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

项目摘要/摘要血管生成是血管形成的过程，其中增殖的内皮细胞（ECS）从预先存在的视频以扩展血管网络。受损或过量血管生成与人类视网膜疾病的发病机理，例如视网膜低血管家族性真实玻璃体病变和高血管糖尿病性视网膜病。 EC主要使用糖酵解途径，该途径在血管生成。最近的研究表明，线粒体活性对于血管生成至关重要，但其机制至关重要并不完全清楚。我们已经研究了三种线粒体蛋白具有不同的血管生成功能活动 - 线粒体转录因子（TFAM），呼吸复合物IV成分（COX10）和A 线粒体氧化还原蛋白硫氧还蛋白2（TRX2）。我们的数据表明：1）在一个中TFAM，COX10或TRX2沉默体外3D发芽测定法减弱了EC发芽，这与降低EC相关，但与 ROS产生或EC凋亡； 2）具有诱导TFAM，COX10或TRX2暴露智障的小鼠视网膜血管的生长在早期不穿透到深plexi中（P5 – P12），这使视网膜衰减发芽与MTROS生产无关。 3）三个突变小鼠发展动静脉畸形（AVM）在微血管中具有增强的动脉化和微型神经瘤形成高级年龄（P12 – P30）。低血脉和微型神经瘤表型类似于年龄的人类的表型视网膜和人类视网膜血管异常，例如外套疾病，Leber的军事动脉瘤和家庭透明性玻璃体病（FEVR）； 4）此外，残留EC的单细胞RNA-seq分析表明这三只突变小鼠的EC簇具有降低的基因表达，在血管生成和代谢中途径但增加了TGFR信号传导。基于这些数据，我们假设线粒体活性在血管生长中调节常见的血管生成和代谢途径（而不是ROS/凋亡）成熟。我们提出以下特定目的：1）确定关键的血管生成和代谢途径在视网膜发芽血管生成中的线粒体功能障碍引起的缺陷中形成以及视网膜功能； 2）定义线粒体功能障碍如何调节TGFR-SMAD2/3 信号传导； 3）确定TGFR-SMAD2/3信号在线粒体功能障碍诱导的作用药物阻断和遗传缺乏症的血管延迟和畸形。我们提出的研究将定义线粒体活性调节正常永久性血管生长和成熟，并将为人类永久性血管疾病提供新颖的模型和治疗干预措施与许多病理并发症有关，例如糖尿病，高血压和衰老，可能导致视力丧失。 1