Abstract 3696: Tumor-intrinsic PD-L1 alters tumor chemokines, NK cell trafficking and function, and renders distant PD-L1 null tumors responsive to αPD-L1
Authors: Clark Curtis A. , Gupta Harshita B. , Padron Alvaro , Zhang Deyi , Hurez Vincent , Turk Mary Jo , Li Rong , Curiel Tyler 
Published: July 2017

Programmed death ligand (PD-L) 1 is expressed on many tumors and inhibits anti-tumor T cells through programmed death (PD)-1. Tumor PD-L1 predicts αPD-L1 treatment effects, but mechanism(s) for PD-L1- tumor response to αPD-L1 are unclear. Our studies suggest tumor-intrinsic PD-L1 signals and spatially varied PD-L1 expression may contribute to response deviation. We used PD-L1+ B16 melanoma (ctrl) and made PD-L1KO by CRISPR. αPD-L1 slowed ctrl but not PD-L1KO B16 growth in mice as expected, but PD-L1KO also responded to αPD-L1 if ctrl B16 was on the trans flank. αPD-L1 elicited similar CD3+ T cell infiltration into ctrl vs. PD-L1KO tumors, but without detectable B16-specific T cell increase. CD11b+ cell infiltration was similar in ctrl and PD-L1KO. Strikingly, NKp46+ and NK1.1+ natural killer (NK) cells infiltrated PD-L1KO &gt; ctrl. NK cells increased significantly (~2-fold) after αPD-L1 in PD-L1KO vs. ctrl, along with NK effector functions (e.g., IFN-γ). Tumor PD-L1 altered tumor chemokines (e.g., CXCL12, CCL2) that could explain trafficking. CD4+ and CD8+ T cells chemotaxed to PD-L1KO slightly &gt; ctrl (with slightly &gt; PD-1+ T cells) in transwells, but NK cells migrated ~2-fold more to PD-L1KO vs. ctrl (with increased activation (e.g., PD-1, CD69), also seen in vivo). We observed that tumor β2 microglobulin (B2M) expression was suppressed by tumor PD-L1. Tumor PD-L1 alteration of immune infiltrates by altering chemokines is a novel mechanism for PD-L1 TIL control. These data also indicate that tumor PD-L1 blunts NK cell infiltration in B16 and reduces B2M, which could boost NK cell anti-tumor activity. These changes correlate with rescued αPD-L1 response of PD-L1KO B16. As αPD-L1 did not increase B16-specific T cells in PD-L1low, we hypothesize that rescue of αPD-L1 response is in part NK cell-mediated. Preliminary in vivo NK cell depletion + αPD-L1 experiments implicate a role for NK cells in PD-L1KO tumor response, but more work is needed to identify mechanisms. Effects extend beyond B16 as ctrl (PD-L1+) MB49 bladder cancer cells in trans elicited αPD-L1 response of PD-L1KO MB49 tumors. Mouse breast and ovarian cancers, and human ovarian and bladder cells exhibit similar PD-L1-intrinsic effects. Finally, αPD-L1 directly altered tumor cell chemokine production in vitro, suggesting additional, novel αPD-L1 treatment mechanisms requiring more study. Our models are useful to understand αPD-L1 (and likely αPD-1) responses based on tumor PD-L1 expression and will help define strategies to improve responses in PD-L1low tumors, and possibly poorly responsive PD-L1+ tumors.

Citation Format: Curtis A. Clark, Harshita B. Gupta, Alvaro Padron, Deyi Zhang, Vincent Hurez, Mary Jo Turk, Rong Li, Tyler Curiel. Tumor-intrinsic PD-L1 alters tumor chemokines, NK cell trafficking and function, and renders distant PD-L1 null tumors responsive to αPD-L1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3696. doi:10.1158/1538-7445.AM2017-3696