Fibrosis-Associated Urinary Gene Transcripts for LUTS Detection and Treatment

描述(申请人提供)：下尿路症状(LUTS)是一种进行性疾病，以尿频和尿急、尿流力下降、膀胱排空不全和夜尿为特征，可发展为膀胱功能障碍和尿潴留。对于数以百万计的老年美国男性来说，LUTS是一个昂贵且潜在的严重医疗问题。我们实验室最近获得的数据显示，细胞外基质[ECM]沉积和纤维化是一些有LUTS症状的男性前列腺组织的特征。这种纤维化改变增加了尿路周围组织的僵硬，损害了尿路的柔韧性和顺应性，产生了尿路梗阻症状。本申请提供的初步数据显示，从4名有LUTS症状的男性尿液中提纯的总RNA(美国泌尿系统协会症状指数评分为16、19、22和23)进行全转录组扩增，然后进行转录特异性qRT-PCR，结果显示，与4名无LUTS症状的男性(AUASI为0、0、1和3)相比，TGFb转录水平高约10倍，col1和ASMA转录本水平高2-5倍。这些发现表明，利用新颖的全转录组扩增技术，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，我们现在试图验证这样一种假设，即通过中-重度AUASI评分评估的尿路周围前列腺纤维化可以通过尿液生物标记物的存在来检测到，尿液生物标记物包含特定的基因转录本，编码蛋白质，介导和促进组织纤维化。幸运的是，我们拥有从尿液中恢复和扩增RNA的专业知识，以及大量临床上注释良好的人前列腺组织和来自前列腺癌活检阴性且无症状或有LUTS症状的人的尿样的生物库。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的假设：具体目标1：确定自我报告中度/重度LUTS的男性尿液中中介和促进组织纤维化的蛋白质编码基因的RNA转录水平是否一致且显著高于自我报告没有/轻度LUTS的男性。特定目标2：确定特定目标1检查的部分患者的前列腺活检组织中的胶原含量，并将其与纤维化生物标记物水平和AUASI评分相关联。

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.