Abstract 2217: Pegylated arginine deiminase sensitizes ASS1-negative and KRAS mutant non-small cell lung cancer to PD-1 blockade immunotherapy
Authors: Pavlyk Iuliia , Foster Julie , Dexter Katie , Sobasowski Jane , Bomalarski John , Berlato Chiara , Balkwill Frances , Szlosarek Peter W 
Published: September 2020

Abstract
Cancer cells display dysregulated amino acid synthesis underlying a key metabolic hallmark of tumorigenesis that may be exploited therapeutically via specific amino acid deprivation. In particular, we have identified that arginine deprivation therapy with pegylated arginine deiminase (ADI-PEG20), which hydrolyses exogenous arginine to citrulline and ammonia, is clinically active in argininosuccinate synthetase 1 (ASS1)-deficient non-small cell lung cancer (NSCLC) and enhances the activity of the antifolate pemetrexed. Moreover, prior studies have shown that ADI-PEG20 also induces tumoral PD-L1 expression and T cell infiltration and is additive with anti-PD-1/PD-L1 antibodies in melanoma and colorectal murine models. Here, we reverse translated ADI-PEG20 studies from patients with ASS1-deficient NSCLC, co-associating with KRAS mutations, to analyze the impact of arginine deprivation with PD1-based checkpoint immunotherapy using murine NSCLC cell line models. First, we showed the CMT64 cell line, which harbors the KRAS-G12V mutation and is resistant to anti-programmed death antibodies, was ASS1-negative and highly sensitive to ADI-PEG20 in vitro. Next, CMT64 tumor cells were implanted subcutaneously into the right flank of syngeneic immunocompetent C57BL/6 mice. Once tumors reached 80mm3 animals were randomly assigned to four groups and treated with vehicle control (PBS), ADI-PEG20 (12 mg/kg), anti-PD-1 antibodies (10 mg/kg) or the combination of ADI-PEG20 and PD-1 blockade. CMT64 tumors were refractory to PD-1 blockade while ADI-PEG20 monotherapy induced a modest tumor response, with a two-fold decrease in tumor growth compared to control (p=0.0058). In contrast, the combination of ADI-PEG20 and PD-1 blockade elicited robust anti-tumor activity with a five-fold reduction in CMT64 tumor volume compared to control (p=0.0003). Modulation of the tumor microenvironment was observed by fluorescence activated cell sorting. In particular, we noted that tumor-associated macrophages showed higher expression of MHCII upon ADI-PEG20 and anti-PD-1 therapy (p&lt;0.05). Lastly, mice treated with ADI-PEG20 and anti-PD-1 therapy, survived longer than either ADI-PEG20 or anti-PD-1 therapy alone or PBS. In summary, arginine deprivation with ADI-PEG20 and PD-1 blockade is synthetically lethal in aggressive murine NSCLC and warrants further clinical investigation. A new clinical trial evaluating ADI-PEG20 with atezolizumab in combination with pemetrexed and platinum is opening in 2020 in patients with aggressive ASS1-deficient and PD-1/PD-L1 refractory lung adenocarcinoma (ClinicalTrials.gov Identifier: NCT03498222).
Citation Format: Iuliia Pavlyk, Julie Foster, Katie Dexter, Jane Sobasowski, John Bomalarski, Chiara Berlato, Frances Balkwill, Peter W Szlosarek. Pegylated arginine deiminase sensitizes ASS1-negative and KRAS mutant non-small cell lung cancer to PD-1 blockade immunotherapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2217.