Critical role of Jumonji domain of JMJD1C in MLL-rearranged leukemia

Abstract

JMJD1C, a member of the lysine demethylase 3 family, is aberrantly expressed in mixed lineage leukemia (MLL) gene-rearranged (MLLr) leukemias. We have shown previously that JMJD1C is required for self-renewal of acute myeloid leukemia (AML) leukemia stem cells (LSCs) but not normal hematopoietic stem cells. However, the domains within JMJD1C that promote LSC self-renewal are unknown. Here, we used clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein-9 nuclease (Cas9) negative-selection screening and identified a requirement for the catalytic Jumonji (JmjC) domain and zinc finger domain for leukemia cell survival in vitro and in vivo. In addition, we found that histone H3 lysine 36 methylation (H3K36me) is a marker for JMJD1C activity at gene loci. Moreover, we performed single cell transcriptome analysis of mouse leukemia cells harboring a single guide RNA (sgRNA) against the JmjC domain and identified increased activation of RAS/MAPK and the JAK-STAT pathway in cells harboring the JmjC sgRNA. We discovered that upregulation of interleukin 3 (IL-3) receptor genes mediates increased activation of IL-3 signaling upon JMJD1C loss or mutation. Along these lines, we observed resistance to JMJD1C loss in MLLr AML bearing activating RAS mutations, suggesting that RAS pathway activation confers resistance to JMJD1C loss. Overall, we discovered the functional importance of the JMJD1C JmjC domain in AML leukemogenesis and a novel interplay between JMJD1C and the IL-3 signaling pathway as a potential resistance mechanism to targeting JMJD1C catalytic activity.

Transcription Analysis of a Histones Modifiers Panel Coupled with Tumor Suppressor Genes Displayed Frequent Changes in Acute Myeloid Leukemia-Related Genes

“…Moreover, we found that the expression level of KDM3C was higher in M3 patients in comparison with other subtypes of AML. Previous reports have suggested that via regulation of IL-3/RAS/JAK pathway and also through cooperation with the HOXA9/MEIS1 axis, KDM3C could exert pro-oncogenic function in AML [47]. On the other hand, in contrast to Oncomine databases, which assigned an elevated expression of KDM2B in acute leukemias, we found a normal expression for this gene in the AML patients [19].…”

Small molecular modulators of JMJD1C preferentially inhibit growth of leukemia cells

“…KG‐1 harbors NRAS mutation, P15INK4B methylation, P16INK4A methylation, P53 mutation, RB1 rearrangement and modest JMJD1C expression. One study showed that Ras mutations confer resistance to JMJD1C depletion . However, THP‐1 that harbors MLL‐AF9 fusion, NRAS mutation, P15INK4B deletion, P16INK4A deletion, P53 mutation, RB1 rearrangement and robust JMJD1C expression is also sensitive to JMJD1C depletion and JDI‐4/JDI‐12/JDI‐16 treatment.…”

“…One study showed that Ras mutations confer resistance to JMJD1C depletion. 32 However, THP-1 that harbors MLL-AF9 fusion, NRAS mutation, P15INK4B deletion, P16INK4A deletion, P53 mutation, RB1 rearrangement and robust JMJD1C expression is also sensitive to JMJD1C depletion and JDI-4/JDI-12/JDI-16 treatment. It seems possible that both the Ras mutation and MLL rearrangement confers resistance to JMJD1C depletion.…”

Automated CUT&Tag profiling of chromatin heterogeneity in mixed-lineage leukemia

“…Only ~1% of fusion protein binding sites are shared across leukemia samples, and these shared sites represent 6% of total sequence space (506kb/8377kb) bound by KMT2Ar in any cell type (Supplementary Table 2). The group of 15 genes that is targeted by the fusion protein in all KMT2Ar leukemia samples is highly enriched for master regulators of hematopoiesis as well as genes that are required for KMT2Ar leukemia [24][25][26][27][28] Table 2).…”