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.

肾纤维化的程度是多种进行性肾脏疾病功能结果的最佳预测因子。间质成纤维细胞和成纤维细胞样细胞（FBLC）大概参与了纤维化过程。但是，这种FBLC从未在肾脏中得到很好的表征。因此，在这项研究中，我们使用免疫组织化学以及与表型和功能标记问题的原位杂交对它们进行了表征。 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.为了证明Fsp1^+成纤维细胞在肾纤维发生中是否起着至关重要的作用，我们在FSP1启动子（FSP1.DTK^+）的控制下采用了胸苷激酶（DTK）cDNA的已删除形式的胸苷激酶（DTK）cDNA。由于TK蛋白的转基因FSP1^+成纤维细胞是双阳性的，并且当施用GencyClovir（GCV）时，这些增殖细胞会凋亡，因此我们在其中产生了αGBM肾炎并用GCV治疗它们。在这些FSP1.DTK^+小鼠的αGBM肾肾中，GCV处理显着抑制了FSP1^+成纤维细胞数量的增加以及CTGF和大肠杆菌的产生，从而成功地衰减了肾纤维发生。 FSP1^+被证明是纤维化肾脏中的主要贡献者，而FSP1基因引导的抗纤维化疗法对于各种进行性肾脏疾病似乎很有希望。