Profibrotic immune cells and fibro-adipogenic progenitors contribute to skeletal muscle extracellular matrix remodeling and fibrosis in cancer cachexia.

Cachexia, the loss of skeletal muscle mass and function with cancer, contributes to reduced life quality and worsened survival. Skeletal muscle fibrosis leads to disproportionate muscle weakness; however, the role of infiltrating immune cells and fibro-adipogenic progenitors (FAPs) in cancer-induced muscle fibrosis is not well understood. Using the colon-26 adenocarcinoma (C26) model of cancer cachexia, we sought to examine the changes to skeletal muscle immune cells and FAPs, which contribute to excessive extracellular matrix (ECM) collagen deposition. CD2F1 male mice (n = 35) were implanted with either 106 C26 or CT-26 (weight stable; WS) cells. Skeletal muscle immune cell populations, satellite cells, and FAPs were examined using high-dimensional flow cytometry. Skeletal muscle ECM ultrastructure was assessed via scanning electron microscopy (SEM) of decellularized muscle along with transmission electron microscopy (TEM). Cachectic mice had significant decreases in body weight (-13.4%, P = 0.003) and skeletal muscle mass (-37%, P = 0.006). Cachectic mice had elevated CD45+CD11b+Ly6g+ neutrophils compared with nontumor-bearing controls (128%, P = 0.016) and elevated CD45+CD11b+Ly6g-F480+CD206+MHCII- profibrotic macrophages and increased CD45-Sca1+CD106+CD140a+ FAPs compared with WS (43%, P = 0.014) and controls (59%, P = 0.002) with thickening of the ECM, particularly of the endomysium and perimysium. SEM and TEM analyses also identified clusters of infiltrating cells localized to regions of excessive ECM deposition in cachectic mice that were absent in WS and controls. These data highlight changes to the muscle microenvironment, which contribute to fibrosis and excessive ECM deposition in cancer cachexia. Targeting profibrotic immune cells may represent a promising therapeutic approach to mitigate muscle wasting and dysfunction with cachexia.NEW & NOTEWORTHY Skeletal muscle fibrosis contributes to disproportionate muscle weakness with cancer cachexia. Our novel findings demonstrate an inflammatory and profibrotic cachectic muscle microenvironment, demonstrated by increased fibro-adipogenic progenitor cells, neutrophils, and profibrotic macrophages. Using scanning and transmission electron microscopy, we show expansion and thickening of the extracellular matrix within the endomysium and perimysium. We also identified clusters of infiltrating cells localized to regions of excessive ECM deposition, which were not present in weight-stable tumor-bearing controls.