Targeting FLT3 mutations in AML: review of current knowledge and evidence

Daver, Naval; Schlenk, Richard F.; Russell, Nigel H.; Levis, Mark J.

doi:10.1038/s41375-018-0357-9

Cited by 747 publications

(750 citation statements)

References 91 publications

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“…In particular, AML genomes often harbour hot spot indels whose detection is of clinical importance. These are internal tandem duplications (ITDs) found in FLT3 and KIT [19,20] as well as a 4bp insertion in NPM1. The km algorithm was recently published [15] which performs targeted variant detection.…”

Section: Resultsmentioning

confidence: 99%

Finding a suitable library size to call variants in RNA-seq

Quaglieri

Flensburg

Speed

et al. 2019

Preprint

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Background RNA-Seq allows the study of both gene expression changes and transcribed mutations, providing a highly effective way to gain insight into cancer biology. When planning the sequencing of a large cohort of samples, library size is a fundamental factor affecting both the overall cost and the quality of the results. While several studies analyse the effect that library size has on differential expression analyses, sensitivity analysis for variant detection has received far less attention. Results We simulated shallower sequencing depths by downsampling 45 AML samples that are part of the Leucegene project, which were originally sequenced at high depth. We compared the sensitivity of six methods of recovering validated mutations on the same samples. The methods compared are a combination of three popular callers (MuTect, VarScan, and VarDict) and two filtering strategies. We observed an incremental loss in sensitivity when simulating libraries of 80M, 50M, 40M, 30M and 20M fragments, with the largest loss detected with less than 30M fragments (below 90%). The sensitivity in recovering indels varied markedly between callers, with VarDict showing the highest sensitivity (60%). Single nucleotide variant sensitivity is relatively consistent across methods, apart from MuTect, whose default filters need adjustment when using RNA-Seq. We also analysed 136 RNA-Seq samples from the TCGA-LAML cohort, assessing the change in sensitivity between the initial libraries (average 59M fragments) and after downsampling to 40M fragments. When considering single nucleotide variants in recurrently mutated myeloid genes we found a comparable performance, with a 3% average loss in sensitivity using 40M fragments. Conclusions Between 30M and 40M fragments are needed to recover 90%-95% of the initial variants on recurrently mutated myeloid genes. To extend this result to another cancer type, an exploration of the characteristics of its mutations and gene expression patterns is suggested.

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Section: Resultsmentioning

confidence: 99%

Finding a suitable library size to call variants in RNA-seq

Quaglieri

Flensburg

Speed

et al. 2019

Preprint

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“…These effects are partly mediated by arresting FLT3‐ITD in an under‐glycosylated state and thereby attenuating FLT3‐ITD‐driven AKT and ERK signaling . Because FLT3 plays a very important role in the pathogenesis of AML, various FLT3 inhibitors have been developed . However, their duration of clinical response is short because of the rapid development of resistance …”

Section: Discussionmentioning

confidence: 99%

“…71 Because FLT3 plays a very important role in the pathogenesis of AML, various FLT3 inhibitors have been developed. 72 However, their duration of clinical response is short because of the rapid development of resistance. 73 We found that depletion of fucosylation by Fut8KO in combination with PKC412 efficiently decreases the factor-independent growth of FLT3-WT cells.…”

Section: Discussionmentioning

confidence: 99%

Deficiency of core fucosylation activates cellular signaling dependent on FLT3 expression in a Ba/F3 cell system

et al. 2020

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Fms-like tyrosine kinase 3 (FLT3) is a glycoprotein, that is a member of the class III receptor tyrosine kinase family. Approximately one-third of acute myeloid leukemia (AML) patients have mutations of this gene, and activation of the FLT3 downstream pathway plays an important role in both normal and malignant hematopoiesis.However, the role of N-glycosylation for FLT3 activation remains unclear. In this study, we showed that the N-glycan structures on wild type (WT), internal tandem duplication (ITD), and tyrosine kinase domain (TKD) mutants of FLT3 were different. Interestingly, expression of either WT or mutant FLT3 in Ba/F3 cells, an interleukin-3 (IL-3)-dependent hematopoietic progenitor cell, greatly induced core fucosylation. To elucidate the function of core fucosylation in FLT3-mediated signaling, we used a CRISPR/Cas9 system to establish α1,6-fucosyltransferase (Fut8) knockout (KO) cells. Surprisingly, the Fut8KO resulted in cell proliferation in an IL-3-independent manner in FLT3-WT cells, which was not observed in the parental cells, and suggested that this proliferation is dependent on FLT3 expression. Fut8KO greatly increased cellular tyrosine phosphorylation levels, together with an activation of STAT5, AKT, and ERK signaling, which could be completely neutralized by restoration with Fut8 in the KO cells. Consistently, a tyrosine kinase inhibitor efficiently inhibited cell proliferation induced by Fut8KO or specific fucosylation inhibitor. Additionally, immunostaining with FLT3 showed that the proteins were mainly expressed on the cell surface in the KO cells, which is similar to FLT3-WT cells, but different from the ITD mutant. Finally, we found that Fut8KO could induce dimer-formation in FLT3 without ligand-stimulation. Taken together, the present study clearly defines the regulatory function of core fucosylation in FLT3, which could provide a valuable direction for development of drugs could be effective in the treatment of AML. K E Y W O R D Sacute myeloid leukemia, cellular signaling, core fucosylation, FLT3, Fut8 3240 |

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“…Both types of mutations result in constitutive activation of FLT3 and promote leukemic cell proliferation and survival. The poor outcome associated with FLT3-ITD mutations (3) has generated great interest in the development of specific tyrosine kinase inhibitors (TKI). Recently, an improvement in overall survival was shown for the first time through a randomized phase-3 controlled trial that combined first-generation multi-targeted type II FLT3-TKI midostaurin (PKC412) with standard chemotherapy in both newly diagnosed FLT3-ITD and FLT3-TKD AML, and which led to the approval of midostaurin in this indication in 2017 and its common use since then (4).…”

Section: Introductionmentioning

confidence: 99%

“…g . FLT3-TKD), or micro-environmental factors (3). Although significant efforts are being made to develop more efficient tyrosine kinase inhibitors ( e .…”

Section: Introductionmentioning

confidence: 99%

STAT5-dependent regulation of CDC25A by miR-16 controls proliferation and differentiation in FLT3-ITD acute myeloid leukemia

Sueur

Boutet

Gotanègre

et al. 2019

Preprint

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We recently identified the CDC25A phosphatase as a key actor in proliferation and differentiation in acute myeloid leukemia which expresses the FLT3-ITD mutation. In this paper we demonstrate that CDC25A level is controlled by a complex STAT5/miR-16 transcription and translation pathway working downstream of this receptor. First, we established by CHIP analysis that STAT5 is directly involved in FLT3-ITD-dependent CDC25A gene transcription. In addition, we determined that miR-16 expression is repressed by FLT3-ITD activity, and that STAT5 participates in this repression. In accordance with these results, miR-16 expression was significantly reduced in a panel of AML primary samples carrying the FLT3-ITD mutation when compared with FLT3wt cells. The expression of a miR-16 mimic reduced CDC25A protein and mRNA levels, and RNA interference-mediated down modulation of miR-16 restored CDC25A expression in response to FLT3-ITD inhibition. Finally, decreasing miR-16 expression partially restored the proliferation of cells treated with the FLT3 inhibitor AC220, while the expression of miR-16 mimic stopped this proliferation and induced monocytic differentiation of AML cells. In summary, we identified a FLT3-ITD/STAT5/miR-16/CDC25A axis essential for AML cell proliferation and differentiation.

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Targeting FLT3 mutations in AML: review of current knowledge and evidence

Cited by 747 publications

References 91 publications

Finding a suitable library size to call variants in RNA-seq

Finding a suitable library size to call variants in RNA-seq

Deficiency of core fucosylation activates cellular signaling dependent on FLT3 expression in a Ba/F3 cell system

STAT5-dependent regulation of CDC25A by miR-16 controls proliferation and differentiation in FLT3-ITD acute myeloid leukemia

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