Abrogating TGFβ signaling in TCR-engineered T cells and enhancing antigen processing by tumor cells promotes sustained therapeutic activity in pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDA) is a deadly malignancy with limited effective therapies. Adoptive cell therapy (ACT) is a promising treatment modality for patients with solid tumors but has been limited by the highly fibroinflammatory and immunosuppressive tumor microenvironment (TME). Transforming growth factor-β (TGFβ) participates in the inordinately suppressive TME in PDA. Here, we test the impact of selectiveTgfbr2deletion using CRISPR/Cas9 or genetic approaches in mesothelin (Msln)-specific T cell receptor (TCR) engineered T cells during ACT of PDA. Abrogating TGFβ signaling augmented TCR-engineered T cell accumulation in autochthonous and orthotopic PDA models and promoted terminal effector T cells, although this largely required inclusion of a vaccine at the time of T cell transfer. While loss ofTgfbr2impaired CD103 upregulation, it only modestly impaired donor T cell central, tissue resident, or Tcf1⁺Slamf6⁺stem-like memory T cell formation. These attributes ultimately result in heightened functional capacity and delayed tumor growth. Unexpectedly, however, most tumor-infiltrating engineered T cells failed to differentiate into PD-1⁺Lag3⁺exhausted T cells (T_EX) regardless of TGFβR2 expression and despite abundant Msln protein expression by PDA cells. Forcing Msln epitope processing inKPCtumor cells promoted donor T cell accumulation, acquisition of PD-1 and Lag3, increased IFNγ production by TCR-engineered T cells refractory to TGFβ and bypassed the vaccine requirement for therapeutic efficacy. Thus, promoting increased antigen processing/presentation by tumor cells while abrogatingTgfbr2in engineered T cells can sustain donor T cell function in the suppressive TME and enhance the therapeutic efficacy of ACT. Our study supports pursuit of strategies that modulate tumor intrinsic antigen processing while relieving T cell suppression to safely promote the antitumor activity of TCR-engineered T cells.