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

治疗进展性肾病靶分子的鉴定及其在基因治疗中的应用。

• 批准号: 15590859
• 负责人: HAYASHI Matsuhiko
• 金额: $ 2.11万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590859/
• 关键词: progressive renal diseases; Thy 1.1 nephritis; DNA microarray; NF-κB; Thymosin β-10; angiotensin converting enzyme type 2; NKκB; NFκB

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

抗THY1.1单克隆抗体的单次注射1-22-3，可诱导大鼠可逆肾小球肾炎，而半肾上腺切除术后同样的单一注射会影响不可逆的进行性肾小球肾小球肾炎。为了鉴定新的基因在肾脏疾病的进展中起重要作用，我们制作了这两种大鼠模型，即可逆和不可逆的肾小球肾炎大鼠，并从儿童中提取RNA，以定期进行微阵列分析。 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 were classified into 7 clusters and one cluster contained laminin, collagen type I, KIM-1, and osteopontin, which showed increased during the course of the progression of renal impairment in the irreversible model rats.胸腺素β-10也包括在内……群群和免疫组织学研究表明，胸腺素β-100存在于培养的THP-1细胞中肾脏损伤的过程中，胸腺素β-100，人类巨噬细胞系的肾脏损伤进展，这表明胸腺素β-10的表达与Thppopl thppopl的分化相差而增加。这些结果表明，胸腺素β-10可能在浸润巨噬细胞的激活和诱导间质损伤中起重要作用。我们目前正在研究进行性肾脏疾病治疗的靶基因的可能性。已知大量牛血清白蛋白注射腹腔诱导蛋白尿，从而在该模型中诱导肾脏间质损伤，NF-κB被认为扮演着角色。为了指定肾脏间隙损伤的分子，我们分析了通过注射大量牛血清白蛋白和蛋白尿大鼠诱导的对照蛋白尿大鼠之间的基因表达差异，该基因受到抑制了NF-κB活化层的NF-κB活化层。微阵列分析确定了通过激活NF-κB的激活，造蛋白尿诱导的间质损伤的进行性因素和受保护因素。在此分析中，簇蛋白被鉴定为受保护因素。此外，转化2型酶的血管紧张素随蛋白尿而降低并随着NF-κB的抑制而增加，这表明该酶的降低可能导致肾脏中肾素 - 血管紧张素系统的激活。血管紧张素转化2型酶可能成为进行性肾脏疾病治疗的靶基因。较少的

项目成果

Inhibition of diabetic nephropathy by a decoy peptide corresponding to the "handle" region for nonproteolytic activation of prorenin.

• DOI: 10.1172/jci21398
• 发表时间: 2004-10
• 期刊: The Journal of clinical investigation
• 作者: A. Ichihara;M. Hayashi;Y. Kaneshiro;F. Suzuki;T. Nakagawa;Y. Tada;Yukako Koura;A. Nishiyama;H. Okada;M. Uddin;A. Nabi;Y. Ishida;T. Inagami;T. Saruta

Araki T, Hayashi M, Saruta T.: "Cloning and characterization of a novel gene promoting ureteric bud branching in the metanephros"Kidney International. 64(6). 1968-1977 (2003)

Araki T、Hayashi M、Saruta T.：“促进后肾输尿管芽分支的新基因的克隆和表征”肾脏国际。

Spironolactone in combination with cilazapril ameliorates proteinuria and renal interstitial fibrosis in rats with anti-Thy-i irreversible nephritis.

螺内酯联合西拉普利可改善抗 Thy-i 不可逆性肾炎大鼠的蛋白尿和肾间质纤维化。

• 发表时间: 2004
• 期刊: Hypertension Research 27(12)
• 作者: Asai M;Monkawa T;Marumo T;Fukuda S;Tsuji M;Yoshino J;Kawachi H;Shimizu F;Hayashi M;Saruta T
• 通讯作者: Saruta T

Yoshino J, Monkawa T, Tsuji M, Hayashi M, Saruta T.: "Leukemia inhibitory factor is involved in tubular regeneration after experimental acute renal failure."Journal of the American Society of Nephrology. 14(12). 3090-3101 (2003)

Yoshino J、Monkawa T、Tsuji M、Hayashi M、Saruta T.：“白血病抑制因子参与实验性急性肾衰竭后的肾小管再生。”美国肾脏病学会杂志。