Material-driven fibronectin assembly rescues matrix defects due to mutations in collagen IV in fibroblasts.
Material-driven fibronectin assembly rescues matrix defects due to mutations in collagen IV in fibroblasts.
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DOI：
10.1016/j.biomaterials.2020.120090
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发表时间：
2020-09
期刊：
影响因子：
14
通讯作者：
Salmeron-Sanchez M
中科院分区：
文献类型：
--
作者：
Ngandu Mpoyi E;Cantini M;Sin YY;Fleming L;Zhou DW;Costell M;Lu Y;Kadler K;García AJ;Van Agtmael T;Salmeron-Sanchez M
基底膜 (BM) 是一种特殊的细胞外基质，可为组织提供结构支持并影响细胞行为和信号传导。 COL4A1/COL4A2 是 BM 的主要组成部分，其突变会导致家族性眼、肾和脑血管疾病，包括中风，而这些基因的常见变异是普通人群中脑出血的危险因素。这些表型与基质缺陷相关，这是由于 BM 中突变蛋白的掺入和/或内质网 (ER) 滞留导致的缺失。然而，这些突变对基质硬度的影响、基质对疾病机制的贡献及其对含有 IV 型胶原蛋白突变的细胞生物学的影响仍然知之甚少。为了阐明这一点，我们采用合成聚合物生物界面，即涂有 ECM 蛋白层粘连蛋白或纤连蛋白 (FN) 的聚丙烯酸乙酯 (PEA) 和聚丙烯酸甲酯 (PMA)，来生成受控微环境，并研究它们对含有 COL4A2+/G702D 突变的原代成纤维细胞的细胞表型的影响。在 PEA 上组装的 FN 纳米网络诱导 COL4A2+/G702D 细胞中 IV 型胶原的沉积和组装增加，这与 ER 尺寸减小和 BIP 等蛋白质伴侣水平增加有关，表明细胞的蛋白质折叠能力增加。 PEA 上的 FN 纳米网络还部分挽救了沉积基质和细胞的硬度降低，并通过增加肌动蛋白-肌球蛋白收缩性增强细胞粘附，有效挽救了与 COL4A1/4A2 突变相关的一些细胞表型。 FN 纳米网络增强细胞表型的机制涉及整合素 β1 介导的信号传导。总的来说，这些结果表明生物材料和通过组装的 FN 增强的整合素信号传导能够塑造患者来源细胞中 COL4A2+/G702D 突变的基质和细胞表型。
Basement membranes (BMs) are specialised extracellular matrices that provide structural support to tissues as well as influence cell behaviour and signalling. Mutations in COL4A1/COL4A2, a major BM component, cause a familial form of eye, kidney and cerebrovascular disease, including stroke, while common variants in these genes are a risk factor for intracerebral haemorrhage in the general population. These phenotypes are associated with matrix defects, due to mutant protein incorporation in the BM and/or its absence by endoplasmic reticulum (ER) retention. However, the effects of these mutations on matrix stiffness, the contribution of the matrix to the disease mechanism(s) and its effects on the biology of cells harbouring a collagen IV mutation remain poorly understood. To shed light on this, we employed synthetic polymer biointerfaces, poly(ethyl acrylate) (PEA) and poly(methyl acrylate) (PMA) coated with ECM proteins laminin or fibronectin (FN), to generate controlled microenvironments and investigate their effects on the cellular phenotype of primary fibroblasts harbouring a COL4A2+/G702D mutation. FN nanonetworks assembled on PEA induced increased deposition and assembly of collagen IV in COL4A2+/G702D cells, which was associated with reduced ER size and enhanced levels of protein chaperones such as BIP, suggesting increased protein folding capacity of the cell. FN nanonetworks on PEA also partially rescued the reduced stiffness of the deposited matrix and cells, and enhanced cell adhesion through increased actin-myosin contractility, effectively rescuing some of the cellular phenotypes associated with COL4A1/4A2 mutations. The mechanism by which FN nanonetworks enhanced the cell phenotype involved integrin β1-mediated signalling. Collectively, these results suggest that biomaterials and enhanced integrin signalling via assembled FN are able to shape the matrix and cellular phenotype of the COL4A2+/G702D mutation in patient-derived cells.
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10
作者：
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通讯作者：
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