Whole Genome Sequencing of Spontaneously Occurring Rat Natural Killer Large Granular Lymphocyte Leukemia Identifies JAK1 Somatic Activating Mutation

Abstract

Large granular lymphocyte (LGL) leukemia arises spontaneously in elderly Fischer (F344) rats. This rodent model has been shown to emulate many aspects of the natural killer (NK) variant of human LGL leukemia. Previous transplantation of leukemic material into young F344 rats resulted in several strains of rat NK (RNK) primary leukemic cells. One strain, RNK-16, was adapted into the RNK-16 cell line and established as an aggressive NK-LGL leukemia model. Whole genome sequencing of the RNK-16 cell line identified 255,838 locations where the RNK16 had an alternate allele that was different from F344, including a mutation in Jak1. Functional studies showed Jak1 Y1034C to be a somatic activating mutation that mediated increased STAT signaling, as assessed by phosphoprotein levels. Sanger sequencing of Jak1 in RNK-1, -3, -7, and -16 found only RNK-16 to harbor the Y1034C Jak1 mutation. In vivo studies revealed that rats engrafted with RNK-16 primary material developed leukemia more rapidly than those engrafted with RNK-1, -3, and -7. Additionally, ex vivo RNK-16 spleen cells from leukemic rats exhibited increased STAT1, STAT3, and STAT5 phosphorylation compared to other RNK strains. Therefore, we report and characterize a novel gain-of-function Jak1 mutation in a spontaneous LGL leukemia model that results in increased downstream STAT signaling.

JAK-STAT Signalling Pathway in Cancer

“…While activating mutations in cytokine receptors are not common, or at least not heavily investigated, activating mutations are commonly found in JAK1 and JAK2, which is best highlighted by the literature in myeloproliferative neoplasms and leukaemia. In this issue, a novel JAK1 Y1034C somatic activating mutation was identified from whole genome sequencing of spontaneously occurring rat natural killer large granular lymphocyte leukaemia [ 4 ]. Many oncogenic mutations are located in the JAK pseudokinase domain, which is thought to play an essential role in keeping the kinase inactive in absence of ligand binding to a cytokine receptor.…”

Sorafenib plus memory like natural killer cell combination therapy in hepatocellular carcinoma

“…The HCC cell lines were cultured according to ATCC guidelines in which rat HCC cell lines (McA-RH7777 and N1-S1) were cultured in DMEM and IMDM (Gibco, Waltham, MA), and HepG2 cell line in EMEM (Corning, Manassas, VA). The rat NK cell line (RNK-16) was developed by Wang et al [27] and generously provided by Thomas L. Olson (University of Virginia, Charlottesville, VA). Both effector cell lines were cultured in RPMI medium 1640 (Life Technologies, Waltham, MA).…”