Intracellular vesicle trafficking is an evolutionary conserved process implicated in a great variety of cellular functions and diseases. Pancreatic cancer cells exploit vesicle trafficking via the Golgi apparatus to support cellular flexibility and tumor aggressiveness by relying on vesicle-trafficking proteins such as myoferlin. However, our understanding of myoferlin-dependent vesicle trafficking is limited to cancer cells, while the function of myoferlin in the pancreatic tumor microenvironment, notably cancer-associated fibroblasts (CAFs), has been overlooked.
Here we combine pancreatic adenocarcinoma (PAAD) whole-tumor and single-cell transcriptomic analyses with immunohistochemistry to link stromal myoferlin to tumor aggressiveness. Using 2D and 3D in vitro models of human CAFs, we unveil CAF-specific functions of myoferlin, as MYOF-depleted (MYOFKD) CAFs present impaired activity and reduced extracellular matrix (ECM) production. Analysis of intracellular vesicles in MYOFKD CAFs identifies myoferlin as novel functional member of COP2-coated vesicle trafficking between the endoplasmic reticulum (ER) and Golgi apparatus. Accordingly, MYOFKD causes a TGFß-receptor 1 (TGFBR1) trafficking blockade at the ER/Golgi interface, leading to altered TGFBR1 activation, impaired TGFß signal transduction, loss of ECM production and reduced stroma aggressiveness. Orthotopic transplantation of MYOFKD CAFs with pancreatic cancer cells in mice impairs tumor establishment, while pharmacological targeting of myoferlin reduces tumor desmoplasia in tumor-bearing mice without increasing tumor burden.
Overall, we propose TGFBR1 trafficking, druggable via myoferlin, as novel approach to interfere with signal transduction and to reprogram CAFs. Thus, enabling to efficiently tackle desmoplasia and to control stromal aggressiveness. The translational outlook of these findings is underpinned by the driving role of tumor desmoplasia during therapy resistance and immune exclusion in pancreatic cancer.