DOCSLIB.ORG

The Role of Janus Kinase 3 in CD4+ T Cell Homeostasis and Function: a Dissertation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Role of Janus Kinase 3 in CD4+ T Cell Homeostasis and Function: a Dissertation University of Massachusetts Medical School eScholarship@UMMS GSBS Dissertations and Theses Graduate School of Biomedical Sciences 2004-09-13 The role of Janus Kinase 3 in CD4+ T Cell Homeostasis and Function: A Dissertation Shane Renee Mayack University of Massachusetts Medical School Let us know how access to this document benefits ou.y Follow this and additional works at: https://escholarship.umassmed.edu/gsbs_diss Part of the Amino Acids, Peptides, and Proteins Commons, Cells Commons, Enzymes and Coenzymes Commons, and the Hemic and Immune Systems Commons Repository Citation Mayack SR. (2004). The role of Janus Kinase 3 in CD4+ T Cell Homeostasis and Function: A Dissertation. GSBS Dissertations and Theses. https://doi.org/10.13028/4qq7-k514. Retrieved from https://escholarship.umassmed.edu/gsbs_diss/94 This material is brought to you by eScholarship@UMMS. It has been accepted for inclusion in GSBS Dissertations and Theses by an authorized administrator of eScholarship@UMMS. For more information, please contact [email protected]. THE ROLE OF JANUS KINASE 3 IN CD4+ T CELL HOMEOSTASIS AND FUNCTION A Dissertation Presented BY Shane Renee Mayack Submitted to the Faculty of the University of Massachusetts Graduate School of Biomedical Sciences, Worcester in partial llfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY September 13,2004 Interdisciplinary Graduate Program Department of Pathology Program in ImmunologyiVirology Copyright Information The content of this dissertation has appeared in the following publication: Mayack, S. Berg., L. (2004) Alternative CD4+ T cell differentiation in the absence of Jak3. Submitted for publication. Joonsoo Kang, Chair of Committee Alan Rothman, Member of Committee Robert Woodland, Member of Committee Katherine Luzuriaga, Member of Committee Gordon Freeman, Member of Committee Leslie J. Berg, Dissertation Mentor Anthony Carruthers, Dean of the Graduate School of Biomedical Sciences I would like to thank my longtime boyfnend David who has been by my side throughout my pursuit for higher education. It is him that I have looked to all of these years for advice and inspiration and as a model of honesty, integrity and ambition. It is him that has always, always provided me with a reason to smile. You have enriched every aspect of my life. Thank you for your love and support and the unwavering confidence you have had in me throughout the years. We are a team and I could not have done this without you. I would like to thank my parents. They are truly the most selfless people I have ever encountered. They have both, especially my Mother, sacrificed their entire lives for me and my siblings. For this my gratitude cannot be put into words. In addition, by way of example, they have taught me two very important lessons. The fist was to have an exemplary work ethic even in the face of seemingly little return. The second was to always have a positive attitude, be able to laugh and to persevere even under unpleasant circumstances. Emulating these traits have contributed greatly to my accomplishments in life and are particularly applicable to the many hurdles one faces in science. Thank you for instilling in me that I could achieve anything I wanted to in life. In addition, I am grateful to my mentor Dr. Leslie Berg for welcoming me into the lab without a second thought. Not only has she provided me with sound scientific guidance but also, by allowing me the freedom to pursue the questions of my choosing, has allowed me to gain the confidence in and passion for science that I had temporarily lost throughout my graduate career. Most remarkable though, is the positive and well rounded role model Leslie provides, in general, but particularly for women. She makes balancing a successful career and pursuing a family, or whatever else is important to you, not only seem obtainable but almost effortless. What truly sets Leslie apart as a mentor is her patience and professionalism, unwavering confidence in her students' abilities, and that she always seems to know exactly what to say and how to say it. Thank you for providing a stimulating and challenging yet supportive scientific environment which has allowed me to recognize my own abilities and grow even stronger as a scientist. I would like to acknowledge all of the members of the Berg lab and the entire Pathology department. The openness, collaborative and collegial nature of all of the investigators, post-docs, students and technicians in the Pathology department make it a great place to be a graduate student and significantly contributes to an environment conducive to learning and productivity. I would especially like to thank Sara Gozalo and Dr. Morgan Wallace for helping me get started in the lab. Regina Whitehead for ordering all of my reagents and dealing with any and every issue in a direct and timely fashion. Dr. Joe Maciaszek and Dr. SK Kim for assistance with tail vein injections. I would like to thank Min Shi and Amanda Prince for generating the RT-PCR data presented in this dissertation and for their support and confidence in me. In addition, I would like to thank them both for the opportunity to act (partially) as their mentor when they first joined the lab. I have learned as much from the both of you as I can only hope you have learned from me. I am indebted to the UMASS Sorting Facility, especially Tammy Krumpoch, for isolation of CD4+ CD44-high T cell populations and the UMASS Affyrnetrix Core Facility for performing the microarray hybridizations. I would like to thank Dr. Maria Zapp and Dr. Ellie Kittler for investing their time and resources in me early in my graduate career. During this time, I learned a great deal about science and life. I would like to thank all of the members of my thesis defense committee---Dr. Robert Woodland, Dr. Alan Rothrnan, Dr. Joonsoo Kang, Dr. Katherine Luzuriaga, and Dr. Gordon Freeman for participating in my thesis examination and for all of the time and energy a number of you have spent training me to be a competent scientist. I would especially like to thank Dr. Robert Woodland. At the lowest point in my graduate school career you were there for me and handled a somewhat sensitive matter with the utmost professionalism and concern for my well-being. Your faith in me during that time is the reason I remained in graduate school. Thank You. And, finally, a special thanks to Dr. Joonsoo Kang who has been extremely supportive of all of my research and provided the STATSab-deficient mice for experiments described in ths thesis. In addition, his high expectations provide an extra challenge for all of the students and in the end I think we are all stronger for it. He is an excellent addition to the UMASS community. vii Abstract This dissertation addresses the role for Janus Kinase 3 (Jak3) in CD4+ T cell homeostasis and function. Jak3 is a protein tyrosine kinase whose activity is essential for signals mediated by the yc dependent cytokines IL-2, -4, -7,-9,-15, and -21. Previous data have demonstrated that peripheral CD4+ T cells from Jak3-deficient mice have a memory phenotype and are functionally impaired in both proliferative and IL-2 responses in vitro. Interestingly, Jak3/yc activity has been previously shown to play a role in the prevention of T cell anergy. These studies were initiated to more precisely define the role for Jak31yc cytohnes in the prevention of T cell anergy and the maintenance of functional CD4+ T cell responses. We began to address this question by assessing global gene expression changes between wild type and Jak3-I- CD4+ T cells. These data indicate that Jak3-I- CD4+ T cells have an increase in gene expression levels of inhibitory surface receptors as well as immunosuppressive cytokines. Further analyses confirmed that Jak3-deficient T cells express high levels of PD-1, secrete a Trl-type cytokine profile following direct ex vivo activation, and suppress the proliferation of wild type T cells in vitro. These characteristics indicate that CD4+ Jak3-/- T cells share properties with regulatory T cell subsets that have an important role in peripheral tolerance and the prevention of autoimmunity. We next addressed whether these regulatory characteristics were T cell intrinsic or rather the result of expanding in a Jak3-deficient microenvironment characterized by a number of immune abnormalities and a disrupted splenic architecture. Jak3-I- CD4+ T cells proliferate in vivo in a lymphopenic environment and selectively acquire regulatory T cell characteristics in the absence of any additional activation signals. While the precise mechanism by which Jak3-deficient T cells acquire these characteristics remains unclear, our data indicate that one important component is a T cell-intrinsic requirement for Jak3 signaling. These findings indicate several interesting aspects of T cell biology. First, these studies, demonstrate that the homeostatic proliferation of CD4+ T cells is not dependent on signaling via yc-dependent cytokine receptors. And, second, that the weak activation signals normally associated with homeostatic expansion are sufficient to drive Jak3-1- T cells into a non- conventional differentiation program. Previous data indicate that, for wild type T cells, signaling through both the TCR as well as yc-dependent cytokine receptors promote the homeostatic proliferation of T cells in lymphopenic hosts. Since Jak3-I- T cells are unable to receive these cytokine signals, their proliferation is likely to be wholly dependent on TCR signaling. As a consequence of this TCR signaling, Jak3-I- T cells proliferate, but in addition, are induced to up regulate PD-1 and to selectively activate the IL-10 locus while shutting off the production of IL-2.
Load more

Recommended publications
  • Promoter Janus Kinase 3 Proximal Characterization and Analysis Of
    The Journal of Immunology Characterization and Analysis of the Proximal Janus Kinase 3 Promoter1 Martin Aringer,2*† Sigrun R. Hofmann,2* David M. Frucht,* Min Chen,* Michael Centola,* Akio Morinobu,* Roberta Visconti,* Daniel L. Kastner,* Josef S. Smolen,† and John J. O’Shea3* Janus kinase 3 (Jak3) is a nonreceptor tyrosine kinase essential for signaling via cytokine receptors that comprise the common ␥-chain (␥c), i.e., the receptors for IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21. Jak3 is preferentially expressed in hemopoietic cells and is up-regulated upon cell differentiation and activation. Despite the importance of Jak3 in lymphoid development and immune function, the mechanisms that govern its expression have not been defined. To gain insight into this issue, we set out to characterize the Jak3 promoter. The 5؅-untranslated region of the Jak3 gene is interrupted by a 3515-bp intron. Upstream of this intron and the transcription initiation site, we identified an ϳ1-kb segment that exhibited lymphoid-specific promoter activity and was responsive to TCR signals. Truncation of this fragment revealed that core promoter activity resided in a 267-bp fragment that contains putative Sp-1, AP-1, Ets, Stat, and other binding sites. Mutation of the AP-1 sites significantly diminished, whereas mutation of the Ets sites abolished, the inducibility of the promoter construct. Chromatin immunoprecipitation assays showed that histone acetylation correlates with mRNA expression and that Ets-1/2 binds this region. Thus, transcription factors that bind these sites, especially Ets family members, are likely to be important regulators of Jak3 expression.
    [Show full text]
  • Human and Mouse CD Marker Handbook Human and Mouse CD Marker Key Markers - Human Key Markers - Mouse
    Welcome to More Choice CD Marker Handbook For more information, please visit: Human bdbiosciences.com/eu/go/humancdmarkers Mouse bdbiosciences.com/eu/go/mousecdmarkers Human and Mouse CD Marker Handbook Human and Mouse CD Marker Key Markers - Human Key Markers - Mouse CD3 CD3 CD (cluster of differentiation) molecules are cell surface markers T Cell CD4 CD4 useful for the identification and characterization of leukocytes. The CD CD8 CD8 nomenclature was developed and is maintained through the HLDA (Human Leukocyte Differentiation Antigens) workshop started in 1982. CD45R/B220 CD19 CD19 The goal is to provide standardization of monoclonal antibodies to B Cell CD20 CD22 (B cell activation marker) human antigens across laboratories. To characterize or “workshop” the antibodies, multiple laboratories carry out blind analyses of antibodies. These results independently validate antibody specificity. CD11c CD11c Dendritic Cell CD123 CD123 While the CD nomenclature has been developed for use with human antigens, it is applied to corresponding mouse antigens as well as antigens from other species. However, the mouse and other species NK Cell CD56 CD335 (NKp46) antibodies are not tested by HLDA. Human CD markers were reviewed by the HLDA. New CD markers Stem Cell/ CD34 CD34 were established at the HLDA9 meeting held in Barcelona in 2010. For Precursor hematopoetic stem cell only hematopoetic stem cell only additional information and CD markers please visit www.hcdm.org. Macrophage/ CD14 CD11b/ Mac-1 Monocyte CD33 Ly-71 (F4/80) CD66b Granulocyte CD66b Gr-1/Ly6G Ly6C CD41 CD41 CD61 (Integrin b3) CD61 Platelet CD9 CD62 CD62P (activated platelets) CD235a CD235a Erythrocyte Ter-119 CD146 MECA-32 CD106 CD146 Endothelial Cell CD31 CD62E (activated endothelial cells) Epithelial Cell CD236 CD326 (EPCAM1) For Research Use Only.
    [Show full text]
  • Megakaryopoiesis in Dengue Virus Infected K562 Cell Promotes Viral Replication Which Inhibits 2 Endomitosis and Accumulation of ROS Associated with Differentiation
    bioRxiv preprint doi: https://doi.org/10.1101/2020.06.25.172544; this version posted June 26, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Title: Megakaryopoiesis in Dengue virus infected K562 cell promotes viral replication which inhibits 2 endomitosis and accumulation of ROS associated with differentiation 3 Jaskaran Kaur *1, Yogita Rawat *1, Vikas Sood 2, Deepak Rathore1, Shrikant K. Kumar1, Niraj K. Kumar1 4 and Sankar Bhattacharyya1 5 1 Translational Health Science and Technology Institute, NCR Biotech Science Cluster, PO Box# 4, 6 Faridabad-Gurgaon expressway, Faridabad, Haryana-121001, India 7 2 Department of Biochemistry, School of Chemical and Life Sciences, Jamia Hamdard (Hamdard 8 University) Hamdard Nagar, New Delhi - 110062, India 9 10 *Equal contribution 11 Email for correspondence: [email protected] 12 13 14 Keywords: Dengue virus replication, Megakaryopoiesis, Reactive oxygen species, Endomitosis 15 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.06.25.172544; this version posted June 26, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 16 Abstract: In the human host blood Monocytes and bone marrow Megakaryocytes are implicated as major 17 sites supporting high replication. The human K562 cell line supports DENV replication and represent 18 Megakaryocyte-Erythrocyte progenitors (MEP), replicating features of in vivo Megakaryopoiesis upon 19 stimulation with Phorbol esters. In this article, we report results that indicate the mutual influence of 20 Megakaryopoiesis and DENV replication on each other, through comparison of PMA-induced 21 differentiation of either mock-infected or DENV-infected K562 cells.
    [Show full text]
  • Review Tec Kinases: a Family with Multiple Roles in Immunity
    Immunity, Vol. 12, 373±382, April, 2000, Copyright 2000 by Cell Press Tec Kinases: A Family Review with Multiple Roles in Immunity Wen-Chin Yang,*³§ Yves Collette,*³ inositol phosphates, but they are thought to be relevant Jacques A. NuneÁ s,*³ and Daniel Olive*² for binding of PtdIns lipids to the same sites. In most *INSERM U119 cases, PH domains bind preferentially to PtdIns (4,5)P2 Universite de la Me diterrane e and inositol (1,4,5) P3 (Ins (1,4,5) P3). However, the Btk 13009 Marseille PH domain binds PtdIns (3,4,5)P3 and Ins (1, 3, 4, 5)P4 France the tightest. PtdIns (3, 4, 5)P3, one of the products of the action of PI3K, is thought to act as a second messen- ger to recruit regulatory proteins to the plasma mem- brane via their PH domains (see below). Many of the Antigen receptors on T, B, and mast cells are multimo- mutations in Btk that lead to XLA are point mutations lecular complexes that are activated by interactions with that cluster at one end of the PH domain and could be external signals. These signals are then transmitted to predicted to impair binding to Ins (3,4,5)P (for review regulate gene expression and posttranscriptional modi- 3 see Satterthwaite et al., 1998a) (Figure 1b). Similarly, fications. Nonreceptor tyrosine kinases (NRTK) are key CBA/N xid mice carry an R28C mutation in the Btk PH players that relay and integrate these signals. NRTK are domain. The recent structure of the PH domain from divided into distinct families defined by a prototypic Btk complexed with Ins (1,3,4,5)P4 provides an explana- member: Src, Tec, Syk, Csk, Fes, Abl, Jak, Fak, Ack, tion for several mutations associated with XLA: mis- Brk, and Srm (Bolen and Brugge, 1997).
    [Show full text]
  • Evidence for an Interaction Between the Insulin Receptor and Grb7. a Role for Two of Its Binding Domains, PIR and SH2
    Oncogene (2000) 19, 2052 ± 2059 ã 2000 Macmillan Publishers Ltd All rights reserved 0950 ± 9232/00 $15.00 www.nature.com/onc Evidence for an interaction between the insulin receptor and Grb7. A role for two of its binding domains, PIR and SH2 Anne Kasus-Jacobi1,2,3,Ve ronique Be re ziat1,3, Dominique Perdereau1, Jean Girard1 and Anne-FrancËoise Burnol*,1 1Endocrinologie MeÂtabolisme et DeÂveloppement, CNRS, UPR 1524, 9 rue Jules Hetzel, 92190 Meudon, France The molecular adapter Grb7 is likely to be implicated in new family of adapters has recently emerged, the Grb7 the development of certain cancer types. In this study we family of proteins, comprising Grb7, Grb10 and show that Grb7 binds the insulin receptors, when they Grb14. The members of this family were originally are activated and tyrosine phosphorylated. This interac- cloned by interaction with EGF receptor, using the tion is documented by two-hybrid experiments, GST CORT (cloning of receptor target) system (Daly et al., pull-down assays and in vivo coimmunoprecipitations. In 1996; Margolis, 1994; Ooi et al., 1995), and the use of addition, our results argue in favor of a preferential the yeast two-hybrid technology has emphasized their association between Grb7 and the insulin receptors when implication in signal transduction (Daly, 1998). These compared to other tyrosine kinase receptors like the adapters bind also other tyrosine kinase receptors, like EGF receptor, the FGF receptor and Ret. Interestingly, erbB2, Ret, Elk, PDGF receptors, insulin receptors and Grb7 is not a substrate of the insulin receptor tyrosine IGF-1 receptors (Daly et al., 1996; Dey et al., 1996; kinase activity.
    [Show full text]
  • Expression of the Tumor Necrosis Factor Receptor-Associated Factors
    Expression of the Tumor Necrosis Factor Receptor- Associated Factors (TRAFs) 1 and 2 is a Characteristic Feature of Hodgkin and Reed-Sternberg Cells Keith F. Izban, M.D., Melek Ergin, M.D, Robert L. Martinez, B.A., HT(ASCP), Serhan Alkan, M.D. Department of Pathology, Loyola University Medical Center, Maywood, Illinois the HD cell lines. Although KMH2 showed weak Tumor necrosis factor receptor–associated factors expression, the remaining HD cell lines also lacked (TRAFs) are a recently established group of proteins TRAF5 protein. These data demonstrate that consti- involved in the intracellular signal transduction of tutive expression of TRAF1 and TRAF2 is a charac- several members of the tumor necrosis factor recep- teristic feature of HRS cells from both patient and tor (TNFR) superfamily. Recently, specific members cell line specimens. Furthermore, with the excep- of the TRAF family have been implicated in promot- tion of TRAF1 expression, HRS cells from the three ing cell survival as well as activation of the tran- HD cell lines showed similar TRAF protein expres- ␬ scription factor NF- B. We investigated the consti- sion patterns. Overall, these findings demonstrate tutive expression of TRAF1 and TRAF2 in Hodgkin the expression of several TRAF proteins in HD. Sig- and Reed–Sternberg (HRS) cells from archived nificantly, the altered regulation of selective TRAF paraffin-embedded tissues obtained from 21 pa- proteins may reflect HRS cell response to stimula- tients diagnosed with classical Hodgkin’s disease tion from the microenvironment and potentially (HD). In a selective portion of cases, examination of contribute both to apoptosis resistance and cell HRS cells for Epstein-Barr virus (EBV)–encoded maintenance of HRS cells.
    [Show full text]
  • Lipid-Targeting Pleckstrin Homology Domain Turns Its Autoinhibitory Face Toward the TEC Kinases
    Lipid-targeting pleckstrin homology domain turns its autoinhibitory face toward the TEC kinases Neha Amatyaa, Thomas E. Walesb, Annie Kwonc, Wayland Yeungc, Raji E. Josepha, D. Bruce Fultona, Natarajan Kannanc, John R. Engenb, and Amy H. Andreottia,1 aRoy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011; bDepartment of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115; and cInstitute of Bioinformatics and Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602 Edited by Natalie G. Ahn, University of Colorado Boulder, Boulder, CO, and approved September 17, 2019 (received for review May 3, 2019) The pleckstrin homology (PH) domain is well known for its phos- activation loop phosphorylation site are also controlled by noncatalytic pholipid targeting function. The PH-TEC homology (PHTH) domain domains (17). In addition to the N-terminal PHTH domain, the within the TEC family of tyrosine kinases is also a crucial component TEC kinases contain a proline-rich region (PRR) and Src ho- of the autoinhibitory apparatus. The autoinhibitory surface on the mology 3 (SH3) and Src homology 2 (SH2) domains that impinge PHTH domain has been previously defined, and biochemical investi- on the kinase domain to alter the conformational ensemble and gations have shown that PHTH-mediated inhibition is mutually thus the activation status of the enzyme. A crystal structure of the exclusive with phosphatidylinositol binding. Here we use hydrogen/ BTK SH3-SH2-kinase fragment has been solved (10) showing that deuterium exchange mass spectrometry, nuclear magnetic resonance the SH3 and SH2 domains of BTK assemble onto the distal side of (NMR), and evolutionary sequence comparisons to map where and the kinase domain (the surface opposite the activation loop), how the PHTH domain affects the Bruton’s tyrosine kinase (BTK) stabilizing the autoinhibited form of the kinase in a manner similar domain.
    [Show full text]
  • HCC and Cancer Mutated Genes Summarized in the Literature Gene Symbol Gene Name References*
    HCC and cancer mutated genes summarized in the literature Gene symbol Gene name References* A2M Alpha-2-macroglobulin (4) ABL1 c-abl oncogene 1, receptor tyrosine kinase (4,5,22) ACBD7 Acyl-Coenzyme A binding domain containing 7 (23) ACTL6A Actin-like 6A (4,5) ACTL6B Actin-like 6B (4) ACVR1B Activin A receptor, type IB (21,22) ACVR2A Activin A receptor, type IIA (4,21) ADAM10 ADAM metallopeptidase domain 10 (5) ADAMTS9 ADAM metallopeptidase with thrombospondin type 1 motif, 9 (4) ADCY2 Adenylate cyclase 2 (brain) (26) AJUBA Ajuba LIM protein (21) AKAP9 A kinase (PRKA) anchor protein (yotiao) 9 (4) Akt AKT serine/threonine kinase (28) AKT1 v-akt murine thymoma viral oncogene homolog 1 (5,21,22) AKT2 v-akt murine thymoma viral oncogene homolog 2 (4) ALB Albumin (4) ALK Anaplastic lymphoma receptor tyrosine kinase (22) AMPH Amphiphysin (24) ANK3 Ankyrin 3, node of Ranvier (ankyrin G) (4) ANKRD12 Ankyrin repeat domain 12 (4) ANO1 Anoctamin 1, calcium activated chloride channel (4) APC Adenomatous polyposis coli (4,5,21,22,25,28) APOB Apolipoprotein B [including Ag(x) antigen] (4) AR Androgen receptor (5,21-23) ARAP1 ArfGAP with RhoGAP domain, ankyrin repeat and PH domain 1 (4) ARHGAP35 Rho GTPase activating protein 35 (21) ARID1A AT rich interactive domain 1A (SWI-like) (4,5,21,22,24,25,27,28) ARID1B AT rich interactive domain 1B (SWI1-like) (4,5,22) ARID2 AT rich interactive domain 2 (ARID, RFX-like) (4,5,22,24,25,27,28) ARID4A AT rich interactive domain 4A (RBP1-like) (28) ARID5B AT rich interactive domain 5B (MRF1-like) (21) ASPM Asp (abnormal
    [Show full text]
  • Inhibition of Src Family Kinases and Receptor Tyrosine Kinases by Dasatinib: Possible Combinations in Solid Tumors
    Published OnlineFirst June 13, 2011; DOI: 10.1158/1078-0432.CCR-10-2616 Clinical Cancer Molecular Pathways Research Inhibition of Src Family Kinases and Receptor Tyrosine Kinases by Dasatinib: Possible Combinations in Solid Tumors Juan Carlos Montero1, Samuel Seoane1, Alberto Ocaña2,3, and Atanasio Pandiella1 Abstract Dasatinib is a small molecule tyrosine kinase inhibitor that targets a wide variety of tyrosine kinases implicated in the pathophysiology of several neoplasias. Among the most sensitive dasatinib targets are ABL, the SRC family kinases (SRC, LCK, HCK, FYN, YES, FGR, BLK, LYN, and FRK), and the receptor tyrosine kinases c-KIT, platelet-derived growth factor receptor (PDGFR) a and b, discoidin domain receptor 1 (DDR1), c-FMS, and ephrin receptors. Dasatinib inhibits cell duplication, migration, and invasion, and it triggers apoptosis of tumoral cells. As a consequence, dasatinib reduces tumoral mass and decreases the metastatic dissemination of tumoral cells. Dasatinib also acts on the tumoral microenvironment, which is particularly important in the bone, where dasatinib inhibits osteoclastic activity and favors osteogenesis, exerting a bone-protecting effect. Several preclinical studies have shown that dasatinib potentiates the antitumoral action of various drugs used in the oncology clinic, paving the way for the initiation of clinical trials of dasatinib in combination with standard-of-care treatments for the therapy of various neoplasias. Trials using combinations of dasatinib with ErbB/HER receptor antagonists are being explored in breast, head and neck, and colorectal cancers. In hormone receptor–positive breast cancer, trials using combina- tions of dasatinib with antihormonal therapies are ongoing. Dasatinib combinations with chemother- apeutic agents are also under development in prostate cancer (dasatinib plus docetaxel), melanoma (dasatinib plus dacarbazine), and colorectal cancer (dasatinib plus oxaliplatin plus capecitabine).
    [Show full text]
  • Supplementary Table 1. in Vitro Side Effect Profiling Study for LDN/OSU-0212320. Neurotransmitter Related Steroids
    Supplementary Table 1. In vitro side effect profiling study for LDN/OSU-0212320. Percent Inhibition Receptor 10 µM Neurotransmitter Related Adenosine, Non-selective 7.29% Adrenergic, Alpha 1, Non-selective 24.98% Adrenergic, Alpha 2, Non-selective 27.18% Adrenergic, Beta, Non-selective -20.94% Dopamine Transporter 8.69% Dopamine, D1 (h) 8.48% Dopamine, D2s (h) 4.06% GABA A, Agonist Site -16.15% GABA A, BDZ, alpha 1 site 12.73% GABA-B 13.60% Glutamate, AMPA Site (Ionotropic) 12.06% Glutamate, Kainate Site (Ionotropic) -1.03% Glutamate, NMDA Agonist Site (Ionotropic) 0.12% Glutamate, NMDA, Glycine (Stry-insens Site) 9.84% (Ionotropic) Glycine, Strychnine-sensitive 0.99% Histamine, H1 -5.54% Histamine, H2 16.54% Histamine, H3 4.80% Melatonin, Non-selective -5.54% Muscarinic, M1 (hr) -1.88% Muscarinic, M2 (h) 0.82% Muscarinic, Non-selective, Central 29.04% Muscarinic, Non-selective, Peripheral 0.29% Nicotinic, Neuronal (-BnTx insensitive) 7.85% Norepinephrine Transporter 2.87% Opioid, Non-selective -0.09% Opioid, Orphanin, ORL1 (h) 11.55% Serotonin Transporter -3.02% Serotonin, Non-selective 26.33% Sigma, Non-Selective 10.19% Steroids Estrogen 11.16% 1 Percent Inhibition Receptor 10 µM Testosterone (cytosolic) (h) 12.50% Ion Channels Calcium Channel, Type L (Dihydropyridine Site) 43.18% Calcium Channel, Type N 4.15% Potassium Channel, ATP-Sensitive -4.05% Potassium Channel, Ca2+ Act., VI 17.80% Potassium Channel, I(Kr) (hERG) (h) -6.44% Sodium, Site 2 -0.39% Second Messengers Nitric Oxide, NOS (Neuronal-Binding) -17.09% Prostaglandins Leukotriene,
    [Show full text]
  • Crystal Structure of the Jak3 Kinase Domain in Complex with a Staurosporine Analog
    From www.bloodjournal.org by guest on September 11, 2016. For personal use only. TRANSPLANTATION Crystal structure of the Jak3 kinase domain in complex with a staurosporine analog Titus J. Boggon, Yiqun Li, Paul W. Manley, and Michael J. Eck Jak (Janus kinase) family nonreceptor target for development of lymphocyte- tion of the activation loop. Such a direct tyrosine kinases are central mediators of specific immunosuppressants. Here, we coupling has not been previously ob- cytokine signaling. The Jak kinases ex- present the crystal structure of the Jak3 served in tyrosine kinases and may be hibit distinct cytokine receptor associa- kinase domain in complex with stauro- unique to Jak kinases. The crystal struc- tion profiles and so transduce different sporine analog AFN941. The kinase do- ture provides a detailed view of the Jak3 signals. Jak3 expression is limited to the main is in the active conformation, with active site and will facilitate computa- immune system, where it plays a key role both activation loop tyrosine residues tional and structure-directed approaches in signal transduction from cytokine re- phosphorylated. The phosphate group on to development of Jak3-specific inhibi- ceptors containing the common gamma- pTyr981 in the activation loop is in part tors. (Blood. 2005;106:996-1002) chain, ␥c. Patients unable to signal via ␥c coordinated by an arginine residue in the present with severe combined immunode- regulatory C-helix, suggesting a direct ficiency (SCID). The finding that Jak3 mechanism by which the active position mutations result in SCID has made it a of the C-helix is induced by phosphoryla- © 2005 by The American Society of Hematology Introduction The Janus kinase (Jak) family of cytoplasmic tyrosine kinases are regulation of diverse cell processes, including proliferation, differ- essential for signal transduction from a wide variety of cell-surface entiation, migration, and apoptosis.1,2 receptors.
    [Show full text]
  • Wrangling Phosphoproteomic Data to Elucidate Cancer Signaling Pathways
    University of Montana ScholarWorks at University of Montana Biological Sciences Faculty Publications Biological Sciences 1-3-2013 Wrangling Phosphoproteomic Data to Elucidate Cancer Signaling Pathways Mark L. Grimes University of Montana - Missoula, [email protected] Wan-Jui Lee Laurens van der Maaten Paul Shannon Follow this and additional works at: https://scholarworks.umt.edu/biosci_pubs Part of the Biology Commons Let us know how access to this document benefits ou.y Recommended Citation Grimes, Mark L.; Lee, Wan-Jui; van der Maaten, Laurens; and Shannon, Paul, "Wrangling Phosphoproteomic Data to Elucidate Cancer Signaling Pathways" (2013). Biological Sciences Faculty Publications. 178. https://scholarworks.umt.edu/biosci_pubs/178 This Article is brought to you for free and open access by the Biological Sciences at ScholarWorks at University of Montana. It has been accepted for inclusion in Biological Sciences Faculty Publications by an authorized administrator of ScholarWorks at University of Montana. For more information, please contact [email protected]. Wrangling Phosphoproteomic Data to Elucidate Cancer Signaling Pathways Mark L. Grimes1*, Wan-Jui Lee2, Laurens van der Maaten2, Paul Shannon3 1 Division of Biological Sciences, University of Montana, Missoula, Montana, United States of America, 2 Pattern Recognition and Bioinformatics Group, Delft University of Technology, CD Delft, The Netherlands, 3 Fred Hutchison Cancer Research Institute, Seattle, Washington, United States of America Abstract The interpretation of biological data sets is essential for generating hypotheses that guide research, yet modern methods of global analysis challenge our ability to discern meaningful patterns and then convey results in a way that can be easily appreciated. Proteomic data is especially challenging because mass spectrometry detectors often miss peptides in complex samples, resulting in sparsely populated data sets.
    [Show full text]