Mechanisms of contractile dysfunction in the obstructed bladder: Role of desmin and vimentin

膀胱梗阻收缩功能障碍的机制：结蛋白和波形蛋白的作用

基本信息

批准号：
10522325
负责人：
BOOPATHI ETTICKAN
金额：
$ 49.5万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10522325
关键词：
Address Adenine Nucleotide Translocase Age Animal Model Benign Prostatic Hypertrophy Binding Bladder Bladder Dysfunction Carbachol Cells Complement Complex Data Desmin Diagnosis Disease Elderly man Functional disorder Future Generations Human Incidence Intermediate Filament Proteins Intermediate Filaments Knock-out Knockout Mice Knowledge Lower urinary tract MAPK8 gene MAPK9 gene Maps Measures Mediating Mitochondria Mitochondrial Proteins Molecular Molecular Genetics Mus Muscle Obstruction Operative Surgical Procedures Outer Mitochondrial Membrane Oxygen Consumption Pathogenesis Patients Peptides Pharmaceutical Preparations Phosphorylation Population Prevalence Production Protein Overexpression Proteins Reactive Oxygen Species Rest Role Secondary to Smooth Muscle Smooth Muscle Myocytes Stimulus Tertiary Protein Structure Therapeutic Therapeutic Intervention Vimentin Voltage-Dependent Anion Channel base cost design experimental study genetic approach improved in vivo Model lower urinary tract symptoms mitochondrial creatine kinase mitochondrial dysfunction muscle hypertrophy new therapeutic target novel overexpression pressure side effect small hairpin RNA therapeutic target transcriptome

项目摘要

Abstract Lower urinary tract dysfunction (LUTD) is a common disease whose incidence and prevalence increase as the population ages and the the current therapeutic approaches are ineffective. LUTD is strongly associated with bladder smooth muscle (BSM) hypertrophy secondary to benign prostatic hyperplasia (BPH)-induced partial bladder outlet obstruction (PBOO). The molecular pathogenesis of BPH and BSM dysfunction is poorly understood; filling this gap in our knowledge will likely lead to the identification of new therapeutic targets and more effective LUTD drugs or management strategies. We and others have demonstrated that the contractile dysfunction, increased mitochondrial ROS and reduced ATP levels of BSM in the obstructed bladder is associated with the overexpression of the intermediate filament (IF) proteins desmin and vimentin. Previous studies as well as preliminary data presented herein suggest JNK2 as a major effector of the BSM contractile dysfunction induced by the overexpression of desmin and vimentin and demonstrate that an IFs/JNK2- dependent mechanism contributes to the contractile dysfunction in bladder outlet obstruction. In the current proposal we have identified via microarray significant induction of the mitochondrial protein G0S2 in human and murine BSM strips and cells overexpressing desmin and vimentin. Our additional preliminary data further demonstrate that inhibition of G0S2 expression decreases phospho-JNK levels, increases mitochondrial ATP, and decreases the ROS production in desmin- and vimentin- overexpressing murine and human BSM strips and cells. Further, we demonstrate the interaction of G0S2 with voltage-dependent anion channel (VDAC). VDAC promotes ATP/ADP exchange across the mitochondrial outer membrane in association with adenine nucleotide translocase and mitochondrial creatine kinase. G0S2 binding to VDAC presumably disrupts the ATP/ADP exchange thereby, reducing the mitochondrial ATP level and increasing the mitochondrial ROS production. We hypothesize that mitochondrial G0S2 mediates desmin- and vimentin-induced BSM contractile dysfunction via phospho-JNK2. We further hypothesize that G0S2 interaction with the VDAC promotes the mitochondrial ROS production and the ROS-induced phospho-JNK2 contributes to the contractile dysfunction. Three Specific Aims are designed to address these hypotheses. In Aim 1, we will establish the role of mitochondrial G0S2 in IF protein overexpression-induced BSM contractile dysfunction. In Aim 2 we will determine whether IF protein overexpression-induced BSM contractile dysfunction is due to G0S2 and VDAC interaction. In Aim 3 we will employ in vivo models to establish the role of G0S2 and JNK2 in murine PBOO induced BSM contractile dysfunction: We expect our studies to delineate a mechanism of contractile dysfunction mediated by G0S2 and thus identify therapeutic targets for the treatment of PBOO/LUTD.

抽象的下尿路功能障碍（LUTD）是一种常见病，其发病率和患病率随着年龄的增长而增加。人口年龄和目前的治疗方法无效。 LUTD 与以下因素密切相关：良性前列腺增生（BPH）引起的部分膀胱平滑肌（BSM）肥大膀胱出口梗阻（PBOO）。 BPH 和 BSM 功能障碍的分子发病机制尚不明确明白了；填补我们知识上的空白可能会导致新的治疗靶点的确定和更有效的 LUTD 药物或管理策略。我们和其他人已经证明收缩梗阻膀胱中 BSM 功能障碍、线粒体 ROS 增加和 ATP 水平降低与中间丝 (IF) 蛋白结蛋白和波形蛋白的过度表达有关。以前的研究以及本文提供的初步数据表明 JNK2 是 BSM 收缩的主要效应器结蛋白和波形蛋白过度表达引起的功能障碍，并证明 IFs/JNK2- 依赖性机制导致膀胱出口梗阻的收缩功能障碍。在当前我们通过微阵列鉴定了人类线粒体蛋白 G0S2 的显着诱导作用以及过度表达结蛋白和波形蛋白的鼠 BSM 条带和细胞。我们进一步提供额外的初步数据证明抑制 G0S2 表达会降低磷酸化 JNK 水平，增加线粒体 ATP，并减少结蛋白和波形蛋白过表达的鼠和人 BSM 条中 ROS 的产生和细胞。此外，我们证明了 G0S2 与电压依赖性阴离子通道 (VDAC) 的相互作用。 VDAC 与腺嘌呤相关，促进线粒体外膜上的 ATP/ADP 交换核苷酸转位酶和线粒体肌酸激酶。 G0S2 与 VDAC 的结合可能会破坏 ATP/ADP 交换从而降低线粒体 ATP 水平并增加线粒体 ROS 生产。我们假设线粒体 G0S2 介导结蛋白和波形蛋白诱导的 BSM 收缩通过磷酸-JNK2 的功能障碍。我们进一步假设 G0S2 与 VDAC 的相互作用促进线粒体 ROS 的产生和 ROS 诱导的磷酸化 JNK2 会导致收缩功能障碍。三个具体目标旨在解决这些假设。在目标 1 中，我们将确立以下角色：线粒体 G0S2 在 IF 蛋白过度表达诱导的 BSM 收缩功能障碍中的作用。在目标 2 中，我们将确定 IF 蛋白过度表达诱导的 BSM 收缩功能障碍是否是由 G0S2 和 VDAC 引起的相互作用。在目标 3 中，我们将采用体内模型来确定 G0S2 和 JNK2 在小鼠 PBOO 中的作用诱发 BSM 收缩功能障碍：我们希望我们的研究能够描绘出收缩功能障碍的机制 G0S2 介导的功能障碍，从而确定治疗 PBOO/LUTD 的治疗靶点。