The Extracellular Matrix Environment of Clear Cell Renal Cell Carcinoma Determines Cancer Associated Fibroblast Growth.
The Extracellular Matrix Environment of Clear Cell Renal Cell Carcinoma Determines Cancer Associated Fibroblast Growth.
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肾透明细胞癌的细胞外基质环境决定癌相关成纤维细胞的生长。
DOI：
10.3390/cancers13235873
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发表时间：
2021-11-23
期刊：
影响因子：
5.2
通讯作者：
Oxburgh L
中科院分区：
文献类型：
--
作者：
Bond KH;Chiba T;Wynne KPH;Vary CPH;Sims-Lucas S;Coburn JM;Oxburgh L
透明细胞肾细胞癌（ccRCC）是最常见的肾癌。细胞培养研究有可能解释癌细胞及其支持细胞（基质）之间的相互作用如何决定ccRCC的生长和恶性。肿瘤细胞与来自患者肿瘤的基质一起生长的能力对于这些相互作用的研究至关重要，但传统的培养方法不能提供两种细胞类型的代表。我们假设模拟肿瘤的细胞外环境会促进肿瘤和基质细胞的生长。我们表征了患者ccrcc的细胞外蛋白组成，并定义了一种九组分蛋白混合物来模拟肿瘤微环境。肿瘤细胞、支持基质细胞和免疫细胞的粘附性得到证实。此外，我们发现在我们的蛋白混合物中生长的患者肿瘤细胞保持了肿瘤细胞和癌症相关成纤维细胞（CAFs）的代表，CAFs是一种基质细胞类型，在建立肿瘤微环境中起着决定性的作用，但人们对其知之甚少。该研究证明了CAFs对细胞外蛋白组成的依赖性，并提供了一种研究肿瘤细胞与患者ccrcc分离的CAFs之间相互作用的技术。透明细胞肾细胞癌（ccRCC）是最常见的肾癌，通常由肾上皮细胞的氧感应机制突变引起。由于其假缺氧状态，ccRCC招募了广泛的脉管系统和其他基质成分。传统的细胞培养方法在ccRCC原代培养中不能很好地代表基质细胞类型，我们假设模拟肿瘤的细胞外环境会促进肿瘤和基质细胞的生长。我们采用蛋白质组学方法鉴定了ccRCC细胞外基质（ECM）的成分，发现与健康肾皮质相比，层粘连蛋白、IV型胶原和entactin/nidogen是次要成分。相反，ccRCC ECM主要由胶原VI、纤维连接蛋白和腱素c组成。单细胞表达数据分析表明，癌症相关成纤维细胞是肿瘤ECM产生的主要来源。肿瘤细胞和基质细胞有效地结合九组分ECM混合ccRCC的特征。患者来源的原代肿瘤细胞有效地结合九组分混合物，使我们能够建立肿瘤细胞和基质细胞的混合原代培养。这些微型患者特异性复制品有利于显微镜观察，可用于分析模型肿瘤微环境中细胞之间的相互作用。
Clear cell renal cell carcinoma (ccRCC) is the most common kidney cancer. Cell culture studies have the potential to explain how interactions between cancer cells and their support cells (stroma) determine growth and malignancy of ccRCC. The ability to grow tumor cells together with stroma from patient tumors is critical for studies of these interactions, but conventional culture methods do not provide representation of both cell types. We hypothesized that mimicking the extracellular environment of the tumor would promote growth of both tumor and stromal cells. We characterized the extracellular protein composition of patient ccRCCs and defined a nine-component protein blend to mimic the tumor microenvironment. Adherence of tumor cells, supporting stromal cells, and immune cells was demonstrated. Furthermore, we found that cells from patient tumors grown in our protein blend maintained representation of both tumor cells and cancer-associated fibroblasts (CAFs), a stromal cell type that plays a defining but poorly understood role in establishing the tumor microenvironment. This study demonstrates the dependence of CAFs on the extracellular protein composition and provides a technology to investigate interactions between tumor cells and CAFs isolated from patient ccRCCs. Clear cell renal cell carcinoma (ccRCC) is the most common kidney cancer and is often caused by mutations in the oxygen-sensing machinery of kidney epithelial cells. Due to its pseudo-hypoxic state, ccRCC recruits extensive vasculature and other stromal components. Conventional cell culture methods provide poor representation of stromal cell types in primary cultures of ccRCC, and we hypothesized that mimicking the extracellular environment of the tumor would promote growth of both tumor and stromal cells. We employed proteomics to identify the components of ccRCC extracellular matrix (ECM) and found that in contrast to healthy kidney cortex, laminin, collagen IV, and entactin/nidogen are minor contributors. Instead, the ccRCC ECM is composed largely of collagen VI, fibronectin, and tenascin C. Analysis of single cell expression data indicates that cancer-associated fibroblasts are a major source of tumor ECM production. Tumor cells as well as stromal cells bind efficiently to a nine-component ECM blend characteristic of ccRCC. Primary patient-derived tumor cells bind the nine-component blend efficiently, allowing to us to establish mixed primary cultures of tumor cells and stromal cells. These miniature patient-specific replicas are conducive to microscopy and can be used to analyze interactions between cells in a model tumor microenvironment.
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DOI：
10.1073/pnas.88.14.6028
发表时间：
1991-07-01
影响因子：
11.1
作者：
CORNIL, I;THEODORESCU, D;KERBEL, RS
通讯作者：
KERBEL, RS
影响因子：
82.9
作者：
Choueiri, Toni K.;Kaelin, William G., Jr.
通讯作者：
Kaelin, William G., Jr.
影响因子：
64.5
作者：
BROOKS, PC;MONTGOMERY, AMP;CHERESH, DA
通讯作者：
CHERESH, DA
影响因子：
56.9
作者：
BROOKS, PC;CLARK, RAF;CHERESH, DA
通讯作者：
CHERESH, DA
影响因子：
7.5
作者：
Gimona, Mario;Buccione, Roberto;Linder, Stefan
通讯作者：
Linder, Stefan