Phase separation of SHP2E76K promotes malignant transformation of mesenchymal stem cells by activating mitochondrial complexes
Chen Kan, Zhenya Tan, Liwei Liu, Bo Liu, Li Zhan, Jicheng Zhu, Xiaofei Li, Keqiong Lin, Jia Liu, Yakun Liu, Fan Yang, Mandy Wong, Siying Wang, Hong Zheng
Chen Kan, Zhenya Tan, Liwei Liu, Bo Liu, Li Zhan, Jicheng Zhu, Xiaofei Li, Keqiong Lin, Jia Liu, Yakun Liu, Fan Yang, Mandy Wong, Siying Wang, Hong Zheng
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Research Article Oncology Stem cells

Phase separation of SHP2E76K promotes malignant transformation of mesenchymal stem cells by activating mitochondrial complexes

Abstract

Mesenchymal stem cells (MSCs), suffering from diverse gene hits, undergo malignant transformation and aberrant osteochondral differentiation. Src homology region 2–containing protein tyrosine phosphatase 2 (SHP2), a nonreceptor protein tyrosine phosphatase, regulates multicellular differentiation, proliferation, and transformation. However, the role of SHP2 in MSC fate determination remains unclear. Here, we showed that MSCs bearing the activating SHP2E76K mutation underwent malignant transformation into sarcoma stem-like cells. We revealed that the SHP2E76K mutation in mouse MSCs led to hyperactive mitochondrial metabolism by activating mitochondrial complexes I and III. Inhibition of complexes I and III prevented hyperactive mitochondrial metabolism and malignant transformation of SHP2E76K MSCs. Mechanistically, we verified that SHP2 underwent liquid-liquid phase separation (LLPS) in SHP2E76K MSCs. SHP2 LLPS led to its dissociation from complexes I and III, causing their hyperactivation. Blockade of SHP2 LLPS by LLPS-defective mutations or allosteric inhibitors suppressed complex I and III hyperactivation as well as malignant transformation of SHP2E76K MSCs. These findings reveal that complex I and III hyperactivation driven by SHP2 LLPS promotes malignant transformation of SHP2E76K MSCs and suggest that inhibition of SHP2 LLPS could be a potential therapeutic target for the treatment of activated SHP2–associated cancers.

Authors

Chen Kan, Zhenya Tan, Liwei Liu, Bo Liu, Li Zhan, Jicheng Zhu, Xiaofei Li, Keqiong Lin, Jia Liu, Yakun Liu, Fan Yang, Mandy Wong, Siying Wang, Hong Zheng

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Figure 1

MSCs with the SHP2E76K mutation undergo malignant transformation and model leiomyosarcoma formation.

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MSCs with the SHP2E76K mutation undergo malignant transformation and mod...
(A and B) immunofluorescence (IF) images (A) and statistical analysis (B) of Ki67 staining of WT and SHP2E76K MSCs (n = 5 per group). Scale bar, 200 μm. Data are represented as the mean ± SD. **P < 0.01 (2-tailed unpaired t test). (C and D) Microscopy image (C) and statistical analysis (D) of soft agar colony formation capacity of WT and SHP2E76K MSCs (n = 5 per group). Scale bar, 200 μm. Data are represented as the mean ± SD. ****P < 0.01 (2-tailed unpaired t test). (E) Xenograft tumor transplantation of WT and SHP2E76K MSCs in nude mice (n = 5 per group). (F) Statistical analysis of the tumor incidence of WT and SHP2E76K MSCs in nude mice (n = 5). (G) HE staining and IF staining images of the indicated antibodies in SHP2E76K-induced sarcoma from nude mice. Scale bar, 200 μm. (H) Xenograft tumor transplantation of WT and SHP2E76K MSCs in immunocompetent (C57BL/6) mice. (I) Statistical analysis of the tumor incidence of WT and SHP2E76K MSCs in C57BL/6 mice (n = 4 per group). (J) HE staining and IF staining of the indicated antibodies in SHP2E76K-induced sarcoma from C57BL/6 mice. Scale bar, 200 μm. (K) Representative IF images of WT and SHP2E76K human umbilical cord MSCs in the field of 200×. (L and M) Representative tumor images (L) and statistical analysis (M) of tumor incidence in nude mice with subcutaneous inoculation of WT and SHP2E76K human MSCs (n = 5 per group). (N) Representative HE and immunohistochemical images of tumor developed by SHP2E76K human umbilical cord MSCs. Scale bar, 500 μm.