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Pluripotent Stem Cell Derived 3D Retinas for Studies of Early Onset Retinal Degeneration

多能干细胞衍生的 3D 视网膜用于研究早发性视网膜变性

基本信息

批准号：
10600997
负责人：
KARL J WAHLIN
金额：
$ 39.5万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
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：
2020-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10600997
关键词：
3-Dimensional ARA9 protein Address Affect Age related macular degeneration Alleles Animals Apoptosis Binding Binding Proteins Biological Models Biology Birth Blindness CRISPR/Cas technology Cell Cycle Proteins Cell Death Cell Survival Cells Childhood Clinical Complex Cone Cyclic GMP DNA Repair Data Defect Disease Electrophysiology (science)Experimental Models Eye diseases Fluorescence Freezing Functional disorder Gene Expression Genes Goals Human Inherited Knock-out Knowledge Label Laboratories Lead Leber&apos s amaurosis Measures Modeling Molecular Monitor Morphology Mus Mutation Organoids Pathogenesis Pathway interactions Patients Phenotype Photoreceptors Pluripotent Stem Cells Population Primates Property Proteins RPE65 protein Recombinant adeno-associated virus (rAAV)Recovery of Function Reporter Resources Retina Retinal Degeneration Retinal Diseases Retinal Dystrophy Retinitis Pigmentosa Rod Role Severities Sorting Structure Study models System Testing Time Tissues Variant Vertebrate Photoreceptors Viral Western Blotting Work cell type comparative cone-rod dystrophy disease phenotype disease-in-a-dish early onset gene correction gene therapy genome editing human disease human pluripotent stem cell human stem cells in vivo inherited retinal degeneration innovation innovative technologies insight knockout gene live cell imaging mRNA Expression mutant photoreceptor degeneration polyproline prevent repaired response selective expression stem cell derived tissues stem cells therapy development

项目摘要

SUMMARY Retinal degenerative (RD) diseases, such as Retinitis pigmentosa (RP) and Leber congenital amaurosis (LCA), cause dysfunction and cell death of photoreceptor (PR) cells, ultimately leading to blindness. LCA is the leading cause of inherited childhood blindness resulting in a loss of vision at or soon after birth. Though this is considered to be quite rare, these blinding diseases are devastating for those affected. Current efforts are being made to develop gene-therapies aimed at correcting some of the genes affected in RD and this approach has shown some promise in animals and humans for restoring RPE65 gene expression, but there are many other causes of RD for which there is no cure. In addition, due to the many mutations involved in RD, there are significant gaps in our understanding of how PR loss occurs. To address this, we will use human pluripotent stem cell (PSC) based retinal cell-reporter lines with RD-associated alleles to help explore the mechanisms of PR cell death. Given the typically long period of time required to generate human retinas in the laboratory, the severity and rapid onset of degeneration in LCA makes it an attractive experimental model to study human RD and to develop potential therapies. We will study the aryl hydrocarbon receptor interacting protein-like1 (AIPL1) gene to explore three functional domains that harbor naturally occurring mutations in patients with LCA and cone-rod dystrophy (CORD). A comparative analysis of different mutations might lead to a better understanding of how rods and cones die and greater insight into other more common forms of PR degeneration, such as age- related macular degeneration (AMD). A central hypothesis is that human PSC derived 3D retina organoids with AIPL1 mutations will recapitulate human retinal dystrophy resulting in PR loss. This hypothesis is supported by our recent work, and others, showing that human PSCs can be coaxed into becoming retinal eyecup-like structures with PRs, a laminar morphology and outer segment structures that are similar to an actual retina. This proposal will bridge two innovative technologies; (1) genome-editing to generate genetically matched retinal reporter PSC derived retinas with disease-associated mutations and (2) gene-correction to repair genetic defects and promote PR cell survival. Given the very early onset of LCA it is important to define the appropriate windows of time for such treatment options. Not only will these studies lead to new insights into the biology of RD disease, but could also provide an innovative resource to develop therapies for the treatment of RD.

总结视网膜变性（RD）疾病，如视网膜色素变性（RP）和Leber先天性黑蒙 (LCA)引起感光细胞（PR）的功能障碍和细胞死亡，最终导致失明。LCA 是遗传性儿童失明的主要原因，导致出生时或出生后不久视力丧失。虽然这被认为是非常罕见的，这些致盲疾病对受影响的人来说是毁灭性的。目前的努力正在开发旨在纠正RD中受影响的一些基因的基因疗法，已经在动物和人类中显示出恢复RPE 65基因表达的一些希望，但还有许多其他无法治愈的RD原因。此外，由于RD涉及许多突变，我们对公关损失如何发生的理解存在重大差距。为了解决这个问题，我们将使用人类多能干细胞（PSC）为基础的视网膜细胞报告细胞系与RD相关的等位基因，以帮助探索机制， PR细胞死亡。考虑到在实验室中生成人类视网膜通常需要很长的时间， LCA的严重性和快速变性使其成为研究人类RD的有吸引力的实验模型并开发潜在的治疗方法。我们将研究芳香烃受体相互作用蛋白样1（AIPL 1）基因，以探索LCA患者中天然存在突变的三个功能结构域，视锥-视杆细胞营养不良。对不同突变的比较分析可能会导致更好的理解如何杆和锥死亡和更深入了解其他更常见的形式的PR退化，如年龄- 相关性黄斑变性（AMD）。中心假设是人PSC衍生的3D视网膜类器官具有AIPL 1突变的人将重演导致PR损失的人视网膜营养不良。这种假设是我们最近的工作和其他人的工作支持，表明人类PSC可以被诱导成为视网膜，具有PR的眼罩状结构，层状形态和与实际结构相似的外段结构视网膜。这项提案将连接两项创新技术：（1）基因组编辑，具有疾病相关突变的匹配的视网膜报告子PSC衍生的视网膜，和（2）基因校正，修复基因缺陷，促进PR细胞存活。考虑到LCA的早期发作，这些治疗方案的适当时间窗口。这些研究不仅会带来新的见解研究RD疾病的生物学，但也可以提供一个创新的资源来开发治疗方法， RD的治疗。

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）

Bulk RNA sequencing analysis of developing human induced pluripotent cell-derived retinal organoids.

DOI：
10.1038/s41597-022-01853-x
发表时间：
2022-12-09
期刊：
SCIENTIFIC DATA
影响因子：
9.8
作者：
Agarwal, Devansh;Kuhns, Rian;Dimitriou, Christos N. N.;Barlow, Emmalyn;Wahlin, Karl J. J.;Enke, Ray A. A.
通讯作者：
Enke, Ray A. A.

A Tet-Inducible CRISPR Platform for High-Fidelity Editing of Human Pluripotent Stem Cells.

DOI：
10.3390/genes13122363
发表时间：
2022-12-14
期刊：
Genes
影响因子：
3.5
作者：
通讯作者：

Temporal and Isoform-Specific Expression of CTBP2 Is Evolutionarily Conserved Between the Developing Chick and Human Retina.

DOI：
10.3389/fnmol.2021.773356
发表时间：
2021
期刊：
Frontiers in molecular neuroscience
影响因子：
4.8
作者：
Gage E;Agarwal D;Chenault C;Washington-Brown K;Szvetecz S;Jahan N;Wang Z;Jones MK;Zack DJ;Enke RA;Wahlin KJ
通讯作者：
Wahlin KJ

Human retinal ganglion cell neurons generated by synchronous BMP inhibition and transcription factor mediated reprogramming.

DOI：
10.1038/s41536-023-00327-x
发表时间：
2023-09-29
期刊：
NPJ REGENERATIVE MEDICINE
影响因子：
7.2
作者：
Agarwal, Devansh;Dash, Nicholas;Mazo, Kevin W.;Chopra, Manan;Avila, Maria P.;Patel, Amit;Wong, Ryan M.;Jia, Cairang;Do, Hope;Cheng, Jie;Chiang, Colette;Jurlina, Shawna L.;Roshan, Mona;Perry, Michael W.;Rho, Jong M.;Broyer, Risa;Lee, Cassidy D.;Weinreb, Robert N.;Gavrilovici, Cezar;Oesch, Nicholas W.;Welsbie, Derek S.;Wahlin, Karl J.
通讯作者：
Wahlin, Karl J.

CRISPR Generated SIX6 and POU4F2 Reporters Allow Identification of Brain and Optic Transcriptional Differences in Human PSC-Derived Organoids.

DOI：
10.3389/fcell.2021.764725
发表时间：
2021
期刊：
Frontiers in cell and developmental biology
影响因子：
5.5
作者：
Wahlin KJ;Cheng J;Jurlina SL;Jones MK;Dash NR;Ogata A;Kibria N;Ray S;Eldred KC;Kim C;Heng JS;Phillips J;Johnston RJ Jr;Gamm DM;Berlinicke C;Zack DJ
通讯作者：
Zack DJ