3,739 results on '"Janus Kinase 3"'
Search Results
102. Structural and functional analysis of target recognition by the lymphocyte adaptor protein LNK
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James M. Murphy, Yaoyuan Zhang, Nadia J. Kershaw, Jeffrey J. Babon, Carola G. Vinuesa, Julia I. Ellyard, Rhiannon Morris, and Artem Laktyushin
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Science ,Amino Acid Motifs ,General Physics and Astronomy ,SH2 domain ,Crystallography, X-Ray ,General Biochemistry, Genetics and Molecular Biology ,Article ,src Homology Domains ,Mice ,Receptors, Erythropoietin ,Animals ,Humans ,Receptor ,Phosphotyrosine ,Calcium signaling ,Adaptor Proteins, Signal Transducing ,X-ray crystallography ,chemistry.chemical_classification ,Multidisciplinary ,Binding Sites ,Myeloproliferative Disorders ,Functional analysis ,food and beverages ,Signal transducing adaptor protein ,Janus Kinase 3 ,General Chemistry ,Janus Kinase 2 ,Erythropoietin receptor ,Amino acid ,Cell biology ,Proto-Oncogene Proteins c-kit ,chemistry ,fms-Like Tyrosine Kinase 3 ,Mutation ,Phosphorylation ,Cytokines ,Protein Binding ,Signal Transduction - Abstract
The SH2B family of adaptor proteins, SH2-B, APS, and LNK are key modulators of cellular signalling pathways. Whilst SH2-B and APS have been partially structurally and biochemically characterised, to date there has been no such characterisation of LNK. Here we present two crystal structures of the LNK substrate recognition domain, the SH2 domain, bound to phosphorylated motifs from JAK2 and EPOR, and biochemically define the basis for target recognition. The LNK SH2 domain adopts a canonical SH2 domain fold with an additional N-terminal helix. Targeted analysis of binding to phosphosites in signalling pathways indicated that specificity is conferred by amino acids one- and three-residues downstream of the phosphotyrosine. Several mutations in LNK showed impaired target binding in vitro and a reduced ability to inhibit signalling, allowing an understanding of the molecular basis of LNK dysfunction in variants identified in patients with myeloproliferative disease., LNK is a potent negative regulator of cytokine signaling implicated in blood cells proliferation. Here the authors present structures of the substrate recognition (SH2) domain of LNK in complex with phosphorylated motifs from JAK2 and EPOR; providing insight into its binding specificity and mode of action.
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- 2021
103. Docking and Selectivity Studies of Covalently Bound Janus Kinase 3 Inhibitors.
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Zhong HA and Almahmoud S
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- Janus Kinase 1, Janus Kinase 2 metabolism, Janus Kinases metabolism, Ligands, Signal Transduction, Janus Kinase 3, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors chemistry
- Abstract
The Janus kinases (JAKs) are a family of non-receptor cytosolic protein kinases critical for immune signaling. Many covalently bound ligands of JAK3 inhibitors have been reported. To help design selective JAK inhibitors, in this paper, we used five model proteins to study the subtype selectivity of and the mutational effects on inhibitor binding. We also compared the Covalent Dock programs from the Schrodinger software suite and the MOE software suite to determine which method to use for the drug design of covalent inhibitors. Our results showed that the docking affinity from 4Z16 (JAK3 wild-type model), 4E4N (JAK1), 4D1S (JAK2), and 7UYT (TYK2) from the Schrödinger software suite agreed well with the experimentally derived binding free energies with small predicted mean errors. However, the data from the mutant 5TTV model using the Schrödinger software suite yielded relatively large mean errors, whereas the MOE Covalent Dock program gave small mean errors in both the wild-type and mutant models for our model proteins. The docking data revealed that Leu905 of JAK3 and the hydrophobic residue at the same position in different subtypes (Leu959 of JAK1, Leu932 of JAK2, and Val981 of TYK2) is important for ligand binding to the JAK proteins. Arg911 and Asp912 of JAK3, Asp939 of JAK2, and Asp988 of TYK2 can be used for selective binding over JAK1, which contains Lys965 and Glu966 at the respective positions. Asp1021, Asp1039, and Asp1042 can be utilized for JAK1-selective ligand design, whereas Arg901 and Val981 may help guide TYK2-selective molecule design.
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- 2023
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104. Down-Regulation of Store-Operated Ca2+ Entry and Na+/Ca2+ Exchange in MCF-7 Breast Cancer Cells by Pharmacological JAK3 Inhibition.
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Yan, Jing, Zhang, Bingbing, Hosseinzadeh, Zohreh, and Lang, Florian
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CANCER cells , *GENETICS of breast cancer , *CANCER cell proliferation , *THAPSIGARGIN , *CELL death - Abstract
Background/Aims: Oscillations of cytosolic Ca2+ activity ([Ca2+]i) participate in the orchestration of tumor cell proliferation. [Ca2+]i could be increased by intracellular Ca2+ release followed by store-operated Ca2+-entry (SOCE). [Ca2+]i could be decreased by Ca2+ extrusion via Na+/ Ca2+ exchange. Mechanisms accomplishing SOCE include the pore-forming ion channel unit Orai1 and its regulator STIM1, Na+/Ca2+ exchanger isoforms include NCX1. In MCF-7 breast carcinoma cells Orai1 and NCX1 have previously been shown to be modified by pharmacological inhibition of Janus activated kinase JAK2. The present study explored whether SOCE and Na+/ Ca2+ exchange are similarly sensitive to pharmacological JAK3 inhibition. Methods: MCF- 7 breast carcinoma cells were studied in the absence and presence of the JAK3 inhibitor WHI-P154 (22 μM). [Ca2+]i was estimated from Fura-2-fluorescence, SOCE from increase of [Ca2+]i following Ca2+ re-addition after Ca2+-store depletion with sarcoendoplasmatic Ca2+- ATPase (SERCA) inhibitor thapsigargin (1 μM), and Na+/Ca2+ exchanger activity from increase of [Ca2+]i following extracellular Na+ removal. Transcript levels were quantified with RT-PCR. Results: Addition of ATP (100 μM) was followed by a rapid increase of [Ca2+]i, which was significantly blunted by WHI-P154. Thapsigargin-induced intracellular Ca2+ release was not appreciably influenced by WHI-P154. Subsequent SOCE was, however, significantly blunted by WHI-P154. WHI-P154 further significantly decreased Orai1 transcript levels. The increase of [Ca2+]i following extracellular Na+-removal and the NCX1 transcript levels were similarly decreased by WHI-P154. Conclusions: The JAK3 inhibitor WHI-P154 decreases both, Orai1 and NCX1 transcript levels and thus impairs SOCE and Na+/Ca2+ exchange. [ABSTRACT FROM AUTHOR]
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- 2016
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105. WHI-131 Promotes Osteoblast Differentiation and Prevents Osteoclast Formation and Resorption in Mice.
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Cheon, Yoon-Hee, Baek, Jong Min, Ahn, Sung-Jun, Oh, Jaemin, Kim, Ju-Young, Jun, Hong Young, Lee, Myeung Su, Erkhembaatar, Munkhsoyol, and Kim, Min Seuk
- Abstract
ABSTRACT The small molecule WHI-131 is a potent therapeutic agent with anti-inflammatory, antiallergic, and antileukemic potential. However, the regulatory effects of WHI-131 on osteoblast and osteoclast activity are unclear. We examined the effects of WHI-131 on osteoblast and osteoclast differentiation with respect to bone remodeling. The production of receptor activator of nuclear factor kappa-B ligand (RANKL) by osteoblasts in response to interleukin (IL)-1 or IL-6 stimulation decreased by 56.8% or 50.58%, respectively, in the presence of WHI-131. WHI-131 also abrogated the formation of mature osteoclasts induced by IL-1 or IL-6 stimulation. Moreover, WHI-131 treatment decreased RANKL-induced osteoclast differentiation of bone marrow-derived macrophages, and reduced the resorbing activity of mature osteoclasts. WHI-131 further decreased the mRNA and protein expression levels of c-Fos and nuclear factor of activated T cells, cytoplasmic 1 (NFATc1) by almost twofold, and significantly downregulated the mRNA expression of the following genes: tartrate-resistant acid phosphatase ( TRAP), osteoclast-associated receptor ( OSCAR), DC-STAMP, OC-STAMP, ATP6v0d2, and cathepsin K ( CtsK) compared with the control group. WHI-131 further suppressed the phosphorylation of protein kinase B (Akt) and degradation of inhibitor of kappa B (IκB); Ca
2+ oscillation was also affected, and phosphorylation of the C-terminal Src kinase (c-Src)-Bruton agammaglobulinemia tyrosine kinase (Btk)-phospholipase C gamma 2 (PLCγ2) (c-Src-Btk-PLCg2 calcium signaling pathway) was inhibited following WHI-131 treatment. The Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway was activated by WHI-131, accompanied by phosphorylation of STAT3 Ser727 and dephosphorylation of STAT6. In osteoblasts, WHI-131 caused an approximately fourfold increase in alkaline phosphatase activity and Alizarin Red staining intensity. Treatment with WHI-131 increased the mRNA expression levels of genes related to osteoblast differentiation, and induced the phosphorylation of Akt, p38, and Smad1/5/8. Furthermore, 5-week-old ICR mice treated with WHI-131 exhibited antiresorbing effects in a lipopolysaccharide-induced calvaria bone loss model in vivo and increased bone-forming activity in a calvarial bone formation model. Therefore, the results of this study show that WHI-131 plays a dual role by inhibiting osteoclast differentiation and promoting osteoblast differentiation. Thus, WHI-131 could be a useful pharmacological agent to treat osteoporosis by promoting bone growth and inhibiting resorption. © 2015 American Society for Bone and Mineral Research. [ABSTRACT FROM AUTHOR]- Published
- 2016
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106. Betulin, a Newly Characterized Compound in Acacia Auriculiformis Bark, is a Multi-target Protein Kinase Inhibitor
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Lukman Mustapha, Jacobus P. Petzer, A. A. Ahmadu, Axelle Autret, Mélanie Simoes Eugénio, Blandine Baratte, A Haruna, Morgane Rousselot, Stéphane Bach, Lesetja J. Legoabe, Jeannette Chloë Bulinski, Marie-Thérèse Dimanche-Boitrel, Bilqis A. Lawal, Claire Delehouzé, Aniefiok Udobre, Camilla Triscornia, Thomas Robert, University of Calabar (UNICAL), Kaduna State University, Laboratoire de Biologie Intégrative des Modèles Marins (LBI2M), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), University of Ilorin, University of Uyo, Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Columbia University [New York], Institut de recherche en santé, environnement et travail (Irset), Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Santé et de la Recherche Médicale (INSERM)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Université d'Angers (UA), Biosit : biologie, santé, innovation technologique (SFR UMS CNRS 3480 - INSERM 018), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), North-West University [South Aftrica] (NWU), Nigeria Institution Based Research grant, Tertiary Education Trust Fund (TETFund), French National Agency for Research and Technology (ANRT), Cifre PhD fellowship French National Research Agency (ANR) [2019/0686], Fondation d'Entreprise Grand Ouest, French ANR/Investissements d'Avenir program via the OCEANOMICs project French National Research Agency (ANR) [ANR-11-BTBR-0008], ANR-11-BTBR-0008,OCEANOMICS,Biotechnologies et bioressources pour la valorisation des écosystèmes marins planctoniques(2011), Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), SeaBeLife Biotech, Plate-forme de criblage d'inhibiteurs de protéines kinases=Kinase Inhibitor Specialized Screening facility (KISSf), Fédération de recherche de Roscoff (FR2424), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Station biologique de Roscoff (SBR), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), and Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )
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Models, Molecular ,Casein Kinase 1 epsilon ,Protein Conformation ,[SDV]Life Sciences [q-bio] ,Fusion Proteins, bcr-abl ,Pharmaceutical Science ,Organic chemistry ,Apoptosis ,Analytical Chemistry ,Glycogen Synthase Kinase 3 ,chemistry.chemical_compound ,0302 clinical medicine ,QD241-441 ,GSK-3 ,hemic and lymphatic diseases ,Drug Discovery ,NIMA-Related Kinases ,Proto-Oncogene Proteins c-abl ,Mitogen-Activated Protein Kinase 1 ,0303 health sciences ,Mitogen-Activated Protein Kinase 3 ,ABL ,Gene Expression Regulation, Leukemic ,Chemistry ,Kinase ,Acacia ,Protein kinase inhibitor ,3. Good health ,Biochemistry ,Chemistry (miscellaneous) ,030220 oncology & carcinogenesis ,Plant Bark ,Molecular Medicine ,Casein kinase 1 ,Protein Binding ,Signal Transduction ,medicine.drug_class ,Article ,03 medical and health sciences ,medicine ,Humans ,Physical and Theoretical Chemistry ,Protein kinase A ,030304 developmental biology ,Cell Proliferation ,polypharmacology ,Binding Sites ,Betulin ,Plant Extracts ,allergology ,Janus Kinase 3 ,triterpenoids ,Antineoplastic Agents, Phytogenic ,Vascular Endothelial Growth Factor Receptor-2 ,Triterpenes ,protein kinase inhibitors ,Imatinib mesylate ,Acacia stem bark ,K562 Cells - Abstract
The purpose of this work is to investigate the protein kinase inhibitory activity of constituents from ethyl acetate soluble fraction of Acacia auriculiformis stem bark. Column chromatography, gel filtration and NMR spectroscopy were used to purified and characterized betulin from the extract. Betulin which is a known inducer of apoptosis was screened against a panel of 16 disease-related protein kinases. Betulin was shown to inhibit Abelson murine leukemia viral oncogene homolog 1 (ABL1) kinase, casein kinase 1epsilon (CK1epsilon), glycogen synthase kinase 3alpha/β (GSK-3alpha/β), Janus kinase 3 (JAK3), NIMA Related Kinase 6 (NEK6) and vascular endothelial growth factor receptor 2 kinase (VEGFR2) and with activity in µM range. The effect of betulin on the cell viability of doxorubicin-resistant K562R chronic myelogenous leukemia cells was then verified to underline its putative use as anti-cancer compound. Betulin was shown to modulate the mitogen-activated protein (MAP) kinase pathway similarly to imatinib mesylate, a well-known inhibitor of ABL1 kinase. The interaction of betulin and ABL1 was studied by molecular docking showing an interaction of the inhibitor with the ATP binding pocket. Altogether, these data demonstrate that betulin is a multi-target inhibitor of protein kinases, an activity that can contribute to the anticancer properties of the natural compound and notably for treatment of leukemia.
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- 2021
107. Computational Study on Novel Natural Inhibitors Targeting Janus Kinase 3
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Weihang Li, Dong Wang, Ziyi Ding, Sheng Zhong, Shilei Zhang, Hui Li, Yujia Zou, Bo Wu, and Weiye Li
- Subjects
Computer science ,Janus kinase 3 ,Computational biology - Abstract
Objective: The aim of this study is to screen and identify novel leading compounds 5 which can inhibit protein Janus Kinase 3 (JAK3) from a drug library (ZINC database) 6 to provide precise target therapy for lung cancer. 7Methods: A set of computation-aided structural biology methods and chemical virtual 8 screening techniques were carried out to screen novel inhibitor compounds. Libdock 9 scores for potential inhibitors of JAK3 were calculated using fast docking method-10 virtual screening. Next, ADMET properties (absorption, distribution, metabolism, 11 excretion, and toxicity) were conducted to predict their pharmacological characteristics. 12 The binding affinity as well as the interactions between the candidate compounds and 13 JAK3 were calculated and visualized by precise molecular docking algorithm. 14 Ultimately, molecular dynamics simulation (MD) was performed to estimate the 15 stability of the ligand-JAK3 complex under natural environment. 16Results: After screening, two novel natural compounds, ZINC000014952116 and 17 ZINC00000393864, were finally selected as leading compounds from the ZINC15 18 database, which possessed less Ames mutagenicity, rodent carcinogenicity and 19 developmental toxicity potential than other candidate compounds. Additionally, they 20 didn’t inhibit the activity of CYP2D6. Molecular dynamics simulation analysis showed 21 that ZINC000014952116 and ZINC00000393864 could interact with JAK3 steadily, 22 and their ligand-JAK3 complexes could keep stable under natural situation, and act as 23 regulatory role to JAK3. 24Conclusion: This study analyzed that ZINC000014952116 and ZINC00000393864 25 were ideal natural inhibitors targeting JAK3 from the ZINC15 database, which could 26 also provide more options and resources for other cancer chemotherapy.
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- 2021
108. Selective inhibition of JAK3 signaling is sufficient to reverse alopecia areata
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James Ming Chen, Yuqian Chang, Angela M. Christiano, and Z. Dai
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0301 basic medicine ,Ruxolitinib ,Alopecia Areata ,Pyridines ,Administration, Topical ,T cells ,Autoimmunity ,CD8-Positive T-Lymphocytes ,stat ,03 medical and health sciences ,0302 clinical medicine ,Piperidines ,Nitriles ,medicine ,Animals ,Pyrroles ,Protein Kinase Inhibitors ,Skin ,Inflammation ,Mice, Inbred C3H ,Tofacitinib ,Chemistry ,Macrophages ,JAK-STAT signaling pathway ,Janus Kinase 3 ,General Medicine ,Janus Kinase 1 ,Alopecia areata ,Janus Kinase 2 ,Triazoles ,medicine.disease ,Hair follicle ,030104 developmental biology ,medicine.anatomical_structure ,Pyrimidines ,NK Cell Lectin-Like Receptor Subfamily K ,030220 oncology & carcinogenesis ,Cancer research ,Medicine ,Azetidines ,Cytokines ,Pyrazoles ,Signal transduction ,Isonicotinic Acids ,Janus kinase ,medicine.drug ,Research Article - Abstract
The Janus kinase/signal transducers and activators of transcription (JAK/STAT) are key intracellular mediators in the signal transduction of many cytokines and growth factors. Common γ chain cytokines and interferon-γ that use the JAK/STAT pathway to induce biological responses have been implicated in the pathogenesis of alopecia areata (AA), a T cell–mediated autoimmune disease of the hair follicle. We previously showed that therapeutic targeting of JAK/STAT pathways using the first-generation JAK1/2 inhibitor, ruxolitinib, and the pan-JAK inhibitor, tofacitinib, was highly effective in the treatment of human AA, as well as prevention and reversal of AA in the C3H/HeJ mouse model. To better define the role of individual JAKs in the pathogenesis of AA, in this study, we tested and compared the efficacy of several next-generation JAK-selective inhibitors in the C3H/HeJ mouse model of AA, using both systemic and topical delivery. We found that JAK1-selective inhibitors as well as JAK3-selective inhibitors robustly induced hair regrowth and decreased AA-associated inflammation, whereas several JAK2-selective inhibitors failed to restore hair growth in treated C3H/HeJ mice with AA. Unlike JAK1, which is broadly expressed in many tissues, JAK3 expression is largely restricted to hematopoietic cells. Our study demonstrates inhibiting JAK3 signaling is sufficient to prevent and reverse disease in the preclinical model of AA.
- Published
- 2021
109. JAK3 inhibitor-based immunosuppression in allogeneic islet transplantation in cynomolgus monkeys
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Jongwon Ha, Jiyeon Kim, Byoung Hoon Min, Seong Jun Kang, Il Hee Yoon, Chung Gyu Park, Eung Soo Hwang, Jun Seop Shin, Jong Min Kim, and Hyunwoo Chung
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Graft Rejection ,Male ,0301 basic medicine ,Transplantation Conditioning ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Transplantation, Heterologous ,Drug Evaluation, Preclinical ,Islets of Langerhans Transplantation ,030209 endocrinology & metabolism ,Pharmacology ,Article ,Diabetes Mellitus, Experimental ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Transplantation Immunology ,medicine ,Animals ,Protein Kinase Inhibitors ,Immunosuppression Therapy ,Type 1 diabetes ,geography ,geography.geographical_feature_category ,Tofacitinib ,business.industry ,Graft Survival ,Janus Kinase 3 ,Immunosuppression ,medicine.disease ,Streptozotocin ,Islet ,Tacrolimus ,Calcineurin ,Transplantation ,Macaca fascicularis ,surgical procedures, operative ,030104 developmental biology ,Female ,business ,Immunosuppressive Agents ,medicine.drug - Abstract
Islet transplantation is efficacious to prevent severe hypoglycemia and glycemic liability of selected patients of type 1 diabetes. However, since calcineurin inhibitor (CNI) causes β-cell and nephrotoxicity, alternative drug(s) with similar potency and safety profile to CNI will be highly desirable. Here we tested whether JAK3 inhibitor, tofacitinib could be used instead of tacrolimus in CIT07 immunosuppression regimen in cynomolgus nonhuman primate (NHP) model. Five independent streptozotocin (STZ)-induced diabetic monkeys were transplanted with MHC-mismatched allogeneic islets and three animals were further re-transplanted upon insufficient glycemic control or early islet graft rejection. After islet transplantation, blood glucose levels were quickly stabilized and maximal islet graft survival as measured by serum C-peptide concentration was >330, 98, >134, 31, or 22 days, respectively, after transplantation (median survival day; 98 days). Cellular and humoral immune responses were efficiently suppressed by JAK3 inhibitor-based immunosuppression during the follow-up periods. Although intermittent increases of the genome copy number of cynomolgus cytomegalovirus (CMV) were detected by quantitative real-time PCR analyses, serious infections or posttransplant lymphoproliferative disease (PTLD) was not found in all animals. Taken together, we have shown that JAK3 inhibitor could be used in replacement of tacrolimus in a highly translatable NHP islet transplantation model and these results suggest that JAK3 inhibitor will be potentially incorporated in human allogeneic islet transplantation.
- Published
- 2019
110. Genome profiling revealed the activation of IL2RG/JAK3/STAT5 in peripheral T-cell lymphoma expressing the ITK-SYK fusion gene
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Lei‑Lei Zhang, Lin Liu, Tao Guo, Hua‑Xiong Pan, and Yi‑Xuan Wang
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0301 basic medicine ,Cancer Research ,Oncogene Proteins, Fusion ,Syk ,Apoptosis ,Mice, SCID ,Jurkat cells ,Fusion gene ,IL-2-inducible T-cell kinase-spleen tyrosine kinase ,Mice ,0302 clinical medicine ,Mice, Inbred NOD ,hemic and lymphatic diseases ,STAT5 Transcription Factor ,Tumor Cells, Cultured ,peripheral T-cell lymphoma ,STAT5 ,tofacitinib ,Kinase ,hemic and immune systems ,Articles ,Protein-Tyrosine Kinases ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Female ,Signal transduction ,Tyrosine kinase ,Interleukin Receptor Common gamma Subunit ,T cell ,Biology ,xenograft mouse model ,03 medical and health sciences ,medicine ,JAK/STAT pathway ,Biomarkers, Tumor ,Animals ,Humans ,Syk Kinase ,Protein Kinase Inhibitors ,Cell Proliferation ,Genome, Human ,Gene Expression Profiling ,Tumor Suppressor Proteins ,Janus Kinase 3 ,Lymphoma, T-Cell, Peripheral ,Xenograft Model Antitumor Assays ,030104 developmental biology ,interleukin-2 receptor subunit ,biology.protein ,Cancer research - Abstract
Peripheral T‑cell lymphomas (PTCLs) are heterogeneous malignancies that are types of non‑Hodgkin lymphomas; patients with this disease have poor prognoses. The IL‑2‑inducible T‑cell kinase‑spleen tyrosine kinase (ITK‑SYK) fusion gene, the first recurrent chromosome translocation in PTCL‑not otherwise specified (NOS), can drive cellular transformation and the development of T‑cell lymphoma in mouse models. The aim of the current study was to investigate the signal transduction pathways downstream of ITK‑SYK. The authors constructed a lentiviral vector to overexpress the ITK‑SYK fusion gene in Jurkat cells. By using Signal‑Net and cluster analyses of microarray data, the authors identified the tyrosine‑protein kinase JAK (JAK)3/STAT5 signalling pathway as a downstream pathway of ITK‑SYK, activation of which mediates the effects of ITK‑SYK on tumourigenesis. JAK3‑selective inhibitor tofacitinib abrogated the phosphorylation of downstream signalling molecule STAT5, supressed cell growth, induced cell apoptosis and arrested the cell cycle at the G1/S phase in ITK‑SYK+ Jurkat cells. In a xenograft mouse model, tumour growth was significantly delayed by tofacitinib. Since JAK3 associates with interleukin‑2 receptor subunit γ (IL2RG) only, siRNA‑specific knockdown of IL2RG showed the same effect as tofacitinib treatment in vitro. These results first demonstrated that the activation of the IL2RG/JAK3/STAT5 signalling pathway contributed greatly to the oncogenic progress regulated by ITK‑SYK, supporting further investigation of JAK3 inhibitors for the treatment of PTCLs carrying the ITK‑SYK fusion gene.
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- 2019
111. CD25 and Protein Phosphatase 2A Cooperate to Enhance IL-2R Signaling in Human Regulatory T Cells
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Thomas R. Malek, Ying Ding, Aixin Yu, and George C. Tsokos
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Immunology ,chemical and pharmacologic phenomena ,Lymphocyte Activation ,T-Lymphocytes, Regulatory ,Article ,Cell Line ,Immune tolerance ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Immune Tolerance ,STAT5 Transcription Factor ,Animals ,Humans ,Immunology and Allergy ,Protein Phosphatase 2 ,IL-2 receptor ,Phosphorylation ,STAT5 ,Gene knockdown ,biology ,Chemistry ,Interleukin-2 Receptor alpha Subunit ,Janus Kinase 3 ,Receptors, Interleukin-2 ,hemic and immune systems ,Tyrosine phosphorylation ,Protein phosphatase 2 ,Cell biology ,Killer Cells, Natural ,Gene Expression Regulation ,Gene Knockdown Techniques ,biology.protein ,Signal transduction ,Protein Binding ,Signal Transduction ,030215 immunology - Abstract
Low-dose IL-2 therapy is a direct approach to boost regulatory T cells (Tregs) and promote immune tolerance in autoimmune patients. However, the mechanisms responsible for selective response of Tregs to low-dose IL-2 is not fully understood. In this study we directly assessed the contribution of CD25 and protein phosphatase 2A (PP2A) in promoting IL-2R signaling in Tregs. IL-2–induced tyrosine phosphorylation of STAT5 (pSTAT5) was proportional to CD25 levels on human CD4+ T cells and YT human NK cell line, directly demonstrating that CD25 promotes IL-2R signaling. Overexpression of the PP2A catalytic subunit (PP2Ac) by lentiviral transduction in human Tregs increased the level of IL-2R subunits and promoted tyrosine phosphorylation of Jak3 and STAT5. Interestingly, increased expression of CD25 only partially accounted for this enhanced activation of pSTAT5, indicating that PP2A promotes IL-2R signaling through multiple mechanisms. Consistent with these findings, knockdown of PP2Ac in human Tregs and impaired PP2Ac activity in mouse Tregs significantly reduced IL-2–dependent STAT5 activation. In contrast, overexpression or knockdown of PP2Ac in human T effector cells did not affect IL-2–dependent pSTAT5 activation. Overexpression of PP2Ac in human Tregs also increased the expressions of proteins related to survival, activation, and immunosuppressive function, and upregulated several IL-2–regulated genes. Collectively, these findings suggest that CD25 and PP2A cooperatively enhance the responsiveness of Tregs to IL-2, which provide potential therapeutic targets for low-dose IL-2 therapy.
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- 2019
112. PF-06651600, a Dual JAK3/TEC Family Kinase Inhibitor
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Iain Kilty, Uthpala Seneviratne, M Nusrat Sharif, Gary R. Point, Jean-Baptiste Telliez, Tsung H. Lin, Douglas S. Johnson, Michael I. Jesson, John I. Trujillo, Chuong Nguyen, Hua Xu, Liang Xue, Robert A. Everley, and Atli Thorarensen
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Antigens, Differentiation, T-Lymphocyte ,Gene isoform ,TEC ,CD8-Positive T-Lymphocytes ,Biochemistry ,Serine ,Mice ,Antigens, CD ,Animals ,Humans ,Bruton's tyrosine kinase ,Lectins, C-Type ,Pyrroles ,Kinome ,Protein Kinase Inhibitors ,biology ,Chemistry ,Kinase ,Janus Kinase 3 ,General Medicine ,Protein-Tyrosine Kinases ,Killer Cells, Natural ,Pyrimidines ,Cancer research ,biology.protein ,Molecular Medicine ,CD8 ,Cysteine - Abstract
PF-06651600 was developed as an irreversible inhibitor of JAK3 with selectivity over the other three JAK isoforms. A high level of selectivity toward JAK3 is achieved by the covalent interaction of PF-06651600 with a unique cysteine residue (Cys-909) in the catalytic domain of JAK3, which is replaced by a serine residue in the other JAK isoforms. Importantly, 10 other kinases in the kinome have a cysteine at the equivalent position of Cys-909 in JAK3. Five of those kinases belong to the TEC kinase family including BTK, BMX, ITK, RLK, and TEC and are also inhibited by PF-06651600. Preclinical data demonstrate that inhibition of the cytolytic function of CD8+ T cells and NK cells by PF-06651600 is driven by the inhibition of TEC kinases. On the basis of the underlying pathophysiology of inflammatory diseases such as rheumatoid arthritis, inflammatory bowel disease, alopecia areata, and vitiligo, the dual activity of PF-06651600 toward JAK3 and the TEC kinase family may provide a beneficial inhibitory profile for therapeutic intervention.
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- 2019
113. Aminocyanopyridines as anti‐lung cancer agents by inhibiting the STAT3 pathway
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Haitang Xu, Sensen Qiu, Xiaoying Huang, Ri Cui, Lingyuan Xu, Liangxing Wang, Xu Liu, Fan Shiqian, Chengguang Zhao, Weitao Fu, Lehe Yang, and Liqun Shen
- Subjects
STAT3 Transcription Factor ,0301 basic medicine ,Cancer Research ,Lung Neoplasms ,Pyridines ,Mice, Nude ,Antineoplastic Agents ,Biology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Transcription (biology) ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Phosphorylation ,Lung cancer ,STAT3 ,Molecular Biology ,Gene ,Cell Proliferation ,Mice, Inbred BALB C ,Interleukin-6 ,Janus Kinase 3 ,Janus Kinase 2 ,medicine.disease ,Xenograft Model Antitumor Assays ,Molecular Docking Simulation ,030104 developmental biology ,A549 Cells ,Apoptosis ,030220 oncology & carcinogenesis ,Cancer research ,STAT protein ,biology.protein ,Female ,Nuclear localization sequence ,Signal Transduction - Abstract
Lung cancer is a leading cause of cancer-related death worldwide. Cyanopyridines and aminocyanopyridines with carbon-nitrogen bonds have been proved to exert significant anticancer, antibacterial, and anti-inflammatory effects. In this study, we showed that aminocyanopyridine 3o and 3k displaying potent antitumor activity via inhibiting the signal transducer and activator of transcription 3 (STAT3) pathway. They blocked the constitutive STAT3 phosphorylation in a dose- and time-dependent manner and regulated the transcription of STAT3 target genes encoding apoptosis factors. Most importantly, 3o also inhibited interleukin-6-induced STAT3 activation and nuclear localization. Furthermore, 3o significantly inhibited the tumor growth of H460-derived xenografts. Taken together, these findings suggest that 3o and 3k are promising therapeutic drug candidates for lung cancer by inhibiting persistent STAT3 signaling.
- Published
- 2019
114. Stüve–Wiedemann syndrome: recurrent neonatal infections caused by impairment of JAK/STAT 3 pathway
- Author
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Irsa Rosina-Angelista, Ginette M. Ecury-Goossen, Karolien Van De Maele, Charlotte A. Smulders, Mieke M. van Haelst, Egbert J.W. Redeker, Faculty of Medicine and Pharmacy, Pediatrics, Human genetics, Amsterdam Neuroscience - Complex Trait Genetics, Amsterdam Reproduction & Development (AR&D), Human Genetics, and ACS - Pulmonary hypertension & thrombosis
- Subjects
Adult ,Male ,STAT3 Transcription Factor ,leukemia inhibitory factor receptor ,congenital, hereditary, and neonatal diseases and abnormalities ,Leukemia Inhibitory Factor Receptor alpha Subunit ,Leukemia inhibitory factor receptor ,Osteochondrodysplasias ,medicine.disease_cause ,stat ,Pathology and Forensic Medicine ,03 medical and health sciences ,JAK/STAT 3 pathway ,medicine ,Humans ,Autonomic dysregulation ,Abnormalities, Multiple ,Family ,Genetics(clinical) ,Pediatrics, Perinatology, and Child Health ,Gene ,Genetics (clinical) ,Janus Kinases ,030304 developmental biology ,0303 health sciences ,Mutation ,neonatal infections ,business.industry ,Siblings ,030305 genetics & heredity ,Infant, Newborn ,Stüve-Wiedemann syndrome ,Janus Kinase 3 ,JAK-STAT signaling pathway ,Syndrome ,General Medicine ,medicine.disease ,STUVE-WIEDEMANN SYNDROME ,Pedigree ,Dysplasia ,Pediatrics, Perinatology and Child Health ,Immunology ,Female ,Anatomy ,business ,Exostoses, Multiple Hereditary - Abstract
Stüve-Wiedemann syndrome (OMIM #601559) is a rare, autosomal recessive disorder characterized by skeletal dysplasia, consecutive infections, feeding difficulties and autonomic dysregulation. We present an Afro-Caribbean family with two siblings diagnosed with Stüve-Wiedemann syndrome. The underlying loss-of-function mutation in the leukemia inhibitory factor receptor gene is thought to impair proper functioning of the JAK/STAT 3 pathway. As this affects normal functioning of T-helper cells, these patients are prone to infections with uncommon pathogens as illustrated by this case.
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- 2019
115. Are peptides a solution for the treatment of hyperactivated JAK3 pathways?
- Author
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Claucia Fernanda Volken de Souza, Claudia Monfroni Rocha, Stefan Laufer, Anja Dullius, and Márcia Inês Goettert
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0301 basic medicine ,Immunology ,Disease ,Bioinformatics ,medicine.disease_cause ,Autoimmune Diseases ,03 medical and health sciences ,0302 clinical medicine ,Neoplasms ,Psoriasis ,medicine ,Animals ,Humans ,Pharmacology (medical) ,Pharmacology ,Severe combined immunodeficiency ,Mutation ,business.industry ,Janus Kinase 3 ,Cancer ,medicine.disease ,Clinical trial ,030104 developmental biology ,Drug development ,Cancer cell ,Peptides ,business ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
While the inactivation mutations that eliminate JAK3 function lead to the immunological disorders such as severe combined immunodeficiency, activation mutations, causing constitutive JAK3 signaling, are known to trigger various types of cancer or are responsible for autoimmune diseases, such as rheumatoid arthritis, psoriasis, or inflammatory bowel diseases. Treatment of hyperactivated JAK3 is still an obstacle, due to different sensibility of mutation types to conventional drugs and unwanted side effects, because these drugs are not absolutely specific for JAK3, thus inhibiting other members of the JAK family, too. Lack of information, in which way sole inhibition of JAK3 is necessary for elimination of the disease, calls for the development of isoform-specific JAK3 inhibitors. Beside this strategy, up to date peptides are a rising alternative as chemo- or immunotherapeutics, but still sparsely represented in drug development and clinical trials. Beyond a possible direct inhibition function, crossing the cancer cell membrane and interfering in disease-causing pathways or triggering apoptosis, peptides could be used in future as adjunct remedies to potentialize traditional therapy and preserve non-affected cells. To discuss such feasible topics, this review deals with the knowledge about the structure-function of JAK3 and the actual state-of-the-art of isoform-specific inhibitor development, as well as the function of currently approved drugs or those currently being tested in clinical trials. Furthermore, several strategies for the application of peptide-based drugs for cancer therapy and the physicochemical and structural relations to peptide efficacy are discussed, and an overview of peptide sequences, which were qualified for clinical trials, is given.
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- 2019
116. Design of novel JAK3 Inhibitors towards Rheumatoid Arthritis using molecular docking analysis
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Shreshtha Sharma, Trishang Udhwani, Anuraj Nayarisseri, Palugulla Bhaskar Reddy, Aishwarya Gandhe, Divya Jain, and Sanjeev Kumar Singh
- Subjects
Virtual screening ,Rheumatoid Arthritis ,01 natural sciences ,03 medical and health sciences ,0302 clinical medicine ,food ,BOILED-Egg plot ,Chemistry ,Janus kinase 3 ,JAK 3 inhibitor ,General Medicine ,food.food ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,ADMET ,Biochemistry ,Boiled egg ,030220 oncology & carcinogenesis ,Molecular docking ,Target protein ,Signal transduction ,Janus kinase ,Tyrosine kinase ,PubChem ,Research Article - Abstract
Multiple cytokines play a pivotal role in the pathogenesis of Rheumatoid Arthritis by inducing intracellular signaling and it is known that the members of the Janus kinase (JAK) family are essential for such signal transduction. Janus kinase 3 is a tyrosine kinase that belongs to the Janus family of kinases. Drugs targeting JAK3 in the treatment of Rheumatoid arthritis is relevant. Therefore, it is of interest to design suitable inhibitors for JAK3 dimer using molecular docking with Molegro Virtual Docker. The compound possessing the highest affinity score is subjected to virtual screening to retrieve inhibitors. The compound SCHEMBL19100243 (PubChem CID- 76749591) displays a high affinity with the target protein. The affinity scores of this compound are more than known drugs. ADMET analysis and BOILED Egg plot provide insights into this compound as a potent inhibitor of JAK3.
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- 2019
117. Discovery of an Orally Available Janus Kinase 3 Selective Covalent Inhibitor
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Yiqing Zhou, Xitao Li, Zhengying Pan, Zhong Zhenpeng, and Shi Liyang
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Lipopolysaccharides ,Cell signaling ,Administration, Oral ,01 natural sciences ,Compound 32 ,Cell Line ,Mice ,Structure-Activity Relationship ,03 medical and health sciences ,Adenosine Triphosphate ,Drug Discovery ,medicine ,Animals ,Humans ,Structure–activity relationship ,Protein Kinase Inhibitors ,030304 developmental biology ,chemistry.chemical_classification ,Mice, Inbred ICR ,0303 health sciences ,Kinase ,Janus kinase 3 ,Janus Kinase 3 ,Adenosine ,0104 chemical sciences ,Kinetics ,010404 medicinal & biomolecular chemistry ,Enzyme ,chemistry ,Biochemistry ,Cell culture ,Drug Design ,Leukocytes, Mononuclear ,Cytokines ,Molecular Medicine ,Half-Life ,Signal Transduction ,medicine.drug - Abstract
JAK family kinases are important mediators of immune cell signaling and Janus Kinase 3 (JAK3) has long been indicated as a potential target for autoimmune disorders. Intensive efforts to develop highly selective JAK3 inhibitors have been underway for many years. However, because of JAK3's strong binding preference to adenosine 5'-triphosphate (ATP), a number of inhibitors exhibit large gaps between enzymatic and cellular potency, which hampers efforts to dissect the roles of JAK3 in cellular settings. Using a targeted covalent inhibitor approach, we discovered compound 32, which overcame ATP competition (1 mM) in the enzymatic assay, and demonstrated significantly improved inhibitory activity for JAK3-dependent signaling in mouse CTLL-2 and human peripheral blood mononuclear cells. Compound 32 also exhibited high selectivity within the JAK family and good pharmacokinetic properties. Thus, it may serve as a highly valuable tool molecule to study the overlapping roles of JAK family kinases in complex biological settings. Our study also suggested that for covalent kinase inhibitors, especially those targeting kinases with low Km ATP values, the reversible interactions between molecules and proteins should be carefully optimized to improve the overall potency.
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- 2019
118. Janus kinase-1 and 3 in ankylosing spondylitis
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Chien Chih Lai, Tzu Hao Li, Wei Sheng Chen, Chun Hsiung Chen, Hsien-Tzung Liao, Hung An Chen, Chang-Youh Tsai, and Chung Tei Chou
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Adult ,Male ,CD3 ,CD14 ,Taiwan ,Severity of Illness Index ,Peripheral blood mononuclear cell ,03 medical and health sciences ,0302 clinical medicine ,Humans ,Medicine ,Spondylitis, Ankylosing ,BASDAI ,lcsh:R5-920 ,Ankylosing spondylitis ,biology ,Janus kinase 1 ,business.industry ,Acute-phase protein ,Janus Kinase 3 ,Janus Kinase 1 ,General Medicine ,Middle Aged ,medicine.disease ,C-Reactive Protein ,ROC Curve ,Case-Control Studies ,030220 oncology & carcinogenesis ,Immunology ,Leukocytes, Mononuclear ,biology.protein ,Female ,030211 gastroenterology & hepatology ,lcsh:Medicine (General) ,business ,BASFI ,Biomarkers - Abstract
Background/purpose: To investigate the Janus kinase-1 and 3 (JAK-1 and 3) expression in peripheral blood mononuclear cells (PBMCs) in ankylosing spondylitis (AS). Methods: The levels of JAK-1 and JAK-3 mRNA in PBMCs, CD3+ T cells and CD14+ monocytes were measured by RT-PCR in 52 AS patients and 31 healthy controls (HCs). The demographic features, BASDAI, BASFI, HLA-B27, ESR, CRP and serum immunoglobulin A (IgA) level were recorded and correlated with the JAK-1 & JAK-3 transcripts in patients and HCs as appropriate. Results: JAK-1 and JAK-3 expression in PB CD3+ T cells plus CD14+ monocytes was significantly higher in AS patients than in HCs (p 4) and BASFI (>4) than ESR or CRP in AS patients. Conclusion: In AS, JAK-1 expression in PB cells rather than ESR or CRP might be regarded as a bio-marker for monitoring disease activity and functional index in AS. These findings have also suggested that JAK-1 and JAK-3 expression may play a role in the inflammatory processes in patients with AS. Keywords: Ankylosing spondylitis, Janus kinase (JAK), Acute-phase reactant
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- 2019
119. Transcriptional modulation of the T helper 17/interleukin 17 axis ameliorates renal ischemia-reperfusion injury
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Sang Kyu Ye, Jae Wook Lee, Jung Pyo Lee, Eunjin Bae, Ran Hui Cha, Mi Yeon Yu, Sun Ho Kwon, Joo-Yeon Yoo, Dong Ki Kim, Seung Hee Yang, Yon Su Kim, Dong Jun Park, and Hajeong Lee
- Subjects
STAT3 Transcription Factor ,medicine.medical_specialty ,030232 urology & nephrology ,Inflammation ,030204 cardiovascular system & hematology ,Kidney ,Proinflammatory cytokine ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,medicine ,Animals ,Humans ,STAT3 ,Transplantation ,biology ,business.industry ,Interleukin-17 ,Acute kidney injury ,Janus Kinase 3 ,Janus Kinase 2 ,medicine.disease ,Mice, Inbred C57BL ,Endocrinology ,medicine.anatomical_structure ,Gene Expression Regulation ,Nephrology ,Reperfusion Injury ,biology.protein ,STAT protein ,Cytokines ,Th17 Cells ,Interleukin 17 ,medicine.symptom ,business ,Reperfusion injury - Abstract
BackgroundSignal transducer and activator of transcription 3 (STAT3) is a latent transcription factor critical for T-cell function. Although inhibition of the Janus kinase 2 (JAK2)/STAT3 pathway has been reported to be protective against ischemia-reperfusion injury (IRI), the role of T cell–associated STAT3 in the pathogenesis of renal IRI has not been specifically defined.MethodsWe induced renal IRI in both mice with T cell–specific STAT3 knockout (Lck-Cre;STAT3flox/flox) and wild-type controls (C57BL/6) and assessed renal damage and inflammation at 48 h after IRI. Human proximal tubular epithelial cells grown under hypoxia were treated with a JAK2 inhibitor, caffeic acid 3,4-dihydroxy-phenylethyl ester, to determine the effect of JAK2/STAT3 inhibition on renal epithelia. Independently, we disrupted Cln 3-requiring 9 (Ctr9) to inhibit T helper 17 (Th17) activation via RNA interference and determined if Ctr9 inhibition aggravates renal injury through upregulated Th17 activation.ResultsThe Lck-Cre;STAT3flox/flox mice exhibited significantly reduced kidney damage compared with controls. This protective effect was associated with reduced intrarenal Th17 infiltration and proinflammatory cytokines. Human proximal tubular epithelial cells under hypoxia exhibited significant upregulation of interleukin 17 receptors, and pharmacologic inhibition of JAK2 significantly ameliorated this change. RNA interference with Ctr9 in splenocytes enhanced differentiation into Th17 cells. In vivo knockdown of Ctr9 in mice with renal IRI further aggravated Th17-associated inflammation and kidney injury.ConclusionsSTAT3 in T cells contributes to renal IRI through Th17 activation. Inhibition of Ctr9 further enhances Th17 activation and aggravates kidney injury, further supporting the role of Th17 cells in renal IRI.
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- 2018
120. A benzoxazole derivative PO‐296 inhibits T lymphocyte proliferation by the JAK3/STAT5 signal pathway
- Author
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Hui-Jie Guo, Zou Qiang, Luo Xingyan, Hong Zhou, Yang Liu, Jing Xiong, Li-Mei Li, Shu-Xia Yang, Yantang Wang, Chun-Fen Mo, and Si-Yu Wang
- Subjects
0301 basic medicine ,Cell Survival ,T-Lymphocytes ,Apoptosis ,Enzyme-Linked Immunosorbent Assay ,Lymphocyte Activation ,Biochemistry ,Phosphatidylinositol 3-Kinases ,03 medical and health sciences ,0302 clinical medicine ,STAT5 Transcription Factor ,medicine ,Humans ,Interferon gamma ,Protein kinase A ,Molecular Biology ,Protein kinase B ,Cells, Cultured ,PI3K/AKT/mTOR pathway ,Benzoxazoles ,Interleukin-6 ,Chemistry ,Janus kinase 3 ,Cell Cycle ,Interleukin-17 ,Cell Biology ,T lymphocyte ,030104 developmental biology ,030220 oncology & carcinogenesis ,STAT protein ,Cancer research ,Interleukin-2 ,Phosphorylation ,Interleukin-4 ,medicine.drug - Abstract
Immunosuppressants have shown striking achievements in treating autoimmune diseases in recent years. It is urgent to develop more immunosuppressants to provide more options for patients. PO-296 [2-(6-chlorobenzo[d]oxazol-2-yl)-4,5,6,7-tetrahydro-2H-indazol-3-ol] was identified as a novel benzoxazole derivative. We observed that it exhibits an obvious immunosuppressive activity to T lymphocytes. PO-296 significantly inhibited the proliferation of activated human T lymphocyte without cytotoxicity. Moreover, PO-296 did not affect the expression of cluster of differentiation (CD)-25 or CD69 but induced T lymphocyte cycle arrest in the G0/G1 phase. Furthermore, PO-296 inhibited interleukin (IL)-6, IL-17, and interferon gamma expression but had no effect on IL-2, IL-4, or IL-10. Yet, importantly, PO-296 inhibited the phosphorylation of signal transducer and activator of transcription 5 (STAT5), increased the phosphorylation of p70S6K, but did not affect the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/mitogen-activated protein kinase pathway. In conclusion, these findings indicate that PO-296 inhibits human activated T-lymphocyte proliferation by affecting the janus kinase 3 (JAK3)/STAT5 pathway. PO-296 possesses a potential lead compound for the design and development of new immunosuppressants for the treatment of autoimmune diseases.
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- 2018
121. Endothelial Jak3 expression enhances pro-hematopoietic angiocrine function in mice
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Tyler M. Lu, Shahin Rafii, Raphael Lis, José Gabriel Barcia Durán, David Redmond, Ryan Schreiner, Fuqiang Geng, Jenny Xiang, and Sean Houghton
- Subjects
Male ,0301 basic medicine ,Cell type ,QH301-705.5 ,Medicine (miscellaneous) ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immunity ,medicine ,Animals ,Gene Knock-In Techniques ,Biology (General) ,Haematopoietic stem cells ,Endothelial Cells ,Janus Kinase 3 ,Hematopoietic Stem Cells ,Hematopoiesis ,Cell biology ,Endothelial stem cell ,Haematopoiesis ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Second messenger system ,Female ,Bone marrow ,Stem cell ,General Agricultural and Biological Sciences ,Tyrosine kinase ,Stem-cell niche - Abstract
Jak3 is the only non-promiscuous member of the Jak family of secondary messengers. Studies to date have focused on understanding and targeting the cell-autonomous role of Jak3 in immunity, while functional Jak3 expression outside the hematopoietic system remains largely unreported. We show that Jak3 is expressed in endothelial cells across hematopoietic and non-hematopoietic organs, with heightened expression in the bone marrow. The bone marrow niche is understood as a network of different cell types that regulate hematopoietic function. We show that the Jak3–/– bone marrow niche is deleterious for the maintenance of long-term repopulating hematopoietic stem cells (LT-HSCs) and that JAK3-overexpressing endothelial cells have increased potential to expand LT-HSCs in vitro. This work may serve to identify a novel function for a highly specific tyrosine kinase in the bone marrow vascular niche and to further characterize the LT-HSC function of sinusoidal endothelium., Barcia Durán et al. show that the tyrosine kinase gene Jak3 is expressed in multiple endothelial cell types in the mouse, including in non-hematopoietic organs, with particularly high expression in the bone marrow. Using mice lacking Jak3, they show that Jak3 expressed in the bone marrow niche is important for maintaining long-term repopulating hematopoietic stem cells.
- Published
- 2021
122. Exosomal miR-193b-5p as a regulator of LPS-induced inflammation in dairy cow mammary epithelial cells
- Author
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Jianfa Wang, Tianqi Zhang, Yingying Xie, Xuequan Hu, Rui Wu, Haotian Xu, and Shuai Lian
- Subjects
Lipopolysaccharides ,Inflammation ,Biology ,Exosomes ,Exosome ,Interleukin 22 ,Mammary Glands, Animal ,medicine ,Animals ,Phosphorylation ,Mastitis, Bovine ,Cells, Cultured ,Janus kinase 3 ,NF-kappa B ,Epithelial Cells ,Cell Biology ,General Medicine ,Extracellular vesicle ,NFKBID ,Microvesicles ,Cell biology ,IκBα ,MicroRNAs ,Gene Expression Regulation ,Cytokines ,Cattle ,Female ,medicine.symptom ,Developmental Biology - Abstract
Exosomes are a type of extracellular vesicle that act as shuttles, transporting certain genetic information to other cells. MiRNA cargo within exosomes can regulate gene expression at the transcriptional level. The objective of this study was to investigate the exosomal miRNAs that regulate lipopolysaccharide (LPS)-induced inflammation in dairy cow mammary alveolar (Mac-T) cells. We found two exosome miRNAs upregulated and five exosomal miRNAs downregulated, respectively, in the LPS-stimulated Mac-T cells. MiR-193b-5p was upregulated 6.3-fold in the LPS-stimulated cell-derived exosome. Target prediction results showed that nuclear factor kappa B (NF-κB) inhibitor delta (NFKBID), transforming growth factor-beta 1 induced transcript 1 (TGFB1I1), interleukin 22 (IL-22), TNF receptor superfamily member 11b (TNFRSF11B), and Janus kinase 3 (JAK3) might be the main target genes of miR-193b-5p. After treatment of Mac-T cells with the miR-193b-5p mimic, the phosphorylation levels of inhibitor of nuclear factor-kappa Bα (IκBα) and p65 were upregulated, the level of IL-6 mRNA was upregulated, and IL-1β, TNF-α, and TGF-β mRNA levels were downregulated. After treatment of Mac-T cells with miR-193b-5p inhibitor, the phosphorylation levels of IκBα and p65 were downregulated. In summary, these findings provide strong evidence that exosomal miR-193b-5p could be a regulator of LPS-induced inflammation in Mac-T cells and reveal a new role of exosomal miRNAs in regulating dairy cow mastitis.
- Published
- 2021
123. JAK3 and PI3K mediates the suppressive effects of interferon tau on neutrophil extracellular traps formation during peri-implantation period
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Ajay Kumar Dang and Mohanned Naif Alhussien
- Subjects
MAPK/ERK pathway ,Neutrophils ,p38 mitogen-activated protein kinases ,Immunology ,Pregnancy Proteins ,Extracellular Traps ,Andrology ,Phosphatidylinositol 3-Kinases ,Immune system ,Pregnancy ,Immunology and Allergy ,Animals ,Embryo Implantation ,Protein kinase B ,Protein Kinase Inhibitors ,Insemination, Artificial ,biology ,Chemistry ,Obstetrics and Gynecology ,Janus Kinase 3 ,Neutrophil extracellular traps ,Interferon tau ,Reproductive Medicine ,Myeloperoxidase ,Neutrophil elastase ,Interferon Type I ,biology.protein ,Cattle ,Female ,Signal Transduction - Abstract
Interferon tau (IFNτ) is the main maternal signal for pregnancy in ruminants and modulates the functions of various immune cells, including neutrophils. Neutrophil extracellular traps (NETs) are one of the main defence mechanisms of neutrophils. In this study, we observed higher (p < 0.01) ex-vivo NETs extrusion by blood neutrophils from day 16-18 post artificial insemination (AI) in non-inseminated and inseminated non-pregnant cows compared to pregnant cows. In vitro study also showed that IFNτ hampers NETs formation in dose and time dependent manner. The lowest (p < 0.01) NETs formation and the highest (p < 0.01) mRNA expression (RT-PCR) of IFNτ stimulated genes (ISG15, OAS1, MX1) were observed when neutrophil incubated with 9 ng/mL IFNτ for 3.5 h. Signalling cascades mediating IFNτ impairment of NETs formation were identified using inhibitors of JAK2, JAK3, p38, PI3K/Akt and MAPK/Erk. IFNτ reduced (p < 0.01) the mRNA expression (RT-PCR) and concentration (ELISA) of genes and proteins that mediate NETs formation in blood neutrophils including histones (H1, H2), neutrophil elastase (NE) and myeloperoxidase (MPO). However, the effects of IFNτ on these genes and proteins were eliminated in the presence of JAK3 or PI3K inhibitors. Immunocytochemistry study also showed strong MPO signal in the presence of JAK3 or PI3K inhibitors as compared to positive control (PC, IFNτ alone). The results indicate that IFNτ impairs NETs formation using JAK3 and PI3K and thus essential for successful implantation and establishment of pregnancy in cows.
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- 2021
124. Computational assessment of saikosaponins as adjuvant treatment for COVID-19: molecular docking, dynamics, and network pharmacology analysis
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Shailendra S. Gurav, Satyendra K. Prasad, Saurabh K. Sinha, Rupesh V. Chikhale, Pukar Khanal, Muniappan Ayyanar, Yadu Nandan Dey, Nilambari S. Gurav, Rajesh B. Patil, and Manish M. Wanjari
- Subjects
Bupleurum ,Saikosaponins ,In silico analysis ,Molecular Dynamics Simulation ,010402 general chemistry ,01 natural sciences ,Catalysis ,Inorganic Chemistry ,Molecular dynamics ,Protein Domains ,Drug Discovery ,NADPH-Oxidase-5 ,Humans ,CHEK1 ,Oleanolic Acid ,Physical and Theoretical Chemistry ,Receptor ,Molecular Biology ,chemistry.chemical_classification ,biology ,SARS-CoV-2 ,Interleukin-6 ,010405 organic chemistry ,Chemistry ,Janus kinase 3 ,Organic Chemistry ,Biological activity ,General Medicine ,Saponins ,biology.organism_classification ,COVID-19 Drug Treatment ,0104 chemical sciences ,Molecular Docking Simulation ,Enzyme ,Biochemistry ,Docking (molecular) ,Original Article ,Janus Kinase-3 ,Information Systems - Abstract
Graphic abstract Saikosaponins are major biologically active triterpenoids, usually as glucosides, isolated from Traditional Chinese Medicines (TCM) such as Bupleurum spp., Heteromorpha spp., and Scrophularia scorodonia with their antiviral and immunomodulatory potential. This investigation presents molecular docking, molecular dynamics simulation, and free energy calculation studies of saikosaponins as adjuvant therapy in the treatment for COVID19. Molecular docking studies for 23 saikosaponins on the crystal structures of the extracellular domains of human lnterleukin-6 receptor (IL6), human Janus Kinase-3 (JAK3), and dehydrogenase domain of Cylindrospermum stagnale NADPH–oxidase 5 (NOX5) were performed, and selected protein–ligand complexes were subjected to 100 ns molecular dynamics simulations. The molecular dynamics trajectories were subjected to free energy calculation by the MM-GBSA method. Molecular docking and molecular dynamics simulation studies revealed that IL6 in complex with Saikosaponin_U and Saikosaponin_V, JAK3 in complex with Saikosaponin_B4 and Saikosaponin_I, and NOX5 in complex with Saikosaponin_BK1 and Saikosaponin_C have good docking and molecular dynamics profiles. However, the Janus Kinase-3 is the best interacting partner for the saikosaponin compounds. The network pharmacology analysis suggests saikosaponins interact with the proteins CAT Gene CAT (Catalase) and Checkpoint kinase 1 (CHEK1); both of these enzymes play a major role in cell homeostasis and DNA damage during infection, suggesting a possible improvement in immune response toward COVID-19. Supplementary Information The online version contains supplementary material available at 10.1007/s11030-021-10183-w.
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- 2021
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125. Janus kinase inhibitors for atopic dermatitis: a promising treatment modality
- Author
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Youkyung S. Roh, T. H. Nguyen, A. M. Cartron, Madan M. Kwatra, and Shawn G. Kwatra
- Subjects
Adult ,Ruxolitinib ,Acetonitriles ,Administration, Topical ,Population ,Administration, Oral ,Dermatology ,Severity of Illness Index ,Receptor tyrosine kinase ,Proinflammatory cytokine ,Dermatitis, Atopic ,Pathogenesis ,030207 dermatology & venereal diseases ,03 medical and health sciences ,0302 clinical medicine ,Piperidines ,Nitriles ,medicine ,Humans ,Janus Kinase Inhibitors ,education ,Child ,TYK2 Kinase ,education.field_of_study ,Sulfonamides ,biology ,business.industry ,Janus Kinase 3 ,Atopic dermatitis ,Janus Kinase 1 ,Janus Kinase 2 ,medicine.disease ,Pyridazines ,Pyrimidines ,STAT1 Transcription Factor ,Treatment Outcome ,Tyrosine kinase 2 ,Purines ,030220 oncology & carcinogenesis ,Immunology ,biology.protein ,Quality of Life ,Azetidines ,Pyrazoles ,Safety ,business ,Janus kinase ,Heterocyclic Compounds, 3-Ring ,medicine.drug - Abstract
Atopic dermatitis (AD) is chronic, pruritic, inflammatory skin disease that affects a significant portion of the population in industrialized nations. For nonresponders to conventional therapies, AD can significantly reduce sleep quality and quality of life. AD pathogenesis is multifactorial and involves multiple immune pathways, with recent evidence of T helper (Th)2, Th17 and Th22 axis attenuation in various AD endotypes and racial subtypes. Inhibition of the conserved Janus kinase (JAK) signalling pathway represents a promising therapeutic avenue to reduce the activation of multiple proinflammatory mediators involved in AD pathogenesis. JAK inhibitors exist in both oral and topical forms with variable specificity for the receptor tyrosine kinases JAK1, JAK2, JAK3 and tyrosine kinase 2. Oral formulations include abrocitinib, upadacitinib, baricitinib and gusacitinib, and are most appropriate for patients with moderate to severe AD. Emerging topical formulation in development include ruxolitinib and deglocitinib, which may be used in patients with localized AD and also adjunctively with systemic therapy in patients with more severe disease. With observed rapidity in itch relief and accompanying dramatic reduction in inflammatory lesion count, JAK inhibitors represent a promising new treatment to revolutionize the management of AD.
- Published
- 2021
126. Malignant struma ovarii: next-generation sequencing of six cases revealed Nras, Braf, and Jak3 mutations
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Anna Nelva, Enrico Grosso, Marco Volante, Giuseppe Forte, Emanuela Arvat, Marco Gallo, Maria Scatolini, Mauro Papotti, Daniele Liscia, Francesca Maletta, Flora Cesario, Roberta Poli, and Jasna Metovic
- Subjects
Neuroblastoma RAS viral oncogene homolog ,Proto-Oncogene Proteins B-raf ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,030209 endocrinology & metabolism ,Biology ,Gene mutation ,Thyroid cancer ,GTP Phosphohydrolases ,BRAF mutation ,JAK3 ,Next-generation sequencing ,RAS mutation ,Struma ovarii ,03 medical and health sciences ,Cytokeratin ,0302 clinical medicine ,Endocrinology ,medicine ,Humans ,Ovarian Teratoma ,Thyroid Neoplasms ,Ovarian Neoplasms ,Thyroid ,High-Throughput Nucleotide Sequencing ,Janus Kinase 3 ,Membrane Proteins ,Malignant Struma Ovarii ,medicine.disease ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Mutation ,Cancer research ,Female - Abstract
Struma ovarii (SO) is a highly specialized ovarian teratoma, consisting of thyroid tissue. Rarely, carcinomas histologically identical to their thyroid counterparts may occur, and are comprehensively defined as malignant struma ovarii (MSO). Their optimal management is controversial, and the molecular profile of the malignant counterpart in the ovary is incompletely known. In this study, the clinicopathological and molecular features of six MSO from different Italian Institutions were analysed, to explore genetic profiles of potential therapeutic interest. The histopathological features and immunoprofile (according to the known markers Galectin-3, HBME1, cytokeratin 19 and CD56) were reviewed. In addition, all cases underwent genetic analysis with a next-generation sequencing (NGS) hot spot cancer panel detecting mutations in 50 genes involved in cancerogenesis. RET/PTC rearrangements and TERT promoter alterations were also evaluated. Papillary carcinoma in all similar to its thyroid counterpart was found in five of six cases, including classical (two tumors) and follicular variant (three tumors) types. The last case was a poorly differentiated carcinoma. An activating gene mutation, was detected in five of six cases, including two NRAS, two BRAF, and one JAK3 oncogene mutations. No alterations were found in the other panel genes, nor in TERT promoter, or in RET chromosomal regions. MSO is a rare condition. Papillary carcinoma is the predominant malignant type, sharing both histomorphological and molecular features of its thyroid counterpart. Interestingly, the single case of poorly differentiated carcinoma displayed a JAK3 mutation. The presence of such driving mutation could be of potential interest in guiding postoperative treatment.
- Published
- 2021
127. Activity of JAK/STAT and NF-κB in patients with axial spondyloarthritis.
- Author
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Świerkot, Jerzy, Sokolik, Renata, Czarny, Anna, Zaczyńska, Ewa, Nowak, Beata, Chlebicki, Arkadiusz, Korman, Lucyna, Madej, Marta, Wojtala, Patryk, Lubiński, Łukasz, and Wiland, Piotr
- Abstract
Background: The etiology of axial spondyloarthritis (axSpA) is not fully elucidated. Research continues in determining the mechanisms responsible for initiation of the disease process, its maintenance and development. Objectives: The aim of this study was to evaluate the expression of transcription factors STAT (signal transducer and activator of transcription) and NF-κB (nuclear factor kappa B) as well as Janus kinase3 (JAK3) in the peripheral blood leukocytes. We also analyzed the connection between the degree of activation of transcription factors and the disease activity. Material/Methods: The study involved 46 patients with axSpA and 19 healthy individuals who comprised the control group. The expression of NF-κB, STAT1, STAT3, STAT4, STAT5, STAT6, and JAK3 in peripheral blood leukocytes was assessed. To determine the degree of activation of transcription factors STAT-s and NF-κB and JAK3 kinase, the immunocytochemistry method was used. For location of the factors, the primary monoclonal anti-NF-κB, anti-JAK3 and polyclonal anti-STAT-s antibodies were used (Chemicon International, USA, Abcam, Cambridge, UK), and the set of antibodies Novocastain Super ABC Kit (Novocastra, UK). Results: Expression of STAT1, STAT3, STAT4, STAT5, STAT6, NF-κB and JAK3 was statistically higher in the group of patients with axSpA than in the control group. There was a positive correlation with ESR values and expression of STAT4. There was no correlation between STAT, NF-κB, and JAK3 expression and ASDAS, BASDAI, and BASFI. Nine patients were treated with TNF-α inhibitors. The expression of NF-κB and STAT6 was higher in the group treated with TNF-α inhibitors, even though disease activity in these patients was shown to be lower than in those not receiving such treatment (ASDAS = 1.34±0.51 vs. 3.52±0.90, BASDAI = 2.34±1.92 vs. 5.51±2.41). Conclusions: In the group of patients with axSpA compared with the control group, higher expression of the transcription factors STAT and NF-κB as well as JAK3 was observed. Due to its crucial roles in inflammation and autoimmunity, STAT4 may have promise as an effective therapeutic target for axSpA. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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128. Case Report: Mutations in JAK3 causing severe combined immunodeficiency complicated by disseminated Bacille Calmette-Guérin disease and Pneumocystis pneumonia.
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Pan Y, Pan H, Lian C, Wu B, Lin J, Huang G, and Cui B
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- Female, Humans, Infant, BCG Vaccine adverse effects, Janus Kinase 3, Mutation, Vaccines, Attenuated adverse effects, Pneumonia, Pneumocystis diagnosis, Pneumonia, Pneumocystis drug therapy, Severe Combined Immunodeficiency complications, Severe Combined Immunodeficiency diagnosis, Severe Combined Immunodeficiency genetics
- Abstract
Background: As a form of severe combined immunodeficiency (SCID), Janus kinase 3 ( JAK3 ) deficiency can be fatal during severe infections in children, especially after inoculation of live-attenuated vaccines. We report a unique case of JAK3 deficiency with two compound heterozygous JAK3 mutations complicated by disseminated Bacille Calmette-Guérin (BCG) disease and Pneumocystis pneumonia., Case Description: A 5-month-old Chinese girl presented with recurring fever and productive cough after BCG vaccination and ineffective antibiotic treatment. Chest CT demonstrated bilateral infiltrations, enlarged mediastinal and axillary lymph nodes, and hypoplasia of the thymus. Mycobacterium tuberculosis and Pneumocystis jirovecii were detected from blood samples by sequencing. Acid-fast bacilli were also found from the sputum aspirate and gastric aspirate. Lymphocyte subset analyses indicated T-B+NK- immunodeficiency, and gene sequencing identified two heterozygous missense mutations (one unreported globally) in the Janus homology 7 (JH7) domain of JAK3 . The patient received rifampicin, isoniazid, ethambutol, and trimethoprim/sulfamethoxazole and was discharged after improvements but against advice., Outcome: The patient died at 13 months of age due to severe infections and hepatic damage., Discussion: SCID should be recognized before inoculation of live-attenuated vaccines in children. Newborn screening for SCID is advocated. Further investigations are needed to better understand the pathogenicity of the variants and molecular mechanism of the JH7 domain of JAK3 ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Pan, Pan, Lian, Wu, Lin, Huang and Cui.)
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- 2022
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129. Novel JAK3-specific inhibitor safe and effective in mouse RA model.
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Phillips R
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- Animals, Humans, Mice, Janus Kinase 3, Protein Kinase Inhibitors
- Published
- 2022
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130. Pharmacological MEK inhibition promotes polyclonal T-cell reconstitution and suppresses xenogeneic GVHD
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Akifumi Takaori-Kondo, Kazutaka Kitaura, Seiji Okada, Takero Shindo, Tadasu Shin-I, Hiroyuki Muranushi, Ryuji Suzuki, Shinya Kimura, and Hidekazu Itamura
- Subjects
Pyridones ,T cell ,T-Lymphocytes ,Immunology ,Transplantation, Heterologous ,Clone (cell biology) ,Receptors, Antigen, T-Cell ,Graft vs Host Disease ,chemical and pharmacologic phenomena ,Mice, SCID ,Pyrimidinones ,Biology ,Tacrolimus ,Mice ,Immune system ,immune system diseases ,medicine ,Animals ,Humans ,Protein Kinase Inhibitors ,Cells, Cultured ,Trametinib ,Mice, Knockout ,Mitogen-Activated Protein Kinase Kinases ,MEK inhibitor ,T-cell receptor ,Hematopoietic Stem Cell Transplantation ,Janus Kinase 3 ,medicine.disease ,Clone Cells ,surgical procedures, operative ,Graft-versus-host disease ,medicine.anatomical_structure ,Humanized mouse ,Cancer research ,Immunosuppressive Agents - Abstract
Rapid immune reconstitution without developing graft-versus-host disease (GVHD) is required for the success of allogeneic hematopoietic stem cell transplantation. Here, we analyzed the effects of pharmacological MEK inhibition on human polyclonal T-cell reconstitution in a humanized mouse GVHD model utilizing deep sequencing-based T-cell receptor (TCR) repertoire analysis. GVHD mice exhibited a skewed TCR repertoire with a common clone within target organs. The MEK inhibitor trametinib ameliorated GVHD and enabled engraftment of diverse T-cell clones. Furthermore, trametinib also ameliorated GVHD sparing diverse T cell repertoire, even when it was given from day 15 through 28. Although tacrolimus also reduced development of GVHD, it disturbed diverse T cell reconstitution and resulted in skewed TCR repertoire. Thus, trametinib not only suppresses GVHD-inducing T cells but also promotes human T cell reconstitution in vivo, providing a novel rationale for translational studies targeting human GVHD.
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- 2020
131. LincRNA-Cox2 regulates IL6/JAK3/STAT3 and NF-κB P65 pathway activation in Listeria monocytogenes-infected RAW264.7 cells
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Lin Yuan, Shengxia Chen, Wei Ling, Xugan Jiang, Bing Hu, Ye Lu, and Yurong Zhu
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Microbiology (medical) ,Inflammation ,macromolecular substances ,macrophage ,Microbiology ,NF-κB ,Proinflammatory cytokine ,STAT3 ,03 medical and health sciences ,chemistry.chemical_compound ,Other systems of medicine ,medicine ,Macrophage ,030304 developmental biology ,0303 health sciences ,biology ,030306 microbiology ,Interleukin-6 ,Janus kinase 3 ,NF-kappa B ,Interleukin ,food and beverages ,Janus Kinase 3 ,General Medicine ,lincRNA-Cox2 ,Listeria monocytogenes ,QR1-502 ,Cell biology ,Infectious Diseases ,chemistry ,biology.protein ,STAT protein ,RNA, Long Noncoding ,medicine.symptom ,RZ201-999 - Abstract
Listeria monocytogenes (Lm) can lead to high mortality rates relative to other foodborne pathogens. Lm-induced inflammation is partly characterized by macrophage activation. Long non-coding RNAs (lncRNAs) have important roles in various biological processes. However, it is unknown how lncRNAs regulate the host response to Lm infection. To identify the role of lncRNA in Lm infection, we used in vitro and in vivo models. We found that lincRNA-Cox2 was highly expressed in Lm-infected RAW264.7 cells. LincRNA-Cox2 knockdown resulted in reduced proinflammatory cytokines, apoptosis, migration ability and enhanced phagocytosis of Lm. LincRNA-Cox2 knockdown also reduced the phosphorylation of Janus kinase 3 (JAK3) and signal transducer and activator of transcription (STAT3) and the nuclear translocation of nuclear factor (NF)-κB P65, which are known to be involved in inflammatory responses. Experimentally inhibiting the protein and phosphorylation levels of STAT3 resulted in reduced proinflammatory cytokines and enhanced phagocytosis of Lm by the RAW264.7 cells. Our research suggests that lincRNA-Cox2 plays important roles in inflammation, the phagocytic function and cell migration ability of RAW264.7 cells by activating interleukin (IL)-6/JAK3/STAT3 signaling, and lincRNA-Cox2 also regulates NF-κB P65 nuclear translocation. Our research provides new insights into the regulatory role of lincRNA-Cox2 in Lm infection.
- Published
- 2020
132. SOCS3 overexpression enhances ADM resistance in bladder cancer T24 cells
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M-Z, Li, D-H, Lai, H-B, Zhao, Z, Chen, Q-X, Huang, and J, Situ
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STAT3 Transcription Factor ,Janus Kinase 3 ,Apoptosis ,Janus Kinase 2 ,Up-Regulation ,Proto-Oncogene Proteins c-bcl-2 ,Urinary Bladder Neoplasms ,Doxorubicin ,Drug Resistance, Neoplasm ,Suppressor of Cytokine Signaling 3 Protein ,Cell Line, Tumor ,Humans ,Cell Proliferation ,Signal Transduction - Abstract
JAK-STAT3 signaling pathway widely participates in cell proliferation and apoptosis. Suppressor of cytokine signaling 3 (SOCS3) is a negative regulator of JAK-STAT3. SOCS3 downregulation is associated with drug resistance in breast cancer and leukemia. However, its role in bladder cancer drug resistance is still unclear. This study established ADM resistant bladder cancer cell model to investigate the role of SOCS3-JAK/STAT3 signaling pathway ADM resistance.ADM drug resistant cell line T24/ADM was established. SOCS3, p-JAK2, p-JAK3, and Bcl-2 expressions in T24/ADM, T24, and HBEC cells were compared. Cell proliferation and apoptosis were evaluated by flow cytometry. T24/ADM cells were divided into five groups, including control, pSicoR-blank, pSicoR-SOCS3, FLLL32, and pSicoR-SOCS3 + FLLL32 groups. Cell proliferation was determined by EdU staining.SOCS3 was reduced, while p-JAK2, p-STAT3, and Bcl-2 expressions upregulated in T24 cells compared with HBEC cells. T24/ADM cells exhibited lower SOCS3, higher p-JAK2, p-STAT3, and Bcl-2 levels than T24 cells. Cell apoptosis was higher, whereas cell proliferation was weaker in T24 cells compared with T24/ADM cells. SOCS3 overexpression and/or FLLL32 treatment significantly downregulated p-JAK2, p-STAT3, and Bcl-2 expressions, attenuated cell proliferation, and elevated sensitivity to ADM induced cell apoptosis.SOCS3 reduction was associated with bladder cancer sensitivity to ADM. SOCS3 overexpression decreased JAK-STAT3 signaling pathway activity, declined Bcl-2 expression, inhibited cell proliferation, elevated cell apoptosis, and enhanced ADM sensitivity in T24 cells.
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- 2020
133. Discovery of a Novel Class of Covalent Dual Inhibitors Targeting the Protein Kinases BMX and BTK
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Forster, Michael, Liang, Xiaojun Julia, Schröder, Martin, Gerstenecker, Stefan, Chaikuad, Apirat, Knapp, Stefan, Laufer, Stefan, and Gehringer, Matthias
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Models, Molecular ,tyrosine kinases ,Binding Sites ,Janus kinase 3 ,Molecular Structure ,Molecular Conformation ,Reproducibility of Results ,Protein-Tyrosine Kinases ,Article ,bone marrow tyrosine kinase on chromosome X ,lcsh:Chemistry ,Structure-Activity Relationship ,chemical probes ,lcsh:Biology (General) ,lcsh:QD1-999 ,Bruton’s tyrosine kinase ,Drug Discovery ,Agammaglobulinaemia Tyrosine Kinase ,Humans ,covalent inhibitors ,human activities ,Protein Kinase Inhibitors ,lcsh:QH301-705.5 ,Protein Binding - Abstract
The nonreceptor tyrosine TEC kinases are key regulators of the immune system and play a crucial role in the pathogenesis of diverse hematological malignancies. In contrast to the substantial efforts in inhibitor development for Bruton&rsquo, s tyrosine kinase (BTK), specific inhibitors of the other TEC kinases, including the bone marrow tyrosine kinase on chromosome X (BMX), remain sparse. Here we present a novel class of dual BMX/BTK inhibitors, which were designed from irreversible inhibitors of Janus kinase (JAK) 3 targeting a cysteine located within the solvent-exposed front region of the ATP binding pocket. Structure-guided design exploiting the differences in the gatekeeper residues enabled the achievement of high selectivity over JAK3 and certain other kinases harboring a sterically demanding residue at this position. The most active compounds inhibited BMX and BTK with apparent IC50 values in the single digit nanomolar range or below showing moderate selectivity within the TEC family and potent cellular target engagement. These compounds represent an important first step towards selective chemical probes for the protein kinase BMX.
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- 2020
134. RNA Sequencing Keloid Transcriptome Associates Keloids With Th2, Th1, Th17/Th22, and JAK3-Skewing
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Ning Zhang, Michael M. Espino, Emma Guttman-Yassky, Ester Del Duca, Ana B. Pavel, Jianni Wu, James G. Krueger, Yeriel Estrada, Inna Cueto, and Alyssa Gontzes
- Subjects
0301 basic medicine ,Pathology ,Lymphocyte Activation ,Transcriptome ,Pathogenesis ,Th2 ,030207 dermatology & venereal diseases ,0302 clinical medicine ,Keloid ,T-Lymphocyte Subsets ,Immunology and Allergy ,skin and connective tissue diseases ,keloids ,CCL11 ,Original Research ,medicine.diagnostic_test ,High-Throughput Nucleotide Sequencing ,Middle Aged ,Immunohistochemistry ,Female ,JAK3 ,Adult ,lcsh:Immunologic diseases. Allergy ,medicine.medical_specialty ,Immunology ,Periostin ,03 medical and health sciences ,Th2 Cells ,Biopsy ,medicine ,Humans ,CXCL10 ,Lymphocyte Count ,Aged ,business.industry ,Gene Expression Profiling ,Janus Kinase 3 ,Th1 Cells ,medicine.disease ,030104 developmental biology ,inflammation ,Th17 Cells ,RNA-seq ,immune ,lcsh:RC581-607 ,business ,Biomarkers - Abstract
Keloids are disfiguring, fibroproliferative growths and their pathogenesis remains unclear, inhibiting therapeutic development. Available treatment options have limited efficacy and harbor safety concerns. Thus, there is a great need to clarify keloid pathomechanisms that may lead to novel treatments. In this study, we aimed to elucidate the profile of lesional and non-lesional keloid skin compared to normal skin. We performed gene (RNAseq, qRT-PCR) and protein (immunohistochemistry) expression analyses on biopsy specimens obtained from lesional and non-lesional skin of African American (AA) keloid patients compared to healthy skin from AA controls. Fold-change≥2 and false-discovery rate (FDR)+, CCR9+, and periostin+ immunostaining. In sum, comprehensive molecular profiling demonstrated that both lesional and non-lesional skin show significant immune alternations, and particularly Th2 and JAK3 expression. This advocates for the investigation of novel treatments targeting the Th2 axis and/or JAK/STAT-signaling in keloid patients.
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- 2020
135. Molecular requirements for human lymphopoiesis as defined by inborn errors of immunity
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Erika Della Mina, Antoine Guérin, and Stuart G. Tangye
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0301 basic medicine ,Lymphocyte ,Primary Immunodeficiency Diseases ,Biology ,03 medical and health sciences ,Ikaros Transcription Factor ,Mice ,0302 clinical medicine ,Immune system ,Immunity ,medicine ,Animals ,Humans ,Paired Box Transcription Factors ,Lymphopoiesis ,Lymphocytes ,Progenitor cell ,Precision Medicine ,Janus Kinase 3 ,Cell Differentiation ,Forkhead Transcription Factors ,Cell Biology ,Genetic Therapy ,medicine.disease ,Hematopoietic Stem Cells ,Hematopoiesis ,Haematopoiesis ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Immunology ,Primary immunodeficiency ,Molecular Medicine ,Stem cell ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Hematopoietic stem cells (HSCs) are the progenitor cells that give rise to the diverse repertoire of all immune cells. As they differentiate, HSCs yield a series of cell states that undergo gradual commitment to become mature blood cells. Studies of hematopoiesis in murine models have provided critical insights about the lineage relationships between stem cells, progenitors, and mature cells and these have guided investigations of the molecular basis for these distinct developmental stages. Primary immune deficiencies are caused by inborn errors of immunity that result in immune dysfunction and subsequent susceptibility to severe and recurrent infection(s). Over the last decade there has been a dramatic increase in the number and depth of the molecular, cellular and clinical characterization of such genetically defined causes of immune dysfunction. Patients harboring inborn errors of immunity thus represent a unique resource to improve our understanding of the multi-layered and complex mechanisms underlying lymphocyte development in humans. These breakthrough discoveries not only enable significant advances in the diagnosis of such rare and complex conditions but also provide substantial improvement in the development of personalized treatments. Here, we will discuss the clinical, cellular and molecular phenotypes and treatments of selected inborn errors of immunity that impede, either intrinsically or extrinsically, the development of B- or T-cells at different stages. © AlphaMed Press 2021 SIGNIFICANCE STATEMENT: Hematopoietic stem cells are critical for lifelong production of blood cells. HSCs are distinctively defined by their self-renewal capacity while contributing to the pool of differentiating cells (ie, lymphocytes, neutrophils, and monocytes/macrophages) that ensure immune regulation and protective immunity against pathogen infection. Here, we discuss how the comprehensive study of a growing number of patients harboring rare germline genetic defects that result in immune dysfunctions and increased susceptibility to infectious disease (collectively named primary immunodeficiencies), have enabled a better understanding of the intricate process of lymphocyte development in human, improved diagnosis and facilitated the development of curative rather than supportive treatments for patients with primary immunodeficiencies.
- Published
- 2020
136. Dual inhibitors of Janus kinase 2 and 3 (JAK2/3): designing by pharmacophore- and docking-based virtual screening approach.
- Author
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Jasuja, Haneesh, Chadha, Navriti, Kaur, Maninder, and Silakari, Om
- Abstract
JAK2 and JAK3 are non-receptor protein tyrosine kinases implicated in B-cell- and T-cell-mediated diseases. Both enzymes work via different pathways but are involved in the pathogenesis of common lymphoid-derived diseases. Hence, targeting both Janus kinases together can be a potential strategy for the treatment of these diseases. In the present study, two separate pharmacophore-based 3D-QSAR models ADRR.92 ( $$Q_{\mathrm{test}}^{2} 0.663, R^{2}_{\mathrm{train}} 0.849$$ , F value 219.3) for JAK2 and ADDRR.142 ( $$Q^{2}_{\mathrm{test}}0.655, R_{\mathrm{train}}^{2}$$ 0.869, F value 206.9) for JAK3 were developed. These models were employed for the screening of a PHASE database of approximately 1.5 million compounds; subsequently, the retrieved hits were screened employing docking simulations with JAK2 and JAK3 proteins. Finally, ADME properties of screened dual inhibitors displaying essential interactions with both proteins were calculated to filter candidates with poor pharmacokinetic profiles. These candidates could serve as novel therapeutic agents in the treatment of lymphoid-related diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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137. Alkynyl Benzoxazines and Dihydroquinazolines as Cysteine Targeting Covalent Warheads and Their Application in Identification of Selective Irreversible Kinase Inhibitors
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McAulay, Kirsten, Hoyt, Emily A, Thomas, Morgan, Schimpl, Marianne, Bodnarchuk, Michael S, Lewis, Hilary J, Barratt, Derek, Bhavsar, Deepa, Robinson, David M, Deery, Michael J, Ogg, Derek J, Bernardes, Gonçalo JL, Ward, Richard A, Waring, Michael J, Kettle, Jason G, Hoyt, Emily A [0000-0001-6940-8803], Schimpl, Marianne [0000-0003-2284-5250], Bodnarchuk, Michael S [0000-0002-9172-1203], Lewis, Hilary J [0000-0002-4661-4564], Bernardes, Gonçalo JL [0000-0001-6594-8917], Ward, Richard A [0000-0002-2310-9477], Waring, Michael J [0000-0002-9110-8783], Kettle, Jason G [0000-0001-7373-0758], and Apollo - University of Cambridge Repository
- Subjects
Models, Molecular ,Molecular Structure ,Quinazolines ,Humans ,Janus Kinase 3 ,Protein Kinase Inhibitors ,Benzoxazines - Abstract
With a resurgence in interest in covalent drugs, there is a need to identify new moieties capable of cysteine bond formation that are differentiated from commonly employed systems such as acrylamide. Herein, we report on the discovery of new alkynyl benzoxazine and dihydroquinazoline moieties capable of covalent reaction with cysteine. Their utility as alternative electrophilic warheads for chemical biological probes and drug molecules is demonstrated through site-selective protein modification and incorporation into kinase drug scaffolds. A potent covalent inhibitor of JAK3 kinase was identified with superior selectivity across the kinome and improvements in in vitro pharmacokinetic profile relative to the related acrylamide-based inhibitor. In addition, the use of a novel heterocycle as a cysteine reactive warhead is employed to target Cys788 in c-KIT, where acrylamide has previously failed to form covalent interactions. These new reactive and selective heterocyclic warheads supplement the current repertoire for cysteine covalent modification while avoiding some of the limitations generally associated with established moieties.
- Published
- 2020
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138. The advantages of describing covalent inhibitor in vitro potencies by IC
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Atli, Thorarensen, Paul, Balbo, Mary E, Banker, Robert M, Czerwinski, Max, Kuhn, Tristan S, Maurer, Jean-Baptiste, Telliez, Fabien, Vincent, and Arthur J, Wittwer
- Subjects
Inhibitory Concentration 50 ,Structure-Activity Relationship ,Dose-Response Relationship, Drug ,Molecular Structure ,Humans ,Janus Kinase 3 ,Protein Kinase Inhibitors ,Recombinant Proteins - Abstract
Recent years have seen a resurgence in drug discovery efforts aimed at the identification of covalent inhibitors which has led to an explosion of literature reports in this area and most importantly new approved therapies. These reports and breakthroughs highlight the significant investments made across the industry in SAR campaigns to optimize inhibitors. The potency of covalent inhibitors is generally considered to be more accurately described by the time-independent kinetic parameter k
- Published
- 2020
139. Design, synthesis, biological activity evaluation of 3-(4-phenyl-1H-imidazol-2-yl)-1H-pyrazole derivatives as potent JAK 2/3 and aurora A/B kinases multi-targeted inhibitors
- Author
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Long Meng, Youguang Zheng, Pei Xin, Lin An, Ling Zhang, Jin-An Wang, Chenglin Li, and Ying-Long Miao
- Subjects
Models, Molecular ,Cell cycle checkpoint ,Aurora B kinase ,Antineoplastic Agents ,Apoptosis ,01 natural sciences ,03 medical and health sciences ,Cell Line, Tumor ,Drug Discovery ,Aurora Kinase B ,Humans ,Cytotoxicity ,Protein Kinase Inhibitors ,030304 developmental biology ,Aurora Kinase A ,Cell Proliferation ,Pharmacology ,0303 health sciences ,Janus kinase 2 ,biology ,Dose-Response Relationship, Drug ,010405 organic chemistry ,Chemistry ,Kinase ,Organic Chemistry ,Janus Kinase 3 ,Biological activity ,General Medicine ,Janus Kinase 2 ,Molecular biology ,Amides ,0104 chemical sciences ,Pyrimidines ,biology.protein ,Phosphorylation ,Pyrazoles ,Drug Screening Assays, Antitumor ,K562 cells - Abstract
In this study, a series of 3-(4-phenyl-1H-imidazol-2-yl)-1H-pyrazole derivatives were designed, synthesized, and evaluated for their biological activities. Upon performing kinase assays, most of the compounds exhibited potent inhibition against JAK2/3 and Aurora A/B with the IC50 values ranging from 0.008 to 2.52 μM. Among these derivatives, compound 10e expressed the most moderate inhibiting activities against all the four kinases with the IC50 values of 0.166 μM (JAK2), 0.057 μM (JAK3), 0.939 μM (Aurora A), and 0.583 μM (Aurora B), respectively. Moreover, most of the derived compounds exhibited potent cytotoxicity against human chronic myeloid leukemia cells K562 and human colon cancer cells HCT116, while compound 10e expressed antiproliferative activities against K562 (IC50=6.726 μM). According to western blot analysis, compound 10e down-regulated the phosphorylation of STAT3, STAT5, Aurora A, and Aurora B in a dose-dependent manner in K562 and HCT116 cells. Cell cycle analysis revealed that compound 10e inhibited the proliferation of cells by inducing cell cycle arrest in the G2 phase. The molecular modeling suggested that compound 10e could maintain a binding mode similar to the binding mode of AT9832, a common JAK 2/3 and Aurora A/B kinases multi-target kinase inhibitor. Therefore, compound 10e might be a potential agent for cancer therapy deserving further research.
- Published
- 2020
140. Whole-exome sequencing of T
- Author
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R, El Hawary, S, Meshaal, A A, Mauracher, L, Opitz, D, Abd Elaziz, S, Lotfy, A, Eldash, J, Boutros, N, Galal, J, Pachlopnik Schmid, and A, Elmarsafy
- Subjects
Family Health ,Male ,T-Lymphocytes ,Infant, Newborn ,Infant ,Janus Kinase 3 ,Original Articles ,Pedigree ,Consanguinity ,Mutation ,Exome Sequencing ,Humans ,Egypt ,Female ,Severe Combined Immunodeficiency ,Lymphocyte Count ,Interleukin Receptor Common gamma Subunit - Abstract
Severe combined immunodeficiency (SCID) is fatal if not treated with immune reconstitution. In Egypt, T(‐)B(+) SCID accounts for 38·5% of SCID diagnoses. An accurate genetic diagnosis is essential for choosing appropriate treatment modalities and for offering genetic counseling to the patient’s family. The objectives of this study were to describe the clinical, immunological and molecular characteristics of a cohort of twenty Egyptian patients with T(‐)B(+) SCID. The initial diagnosis (based on clinical features and flow cytometry) was followed by molecular investigation (whole‐exome sequencing). All patients had the classic clinical picture for SCID, including failure to thrive (n = 20), oral candidiasis (n = 17), persistent diarrhea (n = 14), pneumonia (n = 13), napkin dermatitis (n = 10), skin rash (n = 7), otitis media (n = 3) and meningitis (n = 2). The onset of manifestations was at the age of 2·4 ± 1·6 months and diagnosis at the age of 6·7 ± ·5 months, giving a diagnostic delay of 4·3 months. JAK3 gene variants were most frequent (n = 12) with three novel variants identified, followed by IL2Rγ variants (n = 6) with two novel variants. IL7Rα and CD3ε variants were found once, with a novel variant each. T(‐)B(+)NK(−) SCID accounted for approximately 90% of the Egyptian patients with T(‐)B(+)SCID. Of these T(‐)B(+)NK(−) SCID cases, 60% were autosomal recessive syndromes caused by JAK3 mutations and 30% were X‐linked syndromes. It might be useful to sequence the JAK3 gene (i.e. targeted Sanger sequencing) in all T(‐)B(+) SCID patients, especially after X‐linked SCID has been ruled out. Hence, no more than 10% of T(‐)B(+) SCID patients might require next‐generation for a molecular diagnosis.
- Published
- 2020
141. Cryogenic Ion Spectroscopy of a Singly Protonated Peptide DYYVVR: Locating Phosphorylation Sites of a Kinase Domain
- Author
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Shun-ichi Ishiuchi, Masaaki Fujii, Hyuk Kang, Jang Han Kwon, Gyeongok Song, Min Ji Lee, and Kazuya Tsuruta
- Subjects
chemistry.chemical_classification ,Models, Molecular ,Binding Sites ,Chemistry ,Stereochemistry ,Janus kinase 3 ,Spectrum Analysis ,Protonation ,Peptide ,Ion ,Protein kinase domain ,Protein Domains ,Domain (ring theory) ,General Materials Science ,Amino Acid Sequence ,Physical and Theoretical Chemistry ,Tyrosine ,Phosphorylation ,Protons ,Spectroscopy ,Oligopeptides ,Protein Kinases - Abstract
Cryogenic ion spectroscopy (CIS) was applied to singly protonated DYYVVR, a tryptic peptide that contains the two active tyrosine residues (Y980 and Y981) of the Janus kinase 3 (JAK3) kinase domain, together with its point mutants (Y980F and Y981F) and phosphorylated peptides (pY980, pY981, and pY980pY981). The two tyrosine chromophores showed distinguishable UV absorption bands at around 35 200 and 35 450 cm
- Published
- 2020
142. Investigation of Alpinia calcarata constituent interactions with molecular targets of rheumatoid arthritis: docking, molecular dynamics, and network approach
- Author
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Sree Karani Kondapuram, Sanmuga Priya Ekambaram, Thamizharasi Erusappan, and Mohane Selvaraj Coumar
- Subjects
Phytochemicals ,Syk ,Molecular Dynamics Simulation ,010402 general chemistry ,01 natural sciences ,Catalysis ,Mass Spectrometry ,Inorganic Chemistry ,Arthritis, Rheumatoid ,chemistry.chemical_compound ,0103 physical sciences ,Humans ,Syk Kinase ,Physical and Theoretical Chemistry ,Kaempferols ,Flavonoids ,010304 chemical physics ,biology ,Traditional medicine ,Plant Extracts ,Organic Chemistry ,Computational Biology ,Janus Kinase 3 ,Janus Kinase 1 ,biology.organism_classification ,Molecular medicine ,0104 chemical sciences ,Computer Science Applications ,Rhizome ,Galangin ,Molecular Docking Simulation ,Interleukin-1 Receptor-Associated Kinases ,Computational Theory and Mathematics ,chemistry ,Docking (molecular) ,Toll-Like Receptor 8 ,Alpinia ,Matrix Metalloproteinase 2 ,Zingiberaceae ,Quercetin ,Kaempferol ,Chromatography, Liquid - Abstract
Rheumatoid arthritis (RA) is a systemic autoimmune disorder that commonly affects multiple joints of the body. Currently, there is no permanent cure to the disease, but it can be managed with several potent drugs that cause serious side effects on prolonged use. Traditional remedies are considered promising for the treatment of several diseases, particularly chronic conditions, because they have lower side effects compared to synthetic drugs. In folklore, the rhizome of Alpinia calcarata Roscoe (Zingiberaceae) is used as a major ingredient of herbal formulations to treat RA. Phytoconstituents reported in A. calcarata rhizomes are diterpenoids, sesquiterpenoid, flavonoids, phytosterol, and volatile oils. The present study is intended to understand the molecular-level interaction of phytoconstituents present in A. calcarata rhizomes with RA molecular targets using computational approaches. A total of 30 phytoconstituents reported from the plant were used to carry out docking with 36 known targets of RA. Based on the docking results, 4 flavonoids were found to be strongly interacting with the RA targets. Further, molecular dynamics simulation confirmed stable interaction of quercetin with 6 targets (JAK3, SYK, MMP2, TLR8, IRAK1, and JAK1), galangin with 2 targets (IRAK1 and JAK1), and kaempferol (IRAK1) with one target of RA. Moreover, the presence of these three flavonoids was confirmed in the A. calcarata rhizome extract using LC-MS analysis. The computational study suggests that flavonoids present in A. calcarata rhizome may be responsible for RA modulatory activity. Particularly, quercetin and galangin could be potential development candidates for the treatment of RA. Investigation of Alpinia calcarata constituent interactions with molecular targets of rheumatoid arthritis: docking, molecular dynamics, and network approach.
- Published
- 2020
143. Combined anti-fibrotic and anti-inflammatory properties of JAK-inhibitors on macrophages in vitro and in vivo perspectives for scleroderma-associated interstitial lung disease
- Author
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Valérie Lecureur, Marie Lelong, Patrick Jego, Laurent Vernhet, Frédéric Batteux, Claire Piquet-Pellorce, Mohamed Jeljeli, Olivier Fardel, Alice Ballerie, Stéphane Jouneau, Alain Lescoat, Claudie Morzadec, Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), École des Hautes Études en Santé Publique [EHESP] (EHESP), Institut National de la Santé et de la Recherche Médicale, Université de Rennes, Association des sclérodermiques de France, Jonchère, Laurent, Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)
- Subjects
0301 basic medicine ,Anti-Inflammatory Agents ,Biochemistry ,[SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract ,Mice ,0302 clinical medicine ,Piperidines ,Pulmonary fibrosis ,Receptors, Immunologic ,Lung ,Chemistry ,Cell Differentiation ,3. Good health ,Tyrosine kinase 2 ,030220 oncology & carcinogenesis ,Systemic sclerosis ,Female ,Tumor necrosis factor alpha ,medicine.symptom ,[SDV.IMM.IMM] Life Sciences [q-bio]/Immunology/Immunotherapy ,Primary Cell Culture ,Inflammation ,03 medical and health sciences ,macrophage activation ,Nitriles ,medicine ,Animals ,CXCL10 ,Pyrroles ,Protein Kinase Inhibitors ,Pharmacology ,Janus kinase inhibitors ,Scleroderma, Systemic ,Interstitial Lung disease ,Macrophages ,CCL18 ,Janus Kinase 3 ,Janus Kinase 1 ,[SDV.IMM.IMM]Life Sciences [q-bio]/Immunology/Immunotherapy ,Janus Kinase 2 ,medicine.disease ,Chemokine CXCL13 ,Hypochlorous Acid ,Mice, Inbred C57BL ,Pyrimidines ,030104 developmental biology ,Gene Expression Regulation ,Suppressor of Cytokine Signaling 3 Protein ,TLR4 ,Cancer research ,Pyrazoles ,[SDV.MHEP.PSR] Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract ,Lung Diseases, Interstitial ,Janus kinase - Abstract
International audience; Janus kinase (JAK) inhibitors (also termed Jakinibs) constitute a family of small drugs that target various isoforms of JAKs (JAK1, JAK2, JAK3 and/or tyrosine kinase 2 (Tyk2)). They exert anti-inflammatory properties linked, in part, to the modulation of the activation state of pro-inflammatory M1 macrophages. The exact impact of JAK inhibitors on a wider spectrum of activation states of macrophages is however still to be determined, especially in the context of disorders involving concomitant activation of pro-inflammatory M1 macrophages and profibrotic M2 macrophages. This is especially the case in autoimmune pulmonary fibrosis like scleroderma-associated interstitial lung disease (ILD), in which M1 and M2 macrophages play a key pathogenic role. In this study, we directly compared the anti-inflammatory and anti-fibrotic effects of three JAK inhibitors (ruxolitinib (JAK2/1 inhibitor); tofacitinib (JAK3/2 inhibitor) and itacitinib (JAK1 inhibitor)) on five different activation states of primary human monocyte-derived macrophages (MDM). These three JAK inhibitors exert anti-inflammatory properties towards macrophages, as demonstrated by the down-expression of key polarization markers (CD86, MHCII, TLR4) and the limited secretion of key pro-inflammatory cytokines (CXCL10, IL-6 and TNFα) in M1 macrophages activated by IFNγ and LPS or by IFNγ alone. We also highlighted that these JAK inhibitors can limit M2a activation of macrophages induced by IL-4 and IL-13, as notably demonstrated by the down-regulation of the M2a associated surface marker CD206 and of the secretion of CCL18. Moreover, these JAK inhibitors reduced the expression of markers such as CXCL13, MARCO and SOCS3 in alternatively activated macrophages induced by IL-10 and dexamethasone (M2c+dex) or IL-10 alone (M2c MDM). For all polarization states, Jakinibs with inhibitory properties over JAK2 had the highest effects, at both 1 μM or 0.1 μM. Based on these in vitro results, we also explored the effects of JAK2/1 inhibition by ruxolitinib in vivo, on mouse macrophages in a model of HOCl-induced ILD, that mimics scleroderma-associated ILD. In this model, we showed that ruxolitinib significantly prevented the upregulation of pro-inflammatory M1 markers (TNFα, CXCL10, NOS2) and pro-fibrotic M2 markers (Arg1 and Chi3L3). These results were associated with an improvement of skin and pulmonary involvement. Overall, our results suggest that the combined anti-inflammatory and anti-fibrotic properties of JAK2/1 inhibitors could be relevant to target lung macrophages in autoimmune and inflammatory pulmonary disorders that have no efficient disease modifying drugs to date.
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- 2020
144. JANEX-1 improves acute pulmonary embolism through VEGF and FAK in pulmonary artery smooth muscle cells
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Heming Chen, Longfei Pan, Zhuo Peng, Dean Liang, Ruipeng Zhang, Hongyan Tian, and Rui Zhang
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Male ,STAT3 Transcription Factor ,Vascular Endothelial Growth Factor A ,0301 basic medicine ,medicine.medical_specialty ,Common disease ,VEGF receptors ,Myocytes, Smooth Muscle ,Pulmonary Artery ,Vascular Remodeling ,030204 cardiovascular system & hematology ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,Smooth muscle ,Internal medicine ,medicine.artery ,medicine ,Animals ,Phosphorylation ,Lung ,Clinical treatment ,Cell Proliferation ,Original Research ,Platelet-Derived Growth Factor ,biology ,business.industry ,Janus kinase 3 ,Janus Kinase 3 ,JAK Family ,medicine.disease ,Pulmonary embolism ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,Focal Adhesion Protein-Tyrosine Kinases ,Acute Disease ,Pulmonary artery ,Quinazolines ,Cardiology ,biology.protein ,Pulmonary Embolism ,Corrigendum ,business - Abstract
Although clinical treatment has significant progress, acute pulmonary embolism is still a common disease with high morbidity and mortality. Janus Kinase 3, a member of JAK family, has been demonstrated to promote smooth muscle cell proliferation through STAT3. In this work, we explored the effect of JANEX-1 (a specific Janus Kinase 3 inhibitor) on platelet-derived growth factor (PDGF)-induced proliferation-related molecules in pulmonary artery smooth muscle cells (PVSMCs) in vitro and assessed the therapeutic potential of Janus Kinase 3 for vascular remodeling in acute pulmonary embolism mice. The results revealed that Janus Kinase 3 was overexpressed and active in PDGF-induced PVSMCs and acute pulmonary embolism mice, compared to a low expression in normal conditions. JANEX-1, blocking Janus Kinase 3 expression or activity, reduced Janus Kinase 3/STAT3 signaling pathway, VEGF expression, FAK activation, and PDGF-induced proliferation of PVSMCs, while overexpression of VEGF or FAK induced PVSMCs proliferation and resisted the negative effects of JANEX-1. Moreover, JANEX-1 improved right ventricular systolic pressure, survival and lung damage in acute pulmonary embolism-mice, and inhibited the thrombus-induced intimal hyperplasia and the expression of α-SMA, VEGF, and FAK activation under neointimal smooth muscle cells of acute pulmonary embolism mice. In conclusion, the data suggest that JANEX-1 exerts protective effects by inhibiting PVSMCs proliferation and vascular remodeling post-acute pulmonary embolism, in part through Janus Kinase 3/STAT3 signaling pathway-mediated VEGF expression and FAK activation. The data are helpful to elucidate the pharmacological mechanism and potential therapeutic effect of JANEX-1 in APE.Impact statementAccumulating evidence suggests that vascular remodeling due to immoderate proliferation and migration of SMCs is a common process occurring in APE. In this work, we tried to find a breakthrough in the pathological mechanism to alleviate the prognosis of APE by improving SMCs proliferation and explored the effect of JANEX-1 on PDGF-induced proliferation-related molecules in PVSMCs and assessed the therapeutic potential of JAK3 for vascular remodeling in APE mice. We demonstrated that JANEX-1, blocking JAK3 expression or activity, reduced JAK3/STAT3 signaling pathway, VEGF expression and FAK activation, and PDGF-induced proliferation of PVSMCs. Moreover, JANEX-1 inhibited the thrombus-induced intimal hyperplasia and the expression of VEGF and FAK activation in neointimal SMCs of APE mice. The data are helpful to elucidate the pharmacological mechanism and potential therapeutic effect of JANEX-1 in APE.
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- 2020
145. MKL-1 is a coactivator for STAT5b, the regulator of Treg cell development and function
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Kun Chen, Han Han Li, Jun Wang, Zhang Zijian, Zhang Huimin, Zhou Tong Dai, Tong-Cun Zhang, Yuan Xiang, Leyuan Bao, Chao Jiang Gu, Feng Huang, Xing-Hua Liao, Hui Li, Jia Peng Li, and Wei Guo
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Serum Response Factor ,Small interfering RNA ,CD3 Complex ,Pyridines ,Regulator ,lcsh:Medicine ,T-Lymphocytes, Regulatory ,Biochemistry ,STAT5b ,0302 clinical medicine ,STAT5 Transcription Factor ,Phosphorylation ,Promoter Regions, Genetic ,Regulation of gene expression ,0303 health sciences ,Gene knockdown ,lcsh:Cytology ,food and beverages ,FOXP3 ,Forkhead Transcription Factors ,hemic and immune systems ,Tyrphostins ,Cell biology ,Treg ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Protein Binding ,Signal Transduction ,animal structures ,T cell ,chemical and pharmacologic phenomena ,Biology ,03 medical and health sciences ,Protein Domains ,Coactivator ,medicine ,Humans ,Lymphocyte Count ,lcsh:QH573-671 ,Molecular Biology ,Transcription factor ,030304 developmental biology ,Cell Nucleus ,Purpura, Thrombocytopenic, Idiopathic ,Base Sequence ,Research ,lcsh:R ,Interleukin-2 Receptor alpha Subunit ,MKL-1 ,Janus Kinase 3 ,Cell Biology ,Amides ,Trans-Activators ,Interleukin-2 ,ITP ,rhoA GTP-Binding Protein ,Biomarkers - Abstract
BackgroundFoxp3+CD4+regulatory T cells (Treg) constitutes a key event in autoimmune diseases. STAT5b is the critical link between the IL-2/15 and FOXP3, the master regulator of Treg cells.MethodsThe CD3+T cell and Foxp3+CD4+regulatory T cells were overexpressioned or knockdown MKL-1 and STAT5a and tested for Treg cell development and function. Direct interaction of MKL-1 and STAT5a were analyzed by coimmunoprecipitation assays, Luciferase assay, Immunofluoresence Staining and Yeast two-hybrid screening. The effect of MKL-1 and STAT5a on the Treg genes expression was analyzed by qPCR and western blotting and Flow cytometry.ResultsHowever, the molecular mechanisms mediating STAT5b-dependent Treg genes expression and Treg cell phenotype and function in autoimmune diseases are not well defined. Here, we report that the MKL-1 is a coactivator for the major Treg genes transcription factor STAT5b, which is required for human Treg cell phenotype and function. The N terminus of STAT5b, which contains a basic coiled-coil protein–protein interaction domain, binds the C-terminal activation domain of MKL-1 and enhances MKL-1 mediated transcriptional activation of Treg-specific, CArG containing promoters, including the Treg-specific genes Foxp3. Suppression of endogenous STAT5b expression by specific small interfering RNA attenuates MKL-1 transcriptional activation in cultured human cells. The STAT5b–MKL-1 interaction identifies a role of Treg-specific gene regulation and regulated mouse Treg cell development and function and suggests a possible mechanism for the protective effects of autoimmune disease Idiopathic Thrombocytopenic Purpura (ITP).ConclusionsOur studies demonstrate for the first time that MKL-1 is a coactivator for STAT5b, the regulator of Treg cell development and function.
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- 2020
146. Kinome capture sequencing of high-grade serous ovarian carcinoma reveals novel mutations in theJAK3gene
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Wim Brugman, Marja Nieuwland, Magali Michaut, Nitzan Rosenfeld, Mercedes Jimenez-Linan, Ate G.J. van der Zee, Astrid Bosma, Roelof J.C. Kluin, René Bernards, Steven de Jong, James D. Brenton, Kevin J. W. van der Ven, Lorenza Mittempergher, Anna M. Piskorz, Francesco Marass, G. Bea A. Wisman, James Morris, Targeted Gynaecologic Oncology (TARGON), Guided Treatment in Optimal Selected Cancer Patients (GUTS), Mittempergher, Lorenza [0000-0003-3425-3965], Marass, Francesco [0000-0002-8993-7320], and Apollo - University of Cambridge Repository
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0301 basic medicine ,Epidemiology ,medicine.disease_cause ,Cohort Studies ,Database and Informatics Methods ,0302 clinical medicine ,Mutation frequency ,Frameshift Mutation ,Ovarian Neoplasms ,education.field_of_study ,Mutation ,Multidisciplinary ,BRCA1 Protein ,Cancer Risk Factors ,High-Throughput Nucleotide Sequencing ,Nonsense Mutation ,CANCER ,Ovarian Cancer ,Oncology ,030220 oncology & carcinogenesis ,Medicine ,Female ,JAK3 ,Research Article ,EXPRESSION ,GENES ,Science ,Population ,Nonsense mutation ,Genetic Causes of Cancer ,BEVACIZUMAB ,Biology ,Frameshift mutation ,03 medical and health sciences ,Germline mutation ,medicine ,Genetics ,Point Mutation ,Humans ,education ,Medicine and health sciences ,REPAIR ,Biology and life sciences ,Point mutation ,Cancers and Neoplasms ,Janus Kinase 3 ,DNA ,Cystadenocarcinoma, Serous ,Research and analysis methods ,030104 developmental biology ,Biological Databases ,Medical Risk Factors ,Case-Control Studies ,Mutation Databases ,Cancer research ,Somatic Mutation ,Tumor Suppressor Protein p53 ,CDK12 ,Gynecological Tumors ,Protein Kinases ,GENOMIC INSTABILITY - Abstract
High-grade serous ovarian carcinoma (HGSOC) remains the deadliest form of epithelial ovarian cancer and despite major efforts little improvement in overall survival has been achieved. Identification of recurring "driver" genetic lesions has the potential to enable design of novel therapies for cancer. Here, we report on a study to find such new therapeutic targets for HGSOC using exome-capture sequencing approach targeting all kinase genes in 127 patient samples. Consistent with previous reports, the most frequently mutated gene wasTP53(97% mutation frequency) followed byBRCA1(10% mutation frequency). The average mutation frequency of the kinase genes mutated from our panel was 1.5%. Intriguingly, afterBRCA1,JAK3was the most frequently mutated gene (4% mutation frequency). We tested the transforming properties of JAK3 mutants using the Ba/F3 cell-basedin vitrofunctional assay and identified a novel gain-of-function mutation in the kinase domain ofJAK3(p.T1022I). Importantly, p.T1022IJAK3mutants displayed higher sensitivity to the JAK3-selective inhibitor Tofacitinib compared to controls. For independent validation, we re-sequenced the entireJAK3coding sequence using tagged amplicon sequencing (TAm-Seq) in 463 HGSOCs resulting in an overall somatic mutation frequency of 1%. TAm-Seq screening ofCDK12in the same population revealed a 7% mutation frequency. Our data confirms that the frequency of mutations in kinase genes in HGSOC is low and provides accurate estimates for the frequency ofJAK3andCDK12mutations in a large well characterized cohort. Although p.T1022IJAK3mutations are rare, our functional validation shows that if detected they should be considered as potentially actionable for therapy. The observation ofCDK12mutations in 7% of HGSOC cases provides a strong rationale for routine somatic testing, although more functional and clinical characterization is required to understand which nonsynonymous mutations alterations are associated with homologous recombination deficiency., PLoS ONE, 15 (7), ISSN:1932-6203
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- 2020
147. MicroRNA-495 alleviates ulcerative interstitial cystitis via inactivating the JAK-STAT signaling pathway by inhibiting JAK3
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Wei Huo, Yi Hou, and Hai Li
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Urology ,030232 urology & nephrology ,Cystitis, Interstitial ,03 medical and health sciences ,0302 clinical medicine ,Western blot ,microRNA ,medicine ,Animals ,STAT1 ,STAT3 ,030219 obstetrics & reproductive medicine ,medicine.diagnostic_test ,biology ,business.industry ,Tumor Necrosis Factor-alpha ,Obstetrics and Gynecology ,JAK-STAT signaling pathway ,Interstitial cystitis ,Janus Kinase 3 ,medicine.disease ,Rats ,MicroRNAs ,Cancer research ,biology.protein ,Immunohistochemistry ,Female ,Signal transduction ,business ,Signal Transduction - Abstract
As a notable chronic disorder, the incidence of interstitial cystitis (IC) has been documented to have increased among the female population with activity in microRNA-495 (miR-495) implicated in this disease. The current study was aimed at elucidating the effects associated with miR-495 on the inflammatory response and bladder fibrosis in rats with ulcerative IC via the JAK–STAT pathway by targeting JAK3. Ulcerative IC rat models were established. The targeting relationship between JAK3 and miR-495 was evaluated using luciferase reporter assay. After gain- or loss-of-function assays, mast-cell infiltration was assessed using toluidine blue staining, bladder fibrosis using Masson staining, and NO content using nitrate reductase method. JAK3 protein expression was detected by immunohistochemistry, JAK3, STAT1, STAT3, TGFβ-1, Col-I, Col-III, JAK1, JAK2, p-STAT1, and p-STAT3 expression by RT-qPCR and Western blot analysis, and serum IL-6, IL-8, IL-10, IL-17, and TNF-α levels in rats by ELISA. Following transfection of overexpressed miR-495 or siRNA-JAK3, a diminished degree of mast-cell infiltration, number of mast cells, bladder fibrosis, NO content, JAK3-positive expression, mRNA expression of JAK3, STAT1, STAT3, TGFβ-1, Col-I, Col-III, protein expression of JAK1, JAK2, JAK3, p-STAT1, p-STAT3, and expression of IL-6, IL-8, IL-10, IL-17, and TNF-α were identified. Collectively, our key findings provide evidence supporting the notion that the overexpression of miR-495 ameliorates inflammatory response and bladder fibrosis in ulcerative IC rat models via inactivation of the JAK–STAT signaling pathway by inhibiting JAK3.
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- 2020
148. Q: Is it safe to continue biologic agents during surgery in patients with inflammatory bowel disease?
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Douglas L. Nguyen, Jacqueline Duong, Zain Moosvi, and Alexander Abadir
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medicine.medical_specialty ,medicine.drug_class ,Medication Therapy Management ,Integrin ,Anti-Inflammatory Agents ,Monoclonal antibody ,Inflammatory bowel disease ,Perioperative Care ,03 medical and health sciences ,0302 clinical medicine ,Piperidines ,medicine ,Humans ,Surgical Wound Infection ,In patient ,030212 general & internal medicine ,Biological Products ,Tofacitinib ,biology ,business.industry ,Interleukin ,Janus Kinase 3 ,General Medicine ,medicine.disease ,Inflammatory Bowel Diseases ,Surgery ,Biologic Agents ,Pyrimidines ,Elective Surgical Procedures ,biology.protein ,Tumor necrosis factor alpha ,Risk Adjustment ,Tumor Necrosis Factor Inhibitors ,business - Abstract
Patients with inflammatory bowel disease who are taking monoclonal antibodies against tumor necrosis factor alpha (TNF), interleukins 12 and 23, or integrin can continue taking them around the time of surgery, but small-molecule drugs such as tofacitinib should be withheld. Inflammatory bowel
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- 2020
149. Hair Growth Effect of Emulsion Extracted Brevilin A, a JAK3 Inhibitor, from Centipeda minima
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Tuy An Trinh, Jae Sung Pyo, Won-Yung Lee, Chang-Eop Kim, Eun-Seok Park, Sooyeun Lee, Byoung Ha Kim, Ki Sung Kang, and Dong Hwan Lee
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0301 basic medicine ,Bioengineering ,Pharmacology ,Terminal hair ,Placebo ,lcsh:Chemical technology ,Centipeda minima ,lcsh:Chemistry ,03 medical and health sciences ,0302 clinical medicine ,hair growth ,medicine ,Chemical Engineering (miscellaneous) ,network pharmacology ,lcsh:TP1-1185 ,Receptor ,Janus kinase-signal transducer and activator of transcription signaling pathway ,integumentary system ,Chemistry ,Process Chemistry and Technology ,Janus kinase 3 ,brevilin A ,medicine.disease ,030104 developmental biology ,lcsh:QD1-999 ,030220 oncology & carcinogenesis ,Alopecia universalis ,Emulsion ,Signal transduction - Abstract
Janus kinase 3 (JAK3) inhibitors have been used effectively in the treatment of several cases of alopecia universalis and its variants. Our study aims to evaluate whether the emulsion extract of brevilin A from Centipeda minima (CMX) stimulates hair regrowth in a clinical trial, as a JAK3 inhibitor, combined with network pharmacology-based analysis. CMX showed potent inhibition of JAK3 in a concentration-dependent manner. Significant differences in total hair count, terminal hair count, and anagen hair count from the baseline to 24 weeks were observed between the placebo and CMX subjects. The gene set enrichment analysis showed that the targets of CMX are mainly associated with the JAK-STAT signaling pathway, cytokine&ndash, cytokine receptor interactions, and the MAPK signaling pathway. This study suggests that the medicinal herbal extract CMX is useful in the treatment of mild to moderate vertex balding that contribute to the visible improvements in hair growth observed in treated patients.
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- 2020
150. JAK3 restrains inflammatory responses and protects against periodontal disease through Wnt3a signaling
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Xiaoxian Duan, Huizhi Wang, Junling Ren, Lanhai Lu, Richard J. Lamont, Zhen Gu, Muddasir Mohammed, Shuang Liang, Lan Yakoumatos, David A. Scott, Silvia M. Uriarte, and Huaxin Zhou
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0301 basic medicine ,medicine.medical_treatment ,Inflammation ,Biology ,Protective Agents ,Biochemistry ,Article ,Periodontal pathogen ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Wnt3A Protein ,Bacteroidaceae Infections ,Genetics ,medicine ,Animals ,Bone Resorption ,Molecular Biology ,Porphyromonas gingivalis ,Periodontal Diseases ,Innate immune system ,Janus kinase 3 ,Wnt signaling pathway ,Janus Kinase 3 ,biology.organism_classification ,Cell biology ,Ubiquitin ligase ,Mice, Inbred C57BL ,030104 developmental biology ,Cytokine ,biology.protein ,medicine.symptom ,030217 neurology & neurosurgery ,Signal Transduction ,Biotechnology - Abstract
Homeostasis between pro- and anti- inflammatory responses induced by bacteria is critical for the maintenance of health. In the oral cavity, pro-inflammatory mechanisms induced by pathogenic bacteria are well-established; however, the anti-inflammatory responses that act to restrain innate responses remain poorly characterized. Here, we demonstrate that infection with the periodontal pathogen Porphyromonas gingivalis enhances the activity of Janus kinase 3 (JAK3) in innate immune cells, and subsequently phospho-inactivates Nedd4-2, an ubiquitin E3 ligase. In turn, Wingless-INT (Wnt) 3 (Wnt3) ubiquitination is decreased, while total protein levels are enhanced, leading to a reduction in pro-inflammatory cytokine levels. In contrast, JAK3 or Wnt3a inhibition robustly enhances nuclear factor kappa-light-chain-enhancer of activated B cells activity and the production of pro-inflammatory cytokines in P. gingivalis-stimulated innate immune cells. Moreover, using gain- and loss-of-function approaches, we demonstrate that downstream molecules of Wnt3a signaling, including Dvl3 and β-catenin, are responsible for the negative regulatory role of Wnt3a. In addition, using an in vivo P. gingivalis-mediated periodontal disease model, we show that JAK3 inhibition enhances infiltration of inflammatory cells, reduces expression of Wnt3a and Dvl3 in P. gingivalis-infected gingival tissues, and increases disease severity. Together, our results reveal a new anti-inflammatory role for JAK3 in innate immune cells and show that the underlying signaling pathway involves Nedd4-2-mediated Wnt3a ubiquitination.
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- 2020
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