CLARIN 1 RETINAL FUNCTION AND THERAPEUTIC IMPLICATIONS FOR USH3

CLARIN 1 视网膜功能和 USH3 的治疗意义

• 批准号: 10753724
• 负责人: ASTRA DINCULESCU
• 金额: $ 71.06万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10753724
• 关键词: Acceleration; Address; Adhesions; Affect; Age; Alleles; Animal Model; Apical; Auditory; Automobile Driving; Biological Process; Biology; Blindness; Cells; Cellular Stress; Clarin-1; Cochlea; Code; Complex; Cytoskeleton; Data; Defect; Development; Disease; Early identification; Electroretinography; Event; Exons; Eye; Family suidae; Functional disorder; Genes; Hair; Hair Cells; Heterozygote; Human; Immunofluorescence Immunologic; Inherited; Knowledge; Legal Blindness; Life; Link; Mechanical Stress; Messenger RNA; Modeling; Molecular; Molecular Probes; Morphology; Muller' s cell; Mus; Mutation; Nonsense Codon; Optical Coherence Tomography; Outer Limiting Membrane; Pathologic; Pathology; Patients; Phenotype; Photoreceptors; Phylogenetic Analysis; Physiology; Predisposition; Process; Proteins; Publishing; Rare Diseases; Reporting; Research; Research Personnel; Retina; Retinal Cone; Retinal Diseases; Role; Series; Severities; Signal Pathway; Specificity; Structure; Syndrome; Testing; Therapeutic; Transmission Electron Microscopy; Usher Proteins; Usher Syndrome; Usher Syndrome Type 3; Validation; Vertebrate Photoreceptors; Zebrafish; adeno-associated viral vector; age related; autosome; biological adaptation to stress; cell type; clinically relevant; cohort; deaf; deafness; design; experimental study; in vivo; insertion/deletion mutation; legally blind; mutant; nonhuman primate; novel; photoreceptor degeneration; prevent; retinal progenitor cell; retinal rods; scaffold; selective expression; therapy development; transcriptomics; translational therapeutics

Project Summary/Abstract Mutations in the Clarin1 (CLRN1) gene cause Usher syndrome type 3 (USH3), a devastating orphan disease leading to combined blindness and deafness in humans. Despite its very low levels of expression, the lack of CLRN1 results in progressive degeneration of rod and cone photoreceptors and cochlear hair cells. There is no treatment currently available to prevent vision loss. The lack of animal models that display a retinal phenotype has been a major barrier to understanding the USH3 retinal pathophysiology and developing therapies to prevent the progressive degeneration of the photoreceptor cells. In recent studies, we and others reported the surprising discovery that CLRN1 mRNA is enriched in retinal Müller glia across several vertebrate species. Our combined published results challenge a long-standing photoreceptor-driven degenerative mechanism in USH3 and suggest that pathology arises from disruption of key Müller glia-photoreceptor interactions. The significance of CLRN1 expression in Müller glia and the sequence of pathological events culminating in widespread photoreceptor cell loss are unknown. To address these gaps in our knowledge, and test the hypothesis that USH3 is a Müller glia-driven disease affecting photoreceptors, this project proposes a cross-phylogenetic integration approach with newly developed animal models of USH3, zebrafish and pig. In our preliminary data, we show that our zebrafish model lacking clarin1 displays an age-dependent loss of photoreceptors. We will selectively restore or delete CLRN1 expression specifically in Müller glia and determine whether CLRN1 presence/absence alters photoreceptor function, structure, and survival (Aim1). To identify the molecular and cellular basis of USH3 retinal disease in a large animal model, we generated USH3 pigs with heterozygous, biallelic CLRN1 mutations containing distinct insertions/deletions (indels) in exon 1. Many biallelic heterozygous combinations of USH3 mutant pigs have profound hearing loss. A smaller subset shows alterations in the outer retinal laminae detectable by optical coherence tomography and changes in the scotopic electroretinogram. Therefore, we will compare the development of retinal disease across four independent lines of homozygous biallelic USH3 mutant pigs (same indel) to identify the phenotype-driving mutation and specific cellular and molecular changes in Müller glia and photoreceptors (Aim 2). Successful completion of this project will significantly advance the field by providing new animal models and fundamental knowledge on USH3 disease, to ultimately accelerate the development and validation of treatments to prevent blindness.

项目总结/摘要 Clarin 1（CLRN 1）基因突变导致Usher综合征3型（USH 3），一种毁灭性的孤儿疾病 导致人类失明和失聪。尽管表达水平很低，但缺乏 CLRN 1导致视杆和视锥光感受器以及耳蜗毛细胞的进行性变性。没有 目前可用于预防视力丧失的治疗。缺乏显示视网膜表型的动物模型 一直是理解USH 3视网膜病理生理学和开发治疗方法以预防 感光细胞的逐渐退化。在最近的研究中，我们和其他人报告了令人惊讶的 CLRN 1 mRNA在几种脊椎动物的视网膜Müller神经胶质中富集的发现。我们的联合 发表的结果挑战了USH 3中长期存在的光受体驱动的退行性机制， 表明病理学是由关键的Müller胶质细胞-感光细胞相互作用的破坏引起的。的意义 CLRN 1在Müller胶质细胞中的表达和病理事件的顺序最终导致广泛的 光感受器细胞损失是未知的。为了填补我们知识上的这些空白，并验证假设， USH 3是一种影响光感受器的Müller神经胶质驱动的疾病，该项目提出了一种跨系统发育的 与新开发的USH 3、斑马鱼和猪动物模型的整合方法。在我们的初步数据中， 我们发现缺乏clarin 1的斑马鱼模型显示出光感受器的年龄依赖性损失。我们将 选择性恢复或删除CLRN 1表达，特别是在Müller胶质细胞，并确定CLRN 1是否 存在/不存在会改变感光细胞的功能、结构和存活（Aim 1）。为了鉴定分子和 USH 3视网膜疾病的细胞基础在大型动物模型中，我们产生了具有杂合的USH 3猪， 在外显子1中含有不同插入/缺失（indels）的双等位基因CLRN 1突变。多双等位基因杂合子 USH 3突变猪的组合具有严重的听力损失。一个较小的子集显示了外部的改变， 通过光学相干断层扫描和暗视视网膜电图的变化检测视网膜层。 因此，我们将比较视网膜疾病的发展，在四个独立的系的纯合子， 双等位基因USH 3突变猪（相同插入缺失），以鉴定表型驱动突变和特异性细胞和 Müller神经胶质和光感受器的分子变化（目的2）。该项目的成功完成将 通过提供新的动物模型和USH 3疾病的基础知识， 最终加速预防失明的治疗方法的开发和验证。

项目成果

Gene Therapy in a Large Animal Model of PDE6A-Retinitis Pigmentosa.

• DOI: 10.3389/fnins.2017.00342
• 发表时间: 2017
• 期刊: Frontiers in neuroscience
• 影响因子: 4.3
• 作者: Mowat FM;Occelli LM;Bartoe JT;Gervais KJ;Bruewer AR;Querubin J;Dinculescu A;Boye SL;Hauswirth WW;Petersen-Jones SM
• 通讯作者: Petersen-Jones SM

Retinal Gene Therapy for Usher Syndrome: Current Developments, Challenges, and Perspectives.

• DOI: 10.1097/iio.0000000000000378
• 发表时间: 2021-10-01
• 期刊: International ophthalmology clinics
• 作者: Dinculescu A;Link BA;Saperstein DA
• 通讯作者: Saperstein DA

Systemic Delivery of Genes to Retina Using Adeno-Associated Viruses.

使用腺相关病毒将基因系统性递送至视网膜。

• DOI: 10.1007/978-3-030-27378-1_18
• 发表时间: 2019
• 期刊: Advances in experimental medicine and biology
• 作者: Simpson,ChiabP;Bolch,SusanN;Zhu,Ping;Weidert,Frances;Dinculescu,Astra;Lobanova,EkaterinaS
• 通讯作者: Lobanova,EkaterinaS

A Modified Arrestin1 Increases Lactate Production in the Retina and Slows Retinal Degeneration.

• DOI: 10.1089/hum.2021.272
• 发表时间: 2022-07
• 期刊: HUMAN GENE THERAPY
• 影响因子: 4.2
• 作者: Nelson, Tiffany S.;Simpson, Chiab;Dyka, Frank;Dinculescu, Astra;Smith, W. Clay
• 通讯作者: Smith, W. Clay

Modeling and Preventing Progressive Hearing Loss in Usher Syndrome III.

• DOI: 10.1038/s41598-017-13620-9
• 发表时间: 2017-10-18
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Geng R;Omar A;Gopal SR;Chen DH;Stepanyan R;Basch ML;Dinculescu A;Furness DN;Saperstein D;Hauswirth W;Lustig LR;Alagramam KN
• 通讯作者: Alagramam KN