Understanding Endoplasmic Reticulum-Mitochondrial Cross-Talk in Corneal Endothelial Cells

了解角膜内皮细胞中的内质网-线粒体交互作用

基本信息

批准号：
10550019
负责人：
Varun Kumar
金额：
$ 24.9万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10550019
关键词：
Accounting Address Adenosine Triphosphate Affect Age Antioxidants Apoptosis Apoptotic Award Bioenergetics Biological Availability Biology Blindness Cell Death Cell Line Cellular biology Collaborations Complex Cornea Corneal Endothelium DNA Damage Data Degenerative Disorder Deposition Development Disease Down-Regulation Economics Endoplasmic Reticulum Endothelial Cells Endothelium Extracellular Matrix Extracellular Matrix Proteins Female Fuchs&apos Endothelial Dystrophy Genetic Transcription Glare Goals Human In Vitro Incidence Intervention Investigation Keratoplasty Learning Link Mediating Medical Membrane Potentials Mentors Mitochondria Mitochondrial DNA Morphology Mus NQO1 gene Oxidants Oxidative Stress Oxidoreductase Oxygen Consumption Pathogenesis Pathway interactions Patients Persons Pharmacological Treatment Phase Population Predisposition Production Proteins Quinones Reactive Oxygen Species Reporting Research Activity Research Personnel Resource Development Role Signal Transduction Stress Structure Tissues Training UVA induced Up-Regulation Visual age related antioxidant enzyme biological adaptation to stress career development defined contribution endoplasmic reticulum stress in vitro Model in vivo in vivo Model insight light scattering medical schools mitochondrial dysfunction mitochondrial membrane mouse model novel oxidative DNA damage response social

项目摘要

Project Summary Fuchs endothelial corneal dystrophy (FECD) is a common, genetically complex and age-related degenerative disease affecting approximately 4 % of the U.S.A. population with a higher incidence in females. In FECD, corneal endothelial (CE) cell loss is accompanied by extracellular matrix deposition in the form of guttae. My mentor (Ula V. Jurkunas) has reported explicitly that in FECD, oxidant-antioxidant imbalance due to suboptimum Nrf-2 regulated antioxidant defense, including a decline in its transcriptional target, NAD(P)H quinone dehydrogenase 1 (NQO1), leads to oxidative DNA damage, mitochondrial dysfunction, and apoptosis. My preliminary data also suggests the significant loss of CE in NQO1-/- mice compared to WT in the mouse model of FECD. Alike oxidative stress and mitochondrial damage, my co-mentor (Albert S. Jun) and others have implicated the role of ER stress/ Unfolded protein response (UPR) in the pathogenesis of FECD. My preliminary data also suggests the significant and earlier activation of pro-apoptotic ER stress markers for NQO-/- cell line after UVA-induced FECD model in vitro. However, there has not been any study linking ER and mitochondrial stress under oxidant-antioxidant imbalance for CE in FECD. The objective of this proposal is to define the contribution of ER stress on mitochondrial stress (altered bioenergetics and dynamics) under oxidant-antioxidant imbalance in FECD. During the mentored phase (K99) of the award, I will determine whether UVA induces ER and mitochondrial stress (Aim 1a), ER stress alters mitochondrial bioenergetic (Aim 1b) and dynamics (morphology, fragmentation, translocation) all under oxidant-antioxidant imbalance (Aim 2a). I will learn to induce and quantify ER stress with my co-mentor lab’s collaboration (Albert Jun), analyze mitochondrial bioenergetics and dynamics with mentor Ula Jurkunas and co-mentor Pere Puigserver’s lab, learn in vitro and in vivo model of FECD (Jurkunas’s lab) along with the extensive career development activities offered at Harvard Medical School. During the R00 phase, with mentor and co-mentor’s support, I will perform uncompleted parts of Aim1-2 with the additional investigation of the mechanism of ER mediated-activation of mitochondrial intrinsic apoptotic pathway (Aim 1c) via Ca+2 signaling and microtubular rearrangement (Aim 2b). Collectively, these studies will provide new insights and perspectives into the ER-Mitochondrial cross talk for corneal endothelial biology, which will advance our understanding of FECD pathogenesis. A K99 award will allow me to receive additional training in ER and Mitochondria biology along with novel training in corneal endothelial cell biology. The extensive resources and career development opportunities available at Harvard Medical School, Jurkunas, Puigserver, and Jun’s lab, as well as the research activities planned in the K99 phase, will enable me to achieve the long-term goal of becoming an independent investigator dedicated to the study of ER-Mitochondrial cross talk in corneal endothelial biology.

项目概要福克斯内皮性角膜营养不良 (FECD) 是一种常见的、遗传复杂且与年龄相关的退行性病变该疾病影响约 4% 的美国人口，其中女性发病率较高。角膜内皮（CE）细胞损失伴随着以牙胶形式出现的细胞外基质沉积。导师（Ula V. Jurkunas）明确报告说，在 FECD 中，由于次优而导致的氧化-抗氧化失衡 Nrf-2 调节抗氧化防御，包括其转录靶标 NAD(P)H 醌的下降脱氢酶 1 (NQO1) 会导致 DNA 氧化损伤、线粒体功能障碍和细胞凋亡。初步数据还表明，与 WT 小鼠模型相比，NQO1-/- 小鼠的 CE 显着丧失 FECD 的研究与氧化应激和线粒体损伤一样，我的合作导师 (Albert S. Jun) 和其他人也有类似的研究。我的初步研究表明 ER 应激/未折叠蛋白反应 (UPR) 在 FECD 发病机制中的作用。数据还表明 NQO-/- 细胞系的促凋亡 ER 应激标记物显着且更早地激活然而，目前还没有任何将 ER 和线粒体联系起来的研究。 FECD 中 CE 的氧化-抗氧化失衡下的应激本提案的目的是定义氧化-抗氧化作用下内质网应激对线粒体应激（改变生物能学和动力学）的贡献在该奖项的指导阶段（K99），我将确定 UVA 是否会诱发 ER。和线粒体应激（目标 1a），内质网应激改变线粒体生物能（目标 1b）和动力学（形态、碎片、易位）都在氧化剂-抗氧化剂失衡下（目标 2a）。与我的合作导师实验室 (Albert Jun) 合作量化 ER 压力，分析线粒体生物能学与导师 Ula Jurkunas 和合作导师 Pere Puigserver 的实验室进行动态交流，学习体外和体内模型 FECD（尤尔库纳斯实验室）以及哈佛医学院提供的广泛的职业发展活动。在R00阶段，在导师和共同导师的支持下，我将完成Aim1-2中未完成的部分 ER 介导的线粒体内在凋亡途径激活机制的进一步研究（目标 1c）通过 Ca+2 信号传导和微管重排（目标 2b）总的来说，这些研究将提供新的成果。对角膜内皮生物学的 ER-线粒体串扰的见解和观点，这将推进我们对 FECD 发病机制的了解将使我能够接受 ER 和 ER 方面的额外培训。线粒体生物学以及角膜内皮细胞生物学的新颖培训丰富的资源和知识。哈佛医学院、Jurkunas、Puigserver 和 Jun 实验室提供的职业发展机会，如以及K99阶段计划的研究活动，将使我能够实现以下长期目标：成为一名致力于角膜内质网线粒体串扰研究的独立研究者内皮生物学。