Paracrine enhancement of tumor cell proliferation provides indirect stroma-mediated chemoresistance via acceleration of tumor recovery between chemotherapy cycles.

The ability of tumors to survive therapy is mediated not only by cell-intrinsic but also cell-extrinsic, microenvironmental mechanisms. Across many cancers, including triple-negative breast cancer (TNBC), a high stroma/tumor ratio correlates with poor survival. In many contexts, this correlation can be explained by the direct reduction of therapy sensitivity by stroma-produced paracrine factors through activating pro-survival signaling and stemness. We sought to explore whether this direct effect contributes to the link between stroma and poor responses to chemotherapies in TNBC. Our in vitro studies with panels of TNBC cell line models and stromal isolates failed to detect a direct modulation of chemoresistance. However, we found that fibroblasts often enhance baseline tumor cell proliferation. Consistent with this in vitro observation, we found evidence of stroma-enhanced TNBC cell proliferation in vivo , in xenograft models and patient samples. Based on these observations, we hypothesized an indirect link between stroma and chemoresistance, where stroma-augmented proliferation potentiates the recovery of residual tumors between chemotherapy cycles. To test this hypothesis, we developed a spatial agent-based model of tumor response to repeated dosing of chemotherapy. The model was quantitatively parameterized from histological analyses and experimental studies. We found that even a slight enhancement of tumor cell proliferation within stroma-proximal niches can strongly enhance the ability of tumors to survive multiple cycles of chemotherapy under biologically and clinically feasible parameters. In summary, our study uncovered a novel, indirect mechanism of chemoresistance. Further, our study highlights the limitations of short-term cytotoxicity assays in understanding chemotherapy responses and supports the integration of experimental and in silico modeling.