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

• 批准号: 10443059
• 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
• 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/10443059
• 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' 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; Pharmacology; 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; base; blood vessel development; complex IV; effective therapy; inhibitor; malformation; microscopic imaging; mitochondrial dysfunction; novel; p38 Mitogen Activated Protein Kinase; postnatal; ranpirnase; recruit; respiratory; retina blood vessel structure; single-cell RNA sequencing; transcription factor; transcriptome; vascular abnormality

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

项目总结/摘要 血管生成是血管形成的过程，其中增殖的内皮细胞（EC）从血管中生长出来。 预先存在的血管来扩展血管网络。血管生成受损或过度与 人视网膜疾病如视网膜血管减少性家族性渗出性玻璃体视网膜病变和 糖尿病视网膜病变EC主要使用糖酵解途径，糖酵解途径在 血管生成最近的研究表明，线粒体活性是血管生成的关键，但其机制， 尚不完全清楚。我们研究了三种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组分（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Trx 2在一个细胞中的沉默， 体外3D发芽试验减弱了EC发芽，这与EC增殖减少相关，但与 ROS生成或EC凋亡; 2）Tfam、Cox10或Trx 2可诱导缺失的小鼠表现出延迟的 在早期年龄（P5-P12），视网膜血管生长没有穿透到深丛中，这减弱了视网膜的 出芽与线粒体ROS的产生无关; 3）三种突变小鼠发生动静脉畸形 畸形（AVM），在微血管中动脉化增强和微动脉瘤形成， 高龄（P12-P30）。血管减少和微动脉瘤的表型与老年人相似 视网膜和人类视网膜血管异常，如Coats病、Leber军事动脉瘤和家族性 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，在血管生成和代谢中的基因表达降低， 但增加了TGF β R信号通路。基于这些数据，我们假设线粒体活性 调节血管生长中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 线粒体功能障碍诱导的视网膜出芽血管生成、AVM和微动脉瘤缺陷 2）明确线粒体功能障碍如何调节TGF β R-Smad2/3 3）确定TGF β R-Smad2/3信号在线粒体功能障碍诱导的细胞凋亡中的作用。 药物阻断和遗传缺陷导致的血管发育迟缓和畸形。我们提出的研究 将定义线粒体活动调节正常视网膜血管生长的机制， 成熟，并将为人类视网膜血管疾病提供新的模型和治疗干预 与许多病理并发症相关，如糖尿病、高血压和衰老， 视力丧失 1