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Principal Investigator: Ya-Chieh Hsu
Organization: HARVARD UNIVERSITY
Fiscal Year: 2024
Award: $504,905
Funding agency: National Institute of Arthritis and Musculoskeletal and Skin Diseases
Project Summary
Skin stem cells are heavily influenced by signals from their niches including different fibroblasts populations.
While our ability to isolate and molecularly profile diverse cell types has improved drastically in the past decade,
a major roadblock in identifying key genes driving stem cell-niche interactions is the lengthy process of
generating the genetic models needed (e.g., cell-type specific Cre or CreER, and overexpression or knockout
mouse lines) to test gene functions in a cell-type specific manner in a physiologically relevant context. As such,
while many different cell types have been identified and molecularly profiled, the critical genes that drive many
developmental and regeneration processes remain incompletely understood. This substantial knowledge gap
presents a significant impediment to developing therapies for skin diseases.
To address this gap and to showcase how rapid functional genetics can enable new discoveries in stem cell-
niche interactions, we will first build adeno-associated viral (AAV) toolkits to expand the field’s capacity for rapid
functional genetics in multiple dermal cell types in mice. This aim expands on our current success in using AAVs
to transduce dermal cells, with the goal of building tools that allows all skin researchers to modify gene
expression rapidly in dermal populations such as the dermal fibroblasts and DP. We have recently developed
and conducted SHARE-seq on the skin, a high-throughput single cell sequencing method that simultaneously
measures chromatin accessibility (single cell ATACseq) and gene expression (single cell RNAseq) within the
same cell. SHARE-seq data allow us to computationally infer key regulatory elements (enhancers, promoters)
of signature genes for distinct cell types, which further enables the construction of cell-type specific AAV tools.
We know the proposed strategy is feasible, because we have used it to build tools that can manipulate gene
expression in the arrector pili muscles (APMs), a cell type that currently lacks specific Cre/CreER constructs.
APMs are an emerging niche cell type for hair follicle stem cells (HFSCs). However, the molecular mechanisms
by which APMs regulates HFSC behavior remain poorly understood. In Aim2, we will use our AAV tools to
discover APM-derived secreted factors that regulate HFSC activation and maintenance. Collectively, these
results will provide the skin community with much-needed tools to accelerate research in diverse topics, and
may be relevant for understanding and potentially treating a wide range of alopecia conditions. Since AAVs are
non-toxic and non-immunogenic, and since many key tissue-specific regulatory elements retain their specificity
across species, there is an exciting potential to combine our biological findings with our technical advancements
to develop novel gene therapy strategies to treat these skin diseases in the near future.
Terms: <Ablation><Acceleration><Address><Adeno-Associated Viruses><Agonist><Alopecia><Arrector pili><Automobile Driving><Autoregulation><Baldness><Behavior><Biological><Body Tissues><Candidate Disease Gene><Candidate Gene><Cell Body><Cell Communication and Signaling><Cell Signaling><Cells><Cholinergic Differentiation Factor><Chromatin><Communities><Corium><Cutaneous Disorder><Cutis><D-Factor><DNA Therapy><Data><Data Set><Dependoparvovirus><Dependovirus><Deposit><Deposition><Dermal><Dermatoses><Dermis><Development><Enhancers><Fibroblasts><Future><GDNF><GDNF gene><Gene Expression><Gene Transfer Clinical><Genes><Genetic><Genetic Intervention><Genetic Models><Goals><Hair><Hair Follicle><Hair follicle structure><Homeostasis><Immunofluorescence><Immunofluorescence Immunologic><Intracellular Communication and Signaling><Investigators><Involuntary Muscle><KO mice><Knock-out Mice><Knockout Mice><Knowledge><LIF><LIF gene><Maintenance><Measures><Mediating><Mediator><Methods><Mice><Mice Mammals><Molecular><Molecular Fingerprinting><Molecular Profiling><Murine><Mus><Muscle><Muscle Tissue><Natural regeneration><Nerve><Null Mouse><Organ><Physiologic><Physiological><Physiological Homeostasis><Pilor Erectus><Plasmids><Play><Population><Process><Progenitor Cells><Publications><RNA Seq><RNA sequencing><RNAseq><Regeneration><Regulatory Element><Research><Research Personnel><Researchers><Role><Scientific Publication><Serotyping><Signal Transduction><Signal Transduction Systems><Signaling><Single cell seq><Skin><Skin Diseases><Skin Diseases and Manifestations><Smooth Muscle><Specificity><Testing><Therapeutic><Time><Tissues><Tropism><Viral><Work><Wound Repair><adeno associated virus group><biologic><biological signal transduction><cell behavior><cell type><cellular behavior><cutaneous disease><depository><dermal disease><dermal disorder><develop therapy><developmental><driving><gene function><gene manipulation><gene repair therapy><gene signatures><gene testing><gene therapy><gene-based testing><gene-based therapy><genetic manipulation><genetic signature><genetic testing><genetic therapy><genetically manipulate><genetically perturb><genomic therapy><glial cell-line derived neurotrophic factor><hair erector muscle><improved><in vivo><inhibitor><innervation><innovate><innovation><innovative><insight><intervention development><leukemia inhibitor factor><leukemia inhibitory factor><molecular profile><molecular signature><muscle system><muscular><muscular system><nerve supply><new approaches><new drug treatments><new drugs><new pharmacological therapeutic><new technology><new therapeutics><new therapy><next generation therapeutics><novel><novel approaches><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel strategies><novel strategy><novel technologies><novel therapeutics><novel therapy><overexpress><overexpression><papilla><progenitor biology><progenitor cell biology><progenitor cell niche><progenitor niche><promoter><promotor><regenerate><repair><repaired><repository><scATAC sequencing><scATAC-seq><scRNA-seq><single cell ATAC-seq><single cell ATAC-sequencing><single cell Assay for Transposase Accessible Chromatin sequencing><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell next generation sequencing><single cell sequencing><single cell sequencing assay for transposase accessible chromatin><single cell transcriptomic profiling><single-cell Assay for Transposase-Accessible Chromatin with sequencing><single-cell RNA sequencing><single-cell assay for transposase-accessible chromatin using sequencing><single-cell assay for transposase-accessible chromatin-seq><skin disorder><social role><stem and progenitor biology><stem and progenitor cell niche><stem cell biology><stem cell niche><stem cells><success><therapy development><tongue papilla><tool><transcriptome sequencing><transcriptomic sequencing><treatment development><wound healing><wound recovery><wound resolution>