Analysis of Invasive Activity of CAF Spheroids into Three Dimensional (3D) Collagen Matrices

Abstract

Tumor growth and progression is the result of a complex process controlled not only by malignant cancer cells but also by the surrounding tumor microenvironment (TME). Cancer associated fibroblasts (CAFs), the most abundant cellular component of TME, play an active role in tumor invasion and metastasis by promoting cancer cell invasion through cell-cell interactions and secretion of pro-invasive factors such as extracellular matrix (ECM)-degrading proteases. Due to their tumor-promoting activities, there is an emerging interest in investigating CAFs biology and its potential as drug targets for cancer therapies. Here we describe an easy and highly reproducible quantitative method to analyze CAF invasive activity by forming multicellular spheroids embedded into a three-dimensional (3D) matrix that mimics in vivo ECM. Subsequently, invasion is monitored over time using a time-lapse microscope. We also provide an automated image analysis system that enables the rapid quantification of the spheroid area increase (invasive area) over time. The use of a 96-well plate format with one CAF spheroid per well and the automated analysis provides a method suitable for drug screening test, such as protease inhibitors.

Cancer-associated fibroblast-related gene signatures predict survival and drug response in patients with colorectal cancer

“…Cancer-associated fibroblasts (CAFs) are a special type of fibroblasts that surround tumors and form a key part of the TME. In recent years, CAFs have received increasing attention due to their crucial roles in tumor invasion, angiogenesis, and ECM remodeling by promoting cell-cell interaction and the secretion of proinvasive factors (Villaronga et al, 2018;Bertero et al, 2019). Targeting CAFs by altering their numbers, subtype or biological functionality is emerging as an attractive avenue to improve therapeutic strategies for cancer.…”