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.

项目概要/摘要 Clarin1 (CLRN1) 基因突变导致 3 型亚瑟综合症 (USH3)，这是一种毁灭性的孤儿病尽管其表达水平非常低，但缺乏会导致人类失明和耳聋。 CLRN1 会导致视杆细胞、视锥细胞和耳蜗毛细胞进行性退化。目前可用于预防视力丧失的治疗方法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预防疗法的主要障碍在最近的研究中，我们和其他人报告了令人惊讶的感光细胞的渐进性退化。我们发现 CLRN1 mRNA 在多种脊椎动物的视网膜 Müller 胶质细胞中富集。发表的结果挑战了 USH3 和 USH3 中长期存在的光感受器驱动的退行性机制表明病理学是由关键的穆勒神经胶质细胞-光感受器相互作用的破坏引起的。 CLRN1 在 Müller 胶质细胞中的表达以及最终导致广泛传播的病理事件序列感光细胞损失是未知的。为了解决我们知识中的这些空白，并检验以下假设： USH3 是一种影响光感受器的穆勒神经胶质驱动的疾病，该项目提出了一种跨系统发育的方法与新开发的 USH3、斑马鱼和猪动物模型的整合方法在我们的初步数据中，我们证明，缺乏 clarin1 的斑马鱼模型表现出与年龄相关的光感受器损失。选择性恢复或删除 Müller 胶质细胞中的 CLRN1 表达，并确定 CLRN1 是否存在/不存在会改变光感受器的功能、结构和存活（目的1）。为了在大型动物模型中了解 USH3 视网膜疾病的细胞基础，我们培育了具有杂合子的 USH3 猪，双等位基因 CLRN1 突变在外显子 1 中包含不同的插入/缺失 (indels)。许多双等位基因杂合 USH3 突变猪的组合有严重的听力损失。通过光学相干断层扫描和暗视视网膜电图的变化可检测到视网膜层。因此，我们将比较四个独立的纯合子系的视网膜疾病的发展双等位基因 USH3 突变猪（相同的插入缺失），以识别表型驱动突变和特定的细胞和穆勒神经胶质细胞和光感受器的分子变化（目标 2）将成功完成。通过提供新的动物模型和 USH3 疾病的基础知识，显着推进该领域的发展，最终加速预防失明治疗方法的开发和验证。

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）

The USH3A causative gene clarin1 functions in Müller glia to maintain retinal photoreceptors.

USH3A 致病基因 clarin1 在穆勒神经胶质细胞中发挥作用，维持视网膜感光细胞。

DOI：
发表时间：
2024-03-01
期刊：
影响因子：
0
作者：
Nonarath, Hannah J T;Simpson, Samantha L;Slobodianuk, Tricia L;Collery, Ross F;Dinculescu, Astra;Link, Brian A
通讯作者：
Link, Brian A

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

亚瑟综合症视网膜基因治疗：当前发展、挑战和前景。

DOI：
发表时间：
2021-10-01
期刊：
影响因子：
0
作者：
Dinculescu, Astra;Link, Brian A;Saperstein, David A
通讯作者：
Saperstein, David A

Long-Term Effects of Gene Therapy in a Novel Mouse Model of Human MFRP-Associated Retinopathy.

基因治疗对人类 MFRP 相关视网膜病变新型小鼠模型的长期影响。

DOI：
发表时间：
2019
期刊：
影响因子：
4.2
作者：
Chekuri, Anil;Sahu, Bhubanananda;Chavali, Venkata Ramana Murthy;Voronchikhina, Marina;Soto;Suk, John J;Alapati, Akhila N;Bartsch, Dirk;Ayala;Zenteno, Juan C;Dinculescu, Astra;Jablonski, Monica M;Borooah, Shyama
通讯作者：
Borooah, Shyama

Modeling and Preventing Progressive Hearing Loss in Usher Syndrome III.

亚瑟综合症中进行性听力损失的建模和预防 III。

DOI：
发表时间：
2017
期刊：
影响因子：
4.6
作者：
Geng, Ruishuang;Omar, Akil;Gopal, Suhasini R;Chen, Daniel H;Stepanyan, Ruben;Basch, Martin L;Dinculescu, Astra;Furness, David N;Saperstein, David;Hauswirth, William;Lustig, Lawrence R;Alagramam, Kumar N
通讯作者：
Alagramam, Kumar N

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：
发表时间：
2019
期刊：
影响因子：
0
作者：
Simpson, Chiab P;Bolch, Susan N;Zhu, Ping;Weidert, Frances;Dinculescu, Astra;Lobanova, Ekaterina S
通讯作者：
Lobanova, Ekaterina S