Endoglin (ENG/CD105) is highly expressed in cancer-associated fibroblasts (CAFs) from pancreatic and colorectal cancers and correlates with poor prognosis. Here, we reveal that transforming growth factor–β (TGF-β) signaling drives the transition of normal fibroblasts into an ENG⁺ CAF-like state characterized by extracellular matrix accumulation and impaired NK cell infiltration. Functionally, ENG⁺ fibroblasts promote tumor invasion and contribute to an immunosuppressive tumor microenvironment. To target this stromal barrier, we developed ENG-specific chimeric antigen receptor (CAR)-NK cells using a newly identified single-chain variable fragment. These ENG-CAR-NK cells effectively eliminate ENG⁺ CAFs and enhance cancer killing in 3D spheroid models. In vivo, ENG-CAR-NK cells improve tumor infiltration and anti-tumor activity in models co-engrafted with CAFs and cancer cells. Notably, sequential administration of ENG- and mesothelin-targeted CAR-NK cells synergistically enhances tumor clearance. These findings suggest a stromal-targeting immunotherapy strategy to overcome CAFs-mediated hindrance of infiltration in pancreatic cancer.