Abstract 74: Osteoblasts efferocytosis in bone marrow microenvironment induces osteoblasts’ senescence and apoptosis

Abstract

Increased cellular apoptosis is commonly seen in myelodysplastic syndrome (MDS), but how clearance of apoptotic cells, especially neutrophils, impacts disease progression is unknown. Moreover, while dysfunction of bone marrow (BM) stromal cell populations has been described in patients with MDS, the mechanisms inducing stromal defects are not well understood. Our laboratory found that BM macrophages are defective in a murine model of MDS (Vav-Nup98 HoxD13, aka NHD13), with decreased efferocytotic rates and increased inflammatory mediators. The increased apoptotic burden and defective macrophages would be expected to recruit non-professional phagocytes (osteoblasts) in the bone and bone marrow. Based on these data, we hypothesize that OBs (osteoblasts) participate in the clearance of apoptotic cells in the bone marrow microenvironment (BMME) to maintain BM homeostasis. However, the increased osteoblastic efferocytosis burden in MDS contributes to OBs’ dysfunction as a mechanism of MDS-dependent BMME disruption that may lead to MDS progression. We isolated from wild-type (B6) mice bone-associated cells (BAC), which are enriched with OBs. In vitro co-culture with labeled human end-stage neutrophils at 1:1 and 1:10 (OB: PMN) increased OB apoptosis, while senescence was increased only in high-dose PMN co-culture. We found that these changes were cell-autonomous. In vivo injection of neutrophils into WT mice showed around 5-10% efferocytosis rate in OBs. In efferocytotic OBs, apoptosis rates were significantly increased. Together, these data show that OBs participate in efferocytosis and that efferocytosis disrupts OBs, causing apoptosis and senescence. To mimic the MDS microenvironment, we used a genetic model where one of the Isocitrate dehydrogenase 2 (IDH2) mutations found in MDS (R140Q) is targeted to hematopoietic cells via the Vav promoter (IDH2R140Q mice) as well as NHD13 mice, representing early and late MDS BMME respectively. We observed that 6-to-8 months old NHD13 mice experienced more end-stage neutrophils[CL1] in the BMME (4.6% rate of end-stage neutrophils in WT compared to 9.5% rate in NHD13 mice), where OB apoptosis was also increased in vivo. In IDH2R140Q mice at 3 months of age, we confirmed the presence of mutated IDH2 in bone marrow cells by increased intracellular levels of the oncometabolite 2-hydroxyglutarate. At this early time point, IDH2R140Q mice did not have increased rates of apoptosis in CD45+ cells, and we found no increase in OB apoptosis. Analysis of IDH2R140Q mice at later time points is ongoing. Our data identify novel mechanisms to explain the relationship between defective myeloid cells and OB apoptosis in MDS that may contribute to the inflammatory MDS microenvironment, impact disease progression, and serve as future therapeutic approaches for MDS patients. Citation Format: Chunmo Chen, Emily R. Quarato, Yuko Kawano, Noah A. Salama, Hiroki Kawano, Michael W. Becker, Jeevisha Bajaj, Laura Calvi. Osteoblasts efferocytosis in bone marrow microenvironment induces osteoblasts’ senescence and apoptosis [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 74.