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

• 批准号: 10706504
• 负责人: BOOPATHI ETTICKAN
• 金额: $ 46.09万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10706504
• 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; Knockout Mice; Knowledge; Lower urinary tract; MAPK8 gene; MAPK9 gene; Maps; Measures; Mediating; Mitochondria; Mitochondrial Proteins; Molecular; 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; 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; voltage-dependent anion channel 2

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 较低的尿路功能障碍（LUTD）是一种常见疾病 人口年龄和当前的治疗方法无效。 Lutd与 膀胱平滑肌（BSM）继发于良性前列腺增生（BPH）诱导的部分肥大 膀胱出口阻塞（PBOO）。 BPH和BSM功能障碍的分子发病机理较差 理解；在我们的知识中填补这一空白可能会导致确定新的治疗靶标和 更有效的LUTD药物或管理策略。我们和其他人证明了收缩 功能障碍，线粒体ROS增加和阻塞膀胱中BSM的ATP水平降低为 与中间丝（IF）蛋白质蛋白质和波形蛋白的过表达有关。以前的 本文介绍的研究以及初步数据表明JNK2是BSM收缩的主要效果因子 Desmin和Vimentin的过表达引起的功能障碍，表明IFS/JNK2-- 依赖机制导致膀胱出口阻塞的收缩功能障碍。在电流中 提案我们通过微阵列确定了人类线粒体蛋白G0S2的显着诱导 和鼠BSM条和过表达脱敏蛋白和波形蛋白的细胞。我们的其他初步数据 证明抑制G0S2表达会降低磷酸-JNK水平，增加线粒体ATP， 并降低脱敏和波形蛋白过表达的鼠和人类BSM条中的ROS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与腺嘌呤联合促进跨线粒体外膜的ATP/ADP交换 核苷酸易位酶和线粒体肌酸激酶。 G0S2与VDAC结合大概会破坏 因此，ATP/ADP交换，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介导了Desmin-和Vimentin诱导的BSM收缩性 通过磷酸-JNK2功能障碍。我们进一步假设G0S2与VDAC相互作用促进了 线粒体ROS的产生和ROS诱导的磷酸-JNK2导致收缩功能障碍。 三个特定的目标旨在解决这些假设。在AIM 1中，我们将确定 IF蛋白过表达诱导的BSM收缩功能障碍的线粒体G0S2。在目标2中，我们将 确定蛋白质过表达是否诱导的BSM收缩功能障碍是由于G0S2和VDAC引起的 相互作用。在AIM 3中，我们将采用体内模型来确定G0S2和JNK2在Murine Pboo中的作用 诱导的BSM收缩功能障碍：我们希望我们的研究能够描述收缩机制 由G0S2介导的功能障碍，从而确定用于治疗PBOO/LUTD的治疗靶标。