Advances of SHP2 modulators in the cancer immunotherapy / 药学学报

Src homology phosphotyrosyl phosphatase 2 (SHP2) is a protein tyrosine phosphatase encoded by PTPN11, which catalyzes the dephosphorylation of protein tyrosine. As a convergence node, SHP2 mediates multiple signaling pathways such as rat sarcoma (RAS)-rapidly accelerated fibrosarcoma (RAF)-mitogen-activated extracellular signal-regulated kinase (MEK)-extracellular regulated protein kinases (ERK), phosphatidylinositol 3-kinase (PI3K)-serine/threonine kinase (AKT), janus kinase (JAK)-signal transducer and activator of transcription (STAT) and programmed death-1 (PD-1)/programmed cell death-ligand 1 (PD-L1). It can not only regulate the growth and proliferation of tumor cells, but also mediate the immune escape of tumor cells by influencing the tumor microenvironment. Given its dual biological functions in tumor immune regulation, SHP2 is a promising target for cancer immunotherapy. To date, several SHP2 allosteric inhibitors have been advanced into clinical trials for tumor immunotherapy with single or combination therapeutic strategies. Additionally, SHP2 activators also showed therapeutic potential in the field of tumor immune modulation. In this paper, we reviewed the dual function of SHP2 in both tumor and immune cells. Besides, the challenges and prospects of SHP2 modulators in cancer immunotherapy were also briefly discussed, aiming to explore new horizon of SHP2 modulators for tumor immunotherapy.

Targeting macrophagic SHP2 for ameliorating osteoarthritis via TLR signaling

Osteoarthritis (OA), in which M1 macrophage polarization in the synovium exacerbates disease progression, is a major cause of cartilage degeneration and functional disabilities. Therapeutic strategies of OA designed to interfere with the polarization of macrophages have rarely been reported. Here, we report that SHP099, as an allosteric inhibitor of src-homology 2-containing protein tyrosine phosphatase 2 (SHP2), attenuated osteoarthritis progression by inhibiting M1 macrophage polarization. We demonstrated that M1 macrophage polarization was accompanied by the overexpression of SHP2 in the synovial tissues of OA patients and OA model mice. Compared to wild-type (WT) mice, myeloid lineage conditional Shp2 knockout (cKO) mice showed decreased M1 macrophage polarization and attenuated severity of synovitis, an elevated expression of cartilage phenotype protein collagen II (COL2), and a decreased expression of cartilage degradation markers collagen X (COL10) and matrix metalloproteinase 3 (MMP3) in OA cartilage. Further mechanistic analysis showed thatSHP099 inhibited lipopolysaccharide (LPS)-induced Toll-like receptor (TLR) signaling mediated by nuclear factor kappa B (NF-κB) and PI3K-AKT signaling. Moreover, intra-articular injection of SHP099 also significantly attenuated OA progression, including joint synovitis and cartilage damage. These results indicated that allosteric inhibition of SHP2 might be a promising therapeutic strategy for the treatment of OA.

Study on the secondary metabolites of grasshopper-derived fungi Arthrinium sp. NF2410 / 中国天然药物

Two new 2-carboxymethyl-3-hexyl-maleic anhydride derivatives, arthrianhydride A (1) and B (2), along with three known compounds 3-5, were isolated from the fermentation broth of a grasshopper-associated fungus Arthrinium sp. NF2410. The structures of new compounds 1 and 2 were determined based on the analysis of the HR-ESI-MS and NMR spectroscopic data. Furthermore, compounds 1 and 2 were evaluated on inhibitory activity against the enzyme SHP2 and both of them showed moderate inhibitory activity against SHP2.

SHP2 inhibition triggers anti-tumor immunity and synergizes with PD-1 blockade

Tyrosine phosphatase SHP2 is a promising drug target in cancer immunotherapy due to its bidirectional role in both tumor growth promotion and T-cell inactivation. Its allosteric inhibitor SHP099 is known to inhibit cancer cell growth both and . However, whether SHP099-mediated SHP2 inhibition retards tumor growth anti-tumor immunity remains elusive. To address this, a CT-26 colon cancer xenograft model was established in mice since this cell line is insensitive to SHP099. Consequently, SHP099 minimally affected CT-26 tumor growth in immuno-deficient nude mice, but significantly decreased the tumor burden in CT-26 tumor-bearing mice with intact immune system. SHP099 augmented anti-tumor immunity, as shown by the elevated proportion of CD8IFN- T cells and the upregulation of cytotoxic T-cell related genes including , which decreased the tumor load. In addition, tumor growth in mice with SHP2-deficient T-cells was markedly slowed down because of enhanced anti-tumor responses. Finally, the combination of SHP099 and anti-PD-1 antibody showed a higher therapeutic efficacy than either monotherapy in controlling tumor growth in two colon cancer xenograft models, indicating that these agents complement each other. Our study suggests that SHP2 inhibitor SHP099 is a promising candidate drug for cancer immunotherapy.

Tumor-targeted therapy based on allosteric inhibitor of tyrosine phosphatase SHP2 / 中国肿瘤生物治疗杂志

@#蛋白质酪氨酸磷酸化对细胞的生命活动至关重要，其调控异常与多种疾病的发生密切相关。在酪氨酸磷酸酶家族 中，SHP2是目前唯一被证实的原癌蛋白，参与调控多个癌症相关过程。其活化突变会导致白血病、黑色素瘤、乳腺癌及肺癌的发 生。2016年以来，随着高特异性、可口服的SHP2新型变构抑制剂成功开发，靶向抑制SHP2在抑制肿瘤生长以及改善肿瘤耐药性 方面逐渐显现出了强大的临床应用潜力，提示SHP2抑制剂有望成为首个靶向酪氨酸磷酸酶的抗肿瘤靶向药物。

Study on influence of the CpG DNA on activation of IFN-γ signaling transduction regulatory proteins / Монголын Анагаах Ухаан

Introduction@#When human body encounters external pathogens primary/innate immunity cells are activated by recognizing them and secondary/adaptive immunity is activated consecutively. In our previous study, we revealed that there is a synergistic action between TLR9 and IFN-γ signaling in the endothelial cells. @*Purpose@#To determine the role of negative and positive regulator proteins on the IFN-γ/TLR9 signaling pathway. @*Methods@#In this study, murine endothelial cell (END-D) culture was used. END-D cells pre-treated with TLR9 ligand CpG DNA and then stimulated with IFN-γ. The negative (SHP-2, SOCS1, PIAS1) and positive (p38) regulator protein expression was detected by Western blotting. @*Results and Conclusion@#Treatment by TLR9 ligand CpG DNA and IFN-γ increased positive regulator p38 phosphorylation in 0.5 hour. CpG DNA inhibited IFN-γ negative regulator PIAS1 protein expression in 6 hour and SOCS1 and SHP-2 expression could not affect in 4 hour.

Effect of 5-FU on the expression of SHP2 gene in colorectal cancer cells RKO / 药学实践杂志

Objective To investigate the effect of 5-FU on the expression of SHP2 gene in colorectal cancer cells RKO . Methods The human colorectal cancer cell line RKO of logarithmic growth phase was cultured 48 h with 5-FU .The expression of SHP2 was observed by immunofluorescence and Western blotting ;siSHP2 which specifically inhibits the expression of SHP2 was transfected into RKO cells and cultured 48 h with 5-FU .The cell absorbance (A) values were measured with CCK8 and apoptosis was determined by flow cytometry .The sensitivity of RKO to 5-FU and the effect of 5-FU on apoptosis of RKO cells were observed .Results The expression of nuclear protein of SHP2 was improved remarkably after colorectal cancer cell RKO was cultured 48 h with 5-FU .The sensitivity of RKO cells to 5-FU and the cell apoptosis rate induced by 5-FU were decreased after siSHP2 transfection .Conclusion 5-FU exerts anti-cancer activity possibly due to its promoting the apoptosis of RKO cells by influencing the expression of SHP2 .

Shp2 regulates cigarette smoke extract-induced epithelial-mesenchymal transition / 中国药理学通报

Aim To explore the effects of Shp2 on cig-arette smoke extract (CSE)-induced epithelial-mesen-chymal transition (EMT). Methods The effects of CSE on TGF-β1 levels in epithelial cells were meas-ured by Q-PCR and ELISA. Immunofluorescent stai-ning was used to assess the expressions of CSE-induced EMT-related markers. The activation of CSE-induced Shp2,Smad2 was investigated by Western blot. Re-sults CSE induced Shp2 phosphorylation in a concen-tration-dependent manner in A549 cells. PHPS1 inhib-ited the increase in mRNA and protein expression of TGF-β1 induced by CSE. PHPS1 regulated the expres-sions of CSE-induced EMT markers (down-regulation of E-cadherin,up-regulation expression of Vimentin and α-SMA). The inhibition of either Shp2 inhibitor or Shp2 siRNA decreased Smad2 phosphorylation induced by CSE. Conclusions CSE initiates EMT through the Shp2 / Smad2 signaling pathway,which is activated by CSE through TGF-β1 generation. It is suggested that Shp2 might be a possible new target for COPD and lung cancer therapy.

Role of protein tyrosine phosphatase SHP-2 in group I metabotropic glutamate receptors-induced allodynia / 中国药理学通报

Aim To investigate whether the pain modi-fication by group I metabotropic glutamate receptors (mGluRs)required the involvement of Src homology-2 domain-containing phosphatase-2 (SHP-2 ).Methods Co-immunoprecipitation was performed to examine the possible interaction between SHP-2 and group I mGluRs in spinal cord dorsal horn of mice.By measur-ing the paw withdrawal thresholds,the effects of SHP-2 inhibitor NSC-87877 or its catalytically inactive SHP-2 (C459S ) mutant on allodynia induced by group I mGluRs agonist DHPG (50 nmol)were observed.Re-sults Anti-mGluR5 antibody was able to co-immuno-precipitate SHP-2 from spinal dorsal horn of mice, while no SHP-2 was precipitated by anti-mGluR1 anti-body.Inactivation of SHP-2 by NSC-87877 (6 nmol) or SHP-2 (C459S ) effectively attenuated allodynia caused by DHPG.Conclusion SHP-2 can physically interact with mGluR5.The activation of SHP-2 may be necessary for group I mGluRs to process the nocicep-tive information.

Anticancer function of Shp2 in lung adenocarcinoma A549 cells and rela-ted molecular mechanisms / 中国病理生理杂志

AIM: To explore the anticancer function of Shp2 in lung adenocarcinoma A549 cells and the related molecular mechanisms.METHODS: The viability and proliferation of A549 cells treated with Shp2 specific inhibi-tor Phps-1 or cisplatin (DDP) were measured by CCK-8 assay and EdU assay.Annexin V-FITC/PI double staining was ap-plied to detect apoptotic rate of A549 cells with different interventions.The protein levels of caspase-3-17p, Bcl-2, Bax, p-STAT3 /STAT3 and p-ERK/ERK were determined by Western blot.RESULTS: Compared with control group, Phps-1 at the concentration of 20 μmol/L significantly increased the viability of A549 cells after 24 h of treatment ( P <0.05). Meanwhile, the proliferation rate of A549 cells in Phps-1 20 μmol/L group was significant increased compared with control group (P <0.05).The apoptotic rate of A549 cells in DDP treatment group decreased from 13.01% ±2.62% to 3.67%±0.93% after adding Phps-1 (P <0.05).Phps-1 down-regulated the protein levels of caspase-3-17p, Bax and p-ERK, but up-regulated p-STAT3.CONCLUSION: Shp2 is a tumor suppressor in A549 cells, which may be associated with the activation of STAT3 signal pathway.

LRRK2 Inhibits FAK Activity by Promoting FERM-mediated Autoinhibition of FAK and Recruiting the Tyrosine Phosphatase, SHP-2

Mutation of leucine-rich repeat kinase 2 (LRRK2) causes an autosomal dominant and late-onset familial Parkinson's disease (PD). Recently, we reported that LRRK2 directly binds to and phosphorylates the threonine 474 (T474)-containing Thr-X-Arg(Lys) (TXR) motif of focal adhesion kinase (FAK), thereby inhibiting the phosphorylation of FAK at tyrosine (Y) 397 residue (pY397-FAK), which is a marker of its activation. Mechanistically, however, it remained unclear how T474-FAK phosphorylation suppressed FAK activation. Here, we report that T474-FAK phosphorylation could inhibit FAK activation via at least two different mechanisms. First, T474 phosphorylation appears to induce a conformational change of FAK, enabling its N-terminal FERM domain to autoinhibit Y397 phosphorylation. This is supported by the observation that the levels of pY397-FAK were increased by deletion of the FERM domain and/or mutation of the FERM domain to prevent its interaction with the kinase domain of FAK. Second, pT474-FAK appears to recruit SHP-2, which is a phosphatase responsible for dephosphorylating pY397-FAK. We found that mutation of T474 into glutamate (T474E-FAK) to mimic phosphorylation induced more strong interaction with SHP-2 than WT-FAK, and that pharmacological inhibition of SHP-2 with NSC-87877 rescued the level of pY397 in HEK293T cells. These results collectively show that LRRK2 suppresses FAK activation through diverse mechanisms that include the promotion of autoinhibition and/or the recruitment of phosphatases, such as SHP-2.

Significance study of SHP2 Expression in brain tissue of bacterial meningitis rat / 中国基层医药

Objective To study the significance of protein tyrosine phosphatase (SHP2)expression in bac-terial meningitis.Methods 90 rats were divided into meningitis group (72)and healthy controls (18)two groups based on the random number table,The SHP2 expression in rat brain tissue at different time points of meningitis group and healthy control group were tested by reverse transcription (RT)a PCR,Western blotting,immunohistochemical methods,then the relationship between SHP2 protein expression and tumor necrosis factor-α(TNF-α),white blood cell (WBC)counts were observed and analyzed.Results The cortical SHP2 mRNA expression of meningitis rat in-cluding (0.035 ±0.020),(0.200 ±0.049),(0.129 ±0.032)and (0.057 ±0.039),were significantly higher than those of the healthy control group (0.031 ±0.028)(F=12.74,P0.05)and 0.77 (t=4.303,P<0.05).Conclusion SHP2 participates in pathophysiology of bacterial meningitis,the main role may be in suppressing inflammation and repairing inflammatory response,it can be used as a reference indicator of condition changes.

Efecto del factor de crecimiento de fibroblastos 21 / Fibroblast growht factor 21 effects over phosphatase SHP-2 expression in Malme-3 cells

Almost 50 percent of Noonan syndrome patients, characterized by short stature, present activating mutations of the citoplasmatic phosphatase SHP-2, which induce hyperactivation of the Ras/MAPK pathway. On the other hand, the fibroblast growth factor 21 (FGF-21), recently suggested as a FGF with “endocrine” function, would affect longitudinal growth inhibiting growth hormone signaling at chondrocytes level. Union and activation of FGF-21 to its receptor is regulated by the co-factor beta Klotho (KLB). Aims: To determine if FGF-21 and/or FGF-21+KLB are able to modify the genetic expression of SHP-2 ina human skin fibroblast cell line (Malme-3). Methods: cells were incubated with or without FGF-21, FGF-21 + KLB. At 12 and 24 hours after induction total RNA was extracted andSHP-2 mRNA levels were determine by quantitative PCR. Expression of GADPH gene was employed for normalization. Results: Incubation with FGF-21 produce a 36 percent (p = < 0,05)increment in SHP-2 expression, which was not modified with KLB co-incubation. Discussion: it is shown by the first time that FGF-21 is able to produce an increase in SHP-2 gene expression in human fibroblast, which was independent of KLB presence...

Síndrome de Noonan / Noonan syndrome

El Síndrome de Noonan fue descrito por Noonan y Ehmke en 1963. La incidencia se ha estimado en 1 de 1000 y 1 de 2500 nacimientos vivos.1 El gen se encuentra localizado en el cromosoma 12q22 y se hereda en forma autosómica dominante y tiene una expresividad muy variable. La principal característica incluye estatura baja, defectos cardiacos, dismorfismo facial entre otros. Estatura La severidad de los síntomas varían mucho en estos pacientes. Lo que no siempre es fácil hacer el diagnóstico en los primeros años, y muchas veces son subdiagnosticados, condición que nos motivo a revisar el caso.

Noonan Syndrome is a relative common autosomic dominant congenital disorder, with an incidence between 1:1,000 and 1:2,500 children worldwide. The gen is in 12q22 chromosome. The principal features include short stature, typical facial dysmorphology and congenital heart disease, among others. The range and severity of features can vary greatly in patients with NS, therefore, establishing a diagnose is difficult. The syndrome is not always identified at an early age, and many times misdiagnosed.

Advance research on protein tyrosine phosphatase SHP-2 and the regulating role of it in osteoclastogenesis / 医学研究生学报

SHP-2 is one of the protein tyrosine phosphatases which plays a role in the progress of dephosphorylation in organ -isms, participating in many kinds of signal transduction pathways .The mutation of SHP-2 is associated with many kinds of malignant diseases .In recent years , scholars have found that SHP-2 is the key factor in osteoclastogenesis , playing a positive role in the progress of reversible phosphorylation of osteoclasts .The objective of this article is to review the molecular biological characteristics of SHP-2, th diseases associated with the mutation of SHP-2 and the effects of SHP-2 in regulation of osteoclastogenesis .

Role of SHP2 in invasion and metastasis of breast cancer / 中国肿瘤临床

Src homology 2 domain-containing phosphatase (SHP2), which is encoded by proto-oncogene PTPN11, is one of the transmembrane protein-tyrosine phosphatases. SHP2 has an important function in signal transduction pathways and activities of cells through the regulation of tyrosine phosphorylation level of intracellular proteins. The status of SHP2 activation is closely connected with the regulation of hormone levels, state of invasion and metastasis of tumor, development and progression of tumor stem cells of breast cancer, as well as signal pathways including Ras/ERK and PI3K/Akt/mTOR. Gene knockout or gene silencing expression helps inhibit tumor growth, irreversibly hindering the ability of the tumor to regain stem cells and disturb the signal pathways of the invasion and metastasis of breast cancer. Recent studies have shown that SHP2 may help in bringing anticancer drugs to a higher level. This arti-cle concentrates on the research progress in relationship of SHP2 with invasion and metastasis of breast cancer.

PTK2 and PTPN11 expression in myelodysplastic syndromes

OBJECTIVE: The aim of this study was to evaluate the expression of protein tyrosine kinase 2 and protein tyrosine phosphatase non-receptor type 11, which respectively encode focal adhesion kinase protein and src homology 2 domain-containing protein-tyrosine phosphatase 2, in hematopoietic cells from patients with myelodysplastic syndromes. METHODS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions were analyzed by quantitative polymerase chain reaction in bone marrow cells from patients with myelodysplastic syndromes and healthy donors. RESULTS: Protein tyrosine kinase 2 and tyrosine phosphatase non-receptor type 11 expressions did not significantly differ between normal cells and myelodysplastic cells. CONCLUSIONS: Our data suggest that despite the relevance of focal adhesion kinase and src homology 2 domain-containing protein-tyrosine phosphatase 2 in hematopoietic disorders, their mRNA expression do not significantly differ between total bone marrow cells from patients with myelodysplastic syndromes and healthy donors. .

Morphological Change and SHP-2 Gene Expression in Auditory Cortical Neuron after Auditory Deprivation in Rat / 听力学及言语疾病杂志

Objective To investigate the morphological change and SHP -2(src -homology domain contai-ning protein tygosine phosphatase type 2) gene expression in auditory cortical neuron after auditory deprivation in rat .Methods A total of 48 SD rats were randomly divided into 2-week group ,4-week group ,6-week group ,8-week group and 4 corresponding control groups with 6 rats in each group .Bilateral cochlear ablation was done to traumatized groups to ensure their postoperative ABR threshold was above 90 dB SPL .Then paraffin sectioning and HE and Nissl staining were used to detect morphological change of auditory cortical neuron ,simultaneously the RT-PCR was used to measure the expression of SHP -2 gene in auditory cortical neuron of each group ,and relative quantitative analysis was used .Results The HE and Nissl staining revealed that apoptotic shape of auditory cortical neuron became serious by time ,with the diversified cellular morphology .The relative quantity of SHP -2 gene showed statistical differences between any two groups by the method of one -way ANOVA ,showing a rising trend by time (P<0 .05) .Conclusion Along with time ,auditory cortex ,auditory deprivation showed increasing neuronic apoptosis and neuronic proliferation .Apoptosis was the final result from the mutual antagonism by the 2 factors .

The expression and significance of protein tyrosine phosphatase SHP2 in juvenile rat bacterial meningitis model / 中华传染病杂志

Objective To explore the expression and significance of protein tyrosine phosphatase SHP2 in experimental bacterial meningitis.Methods A juvenile rat bacterial meningitis model was established by direct intraeisternal injection with Streptococcus pneumoniae. Uninfected control animals were mock-infected with sterile saline.The transcription and expression of SHP2 were detected by reverse transcription-polymerase chain reaction(RT-PCR),Western blot and immunohistochemistry techniques respectively at 1,3,7 and 14 days after infection.White blood cell(WBC)count,concentration of tumor necrosis factor-alpha(TNF-α)in cerebrospinal fluid(CSF)were also measured.Variables that were not normally distributed were compared by the Kruskal-Wallis test.Multiple comparison used Student-Newman-Keuls(SNK).The association between variables was assessed by using the Pearson correlation coefficient.Results Compared with uninfected controls,rats with bacterial meningitis showed a significant upregulation of SHP2 at both mRNA and protein levels(F=12.74,P<0.01;F=198,P<0.01).S HP2 mRNA levels peaked at 3 days after infection increasing more than five fold and remained at high levels at 7 days.In parallel,SHP2 protein levels began to increase at 3 days after infection,reaching a maximal increase of nearly nine fold at 7 days and remained at high levels at 14 days. Immunohistochemical analysis of SHP2 expression in the juvenile rat brain demonstrated that SHP2 labelling cells,identified as neuronal and glial cells,widely distributed in the cerebral cortex and the increased immunoreactive cells around the third ventricle were mainly glial cells.In addition,the protein levels of SHP2 and WBC counts were significantly correlated with each other(r=0.77,P<0.01),while there were no significant correlations between levels of SHP2 and TNF-a (r=0.08,P>0.05).Conclusions SHP2 may participate the pathological progress of the bacterial meningitis,restrating the inflammation and accelerating the renovation,so it can be regard as an index to measure the state of the illness.