Gurrala Rakesh, Byrne C Ethan, Brown Loren M, Tiongco Rafael Felix P, Matossian Margarite D, Savoie Jonathan J, Collins-Burow Bridgette M, Burow Matthew E, Martin Elizabeth C, Lau Frank H
Department of Surgery, Louisiana State University Health Sciences Center New Orleans, New Orleans, LA, United States.
School of Medicine, Tulane University, New Orleans, LA, United States.
Front Bioeng Biotechnol. 2021 Mar 15;9:618448. doi: 10.3389/fbioe.2021.618448. eCollection 2021.
Solid tumor progression is significantly influenced by interactions between cancer cells and the surrounding extracellular matrix (ECM). Specifically, the cancer cell-driven changes to ECM fiber alignment and collagen deposition impact tumor growth and metastasis. Current methods of quantifying these processes are incomplete, require simple or artificial matrixes, rely on uncommon imaging techniques, preclude the use of biological and technical replicates, require destruction of the tissue, or are prone to segmentation errors. We present a set of methodological solutions to these shortcomings that were developed to quantify these processes in cultured, human breast tissue under the influence of breast cancer cells and allow for the study of ECM in primary breast tumors. Herein, we describe a method of quantifying fiber alignment that can analyze complex native ECM from scanning electron micrographs that does not preclude the use of replicates and a high-throughput mechanism of quantifying collagen content that is non-destructive. The use of these methods accurately recapitulated cancer cell-driven changes in fiber alignment and collagen deposition observed by visual inspection. Additionally, these methods successfully identified increased fiber alignment in primary human breast tumors when compared to human breast tissue and increased collagen deposition in lobular breast cancer when compared to ductal breast cancer. The successful quantification of fiber alignment and collagen deposition using these methods encourages their use for future studies of ECM dysregulation in human solid tumors.
实体瘤的进展受到癌细胞与周围细胞外基质（ECM）之间相互作用的显著影响。具体而言，癌细胞驱动的ECM纤维排列和胶原蛋白沉积的变化会影响肿瘤的生长和转移。目前量化这些过程的方法并不完善，需要简单或人工基质，依赖不常见的成像技术，排除了生物和技术重复的使用，需要破坏组织，或者容易出现分割错误。我们提出了一套针对这些缺点的方法学解决方案，这些方案旨在量化在乳腺癌细胞影响下培养的人乳腺组织中的这些过程，并允许对原发性乳腺肿瘤中的ECM进行研究。在此，我们描述了一种量化纤维排列的方法，该方法可以从扫描电子显微镜图像中分析复杂的天然ECM，且不排除重复使用，还描述了一种非破坏性的高通量量化胶原蛋白含量的机制。使用这些方法准确地重现了通过目视检查观察到的癌细胞驱动的纤维排列和胶原蛋白沉积的变化。此外，与人类乳腺组织相比，这些方法成功地识别出原发性人类乳腺肿瘤中纤维排列增加，与导管癌相比，小叶乳腺癌中胶原蛋白沉积增加。使用这些方法成功量化纤维排列和胶原蛋白沉积，鼓励将其用于未来人类实体瘤中ECM失调的研究。
