Identification of target molecule for the treatment of progressive renal diseases and its application for gene therapy.

单次注射抗Thy1.1单克隆抗体1-22-3在大鼠体内诱导可逆性肾小球肾炎，而在半肾切除术后单次注射同样的单克隆抗体诱导不可逆的进行性肾小球肾炎。为了鉴定在肾脏疾病进展中发挥重要作用的新基因，我们制作了可逆和不可逆肾小球肾炎大鼠两种大鼠模型，并定期从肾脏中提取RNA进行微阵列分析。分析两种模型之间基因表达的差异，以增加2倍以上或减少50%以上的差异为显著。根据这一定义，有191个基因出现显著变化，并进行聚类分析。这191个基因被分为7个簇，其中一个簇含有层粘连蛋白、I型胶原、KIM-1和骨桥蛋白，在不可逆模型大鼠肾损害的进展过程中表现出增加。聚类和免疫组织学研究表明，随着肾损害的进展，胸腺蛋白酶β-10存在于间质组织中。在培养的人巨噬细胞系THP-1细胞中，胸腺蛋白酶β-10的表达随着THP-1细胞向巨噬细胞的分化而增加。提示胸腺酶β-10可能在活化浸润性巨噬细胞和诱导间质损伤中起重要作用。我们目前正在研究治疗进行性肾脏疾病的靶基因的可能性。已知腹腔注射大量牛血清白蛋白可诱导蛋白尿，从而引起肾间质损伤。在该模型中，NF-κB被认为起关键作用。为了明确肾间质损伤的分子，我们分析了注射大量牛血清白蛋白诱导的正常蛋白尿大鼠和接受截断型IκBα腺病毒基因转移的蛋白尿大鼠之间的基因表达差异，该基因可以抑制近端小管中NF-κB的激活。微阵列分析通过NF-κB的激活，确定尿路性间质损伤的进行性因素和保护性因素。在本分析中，聚簇蛋白被确定为保护因子。此外，血管紧张素转换酶2型随蛋白尿而降低，随NF-κB抑制而升高，提示该酶的降低可能导致肾素-血管紧张素系统的激活。血管紧张素转换酶2型可能成为治疗进行性肾脏疾病的靶基因。少

Single injection of anti Thy1.1 monoclonal antibody, 1-22-3, induces reversible glomerulonephritis in the rats, whereas the same single injection after heminephrectomy induces irreversible progressive glomerulonephritis. To identify the novel genes, which play important roles in the progression of renal diseases, we made these two rat models, reversible and irreversible glomemlonephritis rats, and extracted RNA from the kidneys for microarray analysis, periodically. The differences of gene expression between two models were analyzed and the differences of more than 2 time increase or 50% reduction were considered as significant By this definition, 191 genes showed significant changes and cluster analysis was performed. These 191 genes were classified into 7 clusters and one cluster contained laminin, collagen type I, KIM-1, and osteopontin, which showed increase during the course of the progression of renal impairment in the irreversible model rats. Thymosin β-10 is also included in th … More is cluster and immunohistological study revealed that thymosin β-10 is present in the interstitial tissues with progression of the renal impairment In the cultured THP-1 cells, human macrophage cell line, it was shown that expression of thymosin β-10 increased with the differentiation of THP-1 cells to macrophage. These results suggest that thymosin β-10 may play important roles in the activation of infiltrated macrophage and induction of interstitial damages. We are currently working on the possibility of the target gene of the therapy for progressive renal diseases.Injection of massive bovine serum albumin into abdominal cavity is known to induce proteinuria and thereby induces renal interstitial impairment In this model, NF-κB is thought to play a pivotal role. To specify the molecules of renal interstitial impairment, we analyzed gene expression differences between control proteinuric rats induced by injection of massive bovine serum albumin and proteinuric rats received adenoviral gene transfer of truncated form IκBα, which inhibits NF-κB activation in the proximal tubules. Microarray analysis identified progressive factors and protective factors of pioteinuria-induced interstitial impairment through the activation of NF-κB. In this analysis, clusterin was identified as protective factor. In addition, angiotensin converting enzyme type 2 showed decrease with proteinuria and increase with inhibition of NF-κB, suggesting that decrease of this enzyme might result in the activation of renin-angiotensin system in the kidney. Angiotensin converting enzyme type 2 may become a target gene of the treatment of progressive renal diseases. Less