Abstract 3135: Single-cell RNA sequencing identifies macrophage-specific expression signatures associated with phagocytosis of multiple myeloma after treatment with cIAP antagonist

Abstract

Abstract Multiple myeloma (MM) is the second most common hematologic cancer, accounting for roughly 2% of all cancer deaths. Unfortunately, MM remains incurable and nearly all patients experience relapse. Recently, we have identified a novel cIAP1 antagonist, LCL161, with remarkable anti-MM effects. Interestingly, unlike in other tumor types, our previous work found that LCL161 does not directly induce apoptosis of MM cells, but rather drives activation and infiltration of macrophages to the tumor bed and phagocytosis of tumor cells. Furthermore, in vitro experiments demonstrate that the anti-MM activity of the agent relies on the presence of the tumor, with no observed increase in MM phagocytosis when macrophages are treated in isolation. However, LCL161 is a strong and indiscriminate transcriptional activator and thus, the molecular mechanisms underlying this immune response in MM remain poorly understood. In an effort to characterize molecular signatures associated with this response, we collected whole spleens from seven WT mice and eleven mice with VK12598 transplanted myeloma for single cell RNA-seq. Of the eleven tumor-bearing mice, three remained untreated, three were treated with LCL161, three with bortezomib, and three with cyclophosphamide. Additionally, one of the WT mice was treated with LCL161. Using the 10X genomics chromium platform, we measured mRNA levels in over 90,000 individual cells. We identified groups of cells with shared expression profiles, corresponding to known cell types, using graph-based clustering. Consistent with our previous work, we observed a marked reduction in tumor cells and activation of the noncanonical NFkB pathway in mice treated with LCL161. Turning our attention to cells identified as monocytes, with a particular focus on putative macrophages, we sought to identify molecular signatures associated with LCL161 treatment. Indeed, using a binomial model we identified over 500 upregulated genes in LCL161-treated macrophages. However, our previous data suggested that the efficacy of LCL161 was dependent on the presence of tumor cells. Thus, we refined these results by comparing the expression profile of macrophages treated with LCL161 in WT and tumor-bearing mice. These contrasts identified dozens of genes that were significantly upregulated in macrophages from LCL161 treated, tumor-bearing mice. Of note, we identified a small number of genes (including Spp1, Irg1, and Lipg) that were active only in macrophages from mice treated with LCL161 and bearing a transplanted tumor and unexpressed in all other cells. We believe these genes—and the pathways in which they are active—hold promise for the development of novel targeted therapies to better treat MM. Citation Format: Nicholas Banovich, Martin Boateng, Marta Chesi, Leif Bergsagel, Jonathan Keats. Single-cell RNA sequencing identifies macrophage-specific expression signatures associated with phagocytosis of multiple myeloma after treatment with cIAP antagonist [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3135.