Transcriptional Regulation of Cone Photoreceptor Genesis

视锥光感受器起源的转录调控

基本信息

批准号：
10665652
负责人：
MARK M EMERSON
金额：
$ 39.25万
依托单位：
CITY COLLEGE OF NEW YORK
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-01-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10665652
关键词：
Age related macular degeneration Benchmarking Binding Biological Assay Blindness CRISPR/Cas technology Cell Therapy Cells Chick Chickens Cone Development Disease Elements Embryo Event Flow Cytometry Gene Expression Generations Genes Genetic Genetic Transcription Genome Goals Human Knowledge LIM Domain Protein Mediating Methods Mission Molecular Muller&apos s cell Mus Mutagenesis Mutation Analysis National Eye Institute Natural regeneration Nucleic Acid Regulatory Sequences Organism Output Phenotype Photoreceptors Play Population Production Proteins Protocols documentation RXRG gene Regulatory Element Reporter Repression Resolution Retina Retinal Cone Retinitis Pigmentosa Rod Role Site Source Specificity Testing Transcriptional Regulation Vertebrate Photoreceptors Vision Visual Visual impairment cell replacement therapy design gene regulatory network human disease in vivo innovation insight loss of function loss of function mutation postnatal prevent programs rational design retinal rods single-cell RNA sequencing stem cells therapy development transcription factor transcriptome transcriptomics translational impact

项目摘要

PROJECT SUMMARY/ABSTRACT Cone photoreceptors are the primary photoreceptor class that mediates high acuity and daylight vision. As such, their loss during the course of human disease greatly impairs visual function. Cell-based therapies to ameliorate these conditions have been proposed, in which introduction of new cones could replace those lost to disease. These cells could be produced exogenously from sources such as stem cells or from resident, in vivo populations such as Müller glia. For both strategies, it is necessary to develop new methods to specifically promote cone formation, and to assess whether de novo formed cones are molecularly equivalent to those formed during normal development. However, a significant gap exists in the current knowledge of gene regula- tory networks that promote cone photoreceptor development. The long-term goal of this project is to identify the gene regulatory networks active in cone genesis, and to devise methods based on this knowledge to gen- erate new cone cells for cell replacement therapy. Recently, transcription factors with enriched expression in early developing cones have been identified, yet the genetic requirement of these factors or how they interact with one another to promote the cone fate is unknown. The central hypothesis of this proposal is that these transcription factors are required for cone photoreceptor genesis and are part of a gene regulatory network with defined regulatory interactions. The rationale to undertake these studies is that elucidation of the normal genetic mechanisms of cone photoreceptor formation will inform the directed formation of cones from stem cells or endogenous retinal cell populations such as Muller glia. Identification of the proteins that have im- portant regulatory functions in cones will allow development of new cone induction protocols with greater and more efficacious production of functional cones. To accomplish this goal, the following three aims are pro- posed: 1) A functional analysis of early cone-enriched transcription factors, 2) Identification of key expression and functional parameters of Sall1, a transcription factor involved in cone induction and rod repression, and 3) Identification of Nrl cis-regulatory elements that repress Nrl expression in cones. The first aim will use an effi- cient gene editing and single cell transcriptomic approach to determine the necessity of newly identified tran- scription factors expressed in early stage cones. The second aim will investigate the endogenous role of Sall1 in cone formation and the extent to which expression of this gene is sufficient to induce cone transcriptional programs. The third aim will identify the transcriptional mechanisms that prevent the rod-specific transcription factor Nrl from being expressed in cones. This approach is innovative because it will investigate the early mo- lecular events in cone formation using high-resolution and robust methods. Completion of the proposed project will result in significant insights into the gene regulatory networks that promote cone formation and will inform the development of rationally designed and effective methods to promote de novo cone formation.

项目摘要/摘要锥形感受器是介导高敏锐和日光视觉的主要光感受器类。作为这样，它们在人类疾病过程中的丧失会大大损害视觉功能。基于细胞的疗法已经提出了改善这些条件，在其中引入新的锥体可以取代那些丢失的人疾病。这些细胞可以是从干细胞或居民等来源外源产生的体内种群，例如müllerglia。对于这两种策略，有必要开发新方法来具体促进锥形形成，并评估从头形成的锥体是否分子等同于那些在正常发育过程中形成。但是，在当前对基因调节剂的知识中存在很大的差距 - 促进锥形光感受器发展的保守网络。该项目的长期目标是确定基因调节网络活跃在锥发生中，并根据这些知识来设计方法食用新的锥细胞用于细胞替代疗法。最近，具有丰富表达的转录因子已经确定了早期发育的锥体，但是这些因素的遗传需求或它们如何相互作用彼此促进锥命命运是未知的。该提议的核心假设是这些转录因子是锥形感受器创世纪所需要的，并且是基因调节网络的一部分与定义的监管相互作用。进行这些研究的理由是阐明正常锥形光感受器形成的遗传机制将告知茎的锥形形成细胞或内源性视网膜细胞种群，例如Muller Glia。鉴定具有影响的蛋白质锥体中的便携式监管功能将允许开发具有更大和更大和功能锥的更有效产生。为了实现这一目标，以下三个目标是摆姿势：1）对早期锥体富集的转录因子的功能分析，2）鉴定关键表达 SALL1的功能参数，锥体诱导和杆表达涉及的转录因子，3） NRL顺式调节元件的鉴定，这些元件抑制了锥体中的NRL表达。第一个目标将使用努力 - CIENT基因编辑和单细胞转录组方法，以确定新近鉴定的tran-的必要条件脚本因素在早期锥体中表达。第二个目标将研究sall1的内源性作用在锥体形成以及该基因表达的程度足以诱导锥体转录程序。第三个目标将确定可防止杆特异性转录的转录机制因子NRL在锥体中表达。这种方法具有创新性使用高分辨率和鲁棒方法在锥形形成中形成的内部事件。拟议项目的完成将为促进锥形形成的基因调节网络提供重大见解，并将告知开发合理设计和有效的方法来促进从头锥形成。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Quantitative analysis of the ThrbCRM1-centered gene regulatory network.

以 ThrbCRM1 为中心的基因调控网络的定量分析。

DOI：
10.1242/bio.039115
发表时间：
2019
期刊：
Biology open
影响因子：
2.4
作者：
Souferi,Benjamin;Emerson,MarkM
通讯作者：
Emerson,MarkM

Early cis-regulatory events in the formation of retinal horizontal cells.

DOI：
10.1016/j.ydbio.2021.03.016
发表时间：
2021-08
期刊：
Developmental biology
影响因子：
2.7
作者：
Schick E;Gonzalez KC;Dutta P;Hossain K;Ghinia Tegla MG;Emerson MM
通讯作者：
Emerson MM

Identification and characterization of early photoreceptor cis-regulatory elements and their relation to Onecut1.

DOI：
10.1186/s13064-018-0121-x
发表时间：
2018-11-22
期刊：
Neural development
影响因子：
3.6
作者：
Jean-Charles N;Buenaventura DF;Emerson MM
通讯作者：
Emerson MM

OTX2 represses sister cell fate choices in the developing retina to promote photoreceptor specification.

OTX2 抑制发育中视网膜中姐妹细胞的命运选择，以促进感光细胞的特化。

DOI：
10.7554/elife.54279
发表时间：
2020
期刊：
eLife
影响因子：
7.7
作者：
GhiniaTegla,MirunaGeorgiana;Buenaventura,DiegoF;Kim,DianaY;Thakurdin,Cassandra;Gonzalez,KevinC;Emerson,MarkM
通讯作者：
Emerson,MarkM

Fate-restricted retinal progenitor cells adopt a molecular profile and spatial position distinct from multipotent progenitor cells.