Investigating the role of mitochondrial dysfunction in the pathogenesis of retinal vascular diseases

研究线粒体功能障碍在视网膜血管疾病发病机制中的作用

基本信息

批准号：
10662463
负责人：
Jenny Huanjiao Zhou
金额：
$ 41.88万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10662463
关键词：
3-Dimensional Age Age related macular degeneration Aging Aneurysm Apoptosis Arteriovenous malformation Attenuated Biochemical Biological Assay Blindness Blood Vessels Canes Cardiovascular Diseases Cell Proliferation Cells Cellular biology Coats&apos disease Cues DNA Sequence Alteration Data Defect Diabetes Mellitus Diabetic Retinopathy Diving Elderly Endothelial Cells Exhibits Eye Familial exudative vitreoretinopathy Feedback Gene Expression Generations Genes Genetic Genetic Transcription Glycolysis Glycolysis Pathway Growth Human Hypertension Hypervascular Impairment In Vitro MAP Kinase Gene MAPK8 gene Mediating Metabolic Pathway Microaneurysm Military Personnel Mitochondria Mitochondrial Proteins Modeling Molecular Mus Mutant Strains Mice Nuclear Oxidation-Reduction Pathogenesis Pathologic Pathway interactions Penetration Pericytes Phenotype Phosphorylation Process Production Proliferating Proteins Respiration Retina Retinal Diseases Role Signal Transduction Stroke Structure TGF-beta type I receptor Therapeutic Intervention Thioredoxin-2 Vascular Diseases Vascular Endothelial Cell aged angiogenesis blood vessel development complex IV effective therapy inhibitor malformation microscopic imaging mitochondrial dysfunction novel p38 Mitogen Activated Protein Kinase pharmacologic postnatal recruit respiratory retina blood vessel structure single-cell RNA sequencing transcription factor transcriptome vascular abnormality

项目摘要

PROJECT SUMMARY/ABSTRACT Angiogenesis is the process of blood vessel formation in which proliferating endothelial cells (ECs) sprout from preexisting vessels to extend a vascular network. Either impaired or excessive angiogenesis is associated with the pathogenesis of human retina diseases such as retinal hypovascular familial exudative vitreoretinopathy and hypervascular diabetic retinopathy. EC primarily uses the glycolysis pathway which is further enhanced during angiogenesis. Recent studies suggest that mitochondrial activity is critical for angiogenesis, but its mechanism is not entirely clear. We have investigated the angiogenic functions of three mitochondrial proteins with distinct activities - mitochondrial transcriptional factor (TFAM), respiratory complex IV component (COX10) and a mitochondrial redox protein thioredoxin 2 (TRX2). Our data show that: 1) silencing of Tfam, Cox10, or Trx2 in an in vitro 3D sprouting assay attenuates EC sprouting, which correlated with reduced EC proliferation, but not with ROS generation or EC apoptosis; 2) mice with an inducible deletion of Tfam, Cox10, or Trx2 exhibit retarded retinal vessel growth without penetration into the deep plexi at early ages (P5–P12), and this attenuated retinal sprouting was not correlated with mtROS production; 3) the three mutant mice develop arteriovenous malformations (AVM) with enhanced arterialization and microaneurysm formation in the microvessels at advanced ages (P12–P30). The hypovasculature and microaneurysm phenotypes resemble that of aged human retinas and human retinal vascular abnormalities such as Coats' disease, Leber's military aneurysms and familial exudative vitreoretinopathy (FEVR); 4) Furthermore, single-cell RNA-seq analyses of retinal ECs suggest that the three mutant mice have common EC clusters with reduced gene expression in angiogenic and metabolic pathways but increased TGFR signaling. Based on these data, we hypothesize that mitochondrial activity regulates common angiogenic and metabolic pathways (rather than ROS/apoptosis) in vascular growth and maturation. We propose the following specific aims: 1) To identify critical angiogenic and metabolic pathways in mitochondrial dysfunction-induced defects in retinal sprouting angiogenesis, AVM and microaneurysm formation as well as in retinal function; 2) To define how mitochondrial dysfunction regulates TGFR-Smad2/3 signaling; 3) To determine the role of the TGFR-Smad2/3 signaling in mitochondrial dysfunction-induced vascular retardation and malformation by pharmacological blockade and genetic deficiency. Our proposed study will define the mechanism by which mitochondrial activities regulate normal retinal vascular growth and maturation, and will provide a novel model and therapeutic intervention for human retinal vascular diseases associated with many pathological complications such as diabetes, hypertension and aging that can result in vision loss. 1

项目概要/摘要血管生成是血管形成的过程，其中增殖的内皮细胞（EC）从血管中萌芽。现有血管的扩张与血管生成受损或过度相关。视网膜缺血管家族性渗出性玻璃体视网膜病变等人类视网膜疾病的发病机制 EC 主要利用糖酵解途径，该途径在糖尿病视网膜病变期间进一步增强。最近的研究表明线粒体活性对于血管生成至关重要，但其机制尚不完全清楚。我们研究了三种具有不同性质的线粒体蛋白的血管生成功能。活性 - 线粒体转录因子 (TFAM)、呼吸复合物 IV 成分 (COX10) 和线粒体氧化还原蛋白硫氧还蛋白 2 (TRX2) 我们的数据表明：1) Tfam、Cox10 或 Trx2 的沉默。体外 3D 发芽测定会减弱 EC 发芽，这与 EC 增殖减少相关，但与 EC 增殖无关 ROS 生成或 EC 凋亡；2) Tfam、Cox10 或 Trx2 诱导性缺失的小鼠表现出迟缓早期（P5-P12）视网膜血管生长但未穿透深部神经丛，并且这种减弱的视网膜发芽与 mtROS 产生无关；3）三种突变小鼠发育出动静脉畸形（AVM），伴有微血管中动脉化增强和微动脉瘤形成高龄（P12-P30）。血管不足和微动脉瘤表型与老年人相似。视网膜和人类视网膜血管异常，例如 Coats 病、Leber 军用动脉瘤和家族性动脉瘤渗出性玻璃体视网膜病变 (FEVR)；4) 此外，视网膜 EC 的单细胞 RNA-seq 分析表明这三种突变小鼠具有共同的 EC 簇，在血管生成和代谢方面的基因表达降低根据这些数据，我们捕获了线粒体活性。调节血管生长和代谢过程中常见的血管生成和代谢途径（而不是 ROS/细胞凋亡）我们提出以下具体目标：1）确定关键的血管生成和代谢途径。线粒体功能障碍引起的视网膜萌芽血管生成缺陷、AVM 和微动脉瘤形成以及视网膜功能；2) 确定线粒体功能障碍如何调节 TGFR-Smad2/3 3) 确定 TGFβR-Smad2/3 信号传导在线粒体功能障碍诱导中的作用我们提出的研究通过药物阻断和遗传缺陷导致血管发育迟缓和畸形。将定义线粒体活动调节正常视网膜血管生长的机制成熟，将为人类视网膜血管疾病提供新的模型和治疗干预与许多病理并发症有关，例如糖尿病、高血压和衰老，这些并发症可能导致视力丧失。 1