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Laying a Foundation for Precision Medicine for Fuchs' Dystrophy

为福克斯营养不良症的精准医学奠定基础

基本信息

批准号：
10487580
负责人：
Venkateswara Vinod Mootha
金额：
$ 38.13万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10487580
关键词：
Affect Age Aging Anterior Antisense Oligonucleotides Apoptosis Aqueous Humor Binding Biology Blindness Blood CUG repeat Cells Cornea Corneal Diseases Corneal Endothelium Corneal edema Country Cultured Cells DNA Repeat Expansion DNA Sequence Data Defect Degenerative Disorder Disease Disease Progression Disease model Dominant-Negative Mutation Edema Endothelial Cells Endothelium Event Extracellular Matrix Extracellular Matrix Proteins Fibrosis Foundations Fuchs&apos Endothelial Dystrophy Future Gene Expression Genes Genetic Genetic Transcription Glaucoma Histologic Immune Inherited Keratoplasty Knock-in Laboratories Lead Length Leukocytes Link Medical Mitotic Modeling Molecular Mus Myotonic Dystrophy Neural Crest Neurodegenerative Disorders Nuclear Oligonucleotides PF4 Gene Pathogenesis Pathway interactions Patients Plant Roots Population Pre-Clinical Model Prevalence Primary Open Angle Glaucoma Proteins Publishing RNA RNA Splicing Reporting Resources Risk Role Safety Sampling Somatic Mutation Specificity Testing Therapeutic Tissues Toxic effect Trabecular meshwork structure Transcript Transcription Alteration Trinucleotide Repeats United States Up-Regulation Wild Type Mouse Work age related base biobank cell type cohort corneal epithelium drug testing gain of function genomic locus in vivo knock-down mouse model multidisciplinary overexpression precision medicine premature prevent recruit senescence single-cell RNA sequencing therapeutic development transcription factor transcriptome transcriptome sequencing

项目摘要

PROJECT SUMMARY Fuchs’ endothelial corneal dystrophy (FECD) is an age-related degenerative disorder resulting in corneal edema and loss of vision. FECD occurs in 4% of whites over the age of 40 years and is the leading indication for corneal transplantation in the U.S. Over 70% of cases are caused by a CTG triplet repeat expansion in the TCF4 gene. Expanded CUG repeat RNA (CUGexp) transcripts expressed from this gene locus accumulate as nuclear foci in the corneal endothelium of patients. The CUGexp foci bind and functionally sequester the splicing factors MBNL1 and MBNL2 to trigger mis-splicing in FECD endothelial tissue. To examine the endothelial cell-type specificity for FECD, we will determine if somatic mutations in the post-mitotic corneal endothelium of FECD patients results in a larger triplet repeat expansion than in their blood. We will examine if other anterior segment cell-types including corneal epithelium, stromal keratocytes, and trabecular meshwork cells are prone to accumulation of CUGexp foci and associated molecular defects. To test the MBNL sequestration hypothesis, we will examine if the knockdown of MBNLs in healthy donor endothelial tissue is sufficient to recapitulate the mis-splicing and upregulation of extracellular matrix (ECM) genes found early in the disease course. In early and late FECD, we observed a marked overexpression of the cochlin gene that produces a secreted ECM protein also capable of recruiting immune cells. The cochlin protein was previously detected in the trabecular meshwork of patients with primary open angle glaucoma (POAG), and our preliminary data indicate that the protein is present in the aqueous humor and trabecular meshwork of FECD subjects. We will examine levels of the cochlin protein in corneal tissue, aqueous humor samples, and trabecular meshwork tissue of FECD patients for its possible contribution to corneal disease findings and the increased risk for glaucoma in these patients. Additionally, we will examine the prevalence of POAG in our large UTSW FECD cohort and determine if there is a correlation with the triplet repeat length. We will use single-cell RNA sequencing combined with IHC to identify the immune cells and their contribution to late-stage FECD. We have characterized the efficacy of 45 antisense oligonucleotides (ASOs) and duplex RNAs that block disease-causing CUGexp foci. Using patient-derived cells and tissue, we will examine their potency based on their ability to block foci formation and their specificity based on transcriptome-wide assessment of on- and off-target events. We will examine delivery, length of action, and safety using wild-type mice. To enable studies of disease biology and in vivo drug testing, we have generated what we believe is the first mouse model with a knock-in of expanded CTG repeats. In preliminary studies, we show that our knock-in strategy successfully recapitulates CUGexp foci formation in corneal endothelium. We will further characterize this murine model by aging these mice to examine for the transcriptional and histologic alterations of FECD disease, and then use these mice to test our lead CUG-repeat targeting oligonucleotides for therapeutic development.

项目总结