Fibrosis-Associated Urinary Gene Transcripts for LUTS Detection and Treatment

描述（由申请人提供）：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、膀胱排空不完全和夜尿为特征，可发展为膀胱功能障碍和尿潴留。对于数以百万计的美国老年男性来说，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LUTS症状男性尿道周围前列腺组织的特征。这种纤维化改变增加了尿道周围组织的硬度，损害了尿道的灵活性和顺应性，产生尿路梗阻症状。本申请提供的初步数据显示，从4名LUTS（美国泌尿协会症状指数评分[AUASIs]）症状的男性尿液中纯化的总RNA为16,19，22和23)，并进行全转录组扩增，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（AUASIs为0,0,1和3）的类似纯化和处理的RNA相比，TGFb转录物水平高出约10倍，COL1和aSMA转录物水平高出2-5倍。这些发现表明，使用新的全转录组扩增技术，可以选择性地在LUTS症状男性尿液中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，即尿道周围前列腺纤维化导致LUTS（由中重度AUASI评分评估）可能通过尿液生物标志物的存在来检测，这些生物标志物包括编码介导和促进组织纤维化的蛋白质的特定基因转录物。幸运的是，我们拥有从尿液中回收和扩增RNA的专业知识，以及大量临床注释良好的人类前列腺组织生物库和来自前列腺癌活检阴性且无LUTS症状或症状的男性的尿液标本。利用这些资源，我们建议通过完成两个特定目标来验证上述假设：特定目标1：确定自报告中度/重度LUTS的男性尿液中介导和促进组织纤维化的基因编码蛋白的RNA转录水平是否持续且显著高于自报告无/轻度LUTS的男性。特异性目的2：测定特异性目的1中检查的一部分患者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。

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.