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：
10597212
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/10597212
关键词：
Accounting 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 Dedications 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 Nuclear 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 Tubular formation 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.

项目摘要 Fuchs内皮角膜营养不良（FECD）是一种常见的，通常复杂且与年龄有关的变性疾病影响了大约4％的美国人口，而女性中较高的疾病。在粪便中角膜内皮（CE）细胞损失是通过细胞外基质沉积以Guttae形式完成的。我的心理（Ula V. Jurkunas）明确地报告了，在FECD中，由于氧化剂 - 抗氧化剂的不平衡 NRF-2调节的抗氧化剂防御，包括其转录靶标下降NAD（P）H Quinone 脱氢酶1（NQO1），导致氧化DNA损伤，线粒体功能障碍和凋亡。我的初步数据还表明，与小鼠模型中的WT相比，NQO1 - / - 小鼠中CE的明显损失 FECD。我的同事（Albert S. Jun）等实施了ER应力/展开的蛋白质反应（UPR）在FECD发病机理中的作用。我的初步数据还表明，NQO - / - 细胞系的促凋亡ER应力标记的显着和更早激活 UVA诱导的FECD模型在体外。但是，没有任何联系ER和线粒体的研究 FECD中CE的氧化剂抗氧化剂失衡下的应力。该提议的目的是定义在氧化剂 - 抗氧化剂下，ER应力对线粒体应激（改变生物能和动力学）的贡献 FECD失衡。在奖励的修订阶段（K99）期间，我将确定UVA是否引起ER 和线粒体应力（AIM 1A），ER应力改变了线粒体生物能（AIM 1B）和动力学（形态学，碎裂，易位）全部在氧化物抗氧化剂失衡下（AIM 2A）。我会学会诱导并通过我的Co-Mentor实验室合作（Albert Jun）量化ER压力，分析线粒体生物能力以及精神上的ula jurkunas和同事Pere Puigserver的实验室的动态，在体外学习和体内模型 FECD（Jurkunas's Lab）以及哈佛医学院提供的广泛的职业发展活动。在R00阶段，在心理和同事的支持下，我将与AIM1-2进行未完成的部分。 ER介导的线粒体内在凋亡途径的机理的额外投资（AIM 1C）通过CA+2信号传导和微管重排（AIM 2B）。总的来说，这些研究将提供新的对角膜内皮生物学的ER-Mitochrial交叉讲座的见解和观点，这将推进我们对FECD发病机理的理解。 K99奖项将使我能够在ER和线粒体生物学以及角膜内皮细胞生物学的新培训。广泛的资源和哈佛医学院，Jurkunas，Puigserver和Jun的实验室可提供职业发展机会以及在K99阶段计划的研究活动，将使我能够实现长期目标成为一名独立研究者内皮生物学。