Investigation of the Mechanisms of Cone Degeneration in Retinitis Pigmentosa

色素性视网膜炎视锥细胞变性机制的研究

• 批准号: 10532024
• 负责人: Yunlu Xue
• 金额: $ 4.05万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10532024
• 关键词: Affect; Award; Blindness; Candidate Disease Gene; Cells; Cessation of life; Color Visions; Cone; Confocal Microscopy; Data; Development; Disease; Electron Transport; Electrophysiology (science); Electroretinography; Energy-Generating Resources; Enzymes; Fluorescence; Gene Expression; Gene Mutation; Genes; Glucose; Glycolysis; Goals; HIF1A gene; Histology; Image; Individual; Inherited; Investigation; Ketones; Life; Light; Link; Lipids; Mediating; Mentors; Mentorship; Metabolic; Mitochondria; Modeling; Molecular Biology; Muller' s cell; Mus; Muscle; Mutation; NADH; NADP; Ophthalmoscopy; Opsin; Patients; Peripheral; Persons; Phase; Proteins; RPE65 protein; Research Personnel; Residual state; Retinal Cone; Retinal Diseases; Retinitis Pigmentosa; Rod; Safety; Scanning; Signal Transduction; Source; Supervision; TXNIP gene; Testing; Therapeutic; Time; Training; Up-Regulation; Vision; adaptive optics; adaptive optics scanning laser ophthalmoscopy; adeno-associated viral vector; base; design; dosage; effective therapy; functional loss; gene therapy; glucose metabolism; glucose uptake; high resolution imaging; imaging platform; in vivo; knock-down; legally blind; loss of function; mouse model; overexpression; preservation; prevent; ratiometric; retinal imaging; retinal rods; sensor; small hairpin RNA; targeted treatment; therapeutic candidate; therapeutic gene; therapeutically effective; transcriptome sequencing; two-photon; visual cycle

PROJECT SUMMARY / ABSTRACT There is currently no cure for retinitis pigmentosa (RP), which is the most prevalent inherited retinal disease (IRD). RP patietns first suffer from nightblindness caused by expression of disease genes in rods, then gradually lose their peripheral daylight vision, mediated by cones, thus forming tunnel vision. They eventually become legally blind with the complete loss of central cone-mediated vision. It is unknown why the cones lose their function and eventually die after rods die. Rescuing cones from the non-autonomous degeneration has been proposed as a generic therapeutic approach to treat RP, regardless of the rod gene mutation. Recently, the applicant found an effective therapeutic gene, Txnip, out of 30 candidates using AAV-mediated gene therapy in RP mouse models. Txnip is excellent in delaying RP cone degeneration after the rods are gone. However, it is unclear why Txnip can rescue the RP cones from degeneration. Therefore, the applicant propose to study the mechanism of Txnip in the mentored phase (K99), during which the applicant will continue his training in molecular biology, and in two-photon adaptive optics scanning light ophthalmoloscopy (2P-AOSLO) that will allow dissecting the downstream targets of this therapeutic candidate. Furthermore, in the independent phase (R00), the applicant propose to investigate in the mechanism of cone functional loss, which occurs abruptly upon rod death, but well before cone degeneration. Specifically, the applicant will apply shRNA knockdown of proposed downstream genes of the rescue gene, co-deliver the knockdown with the rescue target, and examine if the rescue effect on RP cone survival can be abolished (Aim I, K99). The applicant will also examine the proposed metabolite level changes in RP cones with and without this therapeutic gene by imaging of metabolite sensors ex vivo and in vivo (Aim II, K99). Finally, the applicant will investigate the mechanism of the rapid RP cone ERG function loss based on preliminary electrophysiology, RNA-seq and histology data, in the hope of preserving the RP cone function as long as possible before their eventual death (Aim III, R00). Overall, this project is designed to combine molecular biology, high-resolution imaging, and electrophysiology to find a treatment for RP. The mentored phase of this award will be conducted under the supervision of Dr. Constance Cepko (Harvard), and the co-mentorship of Dr. Jennifer Hunter (U Rochester) who is an expert in 2P-AOSLO. This project will also receive material and advisory support from Dr. Gary Yellen (Harvard), who is an expert in genetically encoded metabolite sensors.

项目概要/摘要 目前尚无法治愈色素性视网膜炎（RP），这是最常见的遗传性视网膜疾病 （税务局）。 RP患者首先患有由视杆细胞中疾病基因表达引起的夜盲症，然后逐渐发展 失去由视锥细胞介导的周边日光视觉，从而形成隧道视觉。他们最终成为 法定失明，中央锥体介导的视力完全丧失。目前尚不清楚为什么视锥细胞会失去作用 功能并最终在杆死亡后死亡。将视锥细胞从非自主退化中拯救出来 被提议作为治疗 RP 的通用治疗方法，无论 rod 基因突变如何。最近， 申请人在使用 AAV 介导的基因治疗的 30 个候选基因中发现了有效的治疗基因 Txnip RP 鼠标型号。 Txnip 在延缓视杆细胞消失后 RP 视锥细胞退化方面表现出色。然而，它是 不清楚为什么 Txnip 可以拯救 RP 视锥细胞免于退化。因此，申请人建议研究 Txnip 的指导阶段（K99）机制，在此期间申请人将继续接受培训 分子生物学，以及双光子自适应光学扫描光检眼镜 (2P-AOSLO) 允许剖析该候选治疗药物的下游靶标。此外，在独立阶段 (R00)，申请人提议研究视锥细胞功能丧失的机制，这种丧失突然发生 视杆细胞死亡，但早在视锥细胞退化之前。具体来说，申请人将应用 shRNA 敲低 提出救援基因的下游基因，与救援目标共同传递敲低，并检查 如果可以取消对 RP 锥生存的救援效果（目标 I，K99）。申请人还将审查 通过代谢物成像提出有和没有这种治疗基因的 RP 锥体中代谢水平的变化 离体和体内传感器（Aim II，K99）。最后，申请人将研究快速RP的机制 基于初步电生理学、RNA-seq 和组织学数据的锥体 ERG 功能丧失，希望 在最终死亡之前尽可能长时间地保留 RP 锥体功能（目标 III，R00）。总体而言，这 该项目旨在结合分子生物学、高分辨率成像和电生理学来寻找 RP 治疗。该奖项的指导阶段将在康斯坦斯博士的监督下进行 Cepko（哈佛大学），以及 2P-AOSLO 专家 Jennifer Hunter 博士（罗切斯特大学）的共同导师。 该项目还将获得加里·耶伦博士（哈佛大学）的物质和咨询支持，他是以下领域的专家： 基因编码的代谢传感器。

项目成果

AAV-Txnip prolongs cone survival and vision in mouse models of retinitis pigmentosa.

• DOI: 10.7554/elife.66240
• 发表时间: 2021-04-13
• 期刊: eLife
• 影响因子: 7.7
• 作者: Xue Y;Wang SK;Rana P;West ER;Hong CM;Feng H;Wu DM;Cepko CL
• 通讯作者: Cepko CL

Augmentation of CD47/SIRPα signaling protects cones in genetic models of retinal degeneration.

• DOI: 10.1172/jci.insight.150796
• 发表时间: 2021-08-23
• 期刊: JCI insight
• 影响因子: 8
• 作者: Wang SK;Xue Y;Cepko CL
• 通讯作者: Cepko CL

Engineering adeno-associated viral vectors to evade innate immune and inflammatory responses.

• DOI: 10.1126/scitranslmed.abd3438
• 发表时间: 2021-02-10
• 期刊: Science translational medicine
• 影响因子: 17.1
• 作者: Chan YK;Wang SK;Chu CJ;Copland DA;Letizia AJ;Costa Verdera H;Chiang JJ;Sethi M;Wang MK;Neidermyer WJ Jr;Chan Y;Lim ET;Graveline AR;Sanchez M;Boyd RF;Vihtelic TS;Inciong RGCO;Slain JM;Alphonse PJ;Xue Y;Robinson-McCarthy LR;Tam JM;Jabbar MH;Sahu B;Adeniran JF;Muhuri M;Tai PWL;Xie J;Krause TB;Vernet A;Pezone M;Xiao R;Liu T;Wang W;Kaplan HJ;Gao G;Dick AD;Mingozzi F;McCall MA;Cepko CL;Church GM
• 通讯作者: Church GM