Previous Pilot & Feasibility Grant Awardees

PI: Alan Zhou, MD, Assistant Professor of Dermatology (Medical Dermatology)

Our research seeks to characterize the skin, nasal and stool microbial profiles and skin proteomic profiles of erythroderma.

PI: Guillermo A. Ameer, D.Sc., Daniel Hale Williams Professor of Biomedical Engineering, Surgery

Our research will help understand the interactions between the immobilized A5G81 peptide and ECs using in vitro skin tissue models.

PI: Anna Fishbein, MD, Associate Professor of Pediatrics (Allergy and Immunology)

Our objective is to develop a minimally invasive biomarker of skin circadian rhythm, and generate pilot data to test the mechanistic relevance of the skin transcriptome rhythm in diseased AD skin versus healthy controls.

PI: Tomoko Hayashida, MD, PhD, Research Associate Professor of Pediatrics (Nephrology)

Our research aims to establish a novel therapeutic approach for scleroderma, a devastating skin fibrosis disorder, by targeting precursors of scar forming myofibroblasts.  Specifically,  we will focus on cellular signals mediated by Th2 T cells, that is implicated by our data in regulating transdifferentiation of the precursor. 

PI: Han Peng, PhD, Research Associate Professor of Dermatology

Our research will explore the ability of topical HDL NPs alone and complexed with miRNAs to reduce the deleterious effects of inflammation in skin using a mouse model of psoriasiform dermatitis.

PI: Rui Yi, PhD, Research Professor of Pathology; Professor of Pathology (Experimental Pathology) and Dermatology

Our research will establish an experimental and computational pipeline to study human hair loss disorders. We anticipate to discovering new insights into transcriptomic and transcriptional changes in these disorders, generating new hypothesis for experimental validation, and guiding new designs for diagnosis and potential treatment.

Co-PI: Abdelhak Belmadani, PhD, Research Associate Professor of Pharmacology

Co-PI: Daniela Menichella, MD, PhD, Assistant Professor of Neurology (Neuromuscular) and Pharmacology

PI: Jaehyuk Choi, MD, PhD, Assistant Professor of Dermatology and Biochemistry and Molecular Genetics

PI: Sui Huang, MD, PhD, Associate Professor of Cell and Developmental Biology

PI: Peter Sporn, MD, Professor of Medicine (Pulmonary and Critical Care), Cell and Developmental Biology and Medical Education

PI: Xiaomin Bao, PhD, Assistant Professor in Molecular Biosciences

PI: Melissa Brown, PhD, Professor of Microbiology-Immunology and Pathology

This proposal will incorporate a series of studies involving the proteosome inhibitor Bortezomib (BZ), an anti-cancer drug used in the treatment of multiple myeloma. Unfortunately, it has side effects, particularly in the skin. Brown hypothesizes that this relates to mast cell recruitment to the skin. She therefore will test this hypothesis in aim 1 and will also assess whether mast cell stabilizers might have utility in blocking the toxic effect of BZ. In aim 2, Brown proposes to assay mast cell recruitment to the skin in patients being treated with BZ.

Specific aims:

• To evaluate the role of mast cells in BZ-dependent cutaneous effects (KO mice).
• To assess BZ effects on mast cells recruitment and function in the skin of multiple myeloma patients undergoing BZ treatment

PI: Irina Budunova, MD, PhD, Associate Professor in Dermatology and Urology

PI: Navdeep S. Chandel, PhD, Associate Professor of Medicine

This proposal explores the consequences of mitochondrial loss of function in keratinocytes. The major function of mitochondria in cellular homeostatis has historically been the generation of energy through oxidative phosphorylation. However, mitochondria can also serve as a signaling organelle. The projects described in Chandel's lab are driven by the hypothesis that when cells encounter stress the mitochondria serve as key regulators of biological outcomes that include the induction of adaptive genes, cellular proliferation, senescence and death. One stress studied is how cells respond to decreased oxygen levels (hypoxia). Multi-cellular organisms have evolved multiple mechanisms to respond to hypoxia. Healthy individuals typically encounter hypoxia at high altitudes, where at least three prominent physiological responses take place: neurotransmitter release by the carotid body to increase breathing, pulmonary vascular constriction to shunt blood to better oxygenated regions of the lung and production of the hormone erythropoietin (EPO) in the liver to enhance red blood cell mass and hemoglobin concentration in the blood. At the molecular level, the physiological responses to hypoxia are mediated by the transcription factor hypoxia inducible factor, HIF-1. A recent study demonstrated that when mice with a keratinocyte-specific deletion of HIF-1 were exposed to hypoxia, the predicted increase in plasma EPO levels was blunted and induction of EOP expression in the kidney was lost. This was suprising because it suggested that the keratinocytes were sensing the hypoxia to activate HIF-1 and regulate EPO production in the kidney. But how cells sense the decrease in oxygen to activate HIF dependent gene expression is not fully understood. This study will test whether mitochondria function as oxygen sensors in the keratinocytes to induce HIF-1 to regulate EPO production in mice.

Specific aims:

• To determine whether loss of TFAM in keratinocytes abolishes the hypoxic increase in HIF-1 in vitro
• To determine whether loss of TFAM in keratinocytes blunts the hypoxic increase in plasma EPO levels in vivo

Information generated from the study will:

• Further our understanding of the role(s) of HIF-1 in cutaneous wound healing
• Provide a rationale for the use of mitchondrial targeted antioxidants as therapy to prevent local inflammation in the skin, since HIF-1 impacts on skin inflammation
• Justify the development of novel topical therapies for treating anemia, if the mitochondrial ROS/HIF pathway in the skin can stimulate systemic erythropoietin levels

PI: Jaime Garcia-Añoveros, PhD, Assistant Professor in the Department of Anesthesiology

This proposal begins to investigate the possibility that keratinocytes or other skin cells can function as sensory transducers, independent of nerves, in response to changes in temperature. Thermosensitive channels in skin keratinocytes and/or sensory neurons innervating the skin open in response to heating or cooling and mediate our perception of ambient temperature. Garcia-Añoveros's lab has found that some basal cells of the epidermis, as well as some cells of hair follicles, express the TRPML3 channel. It has also been observed that this channel may open in response to changes in temperature. This study aims at elucidating the function of TRPML3 channels in the epidermis and hair follicle and may lead to further studies of how the skin, including the vasculature and eccrine glands, detect and respond to changes in temperature.

Specific aims:

• To determine the spatial and temporal expression of TRPML3 during epidermal and hair follicle development
• To determine whether the cold-to-warm current that we have detected in keratinocytes is mediated by TRPML3
• To determine the effects of deleting the Trpml3 gene is skin

PI: Jodi L. Johnson, PhD, Research Assistant Professor of Pathology & Dermatology
CO-I: June K. Robinson, MD, Research Assistant Professor of Dermatology

Johnson theorizes that keratinocytes from non-Hispanic white, Hispanic and African-American kidney transplant patients with medium to dark skin color will display differences when compared with keratinocytes from patients with light skin color. Johnson hypothesizes that the differences in keratinocyte biology from kidney transplant patients with different skin types will significantly correspond with each group’s propensity to develop post-transplant squamous cell carcinoma (SCC) and metastatic SCC. The proposed aim of this study is to understand biological differences between keratinocytes from patients with different skin types to elucidate underlying causes of the lower incidence and rate of SCC formation but slightly higher mortality rate in transplant patients. To complete the aim of this project, Robinson will coordinate the clinical collection of biopsies and patient information and Johnson will coordinate the laboratory studies. This project will formally begin a new collaboration between Johnson and Robinson and will advance the SDRC initiative to focus on differences between keratinocytes of Caucasians and individuals with skin of color.

PI: Steven T. Kosak, PhD, Assistant Professor in Cell and Molecular Biology

Nuclear organization is involved in the regulation of gene expression, and dynamic changes to this organization are thought to be necessary to achieve the precise and coordinated modifications in gene expression that occur during normal cell growth. The nucleus is generally studied as an independent structure; however, it is important to remember that within a cell the nucleus must communicate with other organelles and extracellular signals. Furthermore, most cells are not symmetric structures, and cell polarity is involved in many cellular functions including proliferation, differentiation and morphogenesis1. Therefore, in order to fully examine nuclear organization, Kosak and his team will study the nucleus in the context of an asymmetric cell and incorporate the idea of nuclear polarity, which requires defining nuclear organization with respect to structures and processes on the nuclear exterior. With their award, they propose to examine a previously unstudied aspect of nuclear organization in human cells, nuclear polarity, and to determine how regulation of this form of nuclear organization relates to changes in gene expression during epidermal differentiation.

PI: Douglas A. Kuperman, PhD, Assistant Professor in the Department of Medicine, Division of Allergy-Immunology

This study proposes to test the in vivo relevance of IL-4 and IL-13 acting on keratinocytes in the context of atopic dermatitis. Atopic dermatitis occurs as a result of inappropriate immune responses to common elements in the environment. The T-helper type II (Th2) lymphocyte is a critical contributor to the pathologic inflammatory responses observed in these patients. For example, atopic dermatitis is characterized by increased levels of the Th2 cytokines, IL-4 and IL-13, in the skin. When keratincytes are cultured with either IL-4 or IL-13 they respond by producing a variety of pro-inflammatory mediators and they lose expression of filaggrin as well as loricrin, which are important proteins that maintain skin barrier function. However, there have been no studies performed to test the in vivo relevance of IL-4 and IL-13 signal by ligation of a common receptor, IL-4Rα. Kuperman's hypothesis is that IL-4Rα signaling specifically in keratinocytes contributes to inflammation and failure of skin barrier function in a murine model of atopic dermatitis.

Specific aims:

• To generate a mouse strain that is deficient in IL-4Rα exclusively in keratinocytes
• To determine the in vivo requirement of keratinocyte-specific expression of IL-4Rα in the development of skin pathology associated with experimental atopic dermatitis

PI: Benjamin McNeil, PhD, Research Assistant Professor in Medicine (Allergy-Immunology)

PI: Thomas Meade, PhD, Eileen M. Foell Professor in Cancer Research, Professor of Chemistry, Molecular Biosciences, Neurobiology and Radiology

Gli transcription factors (TFs) represent the terminal step in the Hedgehog (Hh) pathway, which regulates many aspects of embryonic morphogenesis. Aberrant signaling of the Hh pathway is known to promote the establishment and progression of a variety of tumors including basal cell carcinoma (BCC) and medulloblastomas. Due to its involvement in cancer, inhibition of the Hh pathway is of great interest in cancer chemotherapy. Meade intends to attach Co (III)-Gli to Au-NPs in order to achieve effective penetration and delivery of the Gli-inhibitor in the skin. This agent would have possible applications in treatment of skin-associated diseases mediated by the Hh pathway.

To assess the feasibility of this concept, he proposes to investigate the following:

• Synthesis and characterization of Co(III)-Gli conjugated Au-NPs (Au-Gli-Co)
• In vitro validation of the efficacy of Au-Gli-Co at inhibiting the Hh pathway by assessing the efficacy of Au-Gli-Co in vitro, examining the direct target of Au-Gli-Co and determining tissue penetration

This project involves the development of a unique TF inhibitor that targets the terminal step of the Hh pathway to treat BCC. It employs a new mechanism for inhibiting TFs that confers high specificity and efficacy toward the target. By incorporating a novel and effective delivery mechanism using Au-NPs, Meade expects Au-Gli-Co to effectively inhibit the Hh pathway. Further, the Au-NP platform will allow improved delivery of the agent through cells and skin and permit targeted localized delivery of the agent using plasmon activation.

PI: Chad A. Mirkin, PhD, Professor of Chemistry, Medicine and Material Sciences and Engineering and Director of the Institute of Nanotechnology

This proposal will test the function of oligomers in keratinocytes, initially to suppress survivin expression in keratinocytes and then to test the functional effect of targeting a common missense mutation of keratin 14 to reverse the keratin filament defect. Controlling gene expression in primary cultured cells, such as human keratinocytes, is difficult because the epidermal barrier precludes the entry of suppressive nucleic acids. Methods to introduce gene regulation agents such as oligonucleotides have met with limited success. The proposed work will develop oligonucleotide-nanoparticle antisense agents as methods for controlling gene expression in keratinocytes. This work will take advantage of newly discovered conjugate properties of the nanomaterials, including enhanced stability, high penetration ability and lack of toxicity. The ability of these agents to work on genes of relevance in skin biology models will be tested.

Specific aims:

• Optimize the keratinocyte and epidermal uptake of oligonucleotide-gold nanoparticle conjugates (DNA-Au NPs) and determine their utility in suppressing gene expression in vitro and in mouse skin
• Investigate the ability of DNA-Au NPs to selectively suppress mutant keratin 14 expression in keratinocytes from a patient with epidermolysis bullosa simplex (Dowling-Meara)

PI: Brian Mitchell, PhD, Assistant Professor of Cell and Molecular Biology

This proposal aims to develop the experimentally pliable skin of Xenopus embryos as a model system for dissecting the molecular mechanisms via which cells break through and re-establish junctional barriers. Xenopus skin is an ideal system for this analysis since it consists of two distinct layers, the outer keratinocytes, and an inner basal layer composed of precursor cells with the ability to differentiate into either multi-ciliated cells or ionocytes. During development, these inner layer cells undergo a stereotyped migration event in which they intercalate into the keratinocyte layer. Mitchell proposes to define the complex cell biological processes that underlie how skin cells migrate in a directed manner, break down cell-cell junctions of the outer keratinocytes and stop their migration and form new cell-cell junctions.

Specific aims:

• To determine how small GTPases regulate cytoskeletal dynamics during migration and the termination of this migration
• To determine the role of the microtubule binding protein CLAMP on the initiation of multi-ciliated cell intercalation
• To determine which genes are upregulated in both multi-ciliated cells and ionocytes during the intercalation of these diverse cell types

PI: William J. Muller, MD, PhD, Assistant Professor of Pediatrics

Evidence from studies in mice and in tissue culture models suggests that the interaction of herpes simplex virus with one of its principal receptors, the herpes virus entry mediator (HVEM), influences early events in innate immune signaling. Our overall hypothesis is that virus interaction with HVEM alters this signaling to promote viral replication and establishment of infection. Using a virus that is altered to abolish interaction with HVEM, we will use molecular techniques to measure the induction and translation of innate genes in human keratinocytes after infection and how this is influenced by HVEM signaling. We will then confirm and extend these results in assays using wild-type virus and cells lacking HVEM signaling; either murine cells with disruptions in the HVEM gene, or human cells with HVEM expression knocked down.

Specific aims:

• Measure the induction and translation of innate genes in human keratinocytes after infection with wildtype HSV-1 and HSV-2, compared to mutants of these viruses unable to engage HVEM
• Measure the induction and translation of innate genes in murine keratinocyte cultures derived from wildtype mice and mice lacking HVEM (Tnfrsf14-/-mice), and in human keratinocyte cultures for which HVEM is knocked down, after infection with wild-type HSV

PI: Bethany E. Perez White, PhD, Research Assistant Professor in Dermatology

PI: Harris Perlman, PhD, Chief of Rheumatology, Mabel Greene Myers Professor of Medicine, and Professor of Medicine (Rheumatology)

PI: Murali Prakriya, PhD, Associate Professor of Molecular Pharmacology and Biological Chemistry

The hypothesis for this study is that ion channel pores called calcium release activated calcium (CRAC) channels serve as a key route of calcium entry in keratinocytes to regulate gene expression and the release of inflammatory mediators. Prakriya aims to test this hypothesis using an integrated approach that merges electrophysiology, Ca2+ imaging, immune assays and mouse genetics. Confirming this hypothesis would resolve important uncertainties about the identity of Ca2+ influx pathways that operate in keratinocytes. Furthermore positive findings will allow the development of novel therapeutic solutions for inflammatory diseases of the skin and lay a sound foundation on which to build and finance the long-term plan for this research.

Specific aims:

• Determine the functional properties and molecular mechanisms of CRAC channels in kertainocytes
• Determine the functional significance of CRAC channels for gene expression and generation of inflammatory mediators in keratinocytes

Collectively, this study will illuminate potential physiological roles of CRAC channels in keratinocytes and provide fresh insights in to the mechanism of cytokine and chemokine production in the skin.

PI: Karla J. F. Satchell, PhD, Professor in Microbiology-Immunology

Rat sarcoma (Ras) oncoprotein is a ubiquitous small GTPase in eukaryotic cells that is a critical node that senses incoming signals and subsequently activates downstream target proteins. These targets include mitogen activated protein kinase (MAPK) signaling cascades that ultimately turn on additional genes directing cell proliferation, differentiation and survival. Ras is also pivotal to the host response to infection coordinating signals from surface receptors that bind bacterial antigens to active cytokine and chemokine expression and macrophage maturation. Ras enzymatic activity is regulated by cycling between an inactive (GDP-bound) state and an active (GTP-bound) state. Constitutively activating mutations that mimic or stabilize the GTP-bound state are found in about 30 percent of human malignancies, inc. skin melanomas. Germline mutations in Ras or genes connected to RAS are also implicated in developmental RASopathies. Despite the potential for this important protein to be a therapeutic target for these diseases, no inhibitors that directly target RAS have been successfully developed for clinical use and RAS has been labeled "undruggable."

PI: Christian Stehlik, PhD, Assistant Professor of Medicine, Rheumatology Division

The goal of this study is to characterize a novel innate immune function of Keratinocytes by testing our hypothesis that select NLRs are able to promote inflammasome activation in Keratinocytes upon sensing pathogen infection of the skin and to subsequently recruit phagocytes. We further hypothesize that dysregulation of this system contributes to the excessive production of IL-1β in inflammatory skin disease.

Specific aims:

• To determine inflammasome activation in Keratinocytes upon bacterial infection
• To determine if select NLRP mutations cause chronic inflammasome activation in Keratinocytes

PI: Chyung-Ru Wang, PhD, Professor in Microbiology-Immunology

The goal of the proposed research is to investigate the potential role of group 1 CD1-restricted autoreactive T cells in the development of chronic skin inflammation. Wang has significant expertise in the field of CD1 biology, which has been the primary focus of her research for past 20 years. Her lab has generated multiple novel mouse models that have contributed significantly to the study of group 1 and group 2 CD1-restricted T cells in the context of autoimmunity and infectious disease. Recently, her lab has generated double transgenic mice that expressed the human group 1 CD1 molecules and TCR specific to group 1 CD1 and species-conserved self lipid antigens. Using this mouse model, they demonstrated that autoreactive group 1 CD1-restricted T cells have similar developmental requirements, surface phenotypes and functional properties to those of CD1d-restricted NKT cells. Interestingly, these mice spontaneously developed dermatitis when they were on the Apo-E deficient background. They have just obtained a R21 to investigate how group 1 CD1 autoreactive T cells contribute to chronic skin inflammation in the animal model. They are very interested to extend their study into patients with chronic skin inflammation, such as atopic dermatitis and psoriasis. They anticipate that these studies may lead to the identification of new biomarkers or new therapeutic targets for treating these diseases.

PI: Liang Zhou, MD, PhD, Assistant Professor of Pathology and Microbiology-Immunology

This study seeks to understand how T helper (Th) 17 and T regulatory (Treg) immune cells are regulated in the skin, both under steady state and psoriatic inflammatory conditions. Zhou hypothesizes that Ikaros functions as a molecular switch to determine the Treg-Th17 balance via specific DNA-binding zinc fingers, which is important for skin inflammation and the pathogenesis of psoriasis. Specifically, Zhou proposes to study the crosstalk between keratinocytes and T cells mediated by Ikaros using both in vitro cell culture systems and an in vivo model of psoriasis.

Specific aims:

• Determine the molecular mechanisms of action of Ikaros in the Treg /Th17 cell differentiation program
• Determine the impact of Ikzf1ΔF4 T cells on keratinocytes
• Determine the in vivo role of Ikaros using a Th17-mediated murine psoriasis model

Zhou proposes that this study will represent a unique interface between immunology and keratinocyte biology, and we are positioned with the necessary tools to understand the role of Ikaros in skin inflammation. The findings from the proposed studies will provide novel mechanistic insights into the function of multiple Th17 and Treg cell transcription factors in immune regulation and may identify new therapeutic targets for psoriasis and other inflammatory skin disorders.