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Acute myeloid leukemia

SHP1 regulates a STAT6–ITGB3 axis in FLT3ITD-positive AML cells

Leukemia volume 34, pages 1444–1449 (2020)Cite this article

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We proceeded to identify underlying mechanisms for this phenotype. SHP1-depleted cells homed normally to the bone marrow (Supplementary Fig. S2A). Interestingly, spontaneous adhesion of SHP1-depleted cells to the culture dishes was observed upon generation of SHP1-deficient cell pools (Supplementary Fig. S2B). We considered that this phenotype was related to the effects of SHP1 on leukemogenesis and therefore analyzed it systematically. A quantitative analysis of the proteome of FLT3ITD-expressing 32D cells harboring a nontargeting or an SHP1-targeting shRNA was performed (data deposited to ProteomeXchange via the PRIDE database, accession number PXD011586). Enrichment analysis of differentially regulated proteins revealed upregulation of numerous cell adhesion and membrane proteins including the integrins alpha V (ITGAV), alpha M (ITGAM), beta 2 (ITGB2), and beta 3 (ITGB3) (Supplementary Fig. S2C and Fig. 1e). Moreover, analyzes by flow cytometry revealed that ITGAM, ITGA5, ITGB2, and ITGB3 were significantly higher expressed at the cell surface of SHP1-depleted cells than on control cells (Supplementary Fig. S2D). ITGB3 attracted our particular attention because it had been implicated earlier in AML biology [4]. The upregulation of ITGB3 was robustly seen with three different shRNAs depleting SHP1 (Supplementary Fig. S3A, B). We assessed the role of SHP1 activity for ITGB3 regulation by restoring in cells with SHP1-knockdown expression of either active SHP1 or the catalytically inactive SHP1 D419A mutant. The consequence for ITGB3 expression was subsequently assessed. As shown in Fig.1f, knockdown of SHP1 increased ITGB3 levels. The reexpression of active SHP1 clearly reduced ITGB3 expression so that its level was no longer significantly different from that in control cells. Recovery was, however, not complete possibly from technical reasons. In contrast, the reexpression of the SHP1 D419A mutant did not reduce ITGB3 levels, but even further enhanced it presumably by a dominant negative effect towards residual endogenous active SHP1.

Our findings indicate a previously unrecognized role of the PTP SHP1 for FLT3ITD-positive leukemia. We show that SHP1 promotes development of a myeoloproliferative disease in two different mouse models. Mechanistically, we have identified STAT6 as a direct substrate of SHP1 in FLT3ITD-positive myeloid cells. The activation of STAT6 by SHP1 depletion in our cell model leads to upregulation of ITGB3, which contributes to disease attenuation. Consequently, STAT6 or ITGB3 depletion accelerated disease development. Inversely, activation of STAT6 by IL4 treatment, or overexpression of ITGB3 both inhibited FLT3ITD-mediated cell transformation.

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Fig. 1: SHP1 depletion compromises FLT3ITD-driven myeoloproliferative disease and induces upregulation of integrin β3.
Fig. 2: FLT3ITD-dependent myeloproliferation is regulated by an SHP1–STAT6–ITGB3 axis.

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Acknowledgements

We thank the Core Facility Flow Cytometry of the FLI—Leibniz Institute on Aging, Jena, for cell sorting, and Drs A. Ullrich and J. Drube for reagents. We are grateful for support by the Deutsche Forschungsgemeinschaft (BO 1043/11) and the German José Carreras Leukämie-Stiftung (grant DJCLS R 13/06) to FDB. FHH was supported by the Thuringian state program ProExzellenz (RegenerAging—FSU-I-03/14) of the Thuringian Ministry for Research (TMWWDG). AKJ was supported by DAAD. AKJ and MM were supported by the Max Planck Society for the Advancement of Science.

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  1. These authors contributed equally: Daniela Reich, Ashok Kumar Jayavelu

Authors and Affiliations

  1. Institute of Molecular Cell Biology, CMB, Jena University Hospital, Jena, Germany

    Daniela Reich, Anne Kresinsky, Jörg P. Müller, Reinhard Bauer, Julia Kallenbach & Frank-D. Böhmer

  2. Leibniz Institute on Aging, Fritz Lipmann Institute, Jena, Germany

    Anne Kresinsky, Tina M. Schnoeder & Florian H. Heidel

  3. Innere Medizin II, Hämatologie und Onkologie, Universitätsklinikum Jena, Jena, Germany

    Tina M. Schnoeder & Florian H. Heidel

  4. Department of Molecular Medicine, Max Planck Institute of Biochemistry, Martinsried, Germany

    Reinhard Fässler

  5. Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany

    Matthias Mann & Ashok Kumar Jayavelu

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Contributions

AKJ, DR, and F-DB developed the concept and wrote the manuscript. AKJ, DR, AK, JPM, RB, JK, TMS, FHH, and F-DB performed experiments. MM and RF provided key technology and supervised experiments.

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Correspondence to Matthias Mann, Frank-D. Böhmer or Ashok Kumar Jayavelu.

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Reich, D., Kresinsky, A., Müller, J.P. et al. SHP1 regulates a STAT6–ITGB3 axis in FLT3ITD-positive AML cells. Leukemia 34, 1444–1449 (2020). https://doi.org/10.1038/s41375-019-0676-5

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