Abstract 207: Ex vivo modeling of multiple myeloma: A novel drug sensitivity screening platform

Abstract

Abstract Multiple Myeloma (MM) is a heterogeneous malignancy characterized by abnormal clonal plasma cell infiltration in the bone marrow. It is the second most common hematologic malignancy, after non-Hodgkin’s Lymphoma. This deadly disease globally affects 1 to 5 people in every 100,000 people each year. After initial diagnosis, the 5-year survival rate is 44% because there is no curative therapy, and most patients will eventually experience relapse, and some become refractory. Large inter-patient and intra-patient genetic heterogeneity limit the identification of universal drivers of MM. However, several oncogenic dependencies such as primary events related to driver gene mutations and primary translocations can be targeted for better treatment strategies for R/R multiple myeloma. To establish a robust drug sensitivity screening platform, both MM cell lines and cryopreserved primary MM patient samples were used. MM cell lines screen with H929, MM1R and RPMI-8226 was performed to narrow down optimal concentrations of investigative test agents. During assay development, a panel of FDA approved agents for relapsed MM including panobinostat, bortezomib, melphalan, omacetaxine, and selinexor were tested. The cells were seeded at 50,000 cells per well in triplicates in a 96-well plate on day 0. The test agents were added in 9 concentrations with the top concentration being 25 uM with 2-fold dilutions. After 6- days of incubation, a luminescent cell viability assay was performed to calculate relative viability with drug treatment and IC50 was computed by fitting data using a standard four-parameter logistic model. For primary multiple myeloma samples, bone marrow aspirates were collected from Multiple Myeloma patients procured from different providers. BM mononuclear (BMMNC) cells were separated by Ficoll-Hypaque density sedimentation. Unfractionated BMMNC cells were used in these assays to preserve intra-tumor heterogeneity. Primary cells were seeded in triplicates per dose point on day 0 in Champions’ proprietary media. The test agents were also added on day 0 and were incubated for 6 days. On Day 6, the relative cell viability was measured based on Cell Titer Glo readout. The cryopreserved primary MM patient samples were viable in culture for the entire assay duration validating assay feasibility. A panel of FDA approved, standard of care drugs were tested in numerous clinically well-annotated and diverse MM patient models. Primary model selection was based on disease burden including plasma cell count from Core Biopsy and BM aspirates. Multiparametric flow cytometric immunophenotyping was also performed in these primary samples using monoclonal antibodies against CD56, CD19, CD117, CD27, CD138, and CD38. For the first time, we demonstrate the feasibility of a novel, ex-vivo drug sensitivity screening platform with cryopreserved, primary multiple myeloma cells from diverse, clinically well-annotated patient samples. Citation Format: Vaishnavi Sambandam, Sharvari Inamdar, Haoting Hsu, Brandon Walling, Paolo Schiviani, Abhay Andar, Marianna Zipeto, Michael Ritchie, Karin Abarca Heidemann, Maria Mancini. Ex vivo modeling of multiple myeloma: A novel drug sensitivity screening platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 207.