DOCSLIB.ORG

Transcript Profiling of Different Types of Multiple Sclerosis Lesions Yields

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Transcript Profiling of Different Types of Multiple Sclerosis Lesions Yields Mohan et al. Acta Neuropathologica Communications (2014) 2:168 DOI 10.1186/s40478-014-0168-9 RESEARCH ARTICLE Open Access Transcript profiling of different types of multiple sclerosis lesions yields FGF1 as a promoter of remyelination Hema Mohan1,10†, Anita Friese1†, Stefanie Albrecht2†, Markus Krumbholz1†, Christina L Elliott3, Ariel Arthur3, Ramesh Menon4, Cinthia Farina4, Andreas Junker5, Christine Stadelmann5, Susan C Barnett3, Inge Huitinga6, Hartmut Wekerle7, Reinhard Hohlfeld1,8, Hans Lassmann9, Tanja Kuhlmann2, Chris Linington3 and Edgar Meinl1* Abstract Chronic demyelination is a pathological hallmark of multiple sclerosis (MS). Only a minority of MS lesions remyelinates completely. Enhancing remyelination is, therefore, a major aim of future MS therapies. Here we took a novel approach to identify factors that may inhibit or support endogenous remyelination in MS. We dissected remyelinated, demyelinated active, and demyelinated inactive white matter MS lesions, and compared transcript levels of myelination and inflammation-related genes using quantitative PCR on customized TaqMan Low Density Arrays. In remyelinated lesions, fibroblast growth factor (FGF) 1 was the most abundant of all analyzed myelination-regulating factors, showed a trend towards higher expression as compared to demyelinated lesions and was significantly higher than in control white matter. Two MS tissue blocks comprised lesions with adjacent de- and remyelinated areas and FGF1 expression was higher in the remyelinated rim compared to the demyelinated lesion core. In functional experiments, FGF1 accelerated developmental myelination in dissociated mixed cultures and promoted remyelination in slice cultures, whereas it decelerated differentiation of purified primary oligodendrocytes, suggesting that promotion of remyelination by FGF1 is based on an indirect mechanism. The analysis of human astrocyte responses to FGF1 by genome wide expression profiling showed that FGF1 induced the expression of the chemokine CXCL8 and leukemia inhibitory factor, two factors implicated in recruitment of oligodendrocytes and promotion of remyelination. Together, this study presents a transcript profiling of remyelinated MS lesions and identified FGF1 as a promoter of remyelination. Modulation of FGF family members might improve myelin repair in MS. Keywords: Multiple sclerosis, Remyelination, Demyelination, Fibroblast growth factor Introduction but also compromises axonal survival by enhancing sus- The adult central nervous system contains a large pool of ceptibility to damage by inflammatory mediators [4], and mitotic oligodendroglial progenitor cells (OPCs) which by disrupting trophic support provided by myelinating rapidly differentiate and remyelinate lesions in models of oligodendrocytes [5-7]. The frequently observed failure of toxin or immune mediated demyelination [1]. In MS, remyelination in MS does not appear to be due to an however, this endogenous repair mechanism frequently intrinsic, absolute defect of myelination, because de- and fails, resulting in the formation of chronic demyelinated remyelinated MS lesions are frequently found side by side plaques with glial scars, the pathological hallmark of the in the majority of patients. Furthermore, early MS lesions disease [2,3]. This has profound functional consequences often show signs of remyelination [8-12]. as demyelination not only disrupts saltatory conduction, It is unclear why many MS lesions fail to remyelinate. The presence of OPCs and premyelinating oligodendro- * Correspondence: [email protected] cytes in many demyelinated lesions [13,14] suggests that †Equal contributors this could be caused by failure of OPC to differentiate into 1 Institute of Clinical Neuroimmunology, Ludwig Maximilian University myelinating oligodendrocytes. Overcoming this differenti- Munich, Marchioninistraße 15, D-81377 Munich, Germany Full list of author information is available at the end of the article ation block to enhance remyelination by endogenous OPC © 2014 Mohan et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Mohan et al. Acta Neuropathologica Communications (2014) 2:168 Page 2 of 18 is considered a logical strategy to restore saltatory conduc- contained 12 white matter lesions, two of them with a tion and reduce accumulation of disability due to axonal demyelinated core and a remyelinated rim. As control, we pathology [15,16]. Achieving this goal, however, is compli- used 6 control tissue blocks from 4 healthy subjects with- cated as a multitude of cellular and molecular changes out clinical or histological evidence of CNS disease (details within the MS lesions can influence OPC migration, sur- in Additional file 1: Table S1). vival or differentiation [17-19]. While experimental studies For immunostaining of FGF1 we used 10 formalin demonstrate that many factors can individually influence fixed paraffin embedded (FFPE) tissue blocks from 7 MS OPC migration and/or differentiation [20-32], their rela- patients containing remyelinated, demyelinated inactive tive expression within MS lesions and their significance in and demyelinated active areas as well as and 3 control modulating remyelination remains unclear. Expression tissue blocks from 3 healthy subjects. Most of the patients profiling of MS lesions can provide new insight into had secondary progressive MS. The mean disease dur- pathomechanisms [33-35], but this approach has not been ation was 24 years, the sex (f/m) ratio was 2:1 (details applied to remyelinated lesions yet. in Additional file 1: Table S1). To identify endogenous pathways that might be MS lesions were classified according to defined criteria: exploited to enhance remyelination we analyzed white Active lesions contained abundant macrophages with de- matter lesions and used qPCR to focus on the expression graded myelin products visualized by luxol fast blue (LFB) of genes regulating oligodendrocytes. We compared dis- or oil red O staining. Inactive demyelinated lesions were sected remyelinated lesions, actively demyelinating lesions, sharply demarcated from the normal appearing white inactive demyelinated lesions and control white matter. matter and largely devoid of macrophages. Remyelinated This revealed an important role of the FGF family in the lesions were sharply demarcated from the normal appear- regulation of remyelination. The FGF family is known to ing white matter and identified by a fainter LFB staining. regulate oligodendrocyte biology and myelin thickness Tissue was collected from donors from whom a writ- [36-40], but its role in MS lesion development is un- ten informed consent was provided for brain autopsy clear. We found that among the analyzed myelination- and the use of the material and clinical information for regulating genes FGF1 was the most abundant transcript research purposes. in remyelinated lesions. In two lesions that contained a demyelinated core and a remyelinated rim, the FGF1 Dissection of brain specimens, RNA extraction, cDNA transcript levels were higher in the remyelinated parts synthesis, and quantitative PCR of brain tissue suggesting that the increased availability of FGF1 may Selected areas from tissue blocks were obtained as follows: support remyelination. Cryosections (20 μm) were mounted on PEN slides (P.A. FGF1 has been reported to promote proliferation of glial L.M. Microlaser, Bernried, Germany). To identify demyeli- precursors [41], but had so far not been linked to remyeli- nated, remyelinated, control white matter, and grey matter nation or its failure in MS. We employed a dissociated areas every 6th section (30 μm) was stained with LFB. The myelinating culture system [42-44], a remyelinating slice unstained sections were superimposed on LFB stained culture model [45,46], a pure oligodendrocyte culture, and sections and the lesion areas were marked and macrodis- genome wide expression profiling of astrocytes to deduce sected manually. In total 200–300 μm of each block was the functional relevance. This revealed that FGF1 promotes used for transcript analysis. To check the precision of myelination as well as remyelination, presumably via an dissection, the macrodissected sections were stained indirect mechanism. In astrocytes, FGF1 induced leukemia with LFB. The control tissue samples used for qPCR inhibitory factor (LIF) and the chemokine CXCL8,both exclusively contained white matter. Two MS tissue blocks implicated in the recruitment of oligodendrocytes and the with adjacent de-and remyelinated lesions were analyzed promotion of remyelination. This suggests that also in vivo individually. FGF1 stimulates astrocytes to release factors promoting RNA was obtained from the dissected tissue specimens remyelination and that selective modulation of FGF signal- by guanidinium thiocyanate-phenol-chloroform extraction ing pathways could provide a novel strategy to enhance (TRI® Reagent, SIGMA, Munich, Germany), and cDNA remyelination in MS and other demyelinating diseases. was synthesized using random hexamers (High Capacity cDNA Reverse Transcription kit, Applied Biosystems Materials and methods (ABI,
Load more

Recommended publications
  • Role and Regulation of Pdgfra Signaling in Liver Development and Regeneration
    The American Journal of Pathology, Vol. 182, No. 5, May 2013 ajp.amjpathol.org GROWTH FACTORS, CYTOKINES, AND CELL CYCLE MOLECULES Role and Regulation of PDGFRa Signaling in Liver Development and Regeneration Prince K. Awuah,* Kari N. Nejak-Bowen,* and Satdarshan P.S. Monga*y From the Division of Experimental Pathology,* Department of Pathology, and the Department of Medicine,y University of Pittsburgh, Pittsburgh, Pennsylvania Accepted for publication January 22, 2013. Aberrant platelet-derived growth factor receptor-a (PDGFRa) signaling is evident in a subset of hepato- cellular cancers (HCCs). However, its role and regulation in hepatic physiology remains elusive. In the Address correspondence to a fi Satdarshan P.S. Monga, M.D., current study, we examined PDGFR signaling in liver development and regeneration. We identi ed a a Divisions of Experimental notable PDGFR activation in hepatic morphogenesis that, when interrupted by PDGFR -blocking anti- Pathology, Pathology and body, led to decreased hepatoblast proliferation and survival in embryonic liver cultures. We also identified Medicine, University of Pitts- temporal PDGFRa overexpression, which is regulated by epidermal growth factor (EGF) and tumor necrosis burgh School of Medicine, 200 factor a, and its activation at 3 to 24 hours after partial hepatectomy. Through generation of hepatocyte- Lothrop St., S-422 BST, Pitts- specific PDGFRA knockout (KO) mice that lack an overt phenotype, we show that absent PDGFRa burgh, PA 15261. E-mail: compromises extracelluar signal-regulated kinases and AKT activation 3 hours after partial hepatectomy, [email protected]. which, however, is alleviated by temporal compensatory increases in the EGF receptor (EGFR) and the hepatocyte growth factor receptor (Met) expression and activation along with rebound activation of extracellular signal-regulated kinases and AKT at 24 hours.
    [Show full text]
  • Alternative Splicing Generates an Isoform of the Human Sef Gene with Altered Subcellular Localization and Specificity
    Alternative splicing generates an isoform of the human Sef gene with altered subcellular localization and specificity Ella Preger*, Inbal Ziv*, Ariel Shabtay*, Ifat Sher*, Michael Tsang†, Igor B. Dawid†, Yael Altuvia‡, and Dina Ron*§ *Department of Biology, Technion–Israel Institute of Technology, Haifa 32000, Israel; †Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892; and ‡Department of Molecular Genetics and Biotechnology, Hebrew University–Hadassah Medical School, Jerusalem 91120, Israel Contributed by Igor B. Dawid, December 1, 2003 Receptor tyrosine kinases (RTKs) control a multitude of biological FGFs comprise a family of 22 structurally related polypeptide processes and are therefore subjected to multiple levels of regu- mitogens that control cell proliferation, differentiation, survival, lation. Negative feedback is one of the mechanisms that provide an and migration and play a key role in embryonic patterning effective means to control RTK-mediated signaling. Sef has re- (14–16). They signal via binding and activation of a family of cently been identified as a specific antagonist of fibroblast growth cell-surface tyrosine kinase receptors designated FGF receptors factor (FGF) signaling in zebrafish and subsequently in mouse and 1–4 (FGFR1–FGFR4) (17–20). Activated receptors trigger sev- human. Sef encodes a putative type I transmembrane protein that eral signal transduction cascades including the Ras͞MAPK and antagonizes the Ras͞mitogen-activated protein kinase pathway in the PI3-kinase pathway (15, 21). Depending on the cell type, all three species. Mouse Sef was also shown to inhibit the phos- FGF can also activate other MAPK pathways, such that leading phatidylinositol 3-kinase pathway.
    [Show full text]
  • PDGF-C and PDGF-D Signaling in Vascular Diseases and Animal Models
    Molecular Aspects of Medicine 62 (2018) 1e11 Contents lists available at ScienceDirect Molecular Aspects of Medicine journal homepage: www.elsevier.com/locate/mam PDGF-C and PDGF-D signaling in vascular diseases and animal models * Erika Folestad a, Anne Kunath b, Dick Wågsater€ b, a Division of Vascular Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden b Division of Drug Research, Department of Medical and Health Sciences, Linkoping€ University, Linkoping,€ Sweden article info abstract Article history: Members of the platelet-derived growth factor (PDGF) family are well known to be involved in different Received 31 August 2017 pathological conditions. The cellular and molecular mechanisms induced by the PDGF signaling have Received in revised form been well studied. Nevertheless, there is much more to discover about their functions and some 14 November 2017 important questions to be answered. This review summarizes the known roles of two of the PDGFs, Accepted 22 January 2018 PDGF-C and PDGF-D, in vascular diseases. There are clear implications for these growth factors in several Available online 14 February 2018 vascular diseases, such as atherosclerosis and stroke. The PDGF receptors are broadly expressed in the cardiovascular system in cells such as fibroblasts, smooth muscle cells and pericytes. Altered expression Keywords: Aneurysm of the receptors and the ligands have been found in various cardiovascular diseases and current studies fi Atherosclerosis have shown important implications of PDGF-C and PDGF-D signaling in brosis, neovascularization, Growth factor atherosclerosis and restenosis. Myocardial infarction © 2018 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND Smooth muscle cells license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
    [Show full text]
  • TABLE S1 Complete Overview of Protocols for Induction of Pscs Into
    TABLE S1 Complete overview of protocols for induction of PSCs into renal lineages Ref 2D/ Cell type Protocol Days Growth factors Outcome Other analyses # 3D hiPSC, hESC, Collagen type I tx murine epidydymal fat pads,ex vivo 54 2D 8 Y27632, AA, CHIR, BMP7 IM miPSC, mESC Matrigel with murine fetal kidney hiPSC, hESC, Suspension,han tx murine epidydymal fat pads,ex vivo 54 2D 20 AA, CHIR, BMP7 IM miPSC, mESC ging drop with murine fetal kidney tx murine epidydymal fat pads,ex vivo 55 hiPSC, hESC Matrigel 2D 14 CHIR, TTNPB/AM580, Y27632 IM with murine fetal kidney Injury, tx murine epidydymal fat 57 hiPSC Suspension 2D 28 AA, CHIR, BMP7, TGF-β1, TTNPB, DMH1 NP pads,spinal cord assay Matrigel,membr 58 mNPC, hESC 3D 7 BPM7, FGF9, Heparin, Y27632, CHIR, LDN, BMP4, IGF1, IGF2 NP Clonal expansion ane filter iMatrix,spinal 59 hiPSC 3D 10 LIF, Y27632, FGF2/FGF9, TGF-α, DAPT, CHIR, BMP7 NP Clonal expansion cord assay iMatrix,spinal 59 murine NP 3D 8-19 LIF, Y27632, FGF2/FGF9, TGF- α, DAPT, CHIR, BMP7 NP Clonal expansion cord assay murine NP, Suspension, 60 3D 7-19 BMP7, FGF2, Heparin, Y27632, LIF NP Nephrotoxicity, injury model human NP membrane filter Suspension, Contractility and permeability assay, ex 61 hiPSC 2D 10 AA, BMP7, RA Pod gelatin vivo with murine fetal kidney Matrigel. 62 hiPSC, hESC fibronectin, 2D < 50 FGF2, AA, WNT3A, BMP4, BMP7, RA, FGF2, HGF or RA + VITD3 Pod collagen type I Matrigel, Contractility and uptake assay,ex vivo 63 hiPSC 2D 13 Y27632, CP21R7, BMP4, RA, BMP7, FGF9, VITD3 Pod collagen type I with murine fetal kidney Collagen
    [Show full text]
  • A Novel Fibroblast Growth Factor 1 Variant Reverses Nonalcoholic Fatty Liver Disease in Type 2 Diabetes
    University of Louisville ThinkIR: The University of Louisville's Institutional Repository Electronic Theses and Dissertations 8-2018 A novel fibroblast growth factor 1 variant reverses nonalcoholic fatty liver disease in type 2 diabetes. Qian Lin University of Louisville Follow this and additional works at: https://ir.library.louisville.edu/etd Part of the Endocrine System Diseases Commons, and the Pharmacology Commons Recommended Citation Lin, Qian, "A novel fibroblast growth factor 1 variant reverses nonalcoholic fatty liver disease in type 2 diabetes." (2018). Electronic Theses and Dissertations. Paper 3016. https://doi.org/10.18297/etd/3016 This Doctoral Dissertation is brought to you for free and open access by ThinkIR: The University of Louisville's Institutional Repository. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of ThinkIR: The University of Louisville's Institutional Repository. This title appears here courtesy of the author, who has retained all other copyrights. For more information, please contact [email protected]. A NOVEL FIBROBLAST GROWTH FACTOR 1 VARIANT REVERSES NONALCOHOLIC FATTY LIVER DISEASE IN TYPE 2 DIABETES BY Qian Lin M.S. at Wenzhou Medical University, 2016 A Dissertation Submitted to the Faculty of the School of Medicine of the University of Louisville for the Degree of Doctor of Philosophy in Pharmacology and Toxicology Department of Pharmacology and Toxicology University of Louisville Louisville, Kentucky August 2018 A NOVEL FIBROBLAST GROWTH FACTOR 1 VARIANT REVERSES NONALCOHOLIC FATTY LIVER DISEASE IN TYPE 2 DIABETES BY Qian Lin Dissertation Approved on August 2, 2018 Dissertation Committee: Dr. Yi Tan Dr. Paul Epstein Dr.
    [Show full text]
  • The Roles of Fgfs in the Early Development of Vertebrate Limbs
    Downloaded from genesdev.cshlp.org on September 26, 2021 - Published by Cold Spring Harbor Laboratory Press REVIEW The roles of FGFs in the early development of vertebrate limbs Gail R. Martin1 Department of Anatomy and Program in Developmental Biology, School of Medicine, University of California at San Francisco, San Francisco, California 94143–0452 USA ‘‘Fibroblast growth factor’’ (FGF) was first identified 25 tion of two closely related proteins—acidic FGF and ba- years ago as a mitogenic activity in pituitary extracts sic FGF (now designated FGF1 and FGF2, respectively). (Armelin 1973; Gospodarowicz 1974). This modest ob- With the advent of gene isolation techniques it became servation subsequently led to the identification of a large apparent that the Fgf1 and Fgf2 genes are members of a family of proteins that affect cell proliferation, differen- large family, now known to be comprised of at least 17 tiation, survival, and motility (for review, see Basilico genes, Fgf1–Fgf17, in mammals (see Coulier et al. 1997; and Moscatelli 1992; Baird 1994). Recently, evidence has McWhirter et al. 1997; Hoshikawa et al. 1998; Miyake been accumulating that specific members of the FGF 1998). At least five of these genes are expressed in the family function as key intercellular signaling molecules developing limb (see Table 1). The proteins encoded by in embryogenesis (for review, see Goldfarb 1996). Indeed, the 17 different FGF genes range from 155 to 268 amino it may be no exaggeration to say that, in conjunction acid residues in length, and each contains a conserved with the members of a small number of other signaling ‘‘core’’ sequence of ∼120 amino acids that confers a com- molecule families [including WNT (Parr and McMahon mon tertiary structure and the ability to bind heparin or 1994), Hedgehog (HH) (Hammerschmidt et al.
    [Show full text]
  • Upregulated Expression of FGF13/FHF2 Mediates Resistance To
    OPEN Upregulated expression of FGF13/FHF2 SUBJECT AREAS: mediates resistance to platinum drugs in CANCER THERAPEUTIC RESISTANCE cervical cancer cells CANCER PREVENTION Tomoko Okada1, Kazuhiro Murata1,2, Ryoma Hirose1,3, Chie Matsuda1, Tsunehiko Komatsu2, STRESS SIGNALLING Masahiko Ikekita3, Miyako Nakawatari4, Fumiaki Nakayama4, Masaru Wakatsuki5, Tatsuya Ohno5,6, CHEMOTHERAPY Shingo Kato5,7, Takashi Imai4 & Toru Imamura1,3 Received 1Signaling Molecules Research Group, Biomedical Research Institute, National Institute of Advanced Industrial Science and 27 June 2013 2 Technology (AIST), Tsukuba, Ibaraki 305-8566, Japan, Division of Hematology, 3rd Department of Internal Medicine, Teikyo 3 Accepted University Chiba Medical Center, Ichihara, Chiba 299-0111, Japan, Department of Applied Biological Science, Faculty of Science 4 18 September 2013 and Technology, Tokyo University of Science, Noda, Chiba 278-8510, Japan, Advanced Radiation Biology Research Program, National Institute of Radiological Sciences, Chiba, Chiba 263-8555, Japan, 5Research Center Hospital for Charged Particle Published Therapy, National Institute of Radiological Sciences, Chiba, Chiba 263-8555, Japan, 6Gunma University Graduate School of 11 October 2013 Medicine, Maebashi, Gunma 371-8511, Japan, 7Saitama Medical University International Medical Center, Hidaka, Saitama 350- 1298, Japan. Correspondence and Cancer cells often develop drug resistance. In cisplatin-resistant HeLa cisR cells, fibroblast growth factor 13 requests for materials (FGF13/FHF2) gene and protein expression was strongly upregulated, and intracellular platinum should be addressed to concentrations were kept low. When the FGF13 expression was suppressed, both the cells’ resistance to T.Imamura (imamura- platinum drugs and their ability to keep intracellular platinum low were abolished. Overexpression of [email protected]) FGF13 in parent cells led to greater resistance to cisplatin and reductions in the intracellular platinum concentration.
    [Show full text]
  • Ep 3217179 A1
    (19) TZZ¥ ___T (11) EP 3 217 179 A1 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 13.09.2017 Bulletin 2017/37 G01N 33/68 (2006.01) (21) Application number: 17167637.2 (22) Date of filing: 02.10.2013 (84) Designated Contracting States: • LIU, Xinjun AL AT BE BG CH CY CZ DE DK EE ES FI FR GB San Diego, CA 92130 (US) GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO • HAUENSTEIN, Scott PL PT RO RS SE SI SK SM TR San Diego, CA 92130 (US) • KIRKLAND, Richard (30) Priority: 05.10.2012 US 201261710491 P San Diego, CA 92111 (US) 17.05.2013 US 201361824959 P (74) Representative: Krishnan, Sri (62) Document number(s) of the earlier application(s) in Nestec S.A. accordance with Art. 76 EPC: Centre de Recherche Nestlé 13779638.9 / 2 904 405 Vers-chez-les-Blanc Case Postale 44 (71) Applicant: Nestec S.A. 1000 Lausanne 26 (CH) 1800 Vevey (CH) Remarks: (72) Inventors: This application was filed on 21-04-2017 as a • SINGH, Sharat divisional application to the application mentioned Rancho Santa Fe, CA 92127 (US) under INID code 62. (54) METHODS FOR PREDICTING AND MONITORING MUCOSAL HEALING (57) The present invention provides methods for pre- an individual with a disease such as IBD. Information on dicting the likelihood of mucosal healing in an individual mucosal healing status derived from the use of the with a disease such as inflammatory bowel disease present invention can also aid in optimizing therapy (IBD).
    [Show full text]
  • Type I and II Interferon Are Associated with High Expression of the Hippo Pathway Family Members
    Cel al & lul ic ar in I l m C m f o u n l a o l n o Journal of Clinical & Cellular r g u y o J ISSN: 2155-9899 Immunology Research Article Type I and II Interferon are Associated with High Expression of the Hippo Pathway Family Members Bianca Sciescia1*, Raquel Tognon2, Natalia de Souza Nunes1, Tathiane Maistro Malta1, Fabiani Gai Frantz1, Fabiola Attie de Castro1, Maira da Costa Cacemiro1 1Department of Clinical, Toxicological and Bromatological Analysis, University of Sao Paulo - USP, Ribeirao Preto - SP, Brazil; 2Department of Pharmacy, Federal University of Juiz de Fora, Campus Governador Valadares, Governador Valadares - MG, Brazil ABSTRACT The Hippo pathway plays a regulatory role on inflammation and cell death and proliferation. Here we described a relationship between Hippo pathway components and inflammation in healthy subjects. The plasma levels of cytokines and chemokines were used to define their inflammatory profile and classify them as normal, high and low producers of cytokines. Leukocytes from healthy subjects with inflammatory profile expressed the highest levels of MSTS1/MST2, SAV1, LATS1/LATS2, MOB1A/MOB1B and YAP genes. The group that overexpressed Hippo pathway-related genes secreted more IFN-ϒ and IFN-α2. Keywords: Hippo pathway; Inflammatory process; Healthy subjects; Cytokines; Chemokines ABBREVATIONS: LATS1/LATS2 kinases (large tumor suppressor kinase 1/2) and MOB kinase activator IL: Interleukin; IFN: Interferon; LATS1/LATS2: Large tumor their adapter proteins MOB1A/MOB1B ( 1A/1B yes-associated suppressor kinase 1/2; MCP-1: Monocyte chemoattractant ), and the transcription coactivators YAP ( protein tafazzin protein protein-1; MIP: Macrophage inflammatory protein; MOB1A/ ) and TAZ ( ).
    [Show full text]
  • Regulation of PDGFC Signalling and Extracellular Matrix Composition by FREM1 in Mice
    RESEARCH ARTICLE Disease Models & Mechanisms 6, 1426-1433 (2013) doi:10.1242/dmm.013748 Regulation of PDGFC signalling and extracellular matrix composition by FREM1 in mice Fenny Wiradjaja1, Denny L. Cottle1, Lynelle Jones1 and Ian Smyth1,2,* SUMMARY Fras1-related extracellular matrix protein 1 (FREM1) is required for epidermal adhesion during embryogenesis, and mice lacking the gene develop fetal skin blisters and a range of other developmental defects. Mutations in members of the FRAS/FREM gene family cause diseases of the Fraser syndrome spectrum. Embryonic epidermal blistering is also observed in mice lacking PdgfC and its receptor, PDGFRα. In this article, we show that FREM1 binds to PDGFC and that this interaction regulates signalling downstream of PDGFRα. Fibroblasts from Frem1-mutant mice respond to PDGFC stimulation, but with a shorter duration and amplitude than do wild-type cells. Significantly, PDGFC-stimulated expression of the metalloproteinase inhibitor Timp1 is reduced in cells with Frem1 mutations, leading to reduced basement membrane collagen I deposition. These results show that the physical interaction of FREM1 with PDGFC can regulate remodelling of the extracellular matrix downstream of PDGFRα. We propose that loss of FREM1 function promotes epidermal blistering in Fraser syndrome as a consequence of reduced PDGFC activity, in addition to its stabilising role in the basement membrane. DMM INTRODUCTION The FRAS/FREM family of proteins share characteristic The FRAS/FREM extracellular matrix (ECM) proteins (FRAS1, chondroitin sulphate proteoglycan (CSPG) core repeats similar FREM1 and FREM2) mediate adhesion between the epidermal to those found in the NG2 proteoglycan (Stallcup, 2002). In this basement membrane and the underlying dermis during embryonic protein, they directly bind platelet-derived growth factor A development (reviewed in Short et al., 2007; Petrou et al., 2008).
    [Show full text]
  • PDGFC (Human) Recombinant Protein
    PDGFC (Human) Recombinant pH 3.0. protein Storage Instruction: Store at -20°C. Reconstitute in water to a concentration of 0.1-1.0 Catalog Number: P6130 mg/mL. Do not vortex. Regulation Status: For research use only (RUO) For extended storage, it is recommended to further dilute in a buffer containing a carrier protein (example 0.1% Product Description: Human PDGFC (Q9NRA1) partial BSA) and store in working aliquots at -20°C to -80°C. recombinant protein expressed in Escherichia coli. Entrez GeneID: 56034 Sequence: MVVDLNLLTEEVRLYSCTPRNFSVSIREELKRTDTIFW Gene Symbol: PDGFC PGCLLVKRCGGNCACCLHNCNECQCVPSKVTKKYHE Gene Alias: FALLOTEIN, SCDGF VLQLRPKTGVRGLHKSLTDVALEHHEECDCVCRGSTG G Gene Summary: The protein encoded by this gene is a member of the platelet-derived growth factor family. The Host: Escherichia coli four members of this family are mitogenic factors for Theoretical MW (kDa): 25.0 cells of mesenchymal origin and are characterized by a core motif of eight cysteines. This gene product appears Reactivity: Human to form only homodimers. It differs from the platelet-derived growth factor alpha and beta Applications: Func, SDS-PAGE polypeptides in having an unusual N-terminal domain, (See our web site product page for detailed applications the CUB domain. [provided by RefSeq] information) Protocols: See our web site at http://www.abnova.com/support/protocols.asp or product page for detailed protocols Form: Lyophilized Preparation Method: Escherichia coli expression system Purity: 98% Endotoxin Level: Endotoxin level is <0.1 ng/ug of protein (<1 EU/ug). Activity: Determined by the dose-dependent stimulation of the proliferation of Balb/c 3T3 cells. The expected The ED50 for this effect is 15-20 ng/mL.
    [Show full text]
  • The Hippo–YAP Pathway in Organ Size Control and Tumorigenesis: an Updated Version
    Downloaded from genesdev.cshlp.org on September 26, 2021 - Published by Cold Spring Harbor Laboratory Press REVIEW The Hippo–YAP pathway in organ size control and tumorigenesis: an updated version Bin Zhao,1 Li Li,1 Qunying Lei,2 and Kun-Liang Guan1,3 1Department of Pharmacology and Moores Cancer Center, University of California at San Diego, La Jolla, California 92093, USA; 2Department of Biological Chemistry, School of Medicine, and Molecular and Cell Biology Laboratory, Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China The Hippo signaling pathway is gaining recognition as The Hippo pathway was named after the Drosophila an important player in both organ size control and Hippo kinase that was discovered using this approach. tumorigenesis, which are physiological and pathological Components of the Hippo pathway are highly conserved processes that share common cellular signaling mecha- in mammals (Fig. 1). Later genetic and biochemical studies nisms. Upon activation by stimuli such as high cell den- gradually shaped the current working model, in which the sity in cell culture, the Hippo pathway kinase cascade mammalian Mst1/2 kinase (Hippo homolog), complexed phosphorylates and inhibits the Yes-associated protein with a scaffold protein, Sav1, phosphorylates and activates (YAP)/TAZ transcription coactivators representing the the Lats1/2 kinase. Lats1/2 is also activated by another major signaling output of the pathway. Altered gene scaffold protein, Mob1 (Fig. 2). These four proteins are expression resulting from YAP/TAZ inhibition affects often referred to as the core components of the Hippo cell number by repressing cell proliferation and promot- pathway. At the upstream, several components have ing apoptosis, thereby limiting organ size.
    [Show full text]