Wang, JiaoToregrosa-Allen, SandraElzey, Bennett D.Utturkar, SagarLanman, Nadia AtallahBernal-Crespo, VictorBehymer, Matthew M.Knipp, Gregory T.Yun, YeonheeVeronesi, Michael C.Sinn, Anthony L.Pollok, Karen E.Brutkiewicz, Randy R.Nevel, Kathryn S.Matosevic, Sandro2023-01-102023-01-102020Wang, J., Toregrosa-Allen, S., Elzey, B. D., Utturkar, S., Lanman, N. A., Bernal-Crespo, V., Behymer, M. M., Knipp, G. T., Yun, Y., Veronesi, M. C., Sinn, A. L., Pollok, K. E., Brutkiewicz, R. R., Nevel, K. S., & Matosevic, S. (2020). Tumor-responsive, multifunctional CAR-NK cells cooperate with impaired autophagy to infiltrate and target glioblastoma (p. 2020.10.07.330043). bioRxiv. https://doi.org/10.1101/2020.10.07.330043https://hdl.handle.net/1805/30894Tumor antigen heterogeneity, a severely immunosuppressive tumor microenvironment (TME) and lymphopenia resulting in inadequate immune intratumoral trafficking have rendered glioblastoma (GBM) highly resistant to therapy. As a result, GBM immunotherapies have failed to demonstrate sustained clinical improvements in patient overall survival (OS). To overcome these obstacles, here we describe a novel, sophisticated combinatorial platform for GBM: the first multifunctional immunotherapy based on genetically-engineered, human NK cells bearing multiple anti-tumor functions, including local tumor responsiveness, that addresses key drivers of GBM resistance to therapy: antigen escape, poor immune cell homing, and immunometabolic reprogramming of immune responses. We engineered dual-specific CAR-NK cells to bear a third functional moiety that is activated in the GBM TME and addresses immunometabolic suppression of NK cell function: a tumor-specific, locally-released antibody fragment which can inhibit the activity of CD73 independently of CAR signaling and decrease the local concentration of adenosine. The multifunctional human NK cells targeted patient-derived GBM xenografts, demonstrated local tumor site specific activity in the tissue and potently suppressed adenosine production. We also unveil a complex reorganization of the immunological profile of GBM induced by inhibiting autophagy. Pharmacologic impairment of the autophagic process not only sensitized GBM to antigenic targeting by NK cells, but promoted a chemotactic profile favorable to NK infiltration. Taken together, our study demonstrates a promising new NK cell-based combinatorial strategy that can target multiple clinically-recognized mechanisms of GBM progression simultaneously.en-USPublisher PolicyTumor-responsiveCAR-NK cellsglioblastomaArticle