Identification of gene regulatory networks that control proliferative and neurogenic competence in mammalian Müller glia

鉴定控制哺乳动物穆勒神经胶质细胞增殖和神经发生能力的基因调控网络

基本信息

批准号：
10636825
负责人：
Seth Blackshaw
金额：
$ 51.19万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10636825
关键词：
Adult CRISPR/Cas technology Catalogs Cell Cycle Cells Chick Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Competence Dedications Degenerative Disorder Disease Family Gene Expression Gene Expression Profile Generations Genes Gliosis Goals Growth Factor Hour Individual Injury Mammals Mediating Molecular Morphology Muller&apos s cell Mus NFIA gene Natural regeneration Neuroglia Photoreceptors Predictive Factor Process Proliferating Radial Repression Rest Retina Retinal Diseases Retinal Dystrophy Retinal Photoreceptors Treatment Factor Work Zebrafish cell type cold blooded vertebrate combinatorial design efficacy validation gene function gene regulatory network hatching insight loss of function morphogens mutant neurogenesis novel strategies overexpression regenerative therapy regenerative treatment response to injury retinal damage retinal neuron retinal progenitor cell retinal rods therapy development transcription factor

项目摘要

Project Summary Müller glia of cold-blooded vertebrates can re-enter the cell cycle and rise to photoreceptors following retinal injury, while mammals have lost this ability. As part of the NEI Audacious Goals Initiative, we have conducted a ü comprehensive analysis of injury-induced changes in gene expression and chromatin accessibility in zebrafish, chick and mouse M ller glia, allowing us to identify both evolutionarily consüerved and species-specific gene regulatory networks that regulate glial reprogramming. This has identified a set of dedicated gene regulatory networks in mice tühat restrict proliferative and neurogenic competence in M ller glia. We aim to use these findings to gain a more complete insight into the molecular mechanisms that regulate neurogenic competence in mammalian M ller glia, and to develop treatments that can maximize generation of glial-derived photoreceptors while simultaneously not depleting the number of existing glia. To do this, we propose to generate individual loss of function mutants of the top candidate negative regulators of proliferative and ü neurogenic competence using AAV-mediated CRISPR/Cas9 gene disruption. We will first validate efficacy of sgRNAs targeting individual TFs, comprehensively profile chanüges in geüne expression in reactive M ller glia following loss of function of these genes, and characterize the fate of M ller glia-derived cells. We will then conduct combinatorial loss of function of negative regulators of M ller glia reprogramming to enhance generation ü of glial-derived retinal progenitor cells in wildtype and Nfia/b/x-deficient mice. Finally, we will combine CRISPR- mediated loss of function analysis with overexpression of Ascl1, Crx and Nrl to enhance generation of M ller glia-derived rod photoreceptors in both wildtype and dystrophic retina. We predict that these studies may substantially advance cell-based regenerative treatments aimed at restoring retinal photoreceptors lost due to blinding diseases.

项目摘要冷血脊椎动物的müller神经胶质可以重新进入细胞周期，并在视网膜后升至感光体受伤，而哺乳动物失去了这种能力。作为NEI大胆目标计划的一部分，我们进行了 ü 斑马鱼中损伤诱导的基因表达变化和染色质可及性的全面分析，雏鸡和小鼠麦芽胶质细胞，使我们能够识别进化论和物种特异性基因调节神经胶质重编程的调节网络。这已经确定了一组专用基因调节小鼠中的网络限制了M ller Glia的增殖和神经源能力。我们的目的是使用这些调查结果可以更完整地了解调节神经源能力的分子机制在哺乳动物的胶质神经胶质中，并开发可以最大程度地生成神经胶质的治疗感光体同时不耗尽现有神经胶质的数量。为此，我们建议产生最高候选人的功能突变体的个人丧失，负面调节因子的增殖和 ü 使用AAV介导的CRISPR/CAS9基因破坏的神经源能力。我们将首先验证效率靶向单个TF的SGRNA，全面介绍了反应性神经胶质中Geüne表达的变化这些基因的功能丧失后，并表征了M ller胶质细胞的命运。然后我们会进行组合丢失的功能的功能负调节剂的功能，以增强产生 ü 野生型和NFIA/B/X缺陷小鼠中神经胶质的视网膜祖细胞的。最后，我们将结合CRISPR- ASCL1，CRX和NRL的过表达介导的功能分析丧失，以增强M ller的产生野生型和营养不良视网膜中的胶质源棒感受器。我们预测这些研究可能基于细胞的基于细胞的基于细胞的再生治疗旨在恢复因视网膜感光体而失去的视网膜感受器盲目的疾病。

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Ectopic insert-dependent neuronal expression of GFAP promoter-driven AAV constructs in adult mouse retina.

DOI：
10.3389/fcell.2022.914386
发表时间：
2022
期刊：
Frontiers in cell and developmental biology
影响因子：
5.5
作者：
通讯作者：

A potential role for somatostatin signaling in regulating retinal neurogenesis.

DOI：
10.1038/s41598-021-90554-3
发表时间：
2021-05-26
期刊：
Scientific reports
影响因子：
4.6
作者：
Weir K;Kim DW;Blackshaw S
通讯作者：
Blackshaw S

IReNA: Integrated regulatory network analysis of single-cell transcriptomes and chromatin accessibility profiles.

DOI：
10.1016/j.isci.2022.105359
发表时间：
2022-11-18
期刊：
ISCIENCE
影响因子：
5.8
作者：
Jiang, Junyao;Lyu, Pin;Li, Jinlian;Huang, Sunan;Tao, Jiawang;Blackshaw, Seth;Qian, Jiang;Wang, Jie
通讯作者：
Wang, Jie

Updates and challenges of axon regeneration in the mammalian central nervous system.

DOI：
10.1093/jmcb/mjaa026
发表时间：
2020-10-01
期刊：
Journal of molecular cell biology
影响因子：
5.5
作者：
Qian C;Zhou FQ
通讯作者：
Zhou FQ

Control of neurogenic competence in mammalian hypothalamic tanycytes.

DOI：
10.1126/sciadv.abg3777
发表时间：
2021-05
期刊：
Science advances
影响因子：
13.6
作者：
Yoo S;Kim J;Lyu P;Hoang TV;Ma A;Trinh V;Dai W;Jiang L;Leavey P;Duncan L;Won JK;Park SH;Qian J;Brown SP;Blackshaw S
通讯作者：
Blackshaw S