Diabetic Retinopathy, Mitochondria Damage and Long Non-coding RNAs

糖尿病视网膜病变、线粒体损伤和长非编码 RNA

• 批准号: 10653935
• 负责人: RENU A. KOWLURU
• 金额: $ 34.65万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10653935
• 关键词: Acceleration; Aging; Apoptosis; Attenuated; Binding; Biochemical; Biogenesis; Biological; Blindness; Blood capillaries; Cells; Characteristics; Chronic Disease; Code; Complex; Complications of Diabetes Mellitus; Cytochromes b; Cytosol; DNA; Data; Defect; Development; Diabetes Mellitus; Diabetic Retinopathy; Diabetic mouse; Digestion; Disease; Down-Regulation; Electron Transport; Electron Transport Complex III; Endothelial Cells; Free Radicals; Fright; Gene Expression; Generations; Genes; Genetic Transcription; Glycine decarboxylase; Goals; Histones; Histopathology; Homeostasis; Human; Human Genome; Hyperglycemia; Impairment; In Vitro; Lead; Messenger RNA; Metabolic; MicroRNAs; Mitochondria; Mitochondrial DNA; Modeling; Molecular; Nuclear; Nucleotides; Open Reading Frames; Pathogenesis; Patients; Play; Porifera; Proteins; RNA; RNA Processing; RNA Sequences; RNase P; Regulation; Retina; Retinal Diseases; Rodent; Role; Superoxides; System; Testing; Therapeutic; Transcript; Translations; Untranslated RNA; Vision; cytochrome c; helicase; in vivo; in vivo Model; innovation; member; mtTF1 transcription factor; new therapeutic target; novel; nuclease; prevent; scaffold; therapeutic target; transcription factor; translational impact; Acceleration; Aging; Apoptosis; Attenuated; Binding; Biochemical; Biogenesis; Biological; Blindness; Blood capillaries; Cells; Characteristics; Chronic Disease; Code; Complex; Complications of Diabetes Mellitus; Cytochromes b; Cytosol; DNA; Data; Defect; Development; Diabetes Mellitus; Diabetic Retinopathy; Diabetic mouse; Digestion; Disease; Down-Regulation; Electron Transport; Electron Transport Complex III; Endothelial Cells; Free Radicals; Fright; Gene Expression; Generations; Genes; Genetic Transcription; Glycine decarboxylase; Goals; Histones; Histopathology; Homeostasis; Human; Human Genome; Hyperglycemia; Impairment; In Vitro; Lead; Messenger RNA; Metabolic; MicroRNAs; Mitochondria; Mitochondrial DNA; Modeling; Molecular; Nuclear; Nucleotides; Open Reading Frames; Pathogenesis; Patients; Play; Porifera; Proteins; RNA; RNA Processing; RNA Sequences; RNase P; Regulation; Retina; Retinal Diseases; Rodent; Role; Superoxides; System; Testing; Therapeutic; Transcript; Translations; Untranslated RNA; Vision; cytochrome c; helicase; in vivo; in vivo Model; innovation; member; mtTF1 transcription factor; new therapeutic target; novel; nuclease; prevent; scaffold; therapeutic target; transcription factor; translational impact

ABSTRACT Retinopathy is one of the most-feared complications of diabetes. In the pathogenesis of this blinding disease, retinal mitochondria become dysfunctional, the electron transport chain (ETC) is compromised, superoxide levels are elevated, and while complex III activity is inhibited, complex I remains unchanged. Mitochondria have their own small DNA (mtDNA), which lacks protective histones, but is packaged into nucleoids that provide some protection and assist in its biogenesis. Diabetes damages mtDNA, impairs its biogenesis, and downregulates gene expression of mtDNA-encoded cytochrome B (CYTB of complex III). Gene expression is also regulated by long noncoding RNAs (LncRNAs), the RNAs with >200 nucleotides and no open reading frame for translation, but they can bind to DNA or RNA, or can act as scaffolds to promote the interaction of proteins. Although majority of the LncRNAs are encoded by nuclear DNA, mtDNA also encodes three LncRNAs, LncND5 and LncND6 for complex I and LncCytB for complex III. Preliminary data show that in hyperglycemic milieu, while LncCytB is downregulated, LncND5 and LncND6 remain unchanged, and nucleoids are decreased and mtDNA sensitivity to nuclease digestion is increased. Based on these, our central hypothesis is that `LncCytB downregulation in diabetes impairs mtDNA nucleoids and attenuates cytochrome B transcription, damaging the mtDNA and the electron transport chain system, and the damaged mitochondria lead to the development of retinopathy'. Aim 1 will investigate the role of LncCytB in nucleoid formation, and the hypothesis predicts that `decrease in LncCytB in diabetes impairs nucleoids, damaging mtDNA integrity and reducing its copy numbers'. Aim 2 will examine the role of LncCytB in the regulation of the ETC, and will test the hypothesis that `downregulation of LncCytB decreases transcription of CYTB, which inhibits the complex III activity and compromises the ETC system'. Aim 3 will investigate the mechanism by which hyperglycemia downregulates LnCytB, and will examine the role of mitochondrial-targeted RNAse P protein 1 in regulation of LncCytB in the mitochondria. The plan will employ in vitro (human retinal endothelial cells) and in vivo (retinal microvessels from rodents) models of diabetic retinopathy, and will utilize fully optimized molecular biological approaches. Our overall goal is to identify novel regulatory mechanisms involved in the pathogenesis of diabetic retinopathy, specifically at the level of mtDNA-encoded LncRNA in mitochondrial homeostasis. The testable central hypothesis is innovative, and has significant translational impact as successful completion of our studies will provide strong background for LncCytB as a potential therapeutic target to prevent the development/ progression of this sight- threatening disease.

抽象的 视网膜病是糖尿病最刺激的并发症之一。在这种盲目疾病的发病机理中， 视网膜线粒体变功能失调，电子传输链（ETC）被损害，超氧化物 水平升高，虽然复杂的III活性受到抑制，但复杂的我保持不变。线粒体 有自己的小型DNA（mtDNA），它缺乏保护性组蛋白，但被包装成核苷中 提供一些保护并协助其生物发生。糖尿病会损害mtDNA，损害其生物发生，并 下调mtDNA编码的细胞色素B（复合物III的CYTB）的基因表达。基因表达是 还由长的非编码RNA（LNCRNA），> 200个核苷酸且无开放式读取的RNA调节 翻译框架，但它们可以与DNA或RNA结合，或者可以充当支架以促进相互作用 蛋白质。尽管大多数LNCRNA是由核DNA编码的，但MTDNA也编码三个 lncrNA，lncnd5和lncnd6，用于复合物I和lnccytb，用于复合物III。初步数据显示 高血糖环境，虽然LNCCYTB被下调，但LNCND5和LNCND6保持不变，并且 核苷降低，mtDNA对核酸酶消化的敏感性增加。基于这些，我们的中心 假设是糖尿病中的lnccytb下调会损害mtDNA核苷，并减弱细胞色素b 转录，损坏mtDNA和电子传输链系统以及损坏的线粒体 导致视网膜病的发展。 AIM 1将研究lnccytb在核苷形成中的作用，该假设预测``''降低了 糖尿病中的lnccytb会损害核苷，损害mtDNA完整性并减少其拷贝数。 AIM 2意志 检查lnccytb在ETC调节中的作用，并将检验以下假设。 lnccytb降低了CYTB的转录，从而抑制复合物III活性并损害ETC 系统'。 AIM 3将研究高血糖下调LNCYTB的机制，并将 检查线粒体靶向的RNase P蛋白1在线粒体中LNCCYTB调节中的作用。 该计划将在体外使用（人类视网膜内皮细胞）和体内（啮齿动物的视网膜微血管） 糖尿病性视网膜病模型，并将利用完全优化的分子生物学方法。我们的总体目标 是为了确定与糖尿病性视网膜病发病机理有关的新型调节机制，特别是在 线粒体稳态中mtDNA编码的lncRNA水平。可检验的中心假设是 创新的，并且随着我们的研究成功完成，具有重大的翻译影响将为强大 LNCCYTB作为防止这种视线发展/进展的潜在治疗靶标的背景 威胁性疾病。