Abstract B062: Low dose dual epigenetic therapy utilizing hypomethylating agents and HDAC inhibitors prolong survival in an orthotopic PDAC model and alter the tumor microenvironment
Authors: Muzyka Stephen , Orlacchio Arturo , Ackermann Amanda , Chinda Yasmine , Shim Yoona , Dolgalev Igor , Gonda Tamas 
Published: September 2024

Background: We have previously studied the effect of hypomethylating agent Decitabine (5- aza-dC; DAC) on pancreatic cancer using the KPC model (KrasG12D/+;Trp53R172H/+;Pdx1- Cre) and observed increased CD4+/CD8+ TILs, greater PD1 expression and slowed tumor growth with increased tumor necrosis. Subsequently we showed a significant survival benefit with DAC+αPD1 compared to either single agent. However, we identified a specific myeloid cell population initially identified by Chi3l3 expression that may be responsible for treatment resistance or escape. Based on prior observations of synergy specifically affecting tumor infiltrating myeloid and lymphoid cells, we focused on identifying a dual epigenetic strategy (a combination of DAC plus histone deacetylase inhibitors (HDACi)) to enhance immunological synergy and test their treatment efficacy with αPD1. Methods: We used a combination primary KPC-derived cell lines, bone marrow (BMDM) and peritoneum derived (PM) myeloid cells and T cells in co-culture systems to assess cytotoxic synergy (Bliss/Lowe consensus) and immune effects (cytokine release, myeloid cell polarization and T cell activation) to evaluate combinations of DAC+HDACi. The KPC orthotopic model relying on US monitoring of tumor size for standardized initiation of therapy was used for survival analysis and profiling in treatment combinations of DAC+HDACi+αPD1. Results: We showed that hypomethylation therapy exerts a primarily paracrine effect on myeloid cells and this polarization results in a decrease in DAC–induced T cell activation. We therefore selected HDAC inhibitors that in combination with DAC alter this paracrine effect. We selected specific HDAC inhibitors based on induction of class of HDAC expression following hypomethylation therapy, highest Bliss synergy scores at low doses and reversal of cytokine release, M2 polarization and T cell inactivation. Three HDAC inhibitors were chosen for in vivo testing, Domatinostat, Pracinostat and Entinostat. Using the treatment paradigm of ultra-low dose DAC+ HDACi (mainly to minimize toxicity) combined with αPD1 in the KPC orthotopic model we show that the triple combination (DAC + Entinostat + αPD1) is well-tolerated and results in a significant survival benefit compared to single and dual agent combinations. In addition, we also demonstrate a reduction in the Chi3l3 expressing myeloid cell population. Conclusions: Our results show that the combination of hypomethylating agent plus HDAC inhibitor has several beneficial effects on the tumor immune response in PDAC. HDAC inhibitors specifically reduce the suppressive myeloid cell population we previously identified after DAC therapy. The combination of dual low dose epigenetic therapy using a hypomethylating agent plus HDAC inhibitor followed by αPD1 results in significant survival benefit in the orthotopic KPC model.

Citation Format: Stephen Muzyka, Arturo Orlacchio, Amanda Ackermann, Yasmine Chinda, Yoona Shim, Igor Dolgalev, Tamas Gonda. Low dose dual epigenetic therapy utilizing hypomethylating agents and HDAC inhibitors prolong survival in an orthotopic PDAC model and alter the tumor microenvironment [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B062.