Association between the Y chromosome and androgens

从临床研究中积累的证据表明，不同病因的肾脏疾病在男性中比年龄匹配的女性更常见。此外，非糖尿病慢性肾脏疾病在男性中的进展速度比女性更快，与血压无关。这些观察表明，遗传和/或荷尔蒙男性特有的因素可能会对人类的肾脏功能和结构产生不利影响。然而，关于Y染色体和雄激素与肾脏表型的关系的联合分析尚未在男性中进行。根据我们的初步研究和以前的报道，我们假设人类Y染色体内的遗传变异至少部分地通过调节循环中雄激素的浓度对男性的肾脏功能和结构产生不利影响。1.利用两个表型良好的波兰男性队列，我们将确定Y染色体非重组区(NRY)的多态与人类雄激素水平和肾功能的关系(特定目标1)。将使用聚合酶链式反应-限制性片段长度多态(PCR-RFLP)方法对NRY内的Hindll(+/-)双等位基因标记进行基因分型。血清游离睾酮、脱氢表雄酮、脱氢表雄酮、硫酸脱氢表雄酮和双氢睾酮的浓度将用放射免疫法进行评估。肾功能将通过内生肌酐清除量和尿白蛋白/肌酐比率进行评估。2.利用因肾癌而行单侧选择性肾切除术的男性未受影响的人肾组织，我们将研究NRY与雄激素水平和肾脏结构的关系(特定目标2)。肾脏结构变化将通过光学显微镜进行评估。增殖细胞核抗原和末端脱氧核苷酸转移酶生物素-dUTP缺口末端标记法分别检测细胞增殖和死亡。细胞外基质沉积将通过免疫组织化学和Western分析I型和IV型胶原的定位和表达来评估。本申请中提出的NRY、雄激素与人类肾脏功能和结构之间的关联的确定将有助于我们更好地理解性别二型性在慢性肾脏疾病的流行和进展中的作用，并为这些疾病过程开发预防和治疗方案。

Accumulating evidence from clinical studies suggest that renal diseases of different etiologies are more prevalent among men than age-matched women. Furthermore, non-diabetic chronic renal diseases progress more rapidly in men than women, independently of blood pressure. These observations suggest that genetic and/or hormonal male-specific factors may adversely affect renal function and structure in humans. However, a joint analysis of the relationships between the Y chromosome and androgens with respect to renal phenotypes has not been performed in men. Based on our preliminary and previously reported studies, we hypothesize that genetic variation within the human Y chromosome acting, at least in part, through modulation of circulating concentrations of androgens have detrimental effects on renal function and structure in men. 1. Using two well-phenotyped cohorts of Polish men, we will determine the association of polymorphisms in the non-recombining region of the Y chromosome (NRY) with androgen levels and renal function in humans (Specific Aim 1). Genotyping of the Hindlll(+/-) bi-allelic marker within the NRY will be performed using the polymerase chain reaction - restriction fragment length polymorphisms (PCR-RFLP) method. Serum concentrations of free testosterone, dehydroepiandrosterone, dehydroepiandrosterone sulfate and dihydrotestosterone will be assessed by radioimmunoassay. Renal function will be evaluated using creatinine clearance and urinary albumin-to-creatinine ratio. 2. Using renal tissue from the unaffected part of the human kidney of men undergoing unilateral elective nephrectomy due to renal cancer, we will examine the association of the NRY with androgen Ievels and renal structure (Specific Aim 2). Renal structural changes will be assessed by light microscopy. Cell proliferation and death will be determined by proliferating cell nuclear antigen and Terminal Deoxynucleotidyl Transferase Biotin-dUTP Nick End labeling (TUNEL), respectively. Extracellular matrix deposition will be assessed by immunohistochemistry and Western analysis of collagen types I and IV localization and expression. Determination of the association among NRY, androgens and human renal function and structure proposed in this application will improve our understanding of sexual dimorphism in prevalence and progression of chronic renal disorders and development of preventive and therapeutic regimens for these disease processes.