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Elucidating the molecular and cellular mechanisms underlying cone survival in the peripheral retina in mouse models of Retinitis Pigmentosa

阐明色素性视网膜炎小鼠模型周边视网膜视锥细胞存活的分子和细胞机制

基本信息

批准号：
10348141
负责人：
Ryoji Amamoto
金额：
$ 7.79万
依托单位：
HARVARD MEDICAL SCHOOL
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：
2021-05-01 至 2022-12-31
项目状态：
已结题

项目摘要

Project Summary Retinitis Pigmentosa (RP) is an inherited retinal disease afflicting 1 in 4,000 people worldwide. The disease progresses initially by rod photoreceptor degeneration caused by mutations in rod-specific genes, although different mutations in different genes converge upon the same rod degeneration phenotype in this disease. However, it is the subsequent cone photoreceptor degeneration that causes loss in daylight color vision and ultimately, diminishing quality of life for most patients. While gene therapy to replace a mutated gene with a functional copy has been successful, given the heterogeneity in mutations and genes, it is difficult to treat all RP cases by targeting the rods. Instead, a generic therapy to preserve the cones upon rod degeneration may lead to a more comprehensive therapeutic option. Despite progress, the molecular mechanism for this secondary cone degeneration remains unclear. In mouse models of RP, the cones in the central retina degenerate after rod death, but interestingly, the cones in the peripheral retina survive long-term. The goal of this proposed research is to understand the molecular and cellular mechanisms underlying peripheral cone survival in mouse models of RP. During the mentored phase of this grant (K99 phase), factors that may be sufficient (Aim 1) and/or necessary (Aim 2) for cone survival will be elucidated by cell type specific RNA sequencing, in vivo retinal electroporation, and temporally-regulated gene deletion. During the independent phase of this grant (R00 phase), the causal relationship between blood-retina-barrier breakdown and cone degeneration will be explored (Aim 3). Completion of the proposed aims will lead to identification of key regulators of cone survival in mouse models of RP. Moreover, we may identify, for the first time, a causal relationship between BRB breakdown and secondary cone death, opening new cellular targets to prevent cone loss in patients with RP. Long-term, the approaches outlined in this grant can become the cornerstone for answering questions regarding how, in general, neurons and other supporting cells degenerate in neurodegenerative disorders across the central nervous system. The mentored phase of this grant is conducted under the guidance of Dr. Constance Cepko, whose lab has developed many techniques over the years to genetically manipulate the retina in vivo and discovered gene regulatory networks underlying retinal cell type specification. The scientific environment that surrounds the Cepko Lab at Harvard Medical School offers valuable opportunities for career development, helping to build a strong foundation for an independent career investigating the molecular mechanisms of retinal degeneration.

项目概要色素性视网膜炎 (RP) 是一种遗传性视网膜疾病，全世界每 4,000 人中就有 1 人患有这种疾病。疾病最初是由于视杆细胞特异性基因突变引起的视杆细胞感光细胞变性而进展，尽管在这种疾病中，不同基因的不同突变会导致相同的视杆细胞退化表型。然而，正是随后的视锥细胞变性导致了日光色觉的丧失和最终，大多数患者的生活质量下降。虽然基因治疗用突变基因替换突变基因功能性拷贝已经成功，考虑到突变和基因的异质性，很难治疗所有的RP 通过瞄准杆的情况。相反，在视杆细胞退化时保留视锥细胞的通用疗法可能会导致以获得更全面的治疗选择。尽管取得了进展，但这种继发性的分子机制视锥细胞退化仍不清楚。在 RP 小鼠模型中，中央视网膜中的视锥细胞在视杆细胞后退化死亡，但有趣的是，视网膜周边的视锥细胞可以长期存活。这项研究的目标目的是了解小鼠模型中外周视锥细胞存活的分子和细胞机制 RP。在本次资助的指导阶段（K99 阶段），可能足够（目标 1）和/或必要的因素（目标 2）将通过细胞类型特异性 RNA 测序、体内视网膜电穿孔来阐明视锥细胞的存活，和时间调控的基因缺失。在本次资助的独立阶段（R00 阶段），因果关系将探讨血液-视网膜屏障破坏与视锥细胞变性之间的关系（目标 3）。完成所提出的目标将导致确定 RP 小鼠模型中视锥细胞存活的关键调节因子。此外，我们可能首次确定 BRB 分解与次级锥体之间的因果关系死亡，开辟新的细胞靶点以防止 RP 患者的视锥细胞丢失。从长远来看，概述的方法这笔赠款可以成为回答有关神经元和其他神经元一般如何运作的问题的基石在中枢神经系统的神经退行性疾病中，支持细胞会退化。受指导的该资助阶段是在 Constance Cepko 博士的指导下进行的，他的实验室开发了许多多年来对体内视网膜进行基因操纵的技术并发现了基因调控网络潜在的视网膜细胞类型规范。哈佛大学 Cepko 实验室周围的科学环境医学院提供宝贵的职业发展机会，有助于为未来的职业生涯奠定坚实的基础独立职业研究视网膜变性的分子机制。