Abstract
SHP2 mediates the activities of multiple receptor tyrosine kinase signaling and its function in endothelial processes has been explored extensively. However, genetic studies on the role of SHP2 in tumor angiogenesis have not been conducted. Here, we show that SHP2 is activated in tumor endothelia. Shp2 deletion and pharmacological inhibition reduce tumor growth and microvascular density in multiple mouse tumor models. Shp2 deletion also leads to tumor vascular normalization, indicated by increased pericyte coverage and vessel perfusion. SHP2 inefficiency impairs endothelial cell proliferation, migration, and tubulogenesis through downregulating the expression of proangiogenic SRY-Box transcription factor 7 (SOX7), whose re-expression restores endothelial function in SHP2-knockdown cells and tumor growth, angiogenesis, and vascular abnormalization in Shp2-deleted mice. SHP2 stabilizes apoptosis signal-regulating kinase 1 (ASK1), which regulates SOX7 expression mediated by c-Jun. Our studies suggest SHP2 in tumor associated endothelial cells is a promising anti-angiogenic target for cancer therapy.
Similar content being viewed by others
Introduction
Tumor growth, similar to that of normal organs, depends greatly on the outgrowth of blood vessels to form neovasculatures that supply raw materials for metabolism. Neovascular structures originate from existing capillaries via angiogenesis1, which is a robust process that involves the establishment of a dynamic balance of pro- and anti-angiogenic factors suspended in the tissue niche. VEGF-A is a critical proangiogenic factor that renders the development of anti-VEGF therapies a promising avenue of research, particularly in the treatment of cancers2. Inhibitors targeting the VEGF pathway have exhibited significant anti-tumor activities in multiple mouse models; however, anti-VEGF therapies have not yet been successfully clinically applied to most cancers in humans3,4. In the absence of VEGF, other proangiogenic factors, including FGF, and ephrin, regulate angiogenesis in anti-VEGF-treated tumors and it has been found that anti-VEGF therapy induces unexpected vessel normalization and extracellular matrix remodeling, with improved vessel perfusion5,6. Vascular abnormalities in cancers include insufficient pericyte coverage, hyperpermeability, and immunosurveillance escape7,8. The challenges faced by novel anti-angiogenesis therapy in cancers have led to rigorous investigations, particularly in the elucidation of the underlying mechanisms.
The protein tyrosine phosphatase SHP2, encoded by PTPN11, is expressed ubiquitously, including in blood vessel cells. As an important participant in growth factor and cytokine signaling, SHP2 is frequently upregulated or mutated in tumors, and its oncogenic behavior has been attributed to abnormal SHP2 expression9,10,11. Structurally, there are two SH2 domains, a protein tyrosine phosphatase (PTP) domain, and a C-terminal tail containing phosphorylatable tyrosine residues. The SH2 domains interact with the PTP domain to form a self-inhibitory intramolecular interaction, and have recently been used to develop allosteric inhibitors12,13. New SHP2 inhibitors have been demonstrated to possess tremendous anti-tumor activity, particularly in combination with other conventional drugs14,15,16,17,18,19,20. Owing to the ubiquitous expression of druggable SHP2, its function in tumor microenvironments has been studied thoroughly. Our group has demonstrated that SHP2 suppressed CXCL9 production in macrophages, which prevented T cell infiltration and promoted tumor growth in miceCells HUVECs were isolated from umbilical cord vein by lavaging with 0.2% (w/v) collagenase solution at 37 °C for 15 min, the cells were collected and suspended in complete M199 medium. HUVECs between passages 4 and 6 were cultured in a complete medium containing 53% M199 (Corning Incorporated), 37% human endothelial serum-free medium (Thermo Fisher Scientific), and 15 μg/mL of endothelial cell growth supplement (Sigma-Aldrich). Human cerebral microvessel endothelial cells (hCMECs) were purchased from Zhejiang Meisen Cell Technology Co., Ltd. Primary mouse lung endothelial cells (MLECs) were isolated from 6- to 8-week-old mice. Briefly, 1 month after tamoxifen administration, mice were anesthetized with 80 mg/kg ketamine and 12 mg/kg xylazine, and then subjected to perfusion by intracardiac injection of PBS (0.1% BSA) for blood removal. Lung tissue was then dissected and cut into 1–2-mm pieces, and digested with a solution containing type-I collagenase (2 mg/mL, Biosharp), dispase (1 mg/mL, Roche), and DNase (10 μg/mL, Roche) in DPBS for 45 min at 37 °C in a rotatory shaker (at 80 rpm). After digestion, the enzymes were neutralized by DMEM plus 20% FBS. A single-cell suspension was prepared using a 40-μm cell strainer. Finally, the cells were further purified using anti-mouse CD31-conjugated magnetic beads (Invitrogen) and maintained in a complete medium containing DMEM with 10% FBS, endothelial cell growth supplement (15 μg/mL, Sigma-Aldrich), 1% nonessential amino acids (Gibco), and heparin (0.1 mg/mL, Solarbio). Mouse Lewis lung cancer (LLC), B16 melanoma cells, and E0771 purchased from ATCC were cultured in DMEM. Non-small cell lung cancer (NSCLC) (H460, A549, H1299) and hepatocellular carcinoma LM3 cells purchased from ATCC were grown in RPMI-1640 (Thermo Fisher Scientific). The media were supplemented with 10% FBS, 100 IU/mL penicillin, and 100 μg/mL streptomycin, and the cells were incubated at 37 °C under 5% CO2. For the double luciferase reporter assay, the promoter fragment containing multiple predicted c-Jun binding sites (nucleotides -2000 to 100 of human SOX7 gene loci) was cloned into a pGL3-basic vector. In 12-well plates, HEK293 cells were transfected with vector or c-Jun for 24 h. The double luciferase reporter assay was conducted following the manual for the Dual-Luciferase reporter gene assay kit (Promega). The reporter gene activity was detected by GloMax® 20/20 Luminometer (Promega) and normalized to the activity of Renilla luciferase. To construct SHP2WT and inhibitor-resistant mutant SHP2 (SHP2T253M/Q257L) LM3 cell lines, SHP2WT and SHP2T253M/Q257L cDNA were constructed into the PLVX-NEO vector and packaged into lentiviruses. LM3 cells were infected with shSHP2-3′utr lentivirus and screened by puromycin. The SHP2 knockdown LM3 cells were then infected with SHP2WT or SHP2T253M/Q257L lentivirus, and neomycin was further used to screen stable transgenic strains. To establish syngeneic mouse tumor models, suspensions of LLC (4 × 105 cells in 100 μL) or B16 (8 × 105 cells in 100 μL) cells were subcutaneously implanted in the dorsal flank of 8- to 10-week-old male mice. For the orthotopic mouse breast tumor model, E0771 cells (8 × 105) were suspended in 100 μL of Matrigel (50% v/v; Corning) in RPMI1640 medium and injected into the mammary fat pads of 8-week-old female mice. The tumor volumes were measured using a digital caliper every 2 days and calculated according to the following formula: V = 0.52 × L × W2 (V, tumor volume; L, longest diameter of the tumor; W, perpendicular diameter of L)69,70. Sixteen or eighteen days after cell injection, tumors were harvested and fixed with 4% paraformaldehyde (PFA) for further histological analysis. Matrigel (500 μL), supplemented with the recombinant mouse FGF2 and VEGF-A (400 ng/mL each) and heparin (50 units/mL) was injected subcutaneously into the flanks of 8- to 10-week-old mice, and they were sacrificed 7 days after injection. The Matrigel plugs were removed, photographed, and fixed in 4% PFA for further analysis. The thoracic aortas were removed from the mice under anesthesia and cut into 1-mm rings. The aortic rings were then placed between two layers of 100 μL growth factor-reduced Matrigel (Corning) supplemented with heparin (20 U/mL, Solarbio), VEGF-A (100 ng/mL, Novoprotein), and FGF2 (100 ng/mL, Novoprotein), and incubated in culture medium (10% FBS, 100 IU/mL penicillin and 100 μg/mL streptomycin). The culture medium was changed every other day. The sprouting area was recorded under a phase-contrast microscope on day 7 and measured using Image J 1.49 v. For western blotting, the total protein concentrations (20 μg) from MLECs or HUVECs were separated using SDS-PAGE gels and transferred onto nitrocellulose membranes (Pall, Port Washington). The primary antibodies used are shown in Supplemental Table 1. For co-immunoprecipitation, cells were harvested and lysed in a cell lysis buffer used for western blotting and IP (Beyotime), protease inhibitor cocktail (Roche), and PhosSTOP (Roche) on ice. Protein G Dynabeads were pretreated with the primary antibody for 10 min at 25 °C and then washed with PBST three times. The cell lysates were then incubated overnight with antibody-conjugated beads at 4 °C. The immunoprecipitates were then used for western blotting by odyssey (version 3.0.29) or LI-COR image studio (version 5.2). For tissue immunofluorescence staining, samples were fixed in 4% PFA, dehydrated in a 30% sucrose solution for 24 h, and embedded using the Tissue-Tek OCT compound. Frozen blocks were cut into 10-μm-thick sections. For cell immunofluorescence staining, endothelial cells were fixed with 4% PFA and permeabilized with 0.5% Triton for 15 min. The samples were blocked with 5% goat or donkey serum in PBST and incubated overnight at 4 °C with primary antibodies (Supplemental Table 1). After performing multiple washes, the samples were incubated for 1 h at room temperature with the secondary antibodies (Supplemental Table 1). The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (Beyotime). The samples were then mounted with fluoromount-G (Southern Biotech), and immunofluorescence images were acquired using a confocal microscope (FV3000, Olympus). The images were further processed using ImageJ 1.49v or OlyVIA VS200. The total RNA was isolated from HUVECs and MLECs by TRIzol and reverse-transcribed to cDNA using the ReverTraAce qPCR RT kit (Toyobo Inc.). Quantitative RT-PCR was conducted using SYBR green dye by CFX96 Touch Real-Time PCR Detection System (Bio-Rad) and the primers listed in Supplemental Table 2 and Supplementary Table 3. Gene expression was calculated using the equation RQ = 2−△△Ct and normalized to GAPDH or 18S RNA values. To measure tumor hypoxia, the mice were injected with Hypoxyprobe-1 (60 mg/kg, Hypoxyprobe) i.p. 1 h before they were sacrificed. Tumors were then removed and embedded in Tissue-Tek OCT. FITC-conjugated mouse anti-pimonidazole monoclonal antibody was applied following the manufacturer’s protocol. To evaluate vascular permeability, 100 μL of FITC-conjugated dextran (25 mg/mL, 70 kDa, Sigma-Aldrich) was intravenously injected and the mice were perfused with 1% PFA to remove circulating dextran after 30 min. Tumors were then dissected and embedded in the Tissue-Tek OCT compound for further analysis. To measure vessel perfusion, DyLight 488-conjugated Tomato lectin (1 mg/mL, 100 μL, Vector Laboratories) was i.v. injected 30 min before sacrifice, and the tumors were harvested for further analysis. Cells were seeded in the 24-well plates at a density of 4 × 104 cells per well. After overnight incubation, the cells were incubated with EdU (Beyotime) for 5 h at 37 °C and then fixed in 4% PFA. After washing with PBS, the cells were treated with 0.5% Triton X-100 for 15 min at room temperature for permeabilization. The cells were incubated and protected from light using a click additive solution and stained with DAPI. Images were obtained using a confocal microscope (FV3000, Olympus). HUVECs or MLECs suspended in the complete medium were seeded in the upper chamber. Four hours later, the medium in the upper chamber was changed to 0.1% FBS in M199, and the lower chamber was filled with 600 μL of complete medium. After incubation for 24 h at 37 °C, non-migrating cells were removed and cells that migrated through the membrane were fixed in 4% PFA and stained with crystal violet solution (Beyotime), after photographed, the crystal violet-stained cells were incubated with 33% acetic acid, and absorbance was detected at 570 nm (SynergyMx M5, Molecular Devices). Cells were seeded in Matrigel-coated u-slides at a density of 1 × 104 cells per well (Ibidi, Germany) and incubated at 37 °C for 4 h and for a further 15 min with calcein (Yeasen). Images were acquired using a fluorescence microscope (IX70, Olympus). Five different fields for each condition were quantified by counting the number of junctions, total tube length, and total branching length using ImageJ 1.49v. NSCLC and matched adjacent normal tissue specimens were collected from the First Affiliated Hospital, College of Medicine, Zhejiang University (Hangzhou, China), and confirmed by pathological diagnosis. This study was approved by the Research Ethics Committee of the First Affiliated Hospital, College of Medicine, Zhejiang University and abided by the Declaration of Helsinki principles. Written informed consent was obtained from all patients prior to the study. Statistical analysis was performed using Prism software (GraphPad Inc, version 6.0 and 8.0). All quantitative data are presented as the mean ± SEM. Unpaired Student’s t-tests were used for comparisons between two groups. Multiple group comparisons were conducted by one-way or two-way ANOVA followed by Tukey’s post hoc tests or multi-comparisons. p < 0.05 was considered statistically significant. Further information on research design is available in the Nature Research Reporting Summary linked to this article.Mouse tumor models
Matrigel plug assay
Mouse aortic ring assay
Co-immunoprecipitation, immunoblotting, and immunofluorescence staining
Quantitative PCR
Pimonidazole staining, vascular leakage, and perfusion assay
EdU incorporation assay
Transwell cell migration assay
Endothelial cell tube formation
NSCLC tissues
Quantification and statistical analysis
Reporting summary
Data availability
Figure 1d, Supplementary Fig. 7f, g were generated from the NCBI’s Gene Expression Omnibus (GEO) database GSE11890442. The transcription factor prediction data were generated by Qiagen Transcript Discovery plugin and from PROMO (http://alggen.lsi.upc.es/cgi-bin/promo_v3/promo/promoinit.cgi?dirDB=TF_8.3) and JASPER (http://jaspar.genereg.net/). The remaining data supporting the findings of this study are available within the article, Supplementary Information, or source data file. Source data are provided with this paper.
References
Kerbel, R. S. Tumor angiogenesis. N. Engl. J. Med. 358, 2039–2049 (2008).
Weis, S. M. & Cheresh, D. A. Tumor angiogenesis: molecular pathways and therapeutic targets. Nat. Med. 17, 1359–1370 (2011).
Ye, W. The complexity of translating anti-angiogenesis therapy from basic science to the clinic. Dev. Cell 37, 114–125 (2016).
Shah, A. A., Kamal, M. A. & Akhtar, S. Tumor angiogenesis and VEGFR-2: mechanism, pathways and current biological therapeutic interventions. Curr. Drug Metab. 22, 50–59 (2020).
Rahbari, N. N. et al. Anti-VEGF therapy induces ECM remodeling and mechanical barriers to therapy in colorectal cancer liver metastases. Sci. Transl. Med. 8, 360ra135 (2016).
Carmeliet, P. & Jain, R. K. Principles and mechanisms of vessel normalization for cancer and other angiogenic diseases. Nat. Rev. Drug Disco. 10, 417–427 (2011).
Mazzone, M. & Bergers, G. Regulation of blood and lymphatic vessels by immune cells in tumors and metastasis. Annu. Rev. Physiol. 81, 535–560 (2019).
Viallard, C. & Larrivee, B. Tumor angiogenesis and vascular normalization: alternative therapeutic targets. Angiogenesis 20, 409–426 (2017).
Rehman, A. U. et al. The landscape of protein tyrosine phosphatase (Shp2) and cancer. Curr. Pharm. Des. 24, 3767–3777 (2018).
Frankson, R. et al. Therapeutic targeting of oncogenic tyrosine phosphatases. Cancer Res. 77, 5701–5705 (2017).
Grossmann, K. S., Rosario, M., Birchmeier, C. & Birchmeier, W. The tyrosine phosphatase Shp2 in development and cancer. Adv. Cancer Res. 106, 53–89 (2010).
Chen, Y. N. et al. Allosteric inhibition of SHP2 phosphatase inhibits cancers driven by receptor tyrosine kinases. Nature 535, 148–152 (2016).
Garcia Fortanet, J. et al. Allosteric inhibition of SHP2: identification of a potent, selective, and orally efficacious phosphatase inhibitor. J. Med. Chem. 59, 7773–7782 (2016).
Fedele, C. et al. SHP2 inhibition prevents adaptive resistance to MEK inhibitors in multiple cancer models. Cancer Disco. 8, 1237–1249 (2018).
Wong, G. S. et al. Targeting wild-type KRAS-amplified gastroesophageal cancer through combined MEK and SHP2 inhibition. Nat. Med. 24, 968–977 (2018).
Ruess, D. A. et al. Mutant KRAS-driven cancers depend on PTPN11/SHP2 phosphatase. Nat. Med. 24, 954–960 (2018).
Mainardi, S. et al. SHP2 is required for growth of KRAS-mutant non-small-cell lung cancer in vivo. Nat. Med. 24, 961–967 (2018).
Dardaei, L. et al. SHP2 inhibition restores sensitivity in ALK-rearranged non-small-cell lung cancer resistant to ALK inhibitors. Nat. Med. 24, 512–517 (2018).
Ahmed, T. A. et al. SHP2 drives adaptive resistance to ERK signaling inhibition in molecularly defined subsets of ERK-dependent tumors. Cell Rep. 26, 65–78 e65 (2019).
Nichols, R. J. et al. RAS nucleotide cycling underlies the SHP2 phosphatase dependence of mutant BRAF-, NF1- and RAS-driven cancers. Nat. Cell Biol. 20, 1064–1073 (2018).
**ao, P. et al. Myeloid-restricted ablation of Shp2 restrains melanoma growth by amplifying the reciprocal promotion of CXCL9 and IFN-gamma production in tumor microenvironment. Oncogene 37, 5088–5100 (2018).
Marasco, M. et al. Molecular mechanism of SHP2 activation by PD-1 stimulation. Sci. Adv. 6, eaay4458 (2020).
Rota, G. et al. Shp-2 is dispensable for establishing T cell exhaustion and for PD-1 signaling in vivo. Cell Rep. 23, 39–49 (2018).
Chemnitz, J. M., Parry, R. V., Nichols, K. E., June, C. H. & Riley, J. L. SHP-1 and SHP-2 associate with immunoreceptor tyrosine-based switch motif of programmed death 1 upon primary human T cell stimulation, but only receptor ligation prevents T cell activation. J. Immunol. 173, 945–954 (2004).
Quintana, E. et al. Allosteric inhibition of SHP2 stimulates antitumor immunity by transforming the immunosuppressive environment. Cancer Res. 80, 2889–2902 (2020).
Wang, Y. et al. SHP2 blockade enhances anti-tumor immunity via tumor cell intrinsic and extrinsic mechanisms. Sci. Rep. 11, 1399 (2021).
Kroll, J. & Waltenberger, J. The vascular endothelial growth factor receptor KDR activates multiple signal transduction pathways in porcine aortic endothelial cells. J. Biol. Chem. 272, 32521–32527 (1997).
Mannell, H. & Krotz, F. SHP-2 regulates growth factor dependent vascular signalling and function. Mini Rev. Med. Chem. 14, 471–483 (2014).
Sinha, S. et al. Dopamine regulates phosphorylation of VEGF receptor 2 by engaging Src-homology-2-domain-containing protein tyrosine phosphatase 2. J. Cell Sci. 122, 3385–3392 (2009).
Mitola, S. et al. Type I collagen limits VEGFR-2 signaling by a SHP2 protein-tyrosine phosphatase-dependent mechanism 1. Circ. Res. 98, 45–54 (2006).
Fraineau, S. et al. The vitamin K-dependent anticoagulant factor, protein S, inhibits multiple VEGF-A-induced angiogenesis events in a Mer- and SHP2-dependent manner. Blood 120, 5073–5083 (2012).
Ukropec, J. A., Hollinger, M. K., Salva, S. M. & Woolkalis, M. J. SHP2 association with VE-cadherin complexes in human endothelial cells is regulated by thrombin. J. Biol. Chem. 275, 5983–5986 (2000).
Wessel, F. et al. Leukocyte extravasation and vascular permeability are each controlled in vivo by different tyrosine residues of VE-cadherin. Nat. Immunol. 15, 223–230 (2014).
Zhang, J. et al. SHP2 protects endothelial cell barrier through suppressing VE-cadherin internalization regulated by MET-ARF1. FASEB J. 33, 1124–1137 (2019).
Lu, Y. et al. Grb-2-associated binder 1 (Gab1) regulates postnatal ischemic and VEGF-induced angiogenesis through the protein kinase A-endothelial NOS pathway. Proc. Natl Acad. Sci. USA 108, 2957–2962 (2011).
Shioyama, W. et al. Docking protein Gab1 is an essential component of postnatal angiogenesis after ischemia via HGF/c-met signaling. Circ. Res. 108, 664–675 (2011).
Zhao, J. et al. Endothelial Grb2-associated binder 1 is crucial for postnatal angiogenesis. Arterioscler Thromb. Vasc. Biol. 31, 1016–1023 (2011).
Luo, M. et al. Annexin A2 supports pulmonary microvascular integrity by linking vascular endothelial cadherin and protein tyrosine phosphatases. J. Exp. Med. 214, 2535–2545 (2017).
Huang, B. et al. Hypoxia-inducible factor-1 drives annexin A2 system-mediated perivascular fibrin clearance in oxygen-induced retinopathy in mice. Blood 118, 2918–2929 (2011).
Fedele, C. et al. SHP2 inhibition diminishes KRASG12C cycling and promotes tumor microenvironment remodeling. J. Exp. Med. 218, e20201414 (2021).
Wang, Y. et al. Targeting the SHP2 phosphatase promotes vascular damage and inhibition of tumor growth. EMBO Mol. Med. 13, e14089 (2021).
McCann, J. V. et al. Endothelial miR-30c suppresses tumor growth via inhibition of TGF-beta-induced Serpine1. J. Clin. Invest. 129, 1654–1670 (2019).
Kim, I. K. et al. Sox7 promotes high-grade glioma by increasing VEGFR2-mediated vascular abnormality. J. Exp. Med. 215, 963–983 (2018).
Yu, L. et al. JAK2 and SHP2 reciprocally regulate tyrosine phosphorylation and stability of proapoptotic protein ASK1. J. Biol. Chem. 284, 13481–13488 (2009).
Gong, H. et al. Shp2 in myocytes is essential for cardiovascular and neointima development. J. Mol. Cell Cardiol. 137, 71–81 (2019).
Klomp, J. et al. Comprehensive transcriptomic profiling reveals SOX7 as an early regulator of angiogenesis in hypoxic human endothelial cells. J. Biol. Chem. 295, 4796–4808 (2020).
Kim, K. et al. SoxF transcription factors are positive feedback regulators of VEGF signaling. Circ. Res. 119, 839–852 (2016).
Ye, X. et al. Norrin, frizzled-4, and Lrp5 signaling in endothelial cells controls a genetic program for retinal vascularization. Cell 139, 285–298 (2009).
Lee, S. H. et al. Notch pathway targets proangiogenic regulator Sox17 to restrict angiogenesis. Circ. Res. 115, 215–226 (2014).
Yang, H. et al. Sox17 promotes tumor angiogenesis and destabilizes tumor vessels in mice. J. Clin. Invest. 123, 418–431 (2013).
Gonzalez-Hernandez, S. et al. Sox17 controls emergence and remodeling of nestin-expressing coronary vessels. Circ. Res. 127, e252–e270 (2020).
Matsui, T. et al. Redundant roles of Sox17 and Sox18 in postnatal angiogenesis in mice. J. Cell Sci. 119, 3513–3526 (2006).
Patel, J. et al. Functional definition of progenitors versus mature endothelial cells reveals key SoxF-dependent differentiation process. Circulation 135, 786–805 (2017).
Young, N. et al. Effect of disrupted SOX18 transcription factor function on tumor growth, vascularization, and endothelial development. J. Natl Cancer Inst. 98, 1060–1067 (2006).
Liu, Y., Yin, G., Surapisitchat, J., Berk, B. C. & Min, W. Laminar flow inhibits TNF-induced ASK1 activation by preventing dissociation of ASK1 from its inhibitor 14-3-3. J. Clin. Invest. 107, 917–923 (2001).
Zhang, R. et al. AIP1 mediates TNF-alpha-induced ASK1 activation by facilitating dissociation of ASK1 from its inhibitor 14-3-3. J. Clin. Invest. 111, 1933–1943 (2003).
Liu, Y. & Min, W. Thioredoxin promotes ASK1 ubiquitination and degradation to inhibit ASK1-mediated apoptosis in a redox activity-independent manner. Circ. Res. 90, 1259–1266 (2002).
Zhang, J., Zhou, H. J., Ji, W. & Min, W. AIP1-mediated stress signaling in atherosclerosis and arteriosclerosis. Curr. Atheroscler. Rep. 17, 503 (2015).
Zhou, H. J. et al. AIP1 mediates vascular endothelial cell growth factor receptor-3-dependent angiogenic and lymphangiogenic responses. Arterioscler Thromb. Vasc. Biol. 34, 603–615 (2014).
Yin, M. et al. ASK1-dependent endothelial cell activation is critical in ovarian cancer growth and metastasis. JCI Insight 2, e91828 (2017).
Tao, B. et al. Myeloid-specific disruption of tyrosine phosphatase Shp2 promotes alternative activation of macrophages and predisposes mice to pulmonary fibrosis. J. Immunol. 193, 2801–2811 (2014).
Tang, K. H. et al. Combined inhibition of SHP2 and CXCR1/2 promotes anti-tumor T cell response in NSCLC. Cancer Discov. https://doi.org/10.1158/2159-8290.CD-21-0369 (2021).
Li, S. et al. SHP2 positively regulates TGFbeta1-induced epithelial-mesenchymal transition modulated by its novel interacting protein Hook1. J. Biol. Chem. 289, 34152–34160 (2014).
Xu, J. et al. Macrophage-restricted Shp2 tyrosine phosphatase acts as a rheostat for MMP12 through TGF-beta activation in the prevention of age-related emphysema in mice. J. Immunol. 199, 2323–2332 (2017).
Zehender, A. et al. The tyrosine phosphatase SHP2 controls TGFbeta-induced STAT3 signaling to regulate fibroblast activation and fibrosis. Nat. Commun. 9, 3259 (2018).
Watanabe, N. et al. BTLA is a lymphocyte inhibitory receptor with similarities to CTLA-4 and PD-1. Nat. Immunol. 4, 670–679 (2003).
Zhang, T. et al. Loss of SHP-2 activity in CD4+ T cells promotes melanoma progression and metastasis. Sci. Rep. 3, 2845 (2013).
Kerr, D. L., Haderk, F. & Bivona, T. G. Allosteric SHP2 inhibitors in cancer: targeting the intersection of RAS, resistance, and the immune microenvironment. Curr. Opin. Chem. Biol. 62, 1–12 (2021).
Hongu, T. et al. Arf6 regulates tumour angiogenesis and growth through HGF-induced endothelial beta1 integrin recycling. Nat. Commun. 6, 7925 (2015).
Kim, C. et al. Vascular RhoJ is an effective and selective target for tumor angiogenesis and vascular disruption. Cancer Cell 25, 102–117 (2014).
Acknowledgements
This work was supported by the Key Research and Development Project of the Ministry of Science and Technology of China (2016YFA0501800 to Y.K.); the National Natural Science Foundation of China (32070952 and 31871399 to H.C., 81873418 to Y.K., and 32000799 to J.Z.), and the Zhejiang Provincial Natural Science Foundation of China (LZ18H020001 to H.C.). We thank Shuangshuang Liu and Qiong Huang from the core facility platform of Zhejiang University School of Medicine for providing technical support. We would like to thank Editage for English language editing.
Author information
Authors and Affiliations
Contributions
Z.X., H.C., and Y.K. designed and analyzed the experiments and wrote the manuscript. X.Z., Z.X., C.G., Q.Y., Yue S., Yi.S., J.Z., J.H., C.Z., and Y.H. performed the experiments. C.G. and Z.X. illustrated the working model and edited the manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Peer review information Nature Communications thanks Carmine Fedele and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
Source data
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
About this article
Cite this article
Xu, Z., Guo, C., Ye, Q. et al. Endothelial deletion of SHP2 suppresses tumor angiogenesis and promotes vascular normalization. Nat Commun 12, 6310 (2021). https://doi.org/10.1038/s41467-021-26697-8
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41467-021-26697-8
- Springer Nature Limited
This article is cited by
-
Engineering nanoparticles-enabled tumor-associated macrophages repolarization and phagocytosis restoration for enhanced cancer immunotherapy
Journal of Nanobiotechnology (2024)
-
VEGF-induced Nrdp1 deficiency in vascular endothelial cells promotes cancer metastasis by degrading vascular basement membrane
Oncogene (2024)
-
Aggresome formation promotes ASK1/JNK signaling activation and stemness maintenance in ovarian cancer
Nature Communications (2024)
-
TCBIR/CD320: a potential therapeutic target upregulated in endothelial cells and associated with immune cell infiltration in liver hepatocellular carcinoma
Discover Oncology (2024)
-
Gastric Cancer Mesenchymal Stem Cells Trigger Endothelial Cell Functional Changes to Promote Cancer Progression
Stem Cell Reviews and Reports (2024)