Effect of fibroblast-specific suicidal-gene against rapidly progressive glomerulonephritis

成纤维细胞特异性自杀基因对急进性肾小球肾炎的作用

• 批准号: 13671127
• 负责人: OKADA Hirokazu
• 金额: $ 1.86万
• 依托单位: Saitama Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671127/
• 关键词: Rapidly progressive glomerulo-nephritis; fibrosis; fibroblast; gene therapy; 急速進行性糸球体腎炎; 腎間質線維化; FSP1; アポトーシス; 抗基底膜抗体腎炎; 間質線維化; トランスジェニックマウス; thymidine kinase; gancyclovir

The extent of renal fibrosis is the best predictor for functional outcomes in a variety of progressive renal diseases. Interstitial fibroblasts and fibroblast-like cells (FbLCs) are presumably involved In the fibrotic process. However, such FbLCs have never been well-characterized in the kidney. Therefore, in this study, we characterized them using immunohistochemistry and in situ hybridization with phenotypic and functional marker probes. In the interstitium of anti-glomerular basement membrane (αGBM) nephritic kidneys, fibroblast-specific protein1 (FSP1)^+ fibroblasts were demonstrated to produce connective tissue growth factor (CTGF) and type I collagen (COLI) whereas vimentin^+ FbLCs synthesized transforming growth factor-β1. To prove whether FSP1^+ fibroblasts played an essential role in renal fibrogenesis, we employed mice transgenic for a deleted form of thymidine kinase (dTK) cDNA under the control of FSP1 promoter (FSP1.dTK^+). Since the transgenic FSP1^+ fibroblasts were dual-positive for TK protein, and these proliferating cells went apoptosis when gancyclovir (GCV) was administered, we generated αGBM nephritis in them and treated them with GCV. In the αGBM nephritic kidney of these FSP1.dTK^+ mice, GCV treatment significantly suppressed the increase in the number of FSP1^+ fibroblasts and the production of CTGF and COLI, resulting in the attenuation of renal fibrogenesis successfully. FSP1^+ were demonstrated to be a master contributor in the fibrosing kidneys, and the FSP1 gene-guided anti-fibrotic therapies seem promising for a variety of progressive renal diseases.

肾纤维化的程度是各种进行性肾脏疾病功能结果的最佳预测指标。间质成纤维细胞和成纤维细胞样细胞 (FbLC) 可能参与纤维化过程。然而，此类 FbLC 从未在肾脏中得到很好的表征。因此，在本研究中，我们使用免疫组织化学和表型和功能标记探针的原位杂交对它们进行了表征。在抗肾小球基底膜 (αGBM) 肾病肾脏的间质中，成纤维细胞特异性蛋白 1 (FSP1)^+ 成纤维细胞被证明能够产生结缔组织生长因子 (CTGF) 和 I 型胶原 (COLI)，而波形蛋白^+ FbLC 合成转化生长因子-β1。为了证明 FSP1^+ 成纤维细胞是否在肾纤维化中发挥重要作用，我们采用了在 FSP1 启动子 (FSP1.dTK^+) 控制下胸苷激酶 (dTK) cDNA 缺失形式的转基因小鼠。由于转基因FSP1^+成纤维细胞对TK蛋白双阳性，并且当施用更昔洛韦(GCV)时这些增殖细胞发生凋亡，因此我们在其中产生αGBM肾炎并用GCV治疗它们。在这些 FSP1.dTK^+ 小鼠的 αGBM 肾病肾中，GCV 治疗显着抑制了 FSP1^+ 成纤维细胞数量的增加以及 CTGF 和 COLI 的产生，成功地减弱了肾纤维化。 FSP1^+ 被证明是肾脏纤维化的主要贡献者，FSP1 基因引导的抗纤维化疗法似乎对多种进行性肾脏疾病有希望。