Abstract A005: Patient-derived cancer associated fibroblasts induce varying rates of colorectal cancer cell invasion using an organ-on-chip model

Abstract Colorectal cancer (CRC) is one of the most common cancers in the United States, with metastatic CRC having an especially poor prognosis. Because of this, significant effort has been put into understanding the metastatic cascade. However, very little is still understood about how the tumor microenvironment (TME) influences this process. Recent studies have implicated cancer associated fibroblasts (CAFs), a major component of the TME, to be involved in tumor cell invasion. CAFs have been reported to be highly heterogeneous, with classification into several subtypes, including inflammatory CAFs (iCAFs) and myofibroblastic CAFs (myCAFs), based on “-omics” based studies. However, there has been limited exploration into the specific functionality of these subtypes in contributing to cancer progression. Therefore, we have worked to develop an organ-on-chip model to study CAF influence on tumor invasion in CRC (CRC-on-chip). This microfluidic device simulates the fluid flow and peristalsis-like motions that occur in the gut and is composed of two extracellular matrix-coated overlapping channels separated by a porous membrane. CAFs and GFP-labelled tumor cells are seeded into the top “epithelial” channel and endothelial cells are seeded in the bottom “vascular” channel. Confocal microscopy is used to track and quantify GFP+ tumor cells intravasating into the vascular channel and determine rates of invasion. The CAFs are derived from our biobank of diverse CRC patient tumor samples. From previous work, we observed patient-specific differences in CAF-induced CRC invasion when exposing human CRC cells on-chip to CAF-conditioned media. Through physical co-cultures, we similarly observed that some CAF lines increased invasion while others did not change invasion. Bulk RNA-sequencing and flow cytometry analysis of the CAF lines revealed varied expression of invasion-related genes and trends toward specific CAF subtype signatures, such as the iCAF and myCAF subtypes. These results suggest that there may be specific CAF phenotypes that induce tumor invasion which could give insight into determining the invasive potential of a primary CRC tumor at diagnosis. Citation Format: Bethany Haliday. Patient-derived cancer associated fibroblasts induce varying rates of colorectal cancer cell invasion using an organ-on-chip model [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr A005.