Dissertations / Theses on the topic 'Connective Tissue Growth Factor ECM'
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1
Tam, Y. Y. A. "Connective tissue growth factor in tissue fibrosis." Thesis, University College London (University of London), 2014. http://discovery.ucl.ac.uk/1448702/.
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Systemic Sclerosis (SSc) is a connective tissue disease characterised by inflammation and autoimmunity, vasculopathy, and interstitial remodelling and fibrosis. This thesis focuses on CTGF (CCN2), a member of the CCN family of matricellular proteins, as elevated CTGF expression is a hallmark of chronic fibrotic diseases such as SSc. In addition to the association of CTGF expression and fibrosis in human disease, experimentally, fibroblast-specific overexpression of CTGF has been shown to induce a fibrotic phenotype, as demonstrated in the Col1a2-CTGF transgenic mice. Prominent features of fibrosis included a thickened dermis, as well as excess collagen deposition in the skin and lung. This CTGF overexpression also provoked changes in the alveolar epithelium. In the lung of Col1a2-CTGF mice, immunostaining revealed a marked increase in the number of cells co-expressing the epithelial marker, TTF-1 and mesenchymal cell markers α-SMA and Snai1, indicative of epithelial-to-mesenchymal transition (EMT)-like changes. This suggested a role for the paracrine effects of CTGF in promoting the phenotypic switching of alveolar epithelial cells. EMT is likely to contribute, at least in part, to the accumulation of interstitial fibroblasts during fibrosis. Complementary in vitro studies in alveolar epithelial cells (AECs) showed that CTGF knockdown using siRNA suppressed TGF-β-induced mesenchymal cell proteins while inducing redistribution of the epithelial cell marker E-cadherin. Immunostaining and Western blotting showed that recombinant CTGF induced EMT-like morphological changes and expression of α-SMA in AECs. Finally, we were interested in whether the reduction or absence of CTGF could abrogate fibrosis. Knockdown of CTGF suppressed the induction of fibrotic proteins in TGF-β-treated control fibroblasts and SSc lung fibroblasts. Deletion of the CTGF gene showed reduced bleomycin-induced pulmonary fibrosis in mice. Overall, these results support that CTGF plays a pivotal role in fibrosis and blocking CTGF activity may be useful as a specific target of attenuating fibrosis in SSc.
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Charrier, Alyssa. "Connective Tissue Growth Factor in Pancreatitis." The Ohio State University, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=osu1366025057.
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Bonniaud, Philippe. "Transforming growth factor-β1, connective tissue growth factor et fibrose pulmonaire." Dijon, 2005. http://www.theses.fr/2005DIJOMU01.
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La fibrose pulmonaire (FP) est une maladie incurable. Nous nous sommes intéressés aux effets pathobiologiques de Transforming growth factor (TGF)βsur la FP. Par adénovecteurs, nous avons transféré à des poumons de rongeurs des cytokines susceptibles d'être impliqués dans la FP-TGF-β1, interleukine (IL)-1β, et connective tissue growth factor (CTGF)-. Nous démontrons que :1) TGFβ est essentiel pour l'initiation et la progression de la FP 2)CTGF est nécessaire mais incapable par lui seul d'induire la progression de la FP. CTGF pourrait être un cofacteur de TGFβ 3)FP et emphysème sont dépendants de Smad3, molécule de signal de TGFβ, démontrant l'importance de TGFβ dans l'équilibre entre accumulation et dégradation de matrice 4)la FP induite par IL-1β est dépendante de TGFβ 5)un inhibiteur spécifique du récepteur ALK5 de TGFβ bloque la FP induite par TGFβ. Conclusion : TGFβet ses voies de signal sont clés dans la FP. Ce travail est une ouverture à des possibilités thérapeutiques.
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Rachfal, Amy Wilson. "Expression and actions of connective tissue growth factor." The Ohio State University, 2004. http://rave.ohiolink.edu/etdc/view?acc_num=osu1069791086.
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Rachfal, Amy Wilson. "Expression and actions of connective tissue growth factor." Connect to this title online, 2003. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1069791086.
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Thesis (Ph. D.)--Ohio State University, 2003.Title from first page of PDF file. Document formatted into pages; contains xx, 186 p.; also includes graphics (some col.) Includes bibliographical references (p. 159-186). Available online via OhioLINK's ETD Center
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Wright, Aleksandra. "The roles and interactions of connective tissue growth factor." Thesis, Imperial College London, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.434988.
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Huang, Bau-Lin. "Connective tissue growth factor gene regulation and function of CTGF /." Diss., Restricted to subscribing institutions, 2009. http://proquest.umi.com/pqdweb?did=2026641211&sid=1&Fmt=2&clientId=1564&RQT=309&VName=PQD.
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Yokoi, Hideki. "Role of connective tissue growth factor in renal tubulointerstitial fibrosis." Kyoto University, 2005. http://hdl.handle.net/2433/144757.
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Gebhardt, Susanne. "Expression, biochemische Charakterisierung und biologische Analyse des CONNECTIVE TISSUE GROWTH FACTOR." kostenfrei, 2008. http://www.opus-bayern.de/uni-wuerzburg/volltexte/2008/2956/.
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Tang, Xiaodi. "The role of connective tissue growth factor (CTGF) in articular cartilage." Thesis, Imperial College London, 2014. http://hdl.handle.net/10044/1/25292.
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Work from our group has identified an important role for the pericellular matrix (PCM) of cartilage in mechanotransduction. This region of the matrix sequesters regulatory molecules such as fibroblastic growth factor 2 (FGF2) and releases them upon mechanical stimulation. By carrying out a proteomic analysis of PCM proteins we identified an additional regulatory molecule, connective tissue growth factor (CTGF). CTGF was of interest to us as this protein is up-regulated in osteoarthritis but its function in chondrocytes is unknown. I confirmed the pericellular localisation of CTGF in cartilage and showed that it binds to heparan sulphate perlecan and like FGF2, CTGF is rapidly released from the PCM upon cartilage injury. In order to determine its function in chondrocytes, I expressed recombinant CTGF and performed a microarray study in isolated human chondrocytes. Four genes were up- regulated by CTGF and all were known to be transforming growth factor beta (TGFβ) inducible. Recombinant CTGF was able to activate SMAD2 in porcine chondrocytes indicating that it can activate the canonical TGFβ signalling pathway. Both CTGF induced gene expression and SMAD2 phosphorylation was abrogated by TGFβ neutralising antibody and the TGFβ receptor inhibitor SB431542, suggesting that the activity of CTGF is TGFβ dependent. Adding exogenous CTGF to rested porcine cartilage increased TGFβ protein accumulation without affecting TGFβ mRNA levels, suggesting that CTGF may control TGFβ protein’s bioavailability. Injured porcine cartilage caused rapid release of endogenous CTGF as well as a delayed accumulation of TGFβ in the conditioned medium (CM). TGFβ proteins are synthesised and secreted in a latent complex associated with the latency- associated peptide (LAP). When I analysed the explantation CM under non-reducing conditions, CTGF resolved at a high molecular weight (~150kDa), and a high molecular weight fraction of this CM contained SMAD2 activity suggesting that CTGF may be in complex with latent TGFβ. This was supported by the identification of LAP in CTGF immunoprecipitates. Injured cartilage from CTGF knockout mice released reduced levels of CTGF but apparently normal levels of TGFβ suggesting that CTGF is not necessary for release of latent TGFβ. However, SMAD2-phosphorylating ability of the medium was compromised indicating that CTGF controls TGFβ bioavailability principally by facilitating activation of latent complex at the surface of the chondrocyte. These data unveil a novel role for CTGF in cartilage through controlling TGFβ bioavailability.
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Watts, Keira Louise. "Differential expression of connective tissue growth factor (CTGF) in fibrogenesis : regulation by simvastatin." Thesis, Keele University, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.398915.
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Connective Tissue Growth Factor (CTGF) is a potent, pro fibrotic mediator, which acts downstream of and in concert with Transforming Growth Factor (TGF)p, to drive fibrogenesis. Significant upregulation of CTGF has been reported in fibrogenic diseases including idiopathic pulmonary fibrosis (IPF). IPF typically features excessive fibroblast proliferation and extracellular matrix (ECM) deposition, features that may be partially due to CTGF over-expression. In this thesis, basal expression of CTGF in response to TGFp (Sng/ml) is confirmed in primary lung fibroblasts (lMR90 and HIPF). Gene expression is studied using competitive RT-PCR; western blotting and immunofluorescence used to measure protein levels. In contrast to IMR90 cells, TGFp significantly induces CTGF (gene and protein) in a timedependent fashion in HIPF cells. The thesis also provides novel insight into elevated CTGF induction and response to TGFp in Rheumatoid Arthritis-derived synovial fibroblasts compared to Osteoarthritis fibroblasts, suggesting that the role of CTGF is not selective to the lung and is implicated in arthropathies. There is currently no effective therapy for aborting fibrogenesis. Simvastatin has been shown to have anti fibrotic potential in renal fibroblasts; however similar action with the context of lung fibrosis has not been documented. Simvastatin is shown to reduce basal gene and protein expression of CTGF in lung fibroblasts and overrides TGFp-induction in a concentration-dependent manner. This modulation of CTGF, which occurs through inhibition of the cholesterol synthesis pathway, is associated with changes in aSMA and collagen gel contraction.Signalling pathways underlying Simvastatin effect on CTGF-TGFB interaction are evaluated using transient reporter transfections of a plasmid construct containing the CTGF promoter linked to luciferase. Simvastatin inhibits CTGF promoter activity in a concentration-dependent fashion inducing an 8.52 fold down regulation over TGFB stimulated at 10)lM concentration. Separate studies explore the role of the small GTPases namely Rho and Ras and show that geranylgeranylpyrophosphate (GGPP), but not farnesylpyrophosphate (FPP) induces CTGF promoter activity following Simvastatin inhibition. In conclusion, this novel data confirms Simvastatin's ability to modulate CTGF expression, and thus fibrogenesis, implicating a critical role for small GTPases, specifically Rho, in CTGF signalling.
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Nishioka, Michiyoshi. "Lipopolysaccharide induced connective tissue growth factor gene expression in human bronchial epithelial cells." Kyoto University, 2010. http://hdl.handle.net/2433/120917.
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Finckenberg, Piet. "Regulation of connective tissue growth factor (CTGF) in hypertension-induced end organ damage." Helsinki : University of Helsinki, 2003. http://ethesis.helsinki.fi/julkaisut/laa/biola/vk/finckenberg/.
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Hora, Caroline. "Connective tissue growth factor, steatosis and fibrosis in patients with chronic hepatitis C /." Bern : [s.n.], 2008. http://www.ub.unibe.ch/content/bibliotheken_sammlungen/sondersammlungen/dissen_bestellformular/index_ger.html.
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Holmes, Alan Matthew. "Regulation of connective tissue growth factor/CCN2 gene expression in systemic sclerosis fibroblasts." Thesis, University College London (University of London), 2007. http://discovery.ucl.ac.uk/1445639/.
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Systemic sclerosis (Scleroderma, SSc) is a chronic, connective tissue disease of unknown etiology, characterised by vascular dysfunction, iriflarnmation and organ fibrosis. Involving both genetic and environmental components, the specific mechanisms which result in fibrosis remain largely unknown. A cardinal feature of SSc is increased synthesis of extracellular matrix (ECM). Dermal fibroblasts cultured from SSc patients maintain many of the abnormal properties seen in vivo, including excess production of collagen type I, and growth factors such as connective tissue growth factor (CTGF/CCN2). CTGF, like many genes dysregulated in SSc, is induced by TGF- p in normal fibroblasts. The overall aim of my studies was to determine the mechanism(s) controlling CTGF over-expression in SSc dermal fibroblasts (SDF). Induction of CTGF by TGF-p was found to be dependent upon elements in the proximal portion of the CTGF promoter, distinct from those of the previously characterised TGF- P response element (TRE). The TRE acts, in NIH/3T3 and HFF cells, as a regulator of basal expression, and is not essential for TGF-P induction of CTGF. Instead TGF-p induces CTGF expression via a Smad3 complex, binding to a bona fide SMAD transcription factor binding site. Over-expression CTGF in SDF is independent of autocrine expression of TGF-P and the SMAD binding element and rather dependent on a functional Sp-binding site. Inhibition of Spl-like DNA binding reduces excessive CTGF expression in SDF. Consistent with this Spl-DNA binding activity is elevated in SDF nuclear extracts. Investigation of the mechanism of elevated Spl-like binding found that SDF exhibited constitunVely active ERK1/2 and JNK1. Inhibition of ERK1/2 repressed elevated Sp-binding and CTGF over-expression observed in SDF. In summary, the data presented in this thesis provide evidence that dysregulation of ERK1/2 in SDF is involved in CTGF over-expression via a Spl-like DNA binding. Thus repression of ERK may represent a candidate in targeting fibrosis in SSc.
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Fan, Vivian H. (Vivian Hanbing). "Polymer-tethered epidermal growth factor as an inductive biomaterial surface for connective tissue progenitors." Thesis, Massachusetts Institute of Technology, 2006. http://hdl.handle.net/1721.1/37959.
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Thesis (Ph. D.)--Massachusetts Institute of Technology, Biological Engineering Division, September 2006."July 2006."
Includes bibliographical references (leaves 123-137).
Connective tissue progenitors (CTP) can act as a pluripotent source of reparative cells during injury and therefore have great potential in regenerative medicine and tissue engineering. However, the response of CTP to most growth factors and cytokines is unknown. Many envisioned applications of CTP, such as treating large defects in bone, involve in vivo implantation of CTP attached to a scaffold, a process that creates an acute inflammatory environment that may be hostile to CTP survival. This project entails the design of a two-component polymeric implant system to aid in the healing process of bony defects by influencing cell behaviors at the implant site through the covalent modification of the implant surface with selected ligands. We investigate cellular responses of CTP on a biomaterial surface covalently modified with epidermal growth factor (EGF) and find that surface-tethered EGF (tEGF) promotes both cell spreading and survival more strongly than saturating concentrations of soluble EGF. By sustaining MEK-ERK signaling, tEGF increases the contact of CTP with an otherwise moderately adhesive synthetic polymer and confers resistance to apoptosis induced by the proinflammatory cytokine, FasL.
(cont.) We confirm that these signaling, spreading, and apoptotic responses are conserved across three sources of CTP: an hTERT-immortalized human mesenchymal stem cell (MSC) line, primary porcine bone-marrow CTP, and primary human bone-marrow-derived CTP. We conclude that tEGF may offer a protective advantage to CTP in vivo during acute inflammatory reactions to tissue engineering scaffolds. The tEGF-modified polymers described here could be used together with structural materials to construct CTP scaffolds for the treatment of hard-tissue lesions, such as large bony defects.
by Vivian H. Fan.
Ph.D.
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Zhang, Xuemei. "Src Kinase Signaling Regulates Connective Tissue Growth Factor (CTGF/CCN2) Induction by Transforming Growth Factor-Beta 1 (TGF-b1) in Osteoblasts." Diss., Temple University Libraries, 2010. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/63095.
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AnatomyPh.D.
Connective tissue growth factor (CTGF/CCN2) is a cysteine rich, extracellular matrix protein that acts as an anabolic growth factor to regulate osteoblast differentiation and function. In osteoblasts, CTGF is induced by transforming growth factor beta 1 (TGF-β1) where it acts as a downstream mediator of TGF-β1 induced extracellular matrix production. The molecular mechanisms that control CTGF induction by TGF-β1 in osteoblasts are not understood. We have previously demonstrated the requirement of Src, Erk and Smad signaling for TGF-β1 induced CTGF promoter activity in primary osteoblasts, however the potential interaction among these signaling pathways in osteoblasts remains unknown. In this study, we demonstrate that CTGF is induced by TGF-β1 in rat osteosarcoma osteoblast like cells (ROS17/2.8). TGF-β1 activates Src and blocking of Src family kinases by PP2 abrogates TGF-β1 induced CTGF up-regulation. Western blot analysis revealed that primary osteoblasts and ROS 17/2.8 cells express not only Src, but also other Src family members, such as Fyn, Yes and Hck. In order to determine whether CTGF up-regulation is controlled by Src or other members, we used either kinase-dead dominant negative Src constructs in primary osteoblasts or Src siRNA in ROS17/2.8 cells to block Src function. Inactivation of Src by both kinase-dead and siRNA prevented TGF-β1 induced CTGF induction, demonstrating that TGF-β1 induced CTGF up-regulation is mediated only by Src not by other members. In addition, we also demonstrated that Erk is activated by TGF-β1 and that blocking of Erk activation using pharmacological inhibitors, PD98059 and U0126, prevents TGF-β1 induced CTGF induction, demonstrating the requirement of Erk for CTGF induction. These results prompted us to further explore the cross-talk between Src, Erk and Smads in ROS17/2.8 cells. Inhibition of Src using PP2 prevented Erk activation, demonstrating that Src is upstream of Erk. To investigate how Src and Erk regulate the canonical TGF-β1 signaling pathway, including Smad2/3 phosphorylation and nuclear translocation of activated Smads, we treated cells with TGF-β1 in the presence or absence of the Src inhibitor, PP2, or the Erk inhibitors, PD98059 or U0126. PP2 pre-treatment prevented the phosphorylation of Smad2/3 at both the SSXS motif and the linker region and consequently blocked their nuclear translocation, demonstrating that Src can regulate Smad signaling. In contrast, the Erk inhibitors did not have any effects on Smad phosphorylation and/or nuclear translocation. To examine whether Erk can modulate Smad signaling indirectly through the activation/ inactivation of required nuclear coactivators/ co-repressors that mediate Smad DNA binding, we used electro-mobility shift assays. These experiments showed that inhibition of Erk activation impaired transcriptional complex formation on the Smad binding element (SBE) and TGF- β responsive element (TRE) of the CTGF promoter, demonstrating that Erk activation is required for SBE and TRE transactivation. Taking together, these data demonstrate that Src is an essential upstream signaling transducer for Erk and Smad signaling in osteoblasts, and that while the Smad and Erk signaling cascades appear to function independent of each other, they are both essential for the formation of a transcriptionally active complex on the CTGF promoter.
Temple University--Theses
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Blalock, Timothy Daniel. "Biochemical characterization and action of connective tissue growth factor and its receptor in corneal scarring." [Gainesville, Fla.] : University of Florida, 2003. http://purl.fcla.edu/fcla/etd/UFE0001161.
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Winn, Simon. "An investigation of the actions of connective tissue growth factor on human renal epithelial cells." Thesis, St George's, University of London, 2018. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.754078.
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Introduction. Connective tissue growth factor (CTGF) is the only CCN member recognized as a mediator of chronic fibrotic disease. Accumulating evidence suggests that CTGF is important as a downstream effector of transforming growth factor (TGFβ) in modulating a sustained pro-fibrotic signal in in vivo models. Moreover, in cultured human proximal tubule epithelial cells (PTECs) CTGF mRNA is readily upregulated by TGFP and the nascent protein accumulates extracellularly. How pro-fibrotic signalling occurs is yet to be ascertained but it likely involves extracellular interactions with secreted CTGF. The tetra-modular structure of CTGF has been shown to possess multiple binding sites for ligands present in the extracellular milieu including matrix proteins, growth factors including the TGFβ superfamily and cell surface receptors. As such, CTGF can behave as a bridge between matrix and cell. Moreover, individual modules possess intrinsic biologic activity. Materials and Methods. Experiments were performed in cultured renal human renal epithelial cells. Recombinant human CTGF protein was generated in-house following plasmid transfection into context relevant PTECs. Immunoblotting and ELISA techniques were used to investigate protein expression in response to incubation with study protein in isolation or in combination with TGFP superfamily members. Protein-protein binding was investigated using surface plasmon resonance (SPR) in order to elucidate how CTGF might regulate TGFβ superfamily cell signalling. Additional experiments with an alternative CCN member, CCN3, were performed. Results. Both full-length and C-terminal CTGF bind to TGFβ and BMP7 and in human renal epithelial cells, this binding modulates the downstream Smad signalling pathways associated with fibrosis. In cultured human podocytes, CTGF drives TGFβ-dependent signalling in the absence of exogenous TGF(3 suggesting activation of latent TGFβ. Moreover, C-terminal CTGF increases the generation of pro-fibrotic markers aSMA and fibronectin, both of which are subsequently blocked by inhibiting the TGFp receptor. Unlike CTGF, an alternative CCN member CCN3 reduces TGFβ-induced signalling in PTECs. Conclusion. CTGF protein has multiple binding sites and modulates the cellular responses of TGFp superfamily members on human renal epithelial cells. Both full-length and C-terminal CTGF bind to TGFβ and BMP-7. CTGF also appears to bind to the receptors of the TGFβ superfamily. C-terminal CTGF has intrinsic profibrotic activity that differs from the full-length protein as demonstrated in cultured human podocytes. The pro-fibrotic activity is suppressed by CCN3 in the CTGF rich environment of the cultured PTEC. The interplay of different CCN proteins in modulating fibrotic pathways in renal epithelial cells warrants further investigation.
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Ayubi, Nawid. "mRNA-Expression vom connective tissue growth factor (CTGF) und hepatocyt growth factor (HGF) nach laserinduzierter Thermotherapie (LITT) und chirurgischer Resektion experimenteller Lebermetastasen." [S.l.] : [s.n.], 2004. http://www.diss.fu-berlin.de/2004/183/index.html.
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Junglas, Benjamin [Verfasser], and Ernst [Akademischer Betreuer] Tamm. "Untersuchungen zur molekularen Funktion des Connective Tissue Growth Factor im Trabekelwerk / Benjamin Junglas. Betreuer: Ernst Tamm." Regensburg : Universitätsbibliothek Regensburg, 2010. http://d-nb.info/1022819542/34.
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Szabo, Z. (Zoltan). "Modulation of connective tissue growth factor and activin receptor 2b function in cardiac hypertrophy and fibrosis." Doctoral thesis, Oulun yliopisto, 2019. http://urn.fi/urn:isbn:9789526223407.
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Abstract The increase of cardiac hemodynamic load that requires increased mechanical performance drives adaptation of the heart to maintain cardiac function. Modification of protein synthesis in cardiomyocytes allows the cells to adapt to the increased load. Cardiomyocyte hypertrophy and activation of cardiac fibroblasts over the long term is maladaptive and leads to heart failure (HF). Members of the transforming growth factor-β (TGF-β) superfamily contribute to the remodeling process. TGF-β1 acts as a paracrine messenger between cardiomyocytes and cardiac fibroblasts. Connective tissue growth factor (CTGF) modulates TGF-β signaling and plays a role in the development of fibrosis. In the current study, we aimed to investigate whether blocking the actions of CTGF could alleviate ischemic injury and reduce cardiac remodeling. We determined whether blocking the action of these ligands would modulate cardiac hypertrophy and fibrosis. In the first study, we found that antagonizing the function of CTGF protected from transverse aortic constriction (TAC) -induced left ventricular remodeling. In the second study in myocardial infarction (MI) model, blocking the function of CTGF resulted in improved post-MI survival and this prevented to the decrease in left ventricular contractile function as compared to the situation in control mice. Treatment with CTGF mAb attenuated the development of dilated cardiomyopathy and limited the increase in cardiomyocyte size and deposition of interstitial fibrosis in a remote area. In the third study, targeting the TGF-β superfamily members myostatin and activins, by administration of a soluble decoy receptor of activin receptor 2B (ACVR2B-Fc) did not affect the extent of MI injury or cardiac remodeling in MI -induced ischemic HF. Understanding the complex and converging pathways regulating cardiac remodeling is a major challenge, but it may allow for opportunities to develop new therapies, new medicines and provide new hope for people with these life-threatening diseasesTiivistelmä Sydämen lisääntynyt kuormitus vaatii lisääntynyttä supistusvoimaa, joka johtaa sydänlihaksen adaptaatioon pumppaustehon ylläpitämiseksi. Alkuvaiheessa sydämen liikakasvu on hyödyllistä, mutta pidempään jatkuessaan se johtaa lopulta pumppaustoiminnan heikkenemiseen ja sydämen vajaatoimintaan. Useiden signalointimekanismien on osoitettu säätelevän sydänlihaksen adaptoitumista patologisille tiloille. Transformoiva kasvutekijä –β (TGF-β) proteiiniperhe säätelee sydämen adaptoitumista sekä vasemman kammion seinämän myötäävyyttä venytykselle. TGF-β1 indusoi supistuskykyisten myofibroblastien muodostumista sekä kollageenin tuotantoa. Runsas kollageenin tuotanto vahvistaa sydämen seinämää ja on tarpeen sydäninfarktivaurion korjaamisessa, mutta pitkään jatkuessaan se heikentää sydämen toimintaa ja altistaa rytmihäiriöille, sydämen vajaatoiminnalle sekä sydänperäiselle äkkikuolemalle. Sidekudoskasvutekijä (CTGF) säätelee TGF-β1:n signalointia ja se osallistuu haavan paranemiseen sekä fibroosiin. Tutkimuksessa selvitettiin, voidaanko sidekudoskasvutekijän tai TGF-β -perheen proteiinien toimintaa estämällä lievittää sydämen vajaatoiminnan kehittymistä. Koetuloksemme osoittavat, että CTGF:n toiminnan estäminen vasta-aineen (mAb) avulla vähentää hemodynaamisen liikakuormituksen indusoimaa vasemman kammion toiminnan heikkenemistä, kammion laajenemista sekä fibroosia. CTGF mAb myös vähentää kuolleisuutta ja estää sydämen toiminnan heikkenemistä sydäninfarktin jälkeen sekä lievittää sydäninfarktin jälkeistä dilatoivan kardiomyopatian kehittymistä. Aktiviinien ja myostatiinin toiminnan esto liukoisen aktiviinireseptori 2B:n (ACVR2B-Fc) avulla sen sijaan ei vaikuta sydäninfarktivaurioon tai iskeemisen vajaatoiminnan kehittymiseen. ACVR2B-Fc kuitenkin lisää luurankolihaksen kasvua, estäen sydämen vajaatoimintaan liittyvää luurankolihaskatoa. Sydämen hypertrofian ja vajaatoiminnan syntymisen kannalta keskeisten signaalinvälitysreittien tunnistaminen ja niiden toiminnan ymmärtäminen auttaisi kehittämään tehokkaampia lääkehoitoja sydänsairauksiin
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Mundy, Christina Maria. "The Interaction Between Connective Tissue Growth Factor and Bone Morphogenetic Protein-2 During Osteoblast Differentiation and Function." Diss., Temple University Libraries, 2014. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/269581.
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Cell BiologyPh.D.
Connective tissue growth factor (CTGF/CCN2) and bone morphogenetic protein (BMP)-2 are both produced and secreted by osteoblasts. Both proteins have been shown to have independent effects in regulating osteoblast proliferation, maturation and mineralization. However, how these two proteins interact during osteoblast differentiation remains unknown. In Chapters 2 and 3, we utilized two cell culture model systems, osteoblasts derived from CTGF knockout (KO) mice and osteoblasts infected with an adenovirus, which over-expresses CTGF (Ad-CTGF), to investigate the effects of CTGF and BMP-2 on osteoblast development and function in vitro. To observe differences in osteoblast maturation and mineralization, we performed alkaline phosphatase (ALP) staining and activity and alizarin red staining, respectively. Contrary to a previously published report, osteoblast maturation and mineralization were similar in osteogenic cultures derived from KO and wild type (WT) calvaria in the absence of BMP-2 stimulation. Interestingly, in KO and WT osteoblast cultures stimulated with BMP-2, the KO osteoblast cultures exhibited increased alkaline phosphatase staining and activity and had larger, fused nodules stained with alizarin red than WT osteoblast cultures. This increase in osteoblast differentiation was accompanied by increased protein levels of phosphorylated Smad 1/5/8 and mRNA expression levels of bone morphogenetic protein receptor Ib. These data confirm enhanced osteoblast maturation and mineralization in BMP-2 induced KO osteoblast cultures. We also examined osteoblast differentiation in cultures that were infected with Ad-CTGF and in control cultures. Continuous over-expression of CTGF resulted in decreased ALP staining and activity, alizarin red staining, and mRNA expression of osteoblast markers in both unstimulated and BMP-2 stimulated cultures. Impaired osteoblast differentiation in cultures over-expressing CTGF was accompanied by decreased protein levels of phosphorylated Smad 1/5/8. In addition to the functional assays that we performed on WT and KO osteoblast cultures, we performed ChIP assays to investigate differences in binding occupancy of transcription factors on the Runx2 and Osteocalcin promoters in BMP-2 induced WT and KO osteoblast cultures. We demonstrate that in BMP-2 induced WT and KO osteoblast cultures, there was greater Smad 1 and JunB occupancy on the Runx2 promoter and Runx2 occupancy on the Osteocalcin promoter in BMP-2 induced KO osteoblast cultures compared to WT cultures. Collectively, the data demonstrate that CTGF acts to negatively regulate BMP-2 induced signaling and osteoblast differentiation. In Chapter 4, we synthesized an active His-tagged BMP-2 recombinant protein to track surface binding of BMP-2 in CTGF WT and KO osteoblasts. We amplified mature BMP-2 in genomic DNA, which was inserted correctly into a pET-28b(+) vector. We ran a SDS-PAGE gel and stained with Coomassie blue to show that we successfully induced BMP-2 in bacteria cells, extracted the protein using urea, and purified and eluted the protein using Nickel charged agarose beads and imidazole elution buffer. Furthermore, by Western blot analysis using anti-His antibody, we confirmed the presence of the His-tag on the BMP-2 protein. Lastly, ALP staining on osteoblast cultures stimulated with our synthesized BMP-2 exhibited increased staining compared to the unstimulated osteoblast cultures, which confirmed the activity of our His-tagged BMP-2 protein. Future studies utilizing this protein will demonstrate that CTGF acts as an extracellular antagonist by limiting the amount of BMP-2 available for receptor binding.
Temple University--Theses
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Pföhler, Johanna [Verfasser]. "In vitro Untersuchungen zur Wirkungsweise eines antifibrotisch wirksamen Antikörpers gegen Connective Tissue Growth Factor in Kombination mit Transforming Growth Factor-beta oder ionisierender Strahlung / Johanna Pföhler." Berlin : Freie Universität Berlin, 2015. http://d-nb.info/107215529X/34.
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Welch, Mathew D. "Molecular mechanism underlying aberrant expression of the connective tissue growth factor in paediatric pre-B cell acute lymphoblastic leukemia." Thesis, Curtin University, 2011. http://hdl.handle.net/20.500.11937/211.
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Acute lymphoblastic leukaemia (ALL) is the most common cancer diagnosed in children aged 1-14 years. There have been vast improvements in clinical outcomes for children diagnosed with ALL with cure rates of up to 90% achievable for some forms of the disease. Despite these successes, some patients still relapse and the prognosis for these individuals is poor, thus there is still a great deal to be learned about the complex biology underlying ALL. Connective tissue growth factor (CTGF/CCN2) is a novel candidate gene in precursor B-cell (pre-B) ALL, and is aberrantly expressed in a high proportion (around 75%) of cases. While the CTGF protein has no known role in lymphocyte biology or haemopoiesis, CTGF gene expression has been associated with a poor outcome in children and adults, particularly in those patients deemed to have a high-risk of relapse. The primary aims of this study were to characterise CTGF expression in paediatric pre-B ALL tumours and cell lines, and to investigate what mechanisms were responsible for its deregulated expression by examining the contribution of both genetic and epigenetic factors.Analysis of a cohort of 73 primary paediatric pre-B ALL specimens, confirmed CTGF was aberrantly expressed in 75% of patients at heterogeneous levels. CTGF expression was found to be associated with prognosis in this cohort, as there was a trend toward lower 5 year relapse free survival (RFS) in patients with CTGFpos ALL (71% RFS in CTGFpos, 83% RFS in CTGFlow/neg), however this did not reach statistical significance (p=0.39). There was no difference in overall 5 year survival (OS) between patients with CTGFpos and CTGFlow/neg ALL. The association between CTGF expression and clinical features recorded at the time of biopsy was undertaken. These features included; age, gender, percentage blasts in bone marrow, peripheral haemoglobin level, spleen or lymph node involvement, or whether the sample was obtained at diagnosis or relapse. Patients with enlarged lymph nodes displayed a lower mean CTGF expression compared to those patients with no lymph node involvement. A similar pattern was observed with patients exhibiting enlargement of the spleen, however this did not reach statistical significance. No other clinical features were associated with CTGF expression.Analysis of global gene expression patterns in three independent paediatric pre-B ALL cohorts (PMH; n=73, Ross; n=118, Kang; n=207), identified five genes that were highly correlated with CTGF expression; SOCS2, MEF2C, ADD3, GSN and DPYSL2. In silico analysis of the 5’ flanking sequences of these genes, as well as CTGF identified predicted binding sites for the two Ikaros family proteins IKAROS and HELIOS, with at least one HELIOS site present in the 5’ flanking sequence of all correlated genes. This suggested a possible role for Ikaros family proteins in modulating CTGF gene expression. Subsequent analysis of a microarray cohort recently characterised for IKAROS gene deletions revealed that mutations or deletions within the IKAROS coding region were significantly associated with higher mean CTGF gene expression, implying that a loss of IKAROS function may promote activation of the CTGF locus. This is the first evidence implicating the Ikaros family of lymphoid transcriptional regulators in deregulation of the CTGF locus.Northern blotting of RNA from B-lineage ALL cell lines uncovered evidence of alternative splicing of CTGF mRNA. Non-canonical transcripts of approximately 1.3kb and 1.6 kb were hybridised by a CTGF-specific probe in extracts from all CTGFpos cell lines. Sequencing of cDNA fragments as well as 5’ and 3’ RACE products from a cell line with the highest level of CTGF expression revealed a number of CTGF transcripts exhibiting internal deletions of exons 2 and 3, as well as truncation of exons 1 and 4. 3’ RACE also identified a 1.3kb transcript that was devoid of 3’ UTR regulatory elements as a result of premature polyadenylation. These findings represent the first direct evidence of alternative splicing of CTGF pre-mRNA in any tissue type and further investigation is warranted to fully characterise these tumour-associated transcripts and their protein coding potential.Structural changes within the genome are common in leukaemia and deletions affecting the long arm of chromosome 6 occur in around 30% of cases of pre-B ALL. To investigate whether deregulation of CTGF expression has a genetic basis, the CTGF locus at 6q23.1 was investigated for structural alterations or mutations. Southern blotting performed in seven B-lineage ALL cell lines (4 CTGFpos, 3 CTGFneg) confirmed that the CTGF locus was not cytogenetically rearranged. Furthermore, analysis of CTGF copy number by qPCR in primary paediatric pre-B ALL specimens (n=17) and B-lineage ALL cell lines (n=7) confirmed that gene amplification could not account for CTGF overexpression. Sequencing of the CTGF promoter and 3’ UTR in three B-lineage ALL cell lines confirmed that there were no mutations affecting these important regulatory regions, although one cell line harboured the rs6918698 -739 C>G SNP, which is predicted to disrupt SP3-mediated repression of CTGF gene expression.Epigenetic regulation of gene expression is frequently altered in neoplasia. The CTGF locus contains a CpG island, and methylation of this region was found to be inversely correlated with CTGF gene expression in B-lineage ALL cell lines, as assessed by both methylation-specific PCR and by bisulfite sequencing. Bisulfite sequencing of primary tumour specimens revealed that hypomethylation of the CTGF locus was a widespread feature of pre-B ALL. By contrast, analysis of primary T-ALL specimens demonstrated extensive methylation at the CTGF locus, indicating that CTGF may be permissive for expression specifically in pre-B ALL.This study has highlighted several novel aspects of CTGF expression in pre-B ALL, including a potential role for Ikaros family proteins in regulating the CTGF locus, and the existence of CTGF mRNA transcripts generated through alternative pre-mRNA splicing. Investigation of mechanisms promoting CTGF gene expression in pre-B ALL revealed that the rs6918698 C>G SNP was present in one pre-B ALL cell line. Hypomethylation of the CTGF locus was a notable feature of primary pre-B ALL specimens and was in contrast to the hypermethylation observed in 2 T-ALL specimens and CD34pos bone marrow cells. These findings will direct future research to elucidate the complex mechanisms regulating CTGF expression in pre-B ALL. It is anticipated that illuminating the role of CTGF in the pathogenesis of ALL may result in significantly improved patient outcomes through the development of targeted and less toxic therapies, and through improved risk-based stratification of patients to ensure those at a high risk of relapse are directed toward an appropriate level of therapeutic intervention.
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Pi, Liya. "The role of connective tissue growth factor (ctgf) in oval cell aided liver regeneration in the 2-aaf/phx model." [Gainesville, Fla.] : University of Florida, 2005. http://purl.fcla.edu/fcla/etd/UFE0010022.
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Thesis (Ph.D.)--University of Florida, 2005.Typescript. Title from title page of source document. Document formatted into pages; contains 162 pages. Includes Vita. Includes bibliographical references.
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Wu, Shumei Jocelyn. "Immunochemical techniques for the detection, isolation and characterization of connective tissue growth factor in diabetic urine and peritoneal dialysis fluid." Thesis, King's College London (University of London), 2012. https://kclpure.kcl.ac.uk/portal/en/theses/immunochemical-techniques-for-the-detection-isolation-and-characterization-of-connective-tissue-growth-factor-in-diabetic-urine-and-peritoneal-dialysis-fluid(c1f3c279-c133-42fa-a733-005deb10ba13).html.
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Connective tissue growth factor (CTGF) has been implicated in the pathogenesis of diabetic nephropathy and has been proposed as a possible specific biomarker for the early detection of fibrosis. CTGF is reported to be a 349-amino acid cysteine-rich, heparin binding monomeric polypeptide that is secreted as a 36-38 kDa protein. To date, the natural form of CTGF has not been successfully purified from clinical samples. Commercial test kits for detecting CTGF by ELISA utilize antibodies raised against recombinant human (rh) CTGF. In the present study, assessment of specific analytical methods for the reliable detection of naturally occurring CTGF has been deployed. In order to achieve this, a peptide sequence to CTGF was synthesized and an antibody raised against this peptide. This antibody was characterized using novel chemistry to improve the specificity of the final assay. The antibody raised against a unique peptide sequence in CTGF, and another to rhCTGF, were used, alongside a commercially available anti-rhCTGF antibody to detect this protein in diabetic urine and peritoneal dialysis fluid. The synthetic peptide antibody was also coupled to Sepharose 4B for the purification of CTGF from clinical samples. ELISA performed on the diabetic urine and peritoneal dialysis fluid showed that the response of the antibody against the synthetic peptide was the highest when compared to the rhCTGF and commercial antibodies. Five peritoneal dialysis fluid and five diabetic urine samples were assayed for CTGF. Diabetic urine was found to be a richer source of CTGF. 2-dimemsional electrophoresis and Western blot analyses on the peritoneal dialysis fluid showed various immunoreactive species to the synthetic peptide antibody at isoelectric point 8.0. It can be concluded that these immunoreactive species may be CTGF bound to other proteins or CTGF complexes that have been formed from the oxidation of the cysteine residues.
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Toda, Naohiro. "Crucial Role of Mesangial Cell-derived Connective Tissue Growth Factor in a Mouse Model of Anti-Glomerular Basement Membrane Glomerulonephritis." Kyoto University, 2018. http://hdl.handle.net/2433/232131.
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Küspert, Sabrina Angela [Verfasser], and Rudolf [Akademischer Betreuer] Fuchshofer. "Die funktionelle Rolle des Wachstumsfaktors Connective Tissue Growth Factor in der Pathogenese des primären Offenwinkelglaukoms / Sabrina Angela Küspert. Betreuer: Rudolf Fuchshofer." Regensburg : Universitätsbibliothek Regensburg, 2015. http://d-nb.info/1065445261/34.
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Lambi, Alex G. "The Intricate Role of Connective Tissue Growth Factor (CTGF/CCN2) in Prenatal Osteogenesis: A Heretofore Oversimplified Dogma of the CCN Field." Diss., Temple University Libraries, 2015. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/233693.
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Cell BiologyPh.D.
Connective tissue growth factor (CTGF/CCN2) is axiomatically necessary for proper skeletal development and function. We need not look further than the studies that have been done to date utilizing mice genetically engineered to lack CTGF production. These CTGF null or knockout (KO) mice fail to form a normal murine skeleton and instead yield one littered with bony dysmorphisms, including incompetent craniofacial development, kinked limb bones, and misshapen ribs that are not conducive to proper respiratory function. As a result, the global lack of CTGF is incompatible with postnatal life. A closer look at several sites demonstrated defects in physiologic processes necessary for bone formation - angiogenesis, chondrogenesis, and osteogenesis. Therefore, the dogma in the CCN protein field to date has been that systemic ablation of CTGF production in vivo results in global defects in bone development. We believe this dogma is an oversimplification of the role of CTGF on skeletal development. Our initial impetus leading us to this belief was the gross identification of the specific skeletal sites malformed in CTGF KO mice, in particular the bones of the limbs. While in the lower limb of CTGF KO mice the tibiae and fibulae are misshapen, the adjacent femora and digits are phenotypically normal. The same is true for the upper limb, in which the radii and ulnae are phenotypically abnormal while the humeri and digits are normal. Therefore, we believe that the role of CTGF in skeletogenesis is site-specific such that its loss affects local skeletal patterning and/or mechanobiological cues resulting in the unique phenotype seen in CTGF KO mice. The research of this dissertation constitutes a comprehensive skeletal analysis of CTGF KO mice and in so doing we determined the extent and location of skeletal abnormalities. We found skeletal site-specific changes in growth plate organization, bone microarchitecture and shape and gene expression levels in CTGF KO compared to wild-type (WT) mice. Growth plate malformations included reduced proliferation zone and increased hypertrophic zone lengths. Appendicular skeletal sites demonstrated decreased metaphyseal trabecular bone, while having increased mid-diaphyseal bone and osteogenic expression markers. Axial skeletal analysis showed decreased bone in caudal vertebral bodies, mandibles, and parietal bones in CTGF KO mice, with decreased expression of osteogenic markers. Analysis of skull phenotypes demonstrated global and regional differences in CTGF KO skull shape resulting from allometric (size-based) and non-allometric shape changes. Localized differences in skull morphology included increased skull width and decreased skull length. We further continued the skeletal characterization of CTGF KO bones with an analysis of bone cell ultrastructure and matrix composition. These studies demonstrated that, while CTGF is not necessary for complete morphologic maturation of bone cells, global ablation results in ultrastructural features not commonly seen in WT bones. Our findings include drastically dilated rough endoplasmic reticulum (RER) in osteoblasts of the tibial diaphyseal region, comprising the phenotypic kink in CTGF KO mice and ultrastructural dysmorphologies of CTGF KO osteoclasts including multi-layered, membranous inclusions, decreased vacuolization and ruffled border extents, and disproportionately large clear zones. Lastly, FT-IR analysis demonstrated heterogeneity in CTGF KO bone composition. The results of this dissertation have revealed a more complex role for CTGF in osteogenesis and have identified potential mechanisms and future research directions to fully understand this intricate story.
Temple University--Theses
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Moroney, James B. "The effect of thyroid hormone-dependent dermal fibroblast proliferation: an investigation of connective tissue growth factor and proliferative cell nuclear antigen." Thesis, Boston University, 2013. https://hdl.handle.net/2144/12164.
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Thesis (M.A.)--Boston UniversityThyroid hormone has significant impact on skin homeostasis and cutaneous wound healing. Previous research has demonstrated both in vitro proliferation of keratinocytes and fibroblasts and in vivo stimulation of epidermal and dermal layers in response to triiodothyronine (T3) administration. However, the physiological mechanism of action involving T3 signaling and the specific intermediate factors of T3-induced cell proliferation are poorly understood. Currently, there is no working model of T3-dependent dermal fibroblast proliferation. In order to gain a more complete understanding of thyroid hormone regulation in wound healing, two known proliferative growth factors, connective tissue growth factor (CTGF) and proliferative cell nuclear antigen (PCNA), were chosen as potential mediators of T3-stimulated fibroblast proliferation. In vitro dermal fibroblast cultures were dosed with one of three experimental T3 concentrations (10-9 M, 10-8 M and 10-7 M) and western blot analysis was conducted to determine whether CTGF and PCNA expression are regulated by T3 stimulation. The results indicated no significant change in CTGF or PCNA expression dependent on T3 concentration. The implications of the findings were addressed and suggestions for future research directions have been proposed. It is still unclear which growth factors are involved in T3-regulated fibroblast proliferation. Once these mediators are identified, it will be possible to construct a mechanism of action to integrate the findings and ultimately develop a complete understanding of cutaneous physiology.
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Friedrich, Emily [Verfasser], and Margarete [Akademischer Betreuer] Goppelt-Strübe. "Regulation der Genexpression von Connective Tissue Growth Factor (CTGF) in humanen Tubulusepithelzellen unter Einfluss von Hypoxie / Emily Friedrich. Gutachter: Margarete Goppelt-Strübe." Erlangen : Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), 2013. http://d-nb.info/1054342210/34.
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LOCATELLI, LUIGI. "Expression of aVB6 integrin by Pkhd1-defective cholangiocytes links enhanced ductal secretion of Macrophage chemokines to progressive portal fibrosis in Congenital Hepatic Fibrosis." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2013. http://hdl.handle.net/10281/41733.
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BACKGROUND AND AIMS: Congenital Hepatic Fibrosis (CHF) is caused by mutations in PKHD1, a gene encoding for fibrocystin, a protein of unknown function, expressed in cholangiocyte cilia and centromers. In CHF, biliary dysgenesis is accompanied by severe progressive portal fibrosis and portal hypertension. The mechanisms responsible for portal fibrosis in CHF are unclear. The αvβ6 integrin mediates local activation of TGFβ1 and is expressed by reactive cholangiocytes during cholestasis. To understand the mechanisms of fibrosis in CHF we studied the expression of αvβ6 integrin and its regulation in Pkhd1del4/del4 mice.
METHODS: In Pkhd1del4/del4 mice we studied, at different ages (1-12 months): a) portal fibrosis (Sirius Red) and portal hypertension (spleen weight/body weight); b) αvβ6 mRNA and protein expression (RT-PCR, IHC); c) α-SMA and TGFβ1 mRNA expression (RT-PCR); d) portal inflammatory infiltrate (IHC for CD45 and FACS analysis of whole liver infiltrate); f) cytokines secretion from cultured monolayers of primary cholangiocytes (Luminex assay); g) cytokine effects on monocyte/macrophage proliferation (MTS assay) and migration (Boyden chamber); h) TGFβ1 and TNFα effects on β6 integrin mRNA expression by cultured cholangiocytes before and after inhibition of the TGFβ receptor type II (TGFβRII); i) TGFβ1 effects on collagen type I (COLL1) mRNA expression by cultured cholangiocytes.
RESULTS: Pkhd1del4/del4 mice showed a progressive increase in αvβ6 integrin expression on biliary cyst epithelia. Expression of αvβ6 correlated with portal fibrosis (r=0.94, p<0.02) and with enrichment of a CD45+ve cell infiltrate in the portal space (r=0.97, p<0.01). Gene expression of TGFβ1 showed a similar age-dependent increase. FACS analysis showed that 50-75% of the CD45+ve cells were macrophages (CD45/CD11b/F4/80+ve). Cultured polarized Pkhd1del4/del4 cholangiocytes secreted from the basolateral side significantly increased amounts of CXCL1 and CXCL10 (p<0.05). Both cytokines were able to stimulate macrophage migration (p<0.05). Basal expression of β6 mRNA by cultured Pkhd1del4/del4 cholangiocytes (0.015±0.002 2^-dCt) was potently stimulated by the macrophage-derived cytokines TGFβ1 (0.017±0.002 2^-dCt, p<0.05) and TNFα (0.018±0.003 2^-dCt, p<0.05). Inhibition of TGFβRII completely blunted TGFβ1 (0.014±0.003 2^-dCt, p<0.05) but not TNFα effects (0.017±0.001 2^-dCt, p=ns) on β6 mRNA. COLL1 mRNA expression by cultured Pkhd1del4/del4 cholangiocytes (0.0009±0.0003 2^-dCt) was further and significantly increased after TGFβ1 stimulation (0.002±0.0005 2^-dCt, p<0.05).
CONCLUSIONS: Pkhd1del4/del4 cholangiocytes possess increased basolateral secretory functions of chemokines (CXCL1, CXCL10) able to orchestrate macrophage homing to the peribiliary microenvironment. In turn, by releasing TGFβ1 and TNFα, macrophages up-regulate αvβ6 integrin in Pkhd1del4/del4 cholangiocytes. αvβ6 integrin activates latent TGFβ1, further increasing the fibrogenic properties of cholangiocytes.
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Gonzales, Christopher R. "3,3′,5′-triido-L-thyronine alters protein kinase B, phosphotase and tensin homolog and connective tissue growth factor expression in human dermal fibroblasts." Thesis, Boston University, 2012. https://hdl.handle.net/2144/12397.
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Thesis (M.A.)--Boston University
PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you.Cutaneous tissue repair is complex and involves a variety of growth factors to regulate a balance of regeneration and fibrosis during healing. This process is divided into three sequential and overlapping phases: the inflammatory phase, the proliferative phase, and the remodeling phase. Fibroblasts are crucial during this process in that they help initiate inflammatory activity, deposit extracellular matrix proteins for granulation tissue and deconstruct granulation tissue to make way for mature scar formation. Previous studies on the effects of 3,3',5'-triiodo-L-thyronine (T3) on skin have revealed that healing tissue responds to T3 by accelerating skin cell proliferation and migration. These findings indicate that T3 offers potential as a therapeutic drug for individuals with extensive cutaneous damage, chronic skin maladies or retarded wound healing. The mechanisms underlying these changes are not clearly understood, however, elucidation of changes in protein expression patterns should be evaluated to appropriately judge the therapeutic potential of T3. This study aims to characterize T3 dose responsive expression of protein kinase B, phosphatase and tensin homolog, connective tissue growth factor and wnt5a. Western blot analysis and immunodetection revealed that wnt5a is not expressed in human dermal fibroblasts. Protein kinase B did not vary significantly with T3 concentration ranging from 1.0 nM-1.0 1µM, F(4,5)= 1.93, p > 0.05, nor did connective tissue growth factor, F(4,5) = 2.16, p > 0.05. In contrast, phosphatase and tensin homolog showed a statistically significant change in expression, F(4, 15) = 4.67, p less than 0.05. The results presented here provide insight into protein pathways and growth factors through which thyroid hormone produces its effects on the various cells of the integument and suggests that phosphatase and tensin homolog (PTEN) expression levels are responsive to varying concentrations of T3. Future studies should further evaluate the role of T3 on its various targets as a therapeutic option for skin disorders.
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Pante, Saskia Veronika [Verfasser]. "Die Rolle von Chemokinrezeptor CXCR4 und Connective Tissue Growth Factor innerhalb der Tumor-Stroma-Interaktion in der akuten myeloischen Leukämie / Saskia Veronika Pante." Mainz : Universitätsbibliothek Mainz, 2014. http://d-nb.info/1052000347/34.
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Kiwanuka, Elizabeth. "CCN2 – Keratinocyte Interactions In Vitro and In Vivo." Doctoral thesis, Uppsala universitet, Plastikkirurgi, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-213566.
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Cutaneous wound healing is a complex process involving the migration of inflammatory cells to the wound site, deposition of extracellular matrix, and the reestablishment of an intact epithelial barrier. Re-epithelialization depends on the proliferation and directional migration of keratinocytes from the wound edges. Initially, keratinocytes migrate over a provisional wound matrix that is rich in fibronectin, and as the wound heals the provisional matrix becomes replaced by one consisting of collagen and proteoglycans. Re-epithelialization is tightly regulated by a variety of peptides such as growth factors, cytokines and proteases, and abnormalities may result in chronic non-healing wounds or hypertrophic scars. CCN2 (Connective Tissue Growth Factor) is a multifunctional protein with effects on cells and their interactions with the connective tissue. CCN2 is expressed in a variety of cell types and regulates numerous cell functions including proliferation, differentiation, adhesion, migration and stimulation of collagen production. While the importance of CCN2 for the fibrotic response has been well studied, its involvement in keratinocyte function has not yet been fully explored. Using an in vivo wound model, the expression of CCN2 was captured at the leading keratinocyte edge during re-epithelialization. In vitro, exogenous addition of CCN2 to human keratinocyte cultures promoted keratinocyte migration. Subsequently, integrin a5b1 was identified as an important mediator of CCN2 enhancement of keratinocyte adhesion to fibronectin. CCN2 activated the FAK-MAPK signaling pathway, and pretreatment with MEK1 specific inhibitor PD98059 markedly reduced CCN2-promoted keratinocyte migration. In vitro, CCN2 expression was induced by TGF-β1. Compared with inhibiting the SMAD pathway, blocking MAPK was more effective in reducing TGF-β1-induced CCN2 mRNA and protein expression. In addition, CCN2-induced keratinocyte spreading required FAK. Treatment with CCN2 led to actin disassembly and altered the activity of the Rho proteins and p190RhoGAP in keratinocytes. Furthermore, Cdc42 mediated CCN2-induced cell polarity. In conclusion, using in vivo and in vitro models, CCN2 was shown to regulate keratinocyte function by promoting keratinocyte adhesion, spreading and migration. A complete understanding of CCN2 expression in keratinocytes is crucial in order to develop novel therapies for wound healing.
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Raut, Vivek P. "METHODS TO QUANTITATIVELY ASSESS THE PERFORMANCE OF CONNECTIVE TISSUE PROGENITOR CELLS IN RESPONSE TO SURFACE MODIFIED BIOMATERIALS." Case Western Reserve University School of Graduate Studies / OhioLINK, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=case1372334668.
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Hendesi, Honey. "CONNECTIVE TISSUE GROWTH FACTOR (CTGF/CCN2) REGULATES OSTEOBLAST CYTOSKELETAL REORGANIZATION AND MOTILITY AND ENHANCES DIFFERENTIATION VIA BINDING TO INTEGRIN RECEPTORS AND ACTIVATION OF DOWNSTREAM SIGNALINGS." Diss., Temple University Libraries, 2014. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/263674.
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Cell BiologyPh.D.
Connective Tissue Growth Factor (CTGF) is a matricellular protein that has been shown to mediate cell adhesion, and as a consequence, it regulates cell proliferation, migration, differentiation and gene transcription. Although previous in vivo and in vitro studies supported the anabolic role of CTGF in skeletogenesis, to date mechanisms of this effect remain unknown. So far, no specific receptor has been identified for CTGF, although previous studies have shown that integrins can serve as functional signaling receptors for CTGF. The CTGF-integrin interaction initiates intracellular signaling cascades that ultimately regulate cell cytoskeleton reorganization, gene transcription and cell function. To study the effect of CTGF on osteoblasts, we first conducted adhesion assays using the MC3T3-E1 osteoblastic cell line. We confirmed that osteoblasts adhere to rCTGF in a concentration-dependent manner and we showed this adhesion has characteristics of integrin mediated adhesions. Next, we used an array of blocking antibodies directed against the individual alpha and beta; integrin subunits that are known to be expressed in osteoblasts. Significant decreases in cell adhesion were observed upon treatment with anti-alpha-v or anti-beta1 blocking antibodies. Subsequent coimmunoprecipitation analyses demonstrated that CTGF interacts with alpha-v and beta1 integrins in osteoblasts. Furthermore, we showed that the specificity of this CTGF-integrin interaction occurs in the C-terminal domain (fourth module) of CTGF. The immunefluorescence staining of cells cultured on substrates of rCTGF, fibronectin (positive control) or BSA (negative control) demonstrated that osteoblast adhesion to rCTGF results in actin cytoskeleton reorganization, focal adhesion formation, enhanced cell spreading and Rac activation. These series of events are necessary for proper cell-matrix interaction and integrins' downstream signaling initiation. Next, through alkaline phosphatase (ALP) staining and activity assays, as well as Alizarin red staining, we demonstrated that osteoblast attachment to CTGF matrix enhances cell maturation, bone nodule formation and matrix mineralization. To investigate whether the effect of CTGF on osteoblast differentiation involves activation of specific signaling molecules, we performed Western blot and chromatin immunoprecipitation (ChIP) assays. Osteoblasts cultured on rCTGF expressed higher levels of both total and phosphorylated forms of focal adhesion kinase (FAK) and extracellular signal-regulated kinase (ERK) compared to the cells cultured on BSA. In addition, these osteoblasts showed an increase in runt-related transcription factor 2 (Runx2) binding to the osteocalcin gene promoter compared to the negative control. These experiments confirmed CTGF's effect on enhancing osteoblast differentiation through regulation of important signaling molecules. In another series of experiments, we used primary osteoblasts isolated from CTGF KO mice, their WT littermates, or WT cells infected to overexpress (OE) CTGF to study the effect of different levels of endogenous CTGF on osteoblast cytoskeleton reorganization and motility. Our assays showed enhanced cell adhesion, spreading and Rac expression in CTGF OE osteoblasts, while in CTGF KO osteoblasts, cell adhesion, spreading and Rac expression were significantly decreased. In contrast, CTGF OE osteoblasts that showed high adhesion and spreading, exhibited diminished cell motility and low levels of RhoA expression, while KO cells migrated quickly and expressed high levels of RhoA. Together, these experiments establish CTGF as an adhesion protein for osteoblasts; they demonstrate that the alpha-v beta1 integrin is a functional signaling receptor for CTGF; they confirm that osteoblast differentiation is enhanced when cultured on CTGF matrix through activation of regulatory signaling molecules; and finally, these experiments establish a role for CTGF in the regulation of small RhoGTPases expression, which in turn implies a significant role for CTGF in cell cytoskeleton reorganization and motility.
Temple University--Theses
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Gao, Helen Guoyi Li. "INCREASED FIBROGENIC PROTEINS FOLLOWING PERSISTENT LOW-GRADE INFLAMMATION IN A RAT MODEL OF LONG-TERM OVERUSE." Master's thesis, Temple University Libraries, 2013. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/238810.
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Biomedical SciencesM.S.
We examined the relationship between grip strength declines and muscle-tendon responses induced by long-term performance of a high-repetition, low-force (HRLF) reaching task in rats. We hypothesized that grip strength declines would correlate with inflammation, fibrosis and degradation in flexor digitorum muscles and tendons. Grip strength declined after training, and further in weeks 18 and 24, in reach limbs of HRLF rats. Flexor digitorum tissues of reach limbs showed low-grade increases in inflammatory cytokines: IL-1beta after training and in week 18, IL-1alpha in week 18, TNF-alpha and IL-6 after training and in week 24, and IL-10 in week 24, with greater increases in tendons than muscles. Similar cytokine increases were detected in serum with HRLF: IL-1alpha and IL-10 in week 18, and TNF-alpha and IL-6 in week 24. Grip strength correlated inversely with IL-6 in muscles, tendons and serum, and TNF-alpha in muscles and serum. Four fibrogenic proteins, TGFB1, CTGF, PDGFab and PDGFbb, and hydroxyproline, a marker of collagen synthesis, increased in serum in HRLF weeks 18 or 24, concomitant with epitendon thickening, increased muscle and tendon TGFB1 and CTGF. A collagenolytic gelatinase, MMP2, increased by week 18 in serum, tendons and muscles of HRLF rats. Grip strength correlated inversely with TGFB1 in muscles, tendons and serum; with CTGF-immunoreactive fibroblasts in tendons; and with MMP2 in tendons and serum. Thus, motor declines correlated with low-grade systemic and musculotendinous inflammation throughout task performance, and increased fibrogenic and degradative proteins with prolonged task performance. Serum TNF-alpha, IL-6, TGFB1, CTGF and MMP2 may serve as serum biomarkers of work-related musculoskeletal disorders, although further studies in humans are needed.
Temple University--Theses
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Kim, Sun Wook. "Modulation of Stem Cell Fate by Electrical Stimulation." University of Cincinnati / OhioLINK, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1383812480.
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Singh, Simranjit [Verfasser], Susanne [Akademischer Betreuer] Lutz, Dörthe [Gutachter] Katschinski, Viacheslav [Gutachter] Nikolaev, and Blanche [Gutachter] Schwappach. "Redox regulation of protein phosphatase-1 and ER stress regulation of connective tissue growth factor in cardiomyocytes / Simranjit Singh ; Gutachter: Dörthe Katschinski, Viacheslav Nikolaev, Blanche Schwappach ; Betreuer: Susanne Lutz." Göttingen : Niedersächsische Staats- und Universitätsbibliothek Göttingen, 2017. http://d-nb.info/1136131604/34.
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Khaghani, Seyed A. "Cell and tissue engineering of articular cartilage via regulation and alignment of primary chondrocyte using manipulated transforming growth factors and ECM proteins. Effect of transforming growth factor-beta (TGF-¿1, 2 and 3) on the biological regulation and wound repair of chondrocyte monolayers with and without presence of ECM proteins." Thesis, University of Bradford, 2010. http://hdl.handle.net/10454/5371.
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Articular cartilage is an avascular and flexible connective tissue found in joints. It produces a cushioning effect at the joints and provides low friction to protect the ends of the bones from wear and tear/damage. It has poor repair capacity and any injury can result pain and loss of mobility. One of the common forms of articular cartilage disease which has a huge impact on patient¿s life is arthritis. Research on cartilage cell/tissue engineering will help patients to improve their physical activity by replacing or treating the diseased/damaged cartilage tissue.
Cartilage cell, called chondrocyte is embedded in the matrix (Lacunae) and has round shape in vivo. The in vitro monolayer culture of primary chondrocyte causes morphological change characterized as dedifferentiation. Transforming growth factor-beta (TGF-¿), a cytokine superfamily, regulates cell function, including differentiation and proliferation. The effect of TGF-¿1, 2, 3, and their manipulated forms in biological regulation of primary chondrocyte was investigated in this work. A novel method was developed to isolate and purify the primary chondrocytes from knee joint of neonate Sprague-Dawley rat, and the effect of some supplementations such as hyaluronic acid and antibiotics were also investigated to provide the most appropriate condition for in vitro culture of chondrocyte cells.
Addition of 0.1mg/ml hyaluronic acid in chondrocyte culture media resulted an increase in primary chondrocyte proliferation and helped the cells to maintain chondrocytic morphology.
TGF-¿1, 2 and 3 caused chondrocytes to obtain fibroblastic phenotype, alongside an increase in apoptosis. The healing process of the wound closure assay of chondrocyte monolayers were slowed down by all three isoforms of TGF-¿. All three types of TGF-¿ negatively affected the strength of chondrocyte adhesion. TGF-¿1, 2 and 3 up regulated the expression of collagen type-II, but decreased synthesis of collagen type-I, Chondroitin sulfate glycoprotein, and laminin. They did not show any significant change in production of S-100 protein and fibronectin. TGF-¿2, and 3 did not change expression of integrin-¿1 (CD29), but TGF-¿1 decreased the secretion of this adhesion protein.
Manipulated TGF-¿ showed huge impact on formation of fibroblast like morphology of chondrocytes with chondrocytic phenotype. These isoforms also decreased the expression of laminin, chondroitin sulfate glycoprotein, and collagen type-I, but they increased production of collagen type-II and did not induce synthesis of fibronectin and S-100 protein. In addition, the strength of cell adhesion on solid surface was reduced by manipulated TGF-¿. Only manipulated form of TGF-¿1 and 2 could increase the proliferation rate. Manipulation of TGF-¿ did not up regulate the expression of integrin-¿1in planar culture system.
The implications of this R&D work are that the manipulation of TGF-¿ by combination of TGF-¿1, 2, and 3 can be utilized in production of superficial zone of cartilage and perichondrium. The collagen, fibronectin and hyaluronic acid could be recruited for the fabrication of a biodegradable scaffold that promotes chondrocyte growth for autologous chondrocyte implantation or for formation of cartilage.
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Khaghani, Seyed Ali. "Cell and tissue engineering of articular cartilage via regulation and alignment of primary chondrocyte using manipulated transforming growth factors and ECM proteins : effect of transforming growth factor-beta (TGF-β1, 2 and 3) on the biological regulation and wound repair of chondrocyte monolayers with and without presence of ECM proteins." Thesis, University of Bradford, 2010. http://hdl.handle.net/10454/5371.
Full textAbstract:
Articular cartilage is an avascular and flexible connective tissue found in joints. It produces a cushioning effect at the joints and provides low friction to protect the ends of the bones from wear and tear/damage. It has poor repair capacity and any injury can result pain and loss of mobility. One of the common forms of articular cartilage disease which has a huge impact on patient's life is arthritis. Research on cartilage cell/tissue engineering will help patients to improve their physical activity by replacing or treating the diseased/damaged cartilage tissue. Cartilage cell, called chondrocyte is embedded in the matrix (Lacunae) and has round shape in vivo. The in vitro monolayer culture of primary chondrocyte causes morphological change characterized as dedifferentiation. Transforming growth factor-beta (TGF-β), a cytokine superfamily, regulates cell function, including differentiation and proliferation. The effect of TGF-β1, 2, 3, and their manipulated forms in biological regulation of primary chondrocyte was investigated in this work. A novel method was developed to isolate and purify the primary chondrocytes from knee joint of neonate Sprague-Dawley rat, and the effect of some supplementations such as hyaluronic acid and antibiotics were also investigated to provide the most appropriate condition for in vitro culture of chondrocyte cells. Addition of 0.1mg/ml hyaluronic acid in chondrocyte culture media resulted an increase in primary chondrocyte proliferation and helped the cells to maintain chondrocytic morphology. TGF-β1, 2 and 3 caused chondrocytes to obtain fibroblastic phenotype, alongside an increase in apoptosis. The healing process of the wound closure assay of chondrocyte monolayers were slowed down by all three isoforms of TGF-β. All three types of TGF-β negatively affected the strength of chondrocyte adhesion. TGF-β1, 2 and 3 up regulated the expression of collagen type-II, but decreased synthesis of collagen type-I, Chondroitin sulfate glycoprotein, and laminin. They did not show any significant change in production of S-100 protein and fibronectin. TGF-β2, and 3 did not change expression of integrin-β1 (CD29), but TGF-β1 decreased the secretion of this adhesion protein. Manipulated TGF-β showed huge impact on formation of fibroblast like morphology of chondrocytes with chondrocytic phenotype. These isoforms also decreased the expression of laminin, chondroitin sulfate glycoprotein, and collagen type-I, but they increased production of collagen type-II and did not induce synthesis of fibronectin and S-100 protein. In addition, the strength of cell adhesion on solid surface was reduced by manipulated TGF-β. Only manipulated form of TGF-β1 and 2 could increase the proliferation rate. Manipulation of TGF-β did not up regulate the expression of integrin-β1 in planar culture system. The implications of this R&D work are that the manipulation of TGF-β by combination of TGF-β1, 2, and 3 can be utilized in production of superficial zone of cartilage and perichondrium. The collagen, fibronectin and hyaluronic acid could be recruited for the fabrication of a biodegradable scaffold that promotes chondrocyte growth for autologous chondrocyte implantation or for formation of cartilage.
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Cunningham, Janet Lynn. "Tumour Biological Factors Characterizing Metastasizing Serotonin-producing Ileocaecal Carcinoids." Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl. [distributör], 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7906.
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Grasman, Jonathan M. "Designing Fibrin Microthread Scaffolds for Skeletal Muscle Regeneration." Digital WPI, 2015. https://digitalcommons.wpi.edu/etd-dissertations/18.
Full textAbstract:
Volumetric muscle loss (VML) typically results from traumatic incidents; such as those presented from combat missions, where soft-tissue extremity injuries account for approximately 63% of diagnoses. These injuries lead to a devastating loss of function due to the complete destruction of large amounts of tissue and its native basement membrane, removing important biochemical cues such as hepatocyte growth factor (HGF), which initiates endogenous muscle regeneration by recruiting progenitor cells. Clinical strategies to treat these injuries consist of autologous tissue transfer techniques, requiring large amounts of healthy donor tissue and extensive surgical procedures that can result in donor site morbidity and limited functional recovery. As such, there is a clinical need for an off-the-shelf, bioactive scaffold that directs patient’s cells to align and differentiate into muscle tissue in situ. In this thesis, we developed fibrin microthreads, scaffolds composed of aligned fibrin material that direct cell alignment along the longitudinal axis of the microthread structure, with specific structural and biochemical properties to recreate structural cues lost in VML injuries. We hypothesized that fibrin microthreads with an increased resistance to proteolytic degradation and loaded with HGF would enhance the functional, mechanical regeneration of skeletal muscle tissue in a VML injury. We developed a crosslinking strategy to increase fibrin microthread resistance to enzymatic degradation, and increased their tensile strength and stiffness two- to three-fold. This crosslinking strategy enhanced the adsorption of HGF, facilitated its rapid release from microthreads for 2 to 3 days, and increased the chemotactic response of myoblasts twofold in 2D and 3D assays. Finally, we implanted HGF-loaded, crosslinked (EDCn-HGF) microthreads into a mouse model of VML to evaluate tissue regeneration and functional recovery. Fourteen days post-injury, we observed more muscle ingrowth along EDCn-HGF microthreads than untreated controls, suggesting that released HGF recruited additional progenitor cells to the injury site. Sixty days post-injury, EDCn-HGF microthreads guided mature, organized muscle to replace the microthreads in the wound site. Further, EDCn-HGF microthreads restored the contractile mechanical strength of the tissue to pre-injured values. In summary, we designed fibrin microthreads that recapitulate regenerative cues lost in VML injuries and enhance the functional regeneration of skeletal muscle.
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McCosker, Helen Clare. "Prognostic significance of IGF and ECM induced signalling proteins in breast cancer patients." Thesis, Queensland University of Technology, 2012. https://eprints.qut.edu.au/53580/1/Helen_McCosker_Thesis.pdf.
Full textAbstract:
Breast cancer is a leading contributor to the burden of disease in Australia. Fortunately, the recent introduction of diverse therapeutic strategies have improved the survival outcome for many women. Despite this, the clinical management of breast cancer remains problematic as not all approaches are sufficiently sophisticated to take into account the heterogeneity of this disease and are unable to predict disease progression, in particular, metastasis. As such, women with good prognostic outcomes are exposed to the side effects of therapies without added benefit. Furthermore, women with aggressive disease for whom these advanced treatments would deliver benefit cannot be distinguished and opportunities for more intensive or novel treatment are lost. This study is designed to identify novel factors associated with disease progression, and the potential to inform disease prognosis. Frequently overlooked, yet common mediators of disease are the interactions that take place between the insulin-like growth factor (IGF) system and the extracellular matrix (ECM).
Our laboratory has previously demonstrated that multiprotein insulin-like growth factor-I (IGF-I): insulin-like growth factor binding protein (IGFBP): vitronectin (VN) complexes stimulate migration of breast cancer cells in vitro, via the cooperative involvement of the insulin-like growth factor type I receptor (IGF-IR) and VN-binding integrins. However, the effects of IGF and ECM protein interactions on the dissemination and progression of breast cancer in vivo are unknown. It was hypothesised that interactions between proteins required for IGF induced signalling events and those within the ECM contribute to breast cancer metastasis and are prognostic and predictive indicators of patient outcome.
To address this hypothesis, semiquantitative immunohistochemistry (IHC) analyses were performed to compare the extracellular and subcellular distribution of IGF and ECM induced signalling proteins between matched normal, primary cancer, and metastatic cancer among archival formalin-fixed paraffin-embedded (FFPE) breast tissue samples collected from women attending the Princess Alexandra Hospital, Brisbane. Multivariate Cox proportional hazards (PH) regression survival models in conjunction with a modified „purposeful selection of covariates. method were applied to determine the prognostic potential of these proteins.
This study provides the first in-depth, compartmentalised analysis of the distribution of IGF and ECM induced signalling proteins. As protein function and protein localisation are closely correlated, these findings provide novel insights into IGF signalling and ECM protein function during breast cancer development and progression. Distinct IGF signalling and ECM protein immunoreactivity was observed in the stroma and/or in subcellular locations in normal breast, primary cancer and metastatic cancer tissues. Analysis of the presence and location of stratifin (SFN) suggested a causal relationship in ECM remodelling events during breast cancer development and progression. The results of this study have also suggested that fibronectin (FN) and ¥â1 integrin are important for the formation of invadopodia and epithelial-to-mesenchymal transition (EMT) events. Our data also highlighted the importance of the temporal and spatial distribution of IGF induced signalling proteins in breast cancer metastasis; in particular, SFN, enhancer-of-split and hairy-related protein 2 (SHARP-2), total-akt/protein kinase B 1 (Total-AKT1), phosphorylated-akt/protein kinase B (P-AKT), extracellular signal-related kinase-1 and extracellular signal-related kinase-2 (ERK1/2) and phosphorylated-extracellular signal-related kinase-1 and extracellular signal-related kinase-2 (P-ERK1/2).
Multivariate survival models were created from the immunohistochemical data. These models were found to fit well with these data with very high statistical confidence. Numerous prognostic confounding effects and effect modifications were identified among elements of the ECM and IGF signalling cascade and corroborate the survival models. This finding provides further evidence for the prognostic potential of IGF and ECM induced signalling proteins. In addition, the adjusted measures of associations obtained in this study have strengthened the validity and utility of the resulting models.
The findings from this study provide insights into the biological interactions that occur during the development of breast tissue and contribute to disease progression. Importantly, these multivariate survival models could provide important prognostic and predictive indicators that assist the clinical management of breast disease, namely in the early identification of cancers with a propensity to metastasise, and/or recur following adjuvant therapy. The outcomes of this study further inform the development of new therapeutics to aid patient recovery. The findings from this study have widespread clinical application in the diagnosis of disease and prognosis of disease progression, and inform the most appropriate clinical management of individuals with breast cancer.
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He, Hongbin. "Studies on the genetic control of infection and hepatic disease in schistosoma haematobium and schistosoma japonicum infections in human." Thesis, Aix-Marseille 2, 2010. http://www.theses.fr/2010AIX20720.
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La bilharziose reste un problème de santé majeur. L'équipe du Pr Dessein a montré que les infections élevées étaient déterminées par un locus majeur en 5q31 et que des polymorphismes dans un gène à ce locus,IL13, aggravent l'infection. Notre premier objectif était d'évaluer si des variants d'autres gènes de la voie de l'IL13 intervenaient dans le contrôle de l'infection. Nous avons observé une association entre le SNP rs324013, dans le promoteur de STAT6,et les niveaux d'infection à S. haematobium. Ce polymorphisme a un effet additif avec le polymorphisme IL13rs1800925. Ce SN modifie la fixation de facteurs nucléaires au niveau du promoteur de STAT6. L'équipe du Pr Dessein avait également montré que les fibres hépatiques avancées et sévères étaient déterminées par un autre locus majeur localisé en 6q23. Notre deuxième objectif fut d'évaluer dans le laboratoire du Pr Dessein et en étroite collaboration avec le laboratoire du Pr Li(Yueyang Institute of Parasitic disease)deux gènes candidats(IFNGR1 et CTGF) situés dans cette région chromosomique. Nous avons observé une association entre les deux polyporphismes(rs17066192 er rs673156)localisés dans le promoteur du gène. Nous avons observé une association entre les deux polymorphismes(rs17066192 et rs673156)localisés dans le promoteur du gène IFNGR1 et la fibrose hépatique: le génotype rs673156A/A et rs17066192C/C sont associés à un risque 7.3 fois et 1.5 fois plus élevé, respectivement, de fibrose avancée. Nous avons également montré que les variants rs9402373 et rs12526196 du gène CTGF sont indépendamment associés à la fibrose chez les fermiers et pêcheurs chinois infectés par S.japonicum. Sur la population chinoise d'étude, les risques relatifs associés aux polymorphismes rs9402373 et rs12526196 sont de 2.8 et 3Schistosomiasis remains one of the world’s most prevalent diseases. It comprises a group of chronic diseases caused by helminths of the Schistosoma genus. Schistosoma haematobium causes obstructive nephropathy that can be aggravated by urinary bacterial infections. S.japonicum and S.mansoni cause hepatic fibrosis associated with portal blood hypertension, which can be lethal. In previous studies, our laboratory had shown that worm burden in S.haematobium infections were aggravated by IL13 variants and that severe hepatic fibrosis (HF) was controlled by gene(s) located on 6q23. The present study is to further evaluate other IL-13 pathway genes (STAT6) in the control of infection in Malian farmers and to test candidate genes in the 6q23 region in hepatic fibrosis (HF) in S.japonicum infected Chinese fishermen and farmers. First we have developped an improved FTA® technology technique to perform SNP genotyping. This technique allows us to use saliva samples for genotyping SNPs. Subsequently, this improved FTA® technology was used in our study on HF.Our work on a Malian sample infected with S. haematobium indicated that a polymorphism (rs324013) in the promoter of STAT6 gene was associated with the control of S. haematobium infection levels and has an additive effect with IL13rs1800925, a polymorphism previously associated with infection in this same population. Both SNPs modify the binding of nuclear factors to the promoter regions of their respective genes. Thus, both SNPs may play a crucial role in controlling S. haematobium infection levels. In order to study HF in S.japonicum infections, we have participated actively in the study that recruited of a large sample of Chinese fishermen and farmers who had been exposed to the infection for most of their life. HF was evaluated by ultrasound and covariates that could affect HF were evaluated by interviews. Then, we tested two genes (IFNGR1, CTGF) of the 6q23 region that were good candidates for the control of HF on these samples. Both genes encode molecules that were shown in animal and human studies to have strong effect on extracellular matrix proteins deposition and turnover. We found that two polymorphisms (rs17066192 and rs673156) in IFNGR1 promoter were associated with HF: the rs673156A/A genotype was associated with a 7.3-fold increased risk of advanced HF; and rs17066192C/C genotype with a 1.5-fold increased risk of HF. These results must now be confirmed in another population sample. We also found that variants of CTGF rs9402373 and rs12526196 were independently associated with HF in Chinese fishermen and farmers, in Sudanese, and in Brazilians infected with either S. japonicum or S. mansoni. Our results provide additional evidence for a protective role of IL-13 in schistosome infections, and they also demonstrate that TGFβ / CTGF pathway plays a key role in HF and should be targeted by chemotherapy. Ongoing studies evaluate whether CTGF variants could be used in the prognosis of the HF caused by schistosomes and also by other infectious agents
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Lee, Connie Wing-Ching. "Notch-1 and IGF-1 as Survivin Regulatory Pathways in Cancer: A Dissertation." eScholarship@UMMS, 2008. https://escholarship.umassmed.edu/gsbs_diss/377.
Full textAbstract:
The 21st century brought about a dramatic increase in knowledge about genetic and molecular profiles of cancer. This information has validated the complexity of tumor cells and increased awareness of “nodal proteins”, but has yet to advance the development of rational targeted cancer therapeutics. Nodal proteins are critical cellular proteins that collect biological inputs and distribute the information across diverse biological processes. Survivin acts as a nodal protein by interfacing the multiple signals involved in mitosis and apoptosis and functionally integrate proliferation, cell death, and cellular homeostasis. By characterizing survivin as a target of both Type 1 Insulin-like Growth Factor (IGF-1) and Notch developmental signaling, we contribute to the paradigm of survivin as a nodal protein. The two signaling systems, Notch and IGF-1, regulate survivin by two independent mechanisms. Notch activation induces survivin transcription preferentially in basal breast cancer, a breast cancer subtype with poor prognosis and lack of molecular therapies. Activated Notch binds the transcription factor RBP-Jк and drives transcription from the survivin promoter. Notch mediated survivin expression increases cell cycle kinetics promoting tumor proliferation. Inhibition of Notch in a breast xenograft model reduced tumor growth and systemic metastasis. On the other hand, IGF-1 signaling drives survivin protein translation in prostate cancer cells. Binding of IGF-1 to its receptor activates downstream kinases, mammalian target of rapamycin (mTOR) and p70 S6 protein kinase (p70S6K), which modulates survivin mRNA translation to increase the apoptotic threshold. The multiple roles of survivin in tumorigenesis implicate survivin as a rational target for the “next generation” of cancer therapeutics.
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Whiteside, Eliza Jane. "The expression and regulation of metalloproteinases during normal and malignant trophoblast invasion." Thesis, Queensland University of Technology, 2001.
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Been-Ren, Lin. "Connective Tissue Growth Factor and Its Role in Colorectal Cancer." 2006. http://www.cetd.com.tw/ec/thesisdetail.aspx?etdun=U0001-1907200617392400.
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