Fig. 5: JAK2 targeting is responsible for the effects of ruxolitinib on the fitness advantage of NRAS mutant cells and their increased clonogenic potential.
From: JAK2 inhibition mediates clonal selection of RAS pathway mutations in myeloproliferative neoplasms
A qRT-PCR for Nras and Jak2 expression level in murine 32D cells infected with an Empty, a NrasQ61K or a NrasQ61K-shJak2 encoding vector. Statistical significance was determined using a two-tailed Welch’s t-test. Error bars represent the mean of n = 4 biological replicates ± SEM. B Growth inhibition of 32D murine cells expressing an Empty, a NrasQ61K or a NrasQ61K-shJak2 encoding vector 6 days after GFP sorting. Statistical significance was determined using a two-tailed Welch’s t-test. Error bars represent the mean of n = 6 biological replicates ± SEM. C Model of the in vitro NrasQ61K-shJak2 murine myeloid cells competition assay. Created in BioRender59. D Percentage of GFP+_CRIMSON− 32D cells expressing an Empty, a NrasQ61K or a NrasQ61K_shJak2 encoding vector 3 days after sorting. Statistical significance was determined using two-tailed Welch’s t-test. Error bars represent mean of n = 3 biological replicates ± SEM. E–F Colony formation assay of Jak2WT (E) or Jak2V617F (F) C57BL/6 primary bone marrow c-Kit+ murine cells expressing an Empty, a NrasQ61K or a NrasQ61K-shJak2 encoding vector at least 6 days after GFP sorting. Statistical significance was determined using a two-tailed Welch’s t-test. Error bars represent the mean of n = 4 biological replicates ± SEM. G Genetic variants association with resistance to the JAK2 inhibitors ruxolitinib (n = 497), momelotinib (n = 476) and fedratinib (n = 93) according to the Beat AML v2 human primary patient AML dataset. Data is presented as volcano plots for gene variants' effect size (Glass) on the x-axis versus –log10(P-value) on the y-axis. Significance was set at – log10(P-value)>1.3, based on the two-tailed Student t-test with Welch’s correction, with variants having a negative effect size associated with drug sensitivity (red), while those having a positive effect size associated with drug resistance (blue). P-values are reported in the figure. Source data provided as a Source Data file.
