Abstract 1031: PRMT5 inhibition alters mitochondrial dynamics in mantle cell lymphoma, creating vulnerability to BH3 mimetic compounds
Authors: Hinterschied Claire , Brown Fiona , Ravikrishnan Janani , Helmig-Mason JoBeth , Vaddi Kris , Scherle Peggy , Woyach Jennifer , Chen-Kiang Selina , Elemento Oliver , Paik Jihye , Baiocchi Robert 
Published: June 2022

Abstract
Mantle cell lymphoma (MCL) is an aggressive and incurable blood cancer comprising 5% of all non-Hodgkin lymphomas diagnosed annually. The median age of diagnosis is 68yo, and while many patients initially respond to frontline treatment, relapse is common. There is an unmet need to develop novel therapeutic strategies for the treatment of MCL. Our group has identified protein arginine methyltransferase 5 (PRMT5) as a key driver of MCL pathogenesis. PRMT5 symmetrically dimethylates arginine residues on a number of proteins (P53, E2F1, P65) and histones (H4R3, H3R8, H2AR3), which support tumorigenesis. Selectively inhibiting PRMT5 has shown significant anti-tumor activity in preclinical MCL models, and a Phase 1 clinical trial with PRT543 (Prelude), a novel PRMT5 inhibitor, is underway. While exploring pathways that converge with PRMT5 activity as potential avenues for combination treatment, we identified the intrinsic apoptotic pathway as an attractive target in MCL. BCL2 family proteins either promote or inhibit intrinsic apoptosis at the outer mitochondrial membrane through a dynamic set of binding interactions. Prior work has shown PRMT5 inhibition to drive the expression of multiple pro-death BCL2 family gene products (BAX, BAK, and BBC3/PUMA) in MCL. We hypothesized that combining PRMT5 inhibition with BH3 mimetics, compounds that target pro-survival BCL2 proteins, would induce synergistic cell death in MCL. Selective PRMT5 inhibition with PRT382 inhibits the viability of MCL cell lines with an IC50 below 1uM in eight of nine lines (IC50 44.8nM - 1905.5nM). BH3 mimetics navitoclax (BCL2, BCLXL, and BCLw inhibitor), A852 (BCLXL inhibitor), and AMG176 (MCL1 inhibitor) were found to have IC50s below 1uM in five, two, and four of the nine cell lines respectively. We chose six cell lines to test combination treatment ranging in sensitivity to BH3 mimetics and expression of BCL2 family proteins (CCMCL1, Z-138, UPN1, Granta 519, Mino, and Maver1). Synergistic decreases in viability were tested via MTS assay and analyzed with the Loewe model of synergy. Mino, which showed sensitivity to all three mimetics, exhibited a synergistic reduction in viability with combination PRT382 treatment. Granta 519 and Z-138 exhibited a similar effect with the combination of PRT382 and navitoclax or A852. These observations were confirmed through BH3 profiling, supporting MCL cell line dependence on BCL2, BCLXL, BCLw and MCL1, and increased sensitivity to BCL2 family protein targeting with PRMT5 inhibition. Two patient derived xenograft models were tested ex vivo after treatment with 10mg/kg of the PRMT5 inhibitor PRT382 or vehicle. iBH3 flow-based analysis revealed increased sensitivity of ex vivo PDX cells to pan BCL2, BCLXL, and MCL1 inhibition. These results provide rationale for combining BH3 mimetics and PRMT5 inhibition in clinical trials as a novel treatment strategy for MCL.
Citation Format: Claire Hinterschied, Fiona Brown, Janani Ravikrishnan, JoBeth Helmig-Mason, Kris Vaddi, Peggy Scherle, Jennifer Woyach, Selina Chen-Kiang, Oliver Elemento, Jihye Paik, Robert Baiocchi. PRMT5 inhibition alters mitochondrial dynamics in mantle cell lymphoma, creating vulnerability to BH3 mimetic compounds [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1031.