Abstract 2666: HMGA1: An epigenetic switch required for MPN progression by inducing GATA-2 and cell cycle progression through enhancer rewiring

Abstract

Abstract Myeloproliferative neoplasms (MPN) are clonal hematopoietic stem cell (HSC) disorders characterized by hyperactive JAK/STAT signaling and increased risk of transformation to myelofibrosis (MF) and acute myeloid leukemia (AML). However, mechanisms driving progression remain elusive and outcomes are poor after transformation. The High Mobility Group A1 (HMGA1) gene encodes chromatin regulators which are enriched in normal stem cells and aberrantly overexpressed in diverse, refractory tumors. Hmga1 also drives clonal expansion and leukemogenesis in transgenic mice when overexpressed in lymphoid cells. To investigate HMGA1 in MPN, we compared gene expression in CD34+ stem and progenitors from MPN patients and healthy controls. HMGA1 is overexpressed in MPN, with highest levels after transformation to MF or AML. To assess HMGA1 function in MPN progression, we silenced HMGA1 gene expression in two cell lines derived from patients with JAK2V617F mutant MPN after transformation to leukemia. Strikingly, silencing HMGA1 disrupts cell cycle progression, proliferation, and clonogenicity in vitro while preventing leukemia engraftment in immunodeficient mice. To assess HMGA1 in more indolent disease, we crossed a JAK2V617F mouse model of chronic MPN to an Hmga1 deficient background. Loss of one Hmga1 allele was sufficient to prevent progression to MF. Further, Hmga1 heterozygosity mitigates thrombocytosis and splenomegaly, while preventing expansion in long-term HSC, granulocyte-macrophage progenitors, and megakaryocyte-erythroid progenitors. Hmga1 heterozygosity also prolongs survival in a JAK2V617F murine model of fulminant leukemia and early mortality. To define mechanisms underlying HMGA1 in MPN progression, we performed RNA sequencing in human MPN- AML cell lines and identified HMGA1-dependent transcriptional networks involved in cell fate decisions and cell cycle progression, including the master regulator gene, GATA-2. Mechanistically, HMGA1 occupies a developmental GATA-2 enhancer (+9.5) and recruits active histone marks (H3K4me1, H3K4me3) to increase GATA-2 expression. Silencing GATA-2 recapitulates the anti-leukemia phenotypes observed with HMGA1 silencing while GATA-2 re-expression partially rescues pro-leukemogenic phenotypes that occur in MPN-AML cells after HMGA1 depletion. In matched, primary peripheral blood monocytes from patients with MF, HMGA1 and GATA-2 are co-expressed and both become markedly up-regulated after transformation to AML. Epigenetic drugs predicted to target HMGA1 transcriptional networks synergize with JAK inhibitors to disrupt proliferation in MPN-AML cells. Together, our studies reveal a new paradigm whereby HMGA1 up-regulates GATA-2 to drive leukemic transformation in MPN and illuminate HMGA1 networks as novel therapeutic targets required for MPN progression. Citation Format: Liping Li, Jung-Hyun Kim, Wenyan Lu, Donna Marie Williams, Lingling Xian, Joseph Kim, Ophelia Rogers, Raajit K. Rampal, Richard P. Koche, Leslie Cope, Karen Reddy, Daniel R. Matson, Joe Zhao, Jerry L. Spivak, Alison R. Moliterno, Linda Resar. HMGA1: An epigenetic switch required for MPN progression by inducing GATA-2 and cell cycle progression through enhancer rewiring [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2666.