CLARIN 1 RETINAL FUNCTION AND THERAPEUTIC IMPLICATIONS FOR USH3

CLARIN 1 视网膜功能和 USH3 的治疗意义

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目摘要/摘要 Clarin1（CLRN1）基因中的突变导致usher综合征3型（USH3），一种毁灭性的孤儿疾病导致人类的失明和耳聋。尽管表达水平非常低，但缺乏 CLRN1导致杆和锥形光感受器和耳蜗细胞的进行性变性。没有目前可用于防止视力丧失的处理。缺乏显示视网膜表型的动物模型一直是理解USH3视网膜病理生理学和开发疗法以防止的主要障碍感光细胞的进行性变性。在最近的研究中，我们和其他人报告了惊喜发现CLRN1 mRNA在几种脊椎动物物种中富集于视网膜müller神经胶质。我们的结合已发表的结果挑战了USH3和表明病理是由关键的müller胶质受体相互作用的破坏引起的。的意义 Müller神经胶质中的CLRN1表达和病理性事件的序列最终宽度感光细胞损失尚不清楚。在我们的知识中解决这些差距，并检验以下假设。 USH3是一种影响感光体的Müller胶质驱动疾病，该项目提出了跨系统发育与新开发的USH3，斑马鱼和猪的动物模型的整合方法。在我们的初步数据中我们表明，缺乏clarin1的斑马鱼模型显示了感光体的年龄依赖性损失。我们将选择性恢复或删除MüllerGlia中特别删除CLRN1表达，并确定CLRN1是否存在/不存在改变感光体功能，结构和存活（AIM1）。识别分子和在大型动物模型中，USH3视网膜疾病的细胞基础，我们产生了用杂合的USH3猪双质CLRN1突变包含外显子1中包含不同的插入/缺失（indels）。许多双质杂合子 USH3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变化通过光学相干断层扫描和SCOTOPIC电视图的变化可检测的视网膜薄片。因此，我们将比较四个独立的纯合线中残留疾病的发展双重USH3突变猪（相同的indel），以识别表型驱动突变和特定的细胞和 Müller神经胶质和光感受器的分子变化（AIM 2）。成功完成该项目将通过提供新的动物模型和关于USH3疾病的基本知识，可以显着提高该领域的领域，最终加快治疗的发展和验证以防止失明。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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
影响因子：
0
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
影响因子：
0
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