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Elucidating the role of pioneer factors in RPC developmental competence

阐明先驱因素在 RPC 发展能力中的作用

基本信息

批准号：
10477965
负责人：
Patrick Leavey
金额：
$ 4.68万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-29 至 2024-08-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10477965
关键词：
ATAC-seq Address Age related macular degeneration Amacrine Cells Amphibia Automobile Driving Binding Biological Assay Birth Blindness Cell Death Cell Line Cell Therapy Cells Chimeric Proteins Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Competence Complex Computer software Development Disease Electroporation Epigenetic Process Exhibits Factor Analysis Family Fibrinogen Fishes Gene Expression Generations Genes Genomic Segment Genomics Glaucoma Goals Histology In Vitro Individual Location Modeling Molecular Molecular Mechanisms of Action Muller&apos s cell Mus Mutant Strains Mice Neurons Nuclear Translocation Organizational Change Pattern Phenotype Plasmids Play Population Predictive Factor Process Production Proliferating Regenerative capacity Regulation Resolution Retina Retinal Cone Retinal Diseases Retinal Ganglion Cells Role Route Site System Testing Therapeutic Intervention Tissues Validation Work cell fate specification cell type cold blooded vertebrate epigenomics experimental study gain of function gene regulatory network horizontal cell in vivo induced pluripotent stem cell loss of function neurogenesis overexpression pluripotency response to injury retinal progenitor cell single-cell RNA sequencing small molecule transcription factor

项目摘要

PROJECT SUMMARY Temporal patterning drives retinal cellular diversity through complex gene regulatory networks (GRNs). These GRNs are headed by transcription factors (TFs) that promote stage-specific cell birth while repressing GRNs associated with other developmental timepoints. This cross play is especially noticeable when comparing the GRNs controlling generation of early and late-born cell types in retinal progenitor cells (RPCs). Notably, over the course of differentiation, mammalian retinal cells lose the regenerative capacity seen in fish and amphibian models. This loss of regenerative capacity permits for the cell death associated with leading causes of blindness such as age-related macular degeneration and glaucoma. Key to therapeutic interventions is an understanding of how to stimulate the birth of specific cell types to allow for successful regrowth of the cell populations damaged in the course of these diseases. This understanding hinges on identifying which TFs within the GRNs drive cell fate specification and the loss of pluripotency for the early and late-stage RPCs. Because the genomic organizational states between the early and late-stage RPCs is so distinct, I hypothesize that the TFs driving the GRNs act in a pioneering capacity to drive the birth of temporally restricted cell types. If this holds true, the TFs associated with early-born cell types will be able to drive the birth of cones, amacrines, and horizontal cells in late-stage RPCs through the opening of genomic regions inaccessible at that temporal window. Likewise, late-stage TFs will induce the production of bipolar cells and Müller glia in the early RPC population through genomic organizational changes. To address these hypotheses, I propose two Aims. Aim 1: Functional analysis of top candidate transcription factors for temporal patterning regulation. This work will help me narrow down the list of candidate regulators of temporal patterning through validation of their impact on cell fate in gain-of and loss-of-function experiments in the developing mouse retina. Aim 2: Determine whether TFs that regulate transition from early to late-stage states have pioneering activity. I will take established temporal patterning regulators and assay how they control the dynamics of epigenetic modulation. Once systems for in vitro and in vivo characterization have been established, I will take candidates identified in Aim 1 and use this pipeline to phenotype their pioneering activity. Through the establishment of a set of PFs that drive early and late-stage cell fate specification in the retina, I will be able to better address current barriers to successful iPSC-derived cell-based therapeutic approaches to glaucoma and age-related macular degeneration.

项目总结