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1.
Methods Inf Med ; 57(S 01): e46-e49, 2018 07.
Article in English | MEDLINE | ID: mdl-30016817

ABSTRACT

This article is part of the Focus Theme of Methods of Information in Medicine on the German Medical Informatics Initiative. The Medical Informatics Initiative of the German Federal Ministry of Education and Research will make use of the potential of digitalization in the field of medicine in Germany. The aim is to improve the possibilities for medical research and patient care through innovative IT solutions. In an initial step, data integration centres will be set up at university hospitals to ensure the technical and organizational conditions necessary for multi-site exchange of data between health care and clinical and biomedical research. The Federal Ministry of Education and Research will provide a total of around EUR 150 million for this initiative over the next four years.


Subject(s)
Delivery of Health Care , Medical Informatics , Research , Statistics as Topic , Advisory Committees , Delivery of Health Care/economics , Germany , Medical Informatics/economics
2.
JAKSTAT ; 4(1): e1062596, 2015.
Article in English | MEDLINE | ID: mdl-26413425

ABSTRACT

Aberrant activation of oncogenic kinases is frequently observed in human cancers, but the underlying mechanism and resulting effects on global signaling are incompletely understood. Here, we demonstrate that the oncogenic FIP1L1-PDGFRα kinase exhibits a significantly different signaling pattern compared to its PDGFRα wild type counterpart. Interestingly, the activation of primarily membrane-based signal transduction processes (such as PI3-kinase- and MAP-kinase- pathways) is remarkably shifted toward a prominent activation of STAT factors. This diverging signaling pattern compared to classical PDGF-receptor signaling is partially coupled to the aberrant cytoplasmic localization of the oncogene, since membrane targeting of FIP1L1-PDGFRα restores activation of MAPK- and PI3K-pathways. In stark contrast to the classical cytokine-induced STAT activation process, STAT activation by FIP1L1-PDGFRα does neither require Janus kinase activity nor Src kinase activity. Furthermore, we investigated the mechanism of STAT5 activation via FIP1L1-PDGFRα in more detail and found that STAT5 activation does not involve an SH2-domain-mediated binding mechanism. We thus demonstrate that STAT5 activation occurs via a non-canonical activation mechanism in which STAT5 may be subject to a direct phosphorylation by FIP1L1-PDGFRα.

3.
Cell Commun Signal ; 13: 21, 2015 Mar 31.
Article in English | MEDLINE | ID: mdl-25880691

ABSTRACT

BACKGROUND: Gastrointestinal stromal tumours (GIST) are mainly characterised by the presence of activating mutations in either of the two receptor tyrosine kinases c-KIT or platelet-derived growth factor receptor-α (PDGFRα). Most mechanistic studies dealing with GIST mutations have focused on c-KIT and far less is known about the signalling characteristics of the mutated PDGFRα proteins. Here, we study the signalling capacities and corresponding transcriptional responses of the different PDGFRα proteins under comparable genomic conditions. RESULTS: We demonstrate that the constitutive signalling via the oncogenic PDGFRα mutants favours a mislocalisation of the receptors and that this modifies the signalling characteristics of the mutated receptors. We show that signalling via the oncogenic PDGFRα mutants is not solely characterised by a constitutive activation of the conventional PDGFRα signalling pathways. In contrast to wild-type PDGFRα signal transduction, the activation of STAT factors (STAT1, STAT3 and STAT5) is an integral part of signalling mediated via mutated PDGF-receptors. Furthermore, this unconventional STAT activation by mutated PDGFRα is already initiated in the endoplasmic reticulum whereas the conventional signalling pathways rather require cell surface expression of the receptor. Finally, we demonstrate that the activation of STAT factors also translates into a biologic response as highlighted by the induction of STAT target genes. CONCLUSION: We show that the overall oncogenic response is the result of different signatures emanating from different cellular compartments. Furthermore, STAT mediated responses are an integral part of mutated PDGFRα signalling.


Subject(s)
Gastrointestinal Neoplasms/metabolism , Mutation , Neoplasm Proteins/metabolism , Receptor, Platelet-Derived Growth Factor alpha/metabolism , STAT Transcription Factors/metabolism , Signal Transduction , Cell Line, Tumor , Endoplasmic Reticulum/genetics , Endoplasmic Reticulum/metabolism , Endoplasmic Reticulum/pathology , Enzyme Activation/genetics , Gastrointestinal Neoplasms/genetics , Gastrointestinal Neoplasms/pathology , Humans , Neoplasm Proteins/genetics , Receptor, Platelet-Derived Growth Factor alpha/genetics , STAT Transcription Factors/genetics
4.
Cell Signal ; 27(2): 340-52, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25460044

ABSTRACT

The timely orchestration of multiple signalling pathways is crucial for the integrity of an organism and therefore tightly controlled. Gab family proteins coordinate signal transduction at the plasma membrane (PM) by acting as docking platforms for signalling components involved in MAP kinase (MAPK), PI3 kinase (PI3K), phospholipase C (PLC) and Rho family GTPase signalling. The interaction with these components as well as the targeting of the docking platform to the PM underlies complex spatial and temporal regulatory mechanisms. Deregulated Gab1 activation and membrane binding have been observed in some haematopoietic malignancies and solid tumours, thereby contributing, for example, to the development of Philadelphia chromosome-negative myeloproliferative neoplasms and certain lung cancers. Previously, we could demonstrate that the presence of PIP3 in the PM, which is increased in many cancer cells, is not sufficient for constitutive Gab1 membrane recruitment. In addition, MAPK-dependent phosphorylation of Gab1 at serine 552 (Ser552) is vital for Gab1 membrane binding. Here, we confirm our hypothesis that in the absence of MAPK activity an intrinsic part of Gab1 prevents binding to PIP3 at the PM. This epitope of Gab1, which encompasses Ser552, interacts directly with the Gab1 PH domain. Two arginines located in positions +4 and +8 of Ser552 are essential for the interaction with the PH domain, as well as for the inhibition of membrane recruitment of unphosphorylated Gab1. Ser552 phosphorylation is dispensable in respective arginine to alanine mutants of Gab1. Gab1 recruitment to the PM is highly dynamic and continuous PI3K and MAPK activities are both essential for sustained Gab1 membrane localisation. Our data document the existence of a sophisticated and robust control mechanism that prevents Gab1 translocation and signalling complex assembly after the activation of either MAPK or PI3K alone.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Cell Membrane/metabolism , Mitogen-Activated Protein Kinases/metabolism , Adaptor Proteins, Signal Transducing/genetics , Amino Acid Sequence , Androstadienes/pharmacology , Butadienes/pharmacology , Chromones/pharmacology , HEK293 Cells , Humans , Interleukin-6/metabolism , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Molecular Sequence Data , Morpholines/pharmacology , Mutation , Nitriles/pharmacology , Phosphatidylinositol 3-Kinases/metabolism , Phosphoinositide-3 Kinase Inhibitors , Phosphorylation , Protein Binding , Protein Structure, Tertiary , STAT3 Transcription Factor/metabolism , Signal Transduction/drug effects , Tetradecanoylphorbol Acetate/pharmacology , Translocation, Genetic/drug effects , Wortmannin
5.
JAKSTAT ; 2(3): e24574, 2013 Jul 01.
Article in English | MEDLINE | ID: mdl-24069558

ABSTRACT

The identification of a constitutively active JAK2 mutant, namely JAK2-V617F, was a milestone in the understanding of Philadelphia chromosome-negative myeloproliferative neoplasms. The JAK2-V617F mutation confers cytokine hypersensitivity, constitutive activation of the JAK-STAT pathway, and cytokine-independent growth. In this study we investigated the mechanism of JAK2-V617F-dependent signaling with a special focus on the activation of the MAPK pathway. We observed JAK2-V617F-dependent deregulated activation of the multi-site docking protein Gab1 as indicated by constitutive, PI3K-dependent membrane localization and tyrosine phosphorylation of Gab1. Furthermore, we demonstrate that PI3K signaling regulates MAPK activation in JAK2-V617F-positve cells. This cross-regulation of the MAPK pathway by PI3K affects JAK2-V617F-specific target gene induction, erythroid colony formation, and regulates proliferation of JAK2-V617F-positive patient cells in a synergistically manner.

6.
J Cell Mol Med ; 17(2): 265-76, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23301855

ABSTRACT

The Janus kinase 2 mutant V617F occurs with high frequency in myeloproliferative neoplasms. Further mutations affecting the Janus kinase family have been discovered mostly in leukaemias and in myeloproliferative neoplasms. Owing to their involvement in neoplasia, inflammatory diseases and in the immune response, Janus kinases are promising targets for kinase inhibitor therapy in these disease settings. Various quantitative assays including two newly developed screening assays were used to characterize the function of different small-molecule compounds in cells expressing Jak2V617F. A detailed comparative analysis of different Janus kinase inhibitors in our quantitative assays and the subsequent characterization of additional activities demonstrated for the first time that the most potent Jak2 inhibitor in our study, CEP701, also targets Aurora kinases. CEP701 shows a unique combination of both activities which is not found in other compounds also targeting Jak2. Furthermore, colony forming cell assays showed that Janus kinase 2 inhibitors preferentially suppressed the growth of erythroid colonies, whereas inhibitors of Aurora kinases preferentially blocked myeloid colony growth. CEP701 demonstrated a combined suppression of both colony types. Moreover, we show that combined application of a Janus and an Aurora kinase inhibitor recapitulated the effect observed for CEP701 but might allow for more flexibility in combining both activities in clinical settings, e.g. in the treatment of myeloproliferative neoplasms. The newly developed screening assays are high throughput compatible and allow an easy detection of new compounds with Janus kinase 2 inhibitory activity.


Subject(s)
Carbazoles/pharmacology , Cell Proliferation/drug effects , Janus Kinase 2/antagonists & inhibitors , Leukemia, Erythroblastic, Acute/pathology , Mutation/genetics , Protein Serine-Threonine Kinases/antagonists & inhibitors , Aurora Kinases , Blotting, Western , Cell Cycle/drug effects , Cell Differentiation/drug effects , Colony-Forming Units Assay , Flow Cytometry , Furans , Humans , Janus Kinase 2/genetics , Janus Kinase 2/metabolism , Leukemia, Erythroblastic, Acute/drug therapy , Leukemia, Erythroblastic, Acute/metabolism , Phosphorylation/drug effects , Protein Serine-Threonine Kinases/metabolism , Tumor Cells, Cultured
7.
Eur J Cell Biol ; 91(6-7): 486-95, 2012.
Article in English | MEDLINE | ID: mdl-22138086

ABSTRACT

The hallmark of signalling by many cytokines is the activation of the Janus kinase (Jak)/signal transducer and activator of transcription (STAT) pathway. However, cytokines additionally activate other pathways. In past years we realised that these pathways significantly contribute to the physiological functions of IL-6 and pathophysiological functions in the context of many inflammatory and proliferative diseases. Whereas other articles in this issue of the European Journal of Cell Biology focus on STAT activation and its regulation we here aim to summarise our knowledge and some remaining questions on interleukin-6 (IL-6)-induced STAT-independent pathways as well as the cross-talk with the Jak/STAT pathway. In the early stages of studying cytokine signalling we were used to analysing individual signalling pathways. These days we know about the importance of both, the crosstalk between pathways initiated by combinations of cytokines as well as the crosstalk between individual pathways initiated by a single cytokine. Whereas the inter-cytokine crosstalk can be studied relatively easily, more sophisticated experimental approaches are required to elucidate the intra-cytokine crosstalk.


Subject(s)
Interleukin-6/metabolism , Janus Kinase 1/metabolism , STAT1 Transcription Factor/metabolism , Animals , Humans , Mice , Phosphorylation , Signal Transduction
8.
J Cell Sci ; 122(Pt 1): 55-64, 2009 Jan 01.
Article in English | MEDLINE | ID: mdl-19050043

ABSTRACT

Adaptor proteins involved in signal transduction fulfil their cellular functions by bringing signalling molecules together and by targeting these signalling components to defined compartments within the cell. Furthermore, adaptor proteins represent a molecular platform from which different signalling pathways are initiated. Gab1 is an adaptor protein that recruits the p85 subunit of the phosphatidylinositol 3-kinase, the adaptor Grb2, the adaptor and phosphatase SHP2 and the GTPase-activating protein Ras-GAP. Gab1 thus contributes to the activation of the PI3K cascade and the MAPK cascade through many growth factors and cytokines. The recruitment of Gab1 to phosphatidylinositol (3,4,5)-trisphosphate within the plasma membrane by its pleckstrin-homology domain is regarded as a major regulatory step for the activation of Gab1. Here, we present a new more complex mechanism for Gab1 translocation that involves and depends on the activation of ERK. We demonstrate that the presence of PI3K activity in the cell is not sufficient for binding Gab1 to the plasma membrane. Instead, additional MAPK-dependent phosphorylation of Ser551 in Gab1 is crucial for the recruitment of Gab1 to the plasma membrane. This mechanism represents a new mode of regulation for the function of PH domains.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Cell Membrane/metabolism , Signal Transduction/physiology , Adaptor Proteins, Signal Transducing/genetics , Cell Line , Enzyme Activation , Extracellular Signal-Regulated MAP Kinases/genetics , Extracellular Signal-Regulated MAP Kinases/metabolism , Humans , Interleukin-6/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism
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