Pancreatic cancer-associated retinoblastoma 1 dysfunction enables TGF-β to promote proliferation

Pancreatic ductal adenocarcinoma (PDAC) is often associated with overexpression of TGF-beta. Given its tumor suppressor functions, it is unclear whether TGF-beta is a valid therapeutic target for PDAC. Here, we found that proliferating pancreatic cancer cells (PCCs) from human PDAC patients and multiple murine models of PDAC (mPDAC) often exhibit abundant levels of phosphorylated. retinoblastoma 1 (RB) and Smad.2. TGF-beta 1 treatment enhanced proliferation of PCCs isolated from Kras(G12D)-driven mPDAC that lacked RB (KRC cells). This mitogenic effect was abrogated by pharmacological inhibition of type I TGF-beta receptor kinase, combined inhibition of MEK/Src or MEK/PI3K, and restoration of RB expression. TGF-beta 1 promoted epithelial-to-mesenchymal transition (EMT), invasion, Smad2/3 phosphorylation, Src activation, Wnt reporter activity, and Smad-dependent upregulation of Wnt7b in KRC cells. Importantly, TGF-beta 1-induced mitogenesis was markedly attenuated by inhibition of Wnt secretion. In an in vivo syngeneic orthotopic model, inhibition of TGF-beta signaling suppressed KRC cell proliferation, tumor growth, stroma formation, EMT, metastasis, ascites formation, and Wnt7b expression, and markedly prolonged survival. Together, these data indicate that RB dysfunction converts TGF-beta to a mitogen that activates known oncogenic signaling pathways and upregulates Wnt7b, which synergize to promote PCC invasion, survival, and mitogenesis. Furthermore, this study suggests that concomitantly targeting TGF-beta and Wnt7b signaling in PDAC may disrupt these aberrant pathways, which warrants further evaluation in preclinical models.