dr. Tim Spiro's major interest is the study of tumor cell resistance to chemotherapy, and in particular, the role of DNA repair enzymes in the resistance to nitrosoureas and related agents. The nitrosoureas form adducts at the 06 position of guanine in DNA, in a reaction that subsequently results in cytotoxic crosslinks. The DNA repair enzyme alkylguanine DNA alkyltransferase repairs these adducts, preventing crosslink formation. This ubiquitous DNA repair protein is frequently elevated in a broad range of tumors, and Dr. Spiro and his colleagues have developed sensitive assays for the measurement of this DNA repair protein directly in tumor tissue. Dr. Spiro conducts a translational clinical research program which focuses on the development of new anticancer agents effective in the total depletion of alkylguanine DNA alkyltransferase. Direct tissue alkyltransferase measurements have allowed him to develop novel dose escalation strategies that monitor this efficacy endpoint in tumors, and other surrogate marker tissues such as peripheral blood mononuclear cells. Another aspect of his program is the detailed assessment of pharmacokinetics and metabolism, and the study of pharmacokinetic/pharmacodynamic relationships applied to the development of new anticancer drugs. One such drug is 06-benzylguanine, an agent currently under study through the NCI/CTEP.

Dr. Spiro has a specific interest in dermatologic malignancies, cutaneous T-cell lymphoma and malignant melanoma, both of which have been shown to have tumor cell resistance mediated by alkylguanine DNA alkyltransferase. Building upon data already obtained in the laboratory and in a recently completed Phase I clinical trial with 06-benzylguanine, two clinical trials are soon to be initiated. The first trial is a Phase I study of this agent in cutaneous T-cell lymphomas. 06-benzylguanine will be given intravenously at a fixed dose, along with escalating doses of topical BCNU. This study was initiated as a result of multidisciplinary interactions sponsored by Dermatology and the SDRC, and is now NIH-funded. In a separate Phase II study, a fixed combination of 06-benzylguanine and BCNU will be given intravenously to patients with metastatic malignant melanoma.

Two recent representative publications:

1. Spiro TP, Gerson SL, Liu L, Majka S, Haaga J, Hoppel CL, Ingalls ST, Pluda JM, Willson JK. O6-benzylguanine: a clinical trial establishing the biochemical modulatory in tumor tissue for alkyltransferase-directed DNA repair. Cancer Res; 59:2402-2410, 1999.

2. Stefan TL, Ingalls ST, Minkler PE, Willson JK, Gerson SL, Spiro TP, Hoppel CL. Simultaneous determination of O6-benzylguanine and 8-oxo-O6-benzylguan in human plasma by reversed-phase high-performance liquid chromatograph. J Chromatogr B Biomed Sci Appl:289-298, 2000.

Seth R. Stevens, M.D.
Assistant Professor of Dermatology
Director, Translational Research Core
Previous Pilot and Feasibility Study Recipient
Director, Enrichment Program

Dr. Seth Stevens' program focuses on mechanisms of distinct T cell activation induced by two antigen presenting cells in human epidermis, the Langerhans cell of normal skin and macrophages which infiltrate UV-irradiated skin (UV-macrophages). T cell-mediated immunity activated by these two antigen presenting cells is associated with sensitization (Langerhans cells) and tolerization (UV-macrophages). His work has defined a novel, IL-2 receptor-deficient T cell activation state, which has subsequently also been associated with tolerance in other systems. Specific differences in these antigen presenting cells and the T cell differentiation/activation states this work defines is being explored. His work has important implications for defining the effects of UV on skin immunology in normal humans and the etiology of UV-induced skin cancers, phototherapy of inflammatory and malignant skin diseases and photosensitivity disorders.

A second program addresses the development of appropriate, objective skin disease assessment tools to replace the more subjective ones frequently used in such diseases as cutaneous T cell lymphoma. The Severity-Weighted Assessment Tool developed in collaboration with Dr. Kevin Cooper has been adopted by the International Society for Cutaneous Lymphomas as the gold standard in this field. Dr. Stevens has also collaborated with Dr. Chren in optimizing the Skindex outcomes tool for patients with CTCL.

Two recent representative publications:

1. Stevens SR, Kang K, Cooper KD. Atopic dermatitis: introduction and overview. J Cutan Med Surg; 2:S2-2-S2-7, 1999.

2. Kremer IB, Cooper KD, Teunissen MBM and Stevens SR. Low expression of CD40 and B7 on macrophages infiltrating UV-exposed human skin; role in IL-2Ralpha-T cell activation. Eur J Immunol; 28:2936-2946, 1998.

Man-Sun Sy, Ph.D.
Professor of Pathology
Previous Pilot and Feasibility Recipient
Member, Executive Committee

One of the cell surface molecules that has been implicated to play an important role cellular migration is CD44. One of the ligands for CD44 is hyaluronic acid (HA), a polysaccharide present in tissues and in the circulation. Normal human lymphocytes do not bind HA. Since CD44 is expressed on many cell types and HA is present in most tissues and in the circulation, therefore, interactions between CD44 and HA must be regulated. Dr. Sy's laboratory showed that binding activity of CD44 is regulated in some cells, and not all CD44+ cells can bind HA. Binding of CD44 to HA on a human lymphoma, Jurkat, requires cellular activation. Activation of CD44+ Jurkat cell results in the dimerization of CD44 involving a cysteine in the transmembrane domain. Dimerization of CD44 is essential for binding of high levels of HA. We are continuing to investigate the mechanisms regulating CD44 dimerization. Since CD44 are highly expressed on all major cell types in the skin, and skin has the highest concentrations of HA, abnormal interactions between CD44 and HA may have pathological consequence. A better understanding on the molecular mechanisms regulation the interactions between CD44 and HA may be relevant to inflammatory diseases of the skin. In addition, Dr. Sy has extended these findings to melanoma.

Two recent representative publications:

1. Liu D, Liu T, Sy MS. Identification of two regions in the cytoplasmic domain of CD44 through w PMA, calcium, forskolin differentially regulate the binding of CD44 to hyaluronic acid. Cell Immunol; 190:132-140, 1998.

2. Zanusso G, Liu D, Ferrari S, Hegyi I, Yin X, Aguzzi A, Hornemann S, Liemann S, Glockshuber R, Manson JC, Brown P, Petersen RB, Gambetti P, Sy MS. Prior protein expression in different species: analysis with a panel of new m Proc Natl Acad Sci USA;95:8812-8816, 1998.

Magdalena Tary-Lehmann, M.D., Ph.D.
Assistant Professor of Pathology
Member, Immunobiology/Microbiology Research Area

The focus of research in the laboratory of Dr. Tary-Lehmann is on the characterization of T cell responses to tumors, including those of the skin. The primary goal is to understand what are the rules for the induction of anti-tumor T cell responses, what it takes to engage different effector classes of T cell immunity, including proinflammatory T helper I (Thl) and anti-inflammatory Th2, and how these different types of responses affect tumor growth. The classic "two signal" model holds that T lymphocytes require 2 signals to differentiate into effector cells. First, the recognition of antigen in the context of MHC molecules on antigen presenting cells (APC); and second, costimulation via the accessory molecules that are induced by trauma or "danger" on APC. Signal one alone, in absence of signal two, is thought to inactivate the T cell, via the induction of apoptosis or of unresponsiveness (anergy). According to this classical concept, tumors should be non-stimulatory, because most of them do not express and can not be induced to express the costimulatory molecules that provide 4 6 "signal 2". Dr. Tary-Lehmann's studies, using a modified cytokine ELISA spot assay that has highly improved sensitivity, provided data that questions this classic viewpoint. We have shown that most tumors are immunogenic, but induce the non-destructive, Th2 type T cell response. While it has been believed that the immune system fails to reject cancer cells due the lack of an immune response, our data suggest that tumors grow in spite of a T cell response that, however, does not lead to cancer cell rejection and might even protect the cancer cells: Th2 responses are known to protect normal tissues from an autoimmune T cell attack. Currently Dr. Tary-Lehmann is pursuing this model in murine cancer and in human patients by assessing their T cell responses to their own cancer cells. For dermatologically relevant tumors Dr. Tary-Lehmann is collaborating with Dr. Stevens in characterizing the T cell responses to cutaneous T-cell lymphomas (CTCL) and the anti-tumor response of Sezary syndrome patients. In addition, she collaborates with Drs. Stevens, Cooper and McCormick on ELISA spot assessments in UV and psoriasis systems.

Two recent representative publications:

1. Helms T, Boehm BO, Asaad RJ, Trezza RP, Lehmann PV, Tary-Lehmann M. Direct visualization of cytokine-producing recall antigen-specific CD4 memory T cells in healthy individuals and HIV patients. J Immunol;164:3723-3732, 2000.

2. Heeger PS, Greenspan NS, Kuhlenschmidt S, Dejelo C, Hricik DE, Schulak JA, Tary-Lehmann M. Pretransplant frequency of donor-specific, IFN-gamma-producing lymphocy is a manifestation of immunologic memory and correlates with the risk of posttransplant rejection episodes. J Immunol;163:2267-2275, 1999.

Dennis J. Templeton, M.D., Ph.D.
Associate Professor of Pathology
Previous Pilot and Feasibility Study Recipient

Dr. Templeton's laboratory focuses on the transmission of intracellular signals originating in cell stress. Primarily, they are interested in the cascade of protein kinases that serve as mediators of this pathway. The lab was the first to characterize the now broadly known as MEKK>SEK> APK/JNK pathway, and we have continued to study the mechanism of activation and consequences of stress activation. One of Dr. Templeton's major directions led into the control of MEKK1 function by redox mechanisms. This work identified a new class of stress-inhibiting drugs that are all related to the quinone reductase-sulfhydryl "redox cycling" cycle. Other work has examined the role of SAPK activation in control of the cell cycle. Dr. Templeton has found, for example, that SAPK is activated during late G2, and that inhibition of SAPK activation with a dominant inhibitory mutant transduced by an adenovirus prevents cell cycle passage. Future efforts are aimed at understanding the mechanism of this effect.

Dr. Templeton advises Dr. Stevens on his K08 award and Dr. Ahmad on his proposed P&F award in the current application.

Two recent representative publications:

1. Cross JV, Deak JC, Rich EA, Qian Y, Lewis M, Parrott LA, Mochida K, Gustafson D, Vande Pol S, Templeton DJ. Quinone redutase inhibitors block SAPK/JNK and NfkappaB pathways and potentiate apoptosis. J Biol Chem;274:31150-31154, 1999.

2. Parrott LA, Templeton DJ. Osmotic stress inhibits p70/85 S6 kinase through activation of a protein phosphatase. J Biol Chem;274:24731-24736, 1999.

Zahra Toossi, M.D.
Associate Professor of Medicine (Infectious Diseases)
Member, Immunology/Microbiology Research Area

Research in Dr. Toossi's laboratory is focused on the study of immunoregulation during human tuberculosis and on the impact of tuberculosis on HIV disease. These studies have identified transforming growth factor beta (TGF-beta) as a central cytokine in regulation of the immune response in patients with active pulmonary tuberculosis. Furthermore, this cytokine is overproduced by mononuclear phagocytes of the blood of such patients and at sites of active M tuberculosis infection such as the lung. Ongoing studies will identify the mechanisms of increased production and activation of TGF-beta and the suppressive role of TGF-beta in cytokine circuits during tuberculosis. With regard to the impact of tuberculosis on HIV disease, Dr. Toossi has shown that monocytes from patients with tuberculosis are highly susceptible to a productive HIV infection. This property appears to be related to the in vivo activation of these monocytes and their expanded capacity to produce cytokines. Recent studies have shown that there is a spontaneous activation of nuclear factor kappa b and degradation of its cytoplasmic inhibitor in monocytes from patients. Ongoing studies will determine the consequences of cellular activation on the various steps of the HIV life-cycle.

Dr. Toossi is an active collaborator with Drs. Gilliam and Stevens, and is a consultant on Dr. Gilliam's P&F proposal and an investigator on her NIH K08 application.

Two recent representative publications:

1. Toosi Z, Xia L, Wu M, Salvekar A. Transcriptional activation of HIV by Mycobacterium tuberculosis in human monocytes. Clin Exp Imunol;117:324-330, 1999.

2. Wilkinson RJ, Patel P, Llewelyn M, Hirsch CS, Pasvol G, Snounou G, Davidson RN, Toossi Z. Influence of polymorphism in the genes for the interleukin (IL)-1 receptor antagonist and IL-1 beta on tuberculosis. J Exp Med;189:1863-1874, 1999.

Scott B. Vande Pol, M.D., Ph.D.
Assistant Professor of Pathology
Previous Pilot and Feasibility Study Recipient

Dr. Vande Pol's laboratory studies oncogenes found in papillomaviruses, a class of small DNA tumor viruses now known to be the causative agent in a variety of human epithelial cancers of the skin, ano-genital region, and respiratory tract. He has focused his attention on the papillomavirus E6 genes. Two main projects are in progress in the laboratory: studying the mechanism to targeted protein degradation by the human papillomavirus E6 oncoprotein, and elucidating the mechanism of transformation by Bovine Papillomavirus Type 1 (BPV- 1) E6.

The interaction of the human papillomavirus (HPV) E6 oncoprotein with the cellular p53 tumor suppressor is under active investigation. It is known that 16E6 associates with a mammalian protein called E6AP, and that the complex of E6AP and 16E6 can then associate with the p53 tumor suppressor protein, inducing the ubiquitin-dependent degradation of the tri-molecular complex. Using support from the SDRC we have reconstituted the expression of HPV-16 E6, E6AP and p53 in yeast. The system we devised required extensive experimental development and has now allowed the selection of mutant variants of each component in order to genetically define the structural requirements for both formation of the complex and subsequent ubiquitin dependent degradation. We are interested using this system to gain structural insight into the interaction involved and to elucidate factors that modulate the interaction and degradation of p53 by 16E6.

The second project in the lab concerns the transformation mechanism of the BPV-1 E6 oncoprotein. Normal fibroblasts and epithelial cells as well as immortalized but non-transformed cell lines (such as 3T3 cells) require two separate signals from their environment in order to enter the cell cycle and transit the G1/S boundary: first they must be stimulated by growth factors, and second they must be attached or "anchored" to a suitable immobilized substrate. The attachment signal is mediated by integrin receptors on the cell surface interacting with immobilized "matrix" molecules such as fibronectin, collagen or laminin. BPV-1 E6 can associate with an adapter protein called paxillin that is implicated in the regulation of integrin signaling. Dr. Vande Pol's studies in this area are now being coordinated by the SDRC to interact with Drs. Culp and McCormick on fibronectin isoform and integrin/FAK signaling in psoriasis wound healing.

Two recent representative publications:

1. DAS K, Bohl J, Vande Pol SB. Identification of a second transforming function in bovine papillomavirus ty E6 and the role of E6 transactions with paxillin, E6BP, and E6AP. J Virol;74:812-816, 2000.

2. Vande Pol SB, Brown MC, Turner CE. Association of Bovine Papillomavirus Type 1 E6 oncoprotein with the focal adhesion protein paxillin through a conserved protein interaction motif. Oncogene;16:43-52, 1998.

Martina L. Veigl, Ph.D.
Associate Professor
General Medical Sciences - Oncology
and Envirionmental Health Sciences
Co-Director, Cell Culture and Molecular Technology

Dr.Veigl studies the role of hot spots and genomic instability that occurs during pathways of cancer induction. The studies in her laboratory include investigating the effect of low dose radiation on genomic instability in mammallan cells. For almost ten years she has been directing the Tissue Culture Media Facility of CWRU Comprehensive Cancer Center. Most recently she got involved in setting up a DNA Expression Array Core Facility for the Cancer Center. She is highly trained in DNA expression analysis and several other molecular techniques. She will serve as a resource person for DNA Expression Array work to SDRC investigators.

Two recent representative publications:

1. Chen, W-D, Eshleman, J.R., Aminoshariae, M.R., Ma, A-H, Veloso, N., Markowitz, S.D., Sedwick, W.D. and Veigl, M.L. Cytotoxicity and Mutagenicity of Frameshift-Inducing Agent ICR191 in Mismatch Repair-Deficient Colon Cancer Cells. J. Natl Cancer Inst., 92: 480-485, 2000.

2. Ma, A-H, Xia L., Littman, S.J., Swinler, S., Leder, G., Polinkovsky, A., Olechnowicz, J., Kasturi, L., Lutterbaugh, J., Modrich, P., Veigl, M.L., Markowitz, S.D. and Sedwick, W.D. Somatic Mutation of hPMS2 as a Possible Cause of Sporadic Human Colon Cancer with Microsatellite Instability. Oncogene, 19: 2249-2256, 2000.

Aaron Weinberg, DMD, PhD
Associate Professor, Department of Periodontics
School of Dentistry, CWRU

Dr. Aaron Weinberg's background in clinical periodontics and oral microbiology has led him to investigate the dynamics involved in homeostasis between commensal bacteria and host epithelium. Dr. Weinberg's major research interests are in studying cutaneous and oral epithelial cell innate immune responses to microbial challenges, with an emphasis on epithelial cell derived natural antibiotics. These include the b-defensins (human b-defensin 1, and -2) and the cathelin-like antimicrobial peptide (AP), LL-37. Dr. Weinberg's laboratory discovered that human oral epithelial cells express these peptides, and confirmed their expression in skin keratinocytes.

Dr. Weinberg's efforts are focused in four areas. The first involves identifying and characterizing commensal bacterial b-defensin enhancing components. Dr. Weinberg has determined that the cytoplasmic membrane of the oral bacterium Fusobacterium nucleatum, a commensal gram negative species found in healthy mouths, upregulates the expression of both hBD-1 and hBD-2. These findings are novel and set the stage for the isolation and characterization of the organism's b-defensin enhancing component. The second focal area involves studying the regulation of expression of hBD-1 and hBD-2. Dr. Weinberg's laboratory has determined that hBD-1 is regulated post-transcriptionally, while hBD-2 is transcriptionally regulated. In addition, hBD-1 and hBD-2 are differentially regulated by different bacteria and fungi. Involvement of novel epithelial surface "Toll-like receptors," originally found to regulate AP expression in insects and recently identified in human epithelial cells and macrophages, is currently being investigated. An oral squamous cell carcinoma cell line, which does not express b-defensins, is also being investigated. The third focal area incorporates findings by Dr. Weinberg that levels of expression of hBD-1 and hBD-2 vary between human subjects. What this means in terms of predisposition towards infections in people who express these APs at sub-therapeutic levels, is a subject Dr. Weinberg is focusing on as the third area of investigation; i.e., single nucleotide polymorphism analyses of the AP genes. These studies are intended to lead to genome based testing for AP expression. The forth line of investigation involves identifying functional characteristics of the b-defensins and LL-37. Through a recombinant baculovirus system, these APs are being generated and tested for microbicidal properties against a host of gram negative, gram positive, and fungal organisms. In addition, priming epithelial cells to express b-defensins with natural AP enhancers is proving to be effective in minimizing bacterial invasion.

Interactions between Dr. Weinberg and Dr. Kevin Cooper (Director, SDRC), in order to foster interdisciplinary interest in epithelial cell innate defenses, have led to a collaboration between Dr. Weinberg and researchers in the Department of Dermatology (Drs. Ghannoum, Kang, and McCormick). This collaboration focuses on Candida albicans regulation of b-defensins in oral and skin keratinocytes. Recent findings point to differential regulation of these APs by disseminating vs oropharyngeal candidal isolates, as well as between oral and skin epithelial cells

Two recent representative publications:

1. Weinberg A, Krisanaprakornkit S, Dale BA. Epithelial antimicrobial peptides: Review and significance for oral applications. Crit Rev Oral Biol Med. 9:399-414, 1998.

2. Krisanaprakornkit S, Kimball JR, Weinberg A, Darveau RP, Bainbridge BW, Dale BA. Inducible expression of human b-defensin 2 by Fusobacterium nucleatum in oral epithelial cells: Multiple signaling pathways and the role of commensal bacteria in innate immunity and the epithelial barrier. Infect Immun. 68:2907-2915, 2000.

Jean F. Welter, M.D., Ph.D.
Instructor, Department of Orthopaedics
Present Pilot and Feasibility Study Recipient

Dr. Welter's research program is focused on the study of the effects of mechanical forces on cell growth and differentiation. Mechanical forces are important modulators of growth and form in the tissues of the musculoskeletal system, and may influence the development of other tissues as well.

The primary goal of this laboratory is to understand how gene expression is modulated at the transcriptional level in cells of the osteoblast/osteocyte lineage. A second goal is understanding how mechanical deformation influences the recruitment of bone marrow derived mesenchymal stem cells to the osteoblast differentiation pathway.

Finally, Dr. Welter, effective March 1998, received P&F funding from the SDRC to examine the effects of mechanical forces on the differentiation of epidermal keratinocytes. In vivo, keratinocytes are continuously subjected to mechanical deformation, and preliminary in vitro results suggest that cultured keratinocytes are responsive to loads. This has implications for our understanding of epidermal differentiation both in vivo, and in in vitro cell culture models, and may also prove to be important for the study of disease states such as psoriasis, which involves areas of the skin which undergo significant and repeated mechanical stress.

Two recent representative publications:

1. Agarwal C, Efimova T, Welter JF, Crish JF, Eckert RL. CCAAT/enhancer-binding proteins. A role in regulation of human involucrin promoter response to phorbol ester. J. Biol Chem;274:6190-6194, 1999.

2. Efimova T, LaCelle P, Welter JF, Eckert RL. Regulation of human involucrin promoter
activity by a protein kinase C, Ras MEKK1, MEK3, p38/RK, AP1 signal transduction pathway. J Biol Chem;273:24387-24395, 1998.

Christopher C. Whalen, M.D., M.S.
Assistant Professor of Epidemiology & Biostatistics
Member, Immunology/Microbiology Research Area

Dr. Whalen's research interests focus on the control of infectious diseases in human populations, especially tuberculosis. Since 1992, he has been involved with the Uganda-Case Western Reserve University Research Collaboration as a physician-epidemiologist. In this international collaboration, he has evaluated the interaction of tuberculosis and HIV infection at the cellular, clinical and population level. The working hypothesis is that the immune activation induced in response to tuberculosis will serve to enhance viral replication, increase viral load and accelerate the natural history of HIV infection. At the cellular level, he has examined the impact of the host immune response to tuberculosis on HIV viral load and replication. By studying incident cases of tuberculosis which develop from within a large cohort of HIV-infected subjects, he will be able to evaluate the immunologic and virologic events before, during, and after the diagnosis of tuberculosis. At the clinical and population level, he has described the impact of tuberculosis on the clinical course of HIV infection in a series of epidemiologic survival studies.

To date, he has participated in the conduct of three clinical trials relating to tuberculosis in HIV-infected adults. His main role has been in the design, conduct and analysis of these trials. The Research Collaboration has completed a trial comparing the standard treatment regimen for HIV-associated tuberculosis in Uganda, and most neighboring countries of East Africa, with the standard short-course therapy used extensively in the US. Another trial has evaluated the effect of pentoxifylline as adjuvant therapy for HIV-associated tuberculosis. The third trial establishes the efficacy of three regimens for the prevention of tuberculosis in HIV-infected Uganda adults.

In addition to this work, he is interested in the interaction between HIV and endemic viruses in Uganda, such as Epstein-Barr virus and the human herpes virus-8, the etiologic virus for Kaposi's sarcoma. Dr. Whalen is a co-investigator on a proposed study designed to assess ambulatory treatments for HIV-associated lymphoma, and on a project to explore modes of transmission of HHV-8 in Uganda, in connection with the KS initiative of CWSRU, SDRC, Cancer Center and CFAR. Given his extensive experience in international research, especially in East Africa, he plays an important role in studies that involve the Uganda-CWRU Research Collaboration, including the fungal opportunistic infection collaboration between CFAR and SDRC with Dr. Ghannoum.

Two recent representative publications:

1. Wallis RS, Perkins MD, Phillips M, Joloba M, Namale A, Johnson JL, Whalen CC, Teixeira L, Demchuk B, Dietze R, Mugerwa RD, Eisenach K, Ellner JJ. Predicting the Outcome of Therapy for Pulmonary Tuberculosis. Am J Respir Crit Care Med;161:
1076-1080, 2000.

2. Whalen CC, Johnson JL, Okwera A, Hom D, Huebner R, Mugyenyi P, Mugerwa RD, Ellner JJ. A trial of three regimens to prevent tuberculosis in Ugandan adults infected with the human immunodeficiency virus. Uganda-Case Western Reserve University Research Collaboration [see comments]. N Engl J Med 337:801-808, 1997.

Huntington F. Willard, Ph.D.
Professor and Chairman, Department of Genetics
Member, Executive Committee

Research in the Willard laboratory focuses on aspects of the molecular structure and function of human chromosomes and the human genome. The overall goal is to understand developmental and chromosomal mechanisms involved in gene control and/or implicated in genetic disorders, included inherited skin diseases.

Mammalian X chromosome inactivation results in the cis-controlled inactivation of most, but not all, genes along the length of one of the two X chromosomes in females during early embryogenesis. The mechanism of this chromosome-level gene regulation is unknown and has no obvious precedent, but holds significance for the diagnosis of X-linked disease and evaluation of heterozygous carriers, who show variable symptoms depending on the extent of X inactivation. This is especially true of X-linked disorders of the skin, since the skin is an accessible tissue often used in diagnostic studies. Current studies include cloning and characterization of human genes that appear to "escape" inactivation and identification and cloning of the X inactivation center in mouse and humans that controls the cis effect and appears to be required for inactivation to occur.

A second area of research focus involves the development and study of human artificial chromosomes, assembled in the laboratory from individual components of naturally occurring human chromosomes. Artificial chromosome vectors may be useful for non-viral gene therapy applications, as well as for study of genome and chromosome function. Intensive investigation focuses on centromeres of mammalian chromosomes, which are structurally highly complex. The dominant class of DNA at human centromeres is a family of highly repeated, tandemly arrayed satellite DNA, called alpha satellite. Current efforts are directed at determining the cytological and molecular organization of long arrays of alpha satellite, which measure 300-5,000 kb in length, and at establishing functional tests of centromere function in cell culture systems and in living mice, using artificial chromosomes. Approaches include cloning arrays in yeast artificial chromosomes (YACs), introduction of YACs into mammalian cells, pulsed-field gel electrophoresis, fluorescence in situ hybridization, and genetic and sequence analysis of centromeric DNA variation.

Dr. Willard's presence on the Executive Committee imparts excellence, an awareness of skin applications, and advanced technology for SDRC planning.

Two recent representative publications:

1. Carrel L, Cottle AA, Goglin KC, Willard HF. A first-generation X-inactivation profile of the human X chromosome. Proc Natl Acad Sci USA;96:14440-14444, 1999.

2. Carrel L, Willard HF. Heterogeneous Gene epression from the inactive X chromosome: an X-link gene that escapes X inactivation insome human cell lines but is inactivated in others. Proc Natl Acad Sci USA;96:7364-7370, 1999.

James K. V. Willson, M.D.
Professor of Medicine
Director, Ireland Cancer Center at Case Western Reserve University
And University Hospitals of Cleveland
Member, Executive Committee

James Willson, M.D. is Director of the Ireland Cancer Center, a NCI-designated Comprehensive Cancer Center at Case Western Reserve University and University Hospitals of Cleveland. The Skin Diseases Research Center has played the lead role in the development of a cutaneous oncology initiative at CWRU with the focus on multidisciplinary care and comprehensive research on melanoma and cutaneous lymphoma and the planned development of PC4-photodynamic therapy of skin cancer, as well as the Kaposi's Sarcoma initiatives. Dr. Willson has worked with the members of the SDRC to foster this synergy between the Skin Diseases Research Center and the Ireland Cancer Center. A further example of the interactions has been the development of a translational research oncology training program in which Dr. Cooper leads the tumor immunology and immunotherapy program. This translational research oncology training program supports five training positions for physician scientists each year. In addition to this leadership role, Dr. Willson is also recognized for the development of cell line and xenograft models which have been unique resources for studies of the cellular and molecular biology of cancer. He has used his expertise to contribute to the development of the animal experimentation core and will continue to be an advisor to the Skin Diseases Research Center and assist participants in this SDRC with investigations that require immunodeficient animals. Dr. Willson's participation on the Executive Committee ensures that SDRC is fully aware of and synergistic with the Cancer Center's resources.

Two recent representative publications:

1. Spiro TP, Gerson SL, Liu L, Majka S, Haaga J, Hoppel CL, IngallsST, Pluda JM,
Willson JK. O6-benzylguanine: a clinical trial establishing the biochemical modulatory dose in tumor tissue for alkyltransferase-directed DNA repair. Cancer Res;59:2402-2410, 1999.

2. Whitacre CM, Zborowska E, Willson, JKB, Berger NA. Detection of poly (ADP-ribose) polymerase cleavage in response to treatment with topoisomerase I inhibitors: a potential surrogate end point to assess treatment effectiveness. Clin Cancer Res;5:665-672, 1999.