Mitochondrial Dysfunction in Obstructed Bladder: Role of Desmin and Vimentin

膀胱梗阻的线粒体功能障碍：结蛋白和波形蛋白的作用

基本信息

批准号：
9295007
负责人：
BOOPATHI ETTICKAN
金额：
$ 23.95万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-13 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9295007
关键词：
ATP Synthesis Pathway Address Adenine Nucleotide Translocase Adenosine Diphosphate Adenosine Triphosphate Adverse effects Age Bathing Benign Prostatic Hypertrophy Binding Biochemical Bladder Bladder Dysfunction Carbachol Cells Complement Complex Cytometry Cytosol Data Desmin Diabetes Mellitus Diagnosis Disease Elderly man Functional disorder Generations Genus Hippocampus Human Hypertrophy Incidence Intermediate Filament Proteins Intermediate Filaments Knock-out Knockout Mice Lead Lower urinary tract MAPK9 gene Mass Spectrum Analysis Measures Mediating Mediator of activation protein Mitochondria Mitogen-Activated Protein Kinases Molecular Molecular Genetics Movement Mus Muscle Myosin Light Chains Neurogenic Bladder Obstruction Organ Outer Mitochondrial Membrane Oxygen Consumption Pathogenesis Permeability Pharmaceutical Preparations Pharmacology Phosphorylation Phosphorylation Site Physiological Population Prevalence Process Production Protein Overexpression Proteins Reactive Oxygen Species Regulation Respiration Respiratory physiology Rest Role Secondary to Signal Transduction Small Interfering RNA Smooth Muscle Structure Study of magnetics Testing Therapeutic Intervention Tissues Transfection Vimentin Voltage-Dependent Anion Channel base cost design experimental study inhibitor/antagonist lower urinary tract symptoms mitochondrial creatine kinase mitochondrial dysfunction mitochondrial membrane muscle hypertrophy new therapeutic target novel overexpression pressure public health relevance

项目摘要

DESCRIPTION (provided by applicant): Lower urinary tract dysfunction (LUTD) is associated with bladder smooth muscle (BSM) hypertrophy secondary to benign prostatic hyperplasia (BPH)-induced partial bladder outlet obstruction (PBOO), neurogenic bladder, and diabetes, and its incidence and prevalence are increasing as the population ages. Existing therapeutic interventions are neither highly effective nor durable and have side effects of their own. Biochemical studies have shown reduced mitochondrial ATP and increased reactive oxygen species (ROS) production in the hypertrophied BSM of obstructed bladders and this is associated with overexpression of the intermediate filament (IF) proteins, desmin and vimentin which are important for muscle and mitochondrial function. Mitochondrial and muscle pathophysiology, including decreased ATP, increased ROS production and diminished contractility of smooth muscle in LUTD suggests a role for desmin and vimentin overexpression in inducing mitochondrial and muscle dysfunction. However, potential mechanisms by which these IF proteins mediate BSM dysfunction remain unexplored. Our preliminary data demonstrate that desmin and vimentin overexpression in human and murine BSM strips and cells significantly reduces mitochondrial ATP synthesis; increases ROS production and inhibits carbachol-and KCl-mediated contraction as also seen in BSM with PBOO. Overexpression of IF proteins in BSM strips and cells enhances mitogen-activated protein kinase (MAPK)/JNK2 phosphorylation which coincides with its increased mitochondrial localization and phosphorylation of desmin and vimentin as also seen in BSM with PBOO. Mitochondrial desmin and vimentin could potentially function to inhibit the mitochondrial voltage-dependent anion channel (VDAC) permeability via mitochondrial JNK2. Based on our preliminary data, we hypothesize that the activation of JNK2 and its increased mitochondrial localization is involved in desmin- and vimentin-induced BSM contractile and mitochondrial dysfunction. we further hypothesize that JNK2 phosphorylated mitochondrial desmin and vimentin interacts with the VDAC and this interaction inhibits ATP synthesis and increases ROS production. We designed three Specific Aims to address these hypotheses. In Aim 1, we will establish the robustness of increased IF expression in inducing BSM contractile dysfunction, and clarify JNK2 as a critical effector. In Aim 2, we will determine whether IF protein overexpression-induced BSM contractile and mitochondrial dysfunction is due to JNK2 dependent interaction of desmin and vimentin with VDAC. In Aim 3, we will ascertain the role of mitochondrial JNK2 in PBOO- induced BSM contractile and mitochondrial dysfunction. We expect our studies to delineate a mechanism of contractile dysfunction mediated by mitochondrial JNK2 signaling promoting the VDAC interaction with desmin and vimentin, and thus identify novel therapeutic targets for the treatment of PBOO/LUTD.

描述（由申请人提供）：下尿路功能障碍（LUTD）与继发于良性前列腺增生（BPH）引起的部分膀胱出口梗阻（PBOO）、神经源性膀胱和糖尿病的膀胱平滑肌（BSM）肥大相关，及其相关性。随着人口老龄化，现有的治疗干预措施既不高效也不持久，并且其本身具有副作用，生化研究表明线粒体 ATP 减少，活性氧增加。梗阻膀胱肥大的 BSM 中 ROS 的产生，这与中间丝 (IF) 蛋白、结蛋白和波形蛋白的过度表达有关，这些蛋白对肌肉和线粒体功能很重要，包括 ATP 减少、ROS 产生增加。 LUTD 中平滑肌收缩力的减弱表明结蛋白和波形蛋白过度表达在诱导线粒体和肌肉功能障碍中发挥作用，然而，这些 IF 蛋白介导的潜在机制。我们的初步数据表明，人类和小鼠 BSM 条带和细胞中结蛋白和波形蛋白的过度表达显着降低了线粒体 ATP 的合成；增加了 ROS 的产生并抑制了卡巴胆碱和 KCl 介导的收缩，这在 PBOO 过度表达的 BSM 中也可见到。 BSM 条带和细胞中的 IF 蛋白可增强丝裂原激活蛋白激酶 (MAPK)/JNK2 磷酸化，这与其增加的线粒体定位相一致，并且结蛋白和波形蛋白的磷酸化（也见于 BSM 与 PBOO）线粒体结蛋白和波形蛋白可能通过线粒体 JNK2 抑制线粒体电压依赖性阴离子通道 (VDAC) 通透性。并且其增加的线粒体定位参与结蛋白和波形蛋白诱导的 BSM 收缩和线粒体功能障碍，我们进一步发现 JNK2 磷酸化线粒体。结蛋白和波形蛋白与 VDAC 相互作用，这种相互作用会抑制 ATP 合成并增加 ROS 的产生。在目标 1 中，我们将确定 IF 表达增加在诱导 BSM 收缩功能障碍中的稳健性，并阐明 JNK2。在目标 2 中，我们将确定 IF 蛋白过度表达诱导的 BSM 收缩和线粒体功能障碍是否是由于结蛋白和波形蛋白与 JNK2 依赖性相互作用所致。 VDAC。在目标 3 中，我们将确定线粒体 JNK2 在 PBOO 诱导的 BSM 收缩和线粒体功能障碍中的作用，我们希望我们的研究能够阐明线粒体 JNK2 信号传导促进 VDAC 与结蛋白和波形蛋白相互作用介导的收缩功能障碍的机制。从而确定治疗 PBOO/LUTD 的新治疗靶点。