Abstract 3421: Identifying mechanisms of immune evasion in microsatellite instable endometrial cancers
Authors: Melendez Brenda , Hinchcliff Emily , Gokul Nisha , Whitley Elizabeth , Broaddus Russell R. , Schmandt Rosemarie E. , Lu Karen H. , Yates Melinda S. 
Published: September 2020

Inherited or sporadic defects in mismatch repair genes (MLH1, MSH2, MSH6, and PSM2) can lead to microsatellite instable (MSI) tumors, most commonly colon, endometrial or gastric tumors. High mutation rates in MSI tumors have been associated with greater immunogenicity, but these tumors can evade immune response. Recently developed mouse models of MSI endometrial cancer (EC) were used to evaluate immune surveillance mechanisms.

Uterine-targeted MSH2 knockout (PR-Cre+MSH2flox/flox) mice were characterized and determined that 22% of mice develop spontaneous EC by 12-16 months of age. Microsatellite instability was evaluated by PCR. Immune infiltration was quantified by immunohistochemistry (IHC) using FoxP3 (Tregs), CD11b (myeloid cells) and CD8 (tumor infiltrating lymphocytes, TILs) antibodies. Tumors were divided into TILHigh and TILLow (&lt;5 TIL/mm2). Comprehensive transcriptome analysis of tumor and normal controls was performed using the Affymetrix Clariom D assay. A subset of candidate gene changes were validated by RT-PCR and at the protein level by IHC. Cell lines were generated from spontaneous tumors and used for orthotopic syngeneic tumor studies of advanced EC. Immune infiltration in orthotopic tumors was characterized using flow cytometry.

All endometrial tumors from PR-Cre+MSH2flox/flox mice were MSI and included endometrioid, serous, and mixed histologies. Tumors showed varying degrees of immune infiltrate, irrespective of histology. Of 7 tumors, 4 (57%) were TILHigh and 3 (43%) were TILLow. Transcriptome analysis showed TILHigh tumors upregulated innate immunity related pathways including, pattern recognition-associated factors, Type I IFN signaling molecules and CXCL5. Activation of adaptive immunity was observed by an increase in T cell chemoattractants, CXCL9 and CCL21. TILHigh tumors displayed at least a 2-fold increase in MHC class I molecules and CD86 needed for T cell activation. IL1A, CSF1, and TGFB2 (cytokines involved in immunosuppressive cell recruitment and induction) were also highly expressed. IHC confirmed co-localization of TILs with myeloid cells and Tregs in TILHigh tumors. TILLow tumors had decreased expression of several members in the IFN family responsible for initiating innate responses. Primary cell lines from TILHigh and TILLow tumors used in an orthotopic syngeneic mouse model retained the immune profile of the original tumor.

Understanding mechanisms of immune evasion is critical for improving immunotherapy. This model of MSI EC reflects the spectrum of immunogenicity observed in clinical studies. TILHigh tumors express a wide range of factors involved in innate and adaptive anti-tumor responses, while TILLow tumors lack key mediators needed for immune activation. These new models for MSI EC will be essential to test the efficacy of immunotherapies and study mechanisms of resistance.

Citation Format: Brenda Melendez, Emily Hinchcliff, Nisha Gokul, Elizabeth Whitley, Russell R. Broaddus, Rosemarie E. Schmandt, Karen H. Lu, Melinda S. Yates. Identifying mechanisms of immune evasion in microsatellite instable endometrial cancers [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 3421.