Molecular Mechanisms of Renal Fibrosis

描述(申请人提供)：间质纤维化是终末期肾脏疾病的最终常见途径。肾成纤维细胞是这一过程的主要贡献者，但人们对它们来自哪里或如何参与知之甚少。为了帮助研究和更好地了解它们的作用，我们最近发现了一种新的蛋白质，称为成纤维细胞特异性蛋白-I(FSP1)。FSPI属于细胞内钙结合蛋白S100超家族，调节细胞骨架-膜相互作用、细胞生长和分化。FSP1是成纤维细胞的高保真标记物，通过上皮-间充质转化(EMT)过程，在形成成纤维细胞的转化过程中起指导作用。我们计划使用基于FSP1的分子和转基因探针进一步探索肾成纤维细胞与邻近上皮之间的关系。特别是，我们在FSP1启动子中发现了FTS-1/Carg盒结合位点，它不仅赋予成纤维细胞特异性，还识别了一组组成EMT转录组的其他基因。我们鉴定的一种锌指转录因子CBF(Carg Box Bindingfactor)可能是通过这些位点调控EMT的一个很好的候选基因。我们还制作了一只FSP1+/+GFP小鼠，它标记成纤维细胞为绿色，以便更好地识别、分离和绘制它们在发育过程中的命运；FSP1 GFP/GFP敲门小鼠，用于探索FSP1在调节EMT机制中的作用；以及一只FSP1小鼠，用于选择性地解开成纤维细胞中与EMT或细胞间信号转导相关的基因(Tbr2 FLOX/FLOX、BMP4 FLOX/LacZ、cMET FLOX/FLOX和EGFR FLOX/FLOX)。对这些转基因探针的评估可能会通过揭示或选择性干扰器官发生或损伤过程中肾成纤维细胞与其邻近上皮之间的重要信号来为我们未来的工作提供信息。因此，我们的整体方法应该促进对成纤维细胞在健康和疾病中的理解，以及塑造其表型的关键信号。这一理解可能是了解我们如何进一步操纵它们参与肾纤维化的关键。

DESCRIPTION (provided by applicant): Interstitial fibrosis is the final common pathway to end-stage renal disease. Kidney fibroblasts are principal contributors to this process, but little is known about where they come from or how they engage. To help study and better understand their role, we recently discovered a new protein called fibroblast-specific protein-I (FSP1). FSPI belongs to the S100 superfamily of intracellular calcium-binding proteins that regulate cytoskeletal-membrane interactions, cell growth, and differentiation. FSP1 is a high fidelity marker of fibroblasts and plays an instructive role in shaping their conversion from epithelium through a process called epithelial-mesenchymal transition (EMT). We plan to further explore the relationship between renal fibroblasts and adjacent epithelium using FSPl-based molecular and transgenic probes. In particular, we found an FTS-1/CArG box-binding site in the FSP1 promoter that not only confers fibroblast specificity but also identifies a group of other genes comprising the EMT transcriptome. A zinc-finger transcription factor identified by us and called CBF (CArG box binding factor) may be a good candidate for regulating EMT through those sites. We have also made a FSP1 +/+ GFP mouse that marks fibroblasts green to better identify, isolate, and map their fate during development, a FSP1 GFP/GFP knockin mouse to explore the role of FSP1 in regulating the mechanism of EMT, and a FSP1.Cre mouse to selectively unflox genes in fibroblasts whose functions are associated either with EMT or intercellular signaling (Tbr2 flox/flox , BMP4 flox/lacZ, cMet flox/flox, and Egfr flox/flox). The evaluation of these transgenic probes is likely to inform our future work by revealing, or selectively disturbing, important signals between renal fibroblasts and their adjacent epithelium during organogenesis or injury. Thus, our overall approach should advance understanding of fibroblasts in health and disease as well as the critical signals that shape their phenotype. This understanding may be key to learning how we can further manipulate their participation in renal fibrosis.