DIFFERENTIAL GENE EXPRESSION IN POLYCYSTIC KIDNEYS

常染色体显性遗传性多囊肾病（ADPKD）是一种最常见的 常见的，显性遗传性肾脏疾病的人。ADPKD是负责 10%需要长期透析或肾移植的患者。 目前，有50万美国人和全球500万人受到影响 这种紊乱。 每年的医疗费用估计为10亿美元 仅在美国，美元。 ADPKD的确切分子缺陷尚不清楚 除了移植，没有治愈性治疗存在。 这个建议试图应用现代分子生物学和生物化学 研究ADPKD的异常基因表达，制备分子探针， 来定义一组分子参数， ADPKD。 我们的长期目标是完全描述 在ADPKD的缺陷的基础上。 随着ADPKD基因的特征， 可以设计精确的症状前诊断测试。 此外，本发明还提供了一种方法， 可以开发药物和躯体疗法来治疗和 预防ADPKD。 具体而言，我们建议 (1)构建和表征代表正常人和正常人的cDNA文库 和多囊性人类肾脏， (2)进行详细的平行动态cRNA文库杂交分析 正常肾脏与ADPKD肾脏的比较，以建立一个可靠的上限， 差异表达的转录物的数量， (3)进行平行文库消减以分离和表征ADPKD 特异性cDNA克隆， (4)表征从（3）分离的克隆的染色体位置。 不位于人类16号染色体上的克隆不太可能 与ADPKD相关）。 位于16号染色体上的克隆将被 通过细胞遗传学和与已知ADPKD的连锁分析进一步表征 标记， (5)表征相关克隆的组织分布， (6)利用现代合成肽和抗肽抗体技术， 鉴定和表征由所选克隆代表的蛋白质产物 在培养的细胞和组织切片中。

Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common, dominantly inherited kidney diseases of man. ADPKD is responsible for 10% of all patients requiring chronic dialysis or renal transplant. Currently, 500,000 Americans and 5 million people worldwide are affected with this disorder. Annual healthcare cost is estimated at 1 billion dollars in the U.S. alone. The exact molecular defect of ADPKD is unknown and besides transplantation, no curative treatment exists. This proposal attempts to apply modern molecular biology, and biochemistry to study aberrant gene expression in ADPKD, to produce molecular probes and to define the set of molecular parameters that are specifically altered in ADPKD. Our long term goal is to fully characterize the molecular genetics underlying the defect(s) in ADPKD. With the ADPKD gene characterized, accurate presymptomatic diagnostic tests can be designed. In addition, pharmacological and somatic therapies can be developed to treat and to prevent ADPKD. Specifically we propose to (1) construct and characterize cDNA libraries representative of both normal and polycystic human kidneys, (2) perform detailed parallel kinetic cRNA library hybridization analysis of normal versus ADPKD kidney to establish a reliable upper limit on the number of differentially expressed transcripts, (3) perform parallel library subtractions to isolate and characterize ADPKD specific cDNA clones, (4) characterize the chromosomal locations of clones isolated from (3). Clones that are not located on human chromosome 16 are not likely to be relevant to ADPKD). Clones that are located on chromosome 16 will be characterized futher by cytogenetics and linkage analysis to known ADPKD markers, (5) characterize the tissue distribution of relevant clones, (6) use modern synthetic peptide and antipeptide antibody technologies to identify and characterize protein products represented by selected clones in both cultured cells and tissue sections.