Waters John P, Richards Yvonne C, Skepper Jeremy N, Southwood Mark, Upton Paul D, Morrell Nicholas W, Pober Jordan S, Bradley John R
Department of Medicine, University of Cambridge, Addenbrooke's Hospital and NIHR Cambridge Biomedical Research Centre, Cambridge, UK.
Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK.
J Pathol. 2017 Nov;243(3):390-400. doi: 10.1002/path.4960. Epub 2017 Oct 3.
Glomerular scarring, known as glomerulosclerosis, occurs in many chronic kidney diseases and involves interaction between glomerular endothelial cells (GECs), podocytes, and mesangial cells (MCs), leading to signals that promote extracellular matrix deposition and endothelial cell dysfunction and loss. We describe a 3D tri-culture system to model human glomerulosclerosis. In 3D monoculture, each cell type alters its phenotype in response to TGFβ, which has been implicated as an important mediator of glomerulosclerosis. GECs form a lumenized vascular network, which regresses in response to TGFβ. MCs respond to TGFβ by forming glomerulosclerotic-like nodules with matrix deposition. TGFβ treatment of podocytes does not alter cell morphology but increases connective tissue growth factor (CTGF) expression. BMP7 prevents TGFβ-induced GEC network regression, whereas TGFβ-induced MC nodule formation is prevented by SMAD3 siRNA knockdown or ALK5 inhibitors but not BMP7, and increased phospho-SMAD3 was observed in human glomerulosclerosis. In 3D tri-culture, GECs, podocytes, and MCs form a vascular network in which GECs and podocytes interact intimately within a matrix containing MCs. TGFβ treatment induces formation of nodules, but combined inhibition of ALK5 and CTGF is required to prevent TGFβ-induced nodule formation in tri-cellular cultures. Identification of therapeutic targets for glomerulosclerosis depends on the 3D culture of all three glomerular cells. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
肾小球瘢痕形成，即肾小球硬化，发生于多种慢性肾脏疾病中，涉及肾小球内皮细胞（GECs）、足细胞和系膜细胞（MCs）之间的相互作用，从而产生促进细胞外基质沉积以及内皮细胞功能障碍和丧失的信号。我们描述了一种用于模拟人类肾小球硬化的三维三细胞共培养系统。在三维单培养中，每种细胞类型都会响应转化生长因子β（TGFβ）而改变其表型，TGFβ被认为是肾小球硬化的重要介质。GECs形成有腔的血管网络，该网络会因TGFβ而退化。MCs通过形成具有基质沉积的肾小球硬化样结节来响应TGFβ。用TGFβ处理足细胞不会改变细胞形态，但会增加结缔组织生长因子（CTGF）的表达。骨形态发生蛋白7（BMP7）可防止TGFβ诱导的GEC网络退化，而TGFβ诱导的MC结节形成可通过SMAD3小干扰RNA敲低或ALK5抑制剂来阻止，但不能被BMP7阻止，并且在人类肾小球硬化中观察到磷酸化SMAD3增加。在三维三细胞共培养中，GECs、足细胞和MCs形成一个血管网络，其中GECs和足细胞在含有MCs的基质内紧密相互作用。TGFβ处理会诱导结节形成，但在三细胞培养中需要联合抑制ALK5和CTGF才能防止TGFβ诱导的结节形成。确定肾小球硬化的治疗靶点取决于所有三种肾小球细胞的三维培养。版权所有© 2017英国和爱尔兰病理学会。由约翰·威利父子有限公司出版。
