Fibrosis-Associated Urinary Gene Transcripts for LUTS Detection and Treatment

用于 LUTS 检测和治疗的纤维化相关尿液基因转录

• 批准号: 8486921
• 负责人: Jill A. Macoska
• 金额: $ 21.66万
• 依托单位: UNIVERSITY OF MASSACHUSETTS BOSTON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-20 至 2015-05-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8486921
• 关键词: Address; Adverse effects; Age; Aging; American; Biological Markers; Biopsy; Bladder; Bladder Calculi; Bladder Dysfunction; Collagen; Data; Deposition; Detection; Diagnosis; Disease; Disease Progression; Extracellular Matrix; Fibrosis; Gene Expression Profile; Genes; Human; Impairment; Increased frequency of micturition; Kidney; Laboratories; Left; Lower urinary tract; Malignant neoplasm of prostate; Measures; Mediating; Medical; Medicine; Nocturia; Operative Surgical Procedures; Patient Self-Report; Patients; Pharmaceutical Preparations; Population; Prostate; Prostate Ablation; Prostatic; Prostatic Tissue; Proteins; RNA; RNA Processing; Recurrence; Regimen; Research; Resources; Risk; Specimen; Stream; Symptoms; Techniques; Testing; Therapeutic; Tissues; Transcript; Urethra; Urinary Retention; Urinary tract infection; Urine; Urologist; base; biobank; clinical care; cost; flexibility; improved; indexing; innovation; lower urinary tract symptoms; male; men; micturition urgency; new therapeutic target; novel; prevent; public health relevance; standard of care; treatment strategy; urinary; urologic

DESCRIPTION (provided by applicant): Lower Urinary Tract Symptoms (LUTS) is a progressive disorder marked by urinary frequency and urgency, decreased force of stream, incomplete bladder emptying, and nocturia which can progress to bladder dysfunction and urinary retention. LUTS is a costly and potentially critical medical problem for millions of aging American men. Data recently acquired in our laboratory shows that extracellular matrix [ECM] deposition and fibrosis characterize the peri-urethral prostate tissues of some men symptomatic for LUTS. Such fibrotic changes increase peri-urethral tissue stiffness and compromise urethral flexibility and compliance, producing urinary obstructive symptoms. Preliminary data presented in this application shows that total RNA purified from the urine of 4 men symptomatic for LUTS (American Urological Association Symptom Index Scores [AUASIs] of 16, 19, 22 and 23) and subjected to whole transcriptome amplification followed by transcript-specific qRT-PCR demonstrated ~10-fold higher levels of TGFb transcript and 2-5- fold higher levels of COL1 and aSMA transcripts compared to similarly purified and processed RNA from 4 men asymptomatic for LUTS (AUASIs of 0, 0, 1 and 3). These findings show that specific gene transcripts encoding proteins that mediate and promote tissue fibrosis can be selectively detected in the urine of men symptomatic for LUTS using novel whole transcriptome amplification techniques. Based on these findings, we now seek to test the hypothesis that peri-urethral prostatic fibrosis contributing to LUTS as assessed by moderate-severe AUASI scores may be detected by the presence of urinary biomarkers comprising specific gene transcripts encoding proteins that mediate and promote tissue fibrosis. We are fortunate to possess the expertise to recover and amplify RNA from urine as well as a large clinically well-annotated biorepository of human prostate tissues and urine specimens from men who are prostate cancer biopsy negative and are asymptomatic or symptomatic for LUTS. Utilizing these resources, we propose to test the stated hypothesis through the completion of two Specific Aims: SPECIFIC AIM 1: Determine whether RNA transcript levels from genes encoding proteins that mediate and promote tissue fibrosis are consistently and significantly higher in the urine of men self-reporting moderate/severe LUTS compared to men self-reporting absent/mild LUTS. SPECIFIC AIM 2: Determine the collagen content of prostate biopsy tissues from a subset of patients examined in Specific Aim 1 and correlate these with fibrosis biomarker levels and AUASI scores.

描述（由申请人提供）：下尿路症状（LUTS）是一种进行性疾病，其特征是尿频和尿急、尿流力减弱、膀胱排空不完全和夜尿，可进展为膀胱功能障碍和尿潴留。对于数百万美国老年男性来说，下尿路症状是一个代价高昂且潜在严重的医疗问题。我们实验室最近获得的数据表明，细胞外基质 [ECM] 沉积和纤维化是一些有 LUTS 症状的男性尿道周围前列腺组织的特征。这种纤维化变化增加了尿道周围组织的硬度并损害尿道的灵活性和顺应性，产生尿路梗阻症状。本申请中提供的初步数据显示，从 4 名有 LUTS 症状的男性（美国泌尿外科协会症状指数评分 [AUASI] 为 16、19、22 和 23）的尿液中纯化总 RNA，并进行全转录组扩增，然后进行转录特异性扩增与来自 4 种类似纯化和处理的 RNA 相比，qRT-PCR 证明 TGFb 转录物水平高出约 10 倍，COL1 和 aSMA 转录物水平高出 2-5 倍。无 LUTS 症状的男性（AUASI 为 0、0、1 和 3）。这些发现表明，使用新型全转录组扩增技术，可以在有 LUTS 症状的男性尿液中选择性地检测到编码介导和促进组织纤维化的蛋白质的特定基因转录本。基于这些发现，我们现在试图检验以下假设：通过中度至重度 AUASI 评分评估导致 LUTS 的尿道周围前列腺纤维化可以通过尿液生物标志物的存在来检测，这些生物标志物包含编码介导和促进组织纤维化的蛋白质的特定基因转录本。我们很幸运，拥有从尿液中回收和扩增 RNA 的专业知识，以及一个临床注释良好的大型生物储存库，其中包含人类前列腺组织和前列腺癌活检阴性、无症状或有 LUTS 症状的男性的尿液样本。利用这些资源，我们建议通过完成两个具体目标来检验所述假设：具体目标 1：确定编码介导和促进组织纤维化的蛋白质的基因的 RNA 转录水平在男性自身尿液中是否持续且显着较高。报告中度/重度 LUTS 的男性与自我报告无/轻度 LUTS 的男性相比。具体目标 2：确定具体目标 1 中检查的一部分患者的前列腺活检组织的胶原含量，并将其与纤维化生物标志物水平和 AUASI 评分相关联。