Role of Gab2 in mouse myeloid abnormal proliferation induced by SHP-2D61G/+ mutation

AIM: To investigate whether Gab2, the key adapter protein in the SHP-2 signaling pathway, is involved in mouse myeloid abnormal proliferation induced by SHP-2D61G/+ mutation.METHODS: Four kinds of mouse model genotyped as SHP-2+/+, Gab2-/-, SHP-2D61G/+ and SHP-2D61G/+/Gab2-/- were generated from crossbreeding of Gab2-/- mice and SHP-2D61G/+ mice. The mouse spleen size was analyzed. The number of peripheral blood leukocytes was counted by cell counting and the percentage of Mac-1 or Gr-1 positive myeloid cells in the bone marrow was detected by flow cytometry. The proliferation ability of bone marrow hematopoietic stem/progenitor cells in response to cytokines was assayed by colony formation. The expression of p-ERK and p-Akt and the binding capacity of SHP-2 with Gab2 in the bone marrow-derived mast cells stimulated with IL-3 were detected by Western blotting and immunoprecipitation.RESULTS: The phenotype of myeloproliferative disorder, such as enlarged spleen size, increased leukocyte number and high percentage of myeloid cells, in SHP-2D61G/+ mutant mice was found, and was dramatically improved in SHP-2D61G/+/Gab2-/- double mutation mice. Furthermore, compared with SHP-2D61G/+ mutation mice, significantly decreased colony formation ability of the bone marrow cells with IL-3 stimulation was observed in SHP-2D61G/+/Gab2-/- double mutation mice. A reduced phosphorylation level of ERK/Akt, and SHP-2 without binding of Gab2 were found in SHP-2D61G/+/Gab2-/- bone marrow-derived mast cells with IL-3 stimulation.CONCLUSION: Gab2 knockout significantly reduces mouse myeloid abnormal proliferation induced by SHP-2D61G/+ mutation. The molecular mechanism may be associated with reduced binding of SHP-2D61G/+ under Gab2 knockout, and further weakened the activation of downstream signaling pathways of ERK and Akt.