DOCSLIB.ORG

Expression of Recombinant Platelet-Derived Growth Factor A-And B-Chain Homodimers in Rat-1 Cells and Human Fibroblasts Reveals Differences in Protein Processing and Autocrine Effects

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Expression of Recombinant Platelet-Derived Growth Factor A-And B-Chain Homodimers in Rat-1 Cells and Human Fibroblasts Reveals Differences in Protein Processing and Autocrine Effects MOLECULAR AND CELLULAR BIOLOGY. JUly 1988. p. 2753-2762 Vol. 8, No. 7 0270-7306/88/072753-10$02.00/0 Copyright © 1988. American Society for Microbiology Expression of Recombinant Platelet-Derived Growth Factor A- and B-Chain Homodimers in Rat-1 Cells and Human Fibroblasts Reveals Differences in Protein Processing and Autocrine Effects MARGARET BYWATER,' FREDRIK RORSMAN.' ERIK BONGCAM-RUDLOFF, GEORGE MARK,2 ANNET HAMMACHER,3 CARL-HENRIK HELDIN , BENGT WESTERMARKl AND CHRISTER BETSHOLTZ'* Department of Pathology, Univ'ersit' Hospital,' and Ludwvig Institute for Cancer ResearcIh, Uppsala Brancsh,3 Uppsala, Sweden, and Laboratory of Humnan Carcinogenesis, National Canocer Institute, Bethesda, Maryland2 Received 22 December 1987/Accepted 8 April 1988 The autocrine effects of platelet-derived growth factor (PDGF) A- and B-chain homodimers (PDGF-AA and PDGF-BB) on rat-1 cells and human fibroblasts have been investigated by using human PDGF A- and B-chain cDNA clones expressed in a retroviral vector. Infection with replication-defective virus carrying the B-chain cDNA resulted in a phenotypical transformation resembling that induced by simian sarcoma virus. The resulting cells were focus forming in monolayer cultures, grew to high saturation densities, and formed large colonies in soft agar. The PDGF A-chain transfectants showed no transformed morphology and lacked focus-forming activity but grew to high saturation density in monolayer culture and formed small colonies in soft agar. A similar but weaker effect was obtained with an A-chain cDNA variant containing a 69-base-pair insertion in the 3' end of the protein-coding domain. A- and B-chain transfectants released PDGF receptor- competing activity into the medium, but only the medium conditioned by the B-chain transfectants possessed potent mitogenic activity on human fibroblasts. Both types of transfectants had downregulated levels of PDGF receptors; however, the B-chain transfectants were downregulated to significantly lower levels. Metabolic labeling and immunoprecipitations with PDGF antiserum showed that the PDGF B-chain protein was processed to a 24-kilodalton cell-associated and a 30-kilodalton secreted dimeric protein. The A-chain protein was rapidly secreted as a 31-kilodalton dimeric protein. The present study shows a marked difference in the autocrine effects of PDGF-AA and -BB expressed under the control of a retroviral promoter and suggests that different biological properties may be assigned to these two PDGF isoforms. The discovery of a close structural homology between the press). Human osteosarcoma- (18) and melanoma-derived platelet-derived growth factor (PDGF) B-chain and the pre- (44) growth factors have been identified as PDGF-AA, as has dicted protein product of the simian sarcoma virus (SSV) the major part of the activity secreted by human glioma cells oncogene, v-sis (11, 13, 43), provided the first direct evi- (A. Hammacher, M. Nistdr, B. Westermark, and C.-H. dence that the uncontrolled expression of a growth factor Heldin, submitted for publication 2; M. Nistdr, A. Hamma- may alter cell growth patterns in a way that might contribute cher, K. Mellstrom, A. Siegbahn, L. Ronnstrand, B. to malignant transformation. Additional examples of growth Westermark, and C.-H. Heldin, Cell, in press). The v-sis factor genes that under transcriptional control of viral pro- product (32) and porcine PDGF (39) are made up as homo- moters provide a transforming signal to relevant cells include dimers of B-chain homologs (PDGF-BB). the colony-stimulating factors GM-CSF (27) and CSF-l Recent studies have suggested that the various isoforms of (M-CSF) (37), transforming growth factor-Cx (TGF-oL) (15. 36, PDGF may have different functional activities. Thus, PDGF- 42), epidermal growth factor (EGF) (38), and basic fibroblast AA has been shown to possess relatively low mitogenic growth factor (bFGF) (34). Two potential oncogene prod- activity and be devoid of chemotactic activity and ability to ucts, the hst (41) and int-2 (12) proteins, have recently been induce tyrosine phosphorylation in porcine PDGF receptor shown to be homologous to bFGF and may be further preparations and cytoskeletal reorganization in cultured examples of transforming growth factor genes. human fibroblasts (Nist&r et al., in press). Binding studies Human PDGF is made up as a dimer of one B-chain have indicated the existence of at least two types of PDGF disulfide-linked to a second polypeptide, the A-chain (22). receptors, one (type A) which binds all three dimers, and The sequence of human PDGF A-chain cDNA clones another (type B) which binds only PDGF-BB and PDGF-AB showed a 50% amino acid sequence identity between the A- G. A. A. and B-chain precursors (4). Whereas the B-chain gene is (C.-H. Heldin, Backstrom, Ostman, Hammacher, located on human chromosome 22 (10, 40), the A-chain gene L. Ronnstrand, K. Rubin, M. Nistdr, and B. Westermark, is encoded by a distinct gene (35) on human chromosome 7 EMBO J., in press). Thus, the functions specified above (4). which are specific for B-chain containing PDGF dimers All three possible dimeric isoforms of PDGF have been appear to be triggered through the type B receptor. identified. PDGF purified from human platelets is predomi- The different biological properties of PDGF-AA and nantly a heterodimer (PDGF-AB) (A. Hammacher, U. Hell- PDGF-BB are of interest in relation to the differential man, U. Johnsson, A. Ostman, A. Gunnarsson, B. Wester- expression of the PDGF-A and -B genes in human tumor cell mark, A. Wasteson, and C.-H. Heldin, J. Cell. Biochem., in lines (4). The autocrine action of a growth factor is depen- dent not only on its biological activity in the extracellular space, but also on its synthesis rate, secretion, and process- * Corresponding author. ing. In order to experimentally determine and compare the 2753 2754 BYWATER ET AL. MOL. CELL. BIOL. autocrine effects of PDGF-AA and -BB, we have transfected of a heat-denatured radiolabeled DNA probe per ml was rat-1 cells and human fibroblasts by using human PDGF added, and incubation was continued overnight at 42°C. cDNA clones in a retrovirus vector. Differences were re- Probes were radiolabeled according to Feinberg and Vogel- vealed between the two PDGF proteins in the ability to stein (14). The filters were washed in 0.1x SSC-0.1% SDS at induce a transformed phenotype, growth in semisolid me- 60°C for 30 min and exposed at -70°C to Kodak XAR-5 films dium, and downregulation of PDGF receptors. Analysis of for 1 to 3 days. the PDGF A- and B-chain proteins produced by the trans- PDGF receptor measurements and the PDGF radioreceptor fected cells demonstrates differences in processing and con- assay. Pure PDGF-AB was prepared from human platelets as firmed the differences in biological activity of the two described (19). PDGF-AA was purified to apparent homoge- molecules. neity from supernatants of yeast cells transfected with a PDGF A-chain construct (A. Ostman et al., manuscript in MATERIALS AND METHODS preparation). The concentration of PDGF-AA was deter- mined by amino acid composition analysis. Both factors Construction of infectious retrovirus carrying coding se- were labeled with 1251 by the chloramine T method to quence for PDGF-A and PDGF-B. The human PDGF-A specific activities of 40,000 (PDGF-AB) and 50,000 (PDGF- cDNA clones 13.1 (35) and Dl (4) corresponding to a major AA) cpm/,ug. and a minor form of the PDGF-A mRNA, differing only by Binding of [1251]PDGF-AB was performed as described the absence or presence of exon 6 of the human gene (35), (30). Nonspecific binding was determined as the amount of respectively, were inserted into the BainHI site of the bound [125I]PDGF-AB in the presence of a 100-fold molar retroviral vector pLJ (26). This vector maintains the RNA excess of unlabeled PDGF-AB. packaging site qP. The cDNAs were inserted downstream Serum-free conditioned medium was harvested from ex- from the 5' Moloney murine leukemia virus (Mo-MuLV) ponentially growing G418-selected rat-1 cell cultures carry- long terminal repeat (LTR) and upstream from the neomycin ing the various recombinant retrovirus genomes. For assay- resistance gene, with the help of BarmHI linkers. The two ing the content of PDGF receptor-competing activity in cDNA clones, 1,302 and 1,233 bases long, respectively, these media, human diploid fibroblasts were used as test contain the complete protein coding sequence as well as 387 objects (30). Briefly, confluent cultures of fibroblasts were bases of 5' untranslated sequence (both 13.1 and DI) and 278 incubated with conditioned medium for 2 h at 4°C, followed (Dl) and 254 (13.1) bases of 3' untranslated sequence. The by washings and an additional incubation in buffer contain- Dl clone encodes a 211-amino-acid PDGF-A precursor with ing 2 ng of [125I]PDGF-AB per ml for 60 min at 4°C. After a highly basic and charged C-terminus. The 13.1 clone extensive washing, cells were solubilized in a Triton X-100- predicts a 15-amino-acid-shorter precursor lacking the basic containing buffer, and released radioactivity was determined C-terminus. in a Beckmann gamma counter. A human PDGF-B cDNA (sis-5; C. Betsholtz and A. Mitogenic and growth assays. Mitogenic activity of the Johnsson, unpublished) was cleaved with BamHI to gener- serum-free conditioned medium from marker-selected rat-1 ate a 2,082-base fragment containing the entire protein cultures was measured as [3H]thymidine incorporation in coding domain as well as 254 bases of 5' and 1,105 bases of human fibroblasts (3). The PDGF-specific nature of the 3' untranslated sequence, which was inserted into the same mitogenic activities was determined by neutralization with site of pLJ.
Load more

Recommended publications
  • Structural and Functional Properties of Platelet-Derived Growth Factor and Stem Cell Factor Receptors
    Downloaded from http://cshperspectives.cshlp.org/ on September 28, 2021 - Published by Cold Spring Harbor Laboratory Press Structural and Functional Properties of Platelet-Derived Growth Factor and Stem Cell Factor Receptors Carl-Henrik Heldin and Johan Lennartsson Ludwig Institute for Cancer Research, Uppsala University, SE-751 24 Uppsala, Sweden Correspondence: [email protected] The receptors for platelet-derived growth factor (PDGF) and stem cell factor (SCF) are members of the type III class of PTK receptors, which are characterized by five Ig-like domains extracellularly and a split kinase domain intracellularly. The receptors are activated by ligand-induced dimerization, leading to autophosphorylation on specific tyrosine resi- dues. Thereby the kinase activities of the receptors are activated and docking sites for down- stream SH2 domain signal transduction molecules are created; activation of these pathways promotes cell growth, survival, and migration. These receptors mediate important signals during the embryonal development, and control tissue homeostasis in the adult. Their over- activity is seen in malignancies and other diseases involving excessive cell proliferation, such as atherosclerosis and fibrotic diseases. In cancer, mutations of PDGF and SCF receptors— including gene fusions, point mutations, and amplifications—drive subpopulations of cer- tain malignancies, such as gastrointestinal stromal tumors, chronic myelomonocytic leuke- mia, hypereosinophilic syndrome, glioblastoma, acute myeloid leukemia, mastocytosis, and melanoma. he type III tyrosine kinase receptor family with kinases, and a less well-conserved carboxy- Tconsists of platelet-derived growth factor terminal tail. The ligands for these receptors are (PDGF) receptor a and b, stem cell factor all dimeric molecules, and on binding they in- (SCF) receptor (Kit), colony-stimulating fac- duce receptor dimerization.
    [Show full text]
  • Type of the Paper (Article
    Table S1. Gene expression of pro-angiogenic factors in tumor lymph nodes of Ibtk+/+Eµ-myc and Ibtk+/-Eµ-myc mice. Fold p- Symbol Gene change value 0,007 Akt1 Thymoma viral proto-oncogene 1 1,8967 061 0,929 Ang Angiogenin, ribonuclease, RNase A family, 5 1,1159 481 0,000 Angpt1 Angiopoietin 1 4,3916 117 0,461 Angpt2 Angiopoietin 2 0,7478 625 0,258 Anpep Alanyl (membrane) aminopeptidase 1,1015 737 0,000 Bai1 Brain-specific angiogenesis inhibitor 1 4,0927 202 0,001 Ccl11 Chemokine (C-C motif) ligand 11 3,1381 149 0,000 Ccl2 Chemokine (C-C motif) ligand 2 2,8407 298 0,000 Cdh5 Cadherin 5 2,5849 744 0,000 Col18a1 Collagen, type XVIII, alpha 1 3,8568 388 0,003 Col4a3 Collagen, type IV, alpha 3 2,9031 327 0,000 Csf3 Colony stimulating factor 3 (granulocyte) 4,3332 258 0,693 Ctgf Connective tissue growth factor 1,0195 88 0,000 Cxcl1 Chemokine (C-X-C motif) ligand 1 2,67 21 0,067 Cxcl2 Chemokine (C-X-C motif) ligand 2 0,7507 631 0,000 Cxcl5 Chemokine (C-X-C motif) ligand 5 3,921 328 0,000 Edn1 Endothelin 1 3,9931 042 0,001 Efna1 Ephrin A1 1,6449 601 0,002 Efnb2 Ephrin B2 2,8858 042 0,000 Egf Epidermal growth factor 1,726 51 0,000 Eng Endoglin 0,2309 467 0,000 Epas1 Endothelial PAS domain protein 1 2,8421 764 0,000 Ephb4 Eph receptor B4 3,6334 035 V-erb-b2 erythroblastic leukemia viral oncogene homolog 2, 0,000 Erbb2 3,9377 neuro/glioblastoma derived oncogene homolog (avian) 024 0,000 F2 Coagulation factor II 3,8295 239 1 0,000 F3 Coagulation factor III 4,4195 293 0,002 Fgf1 Fibroblast growth factor 1 2,8198 748 0,000 Fgf2 Fibroblast growth factor
    [Show full text]
  • Single-Cell Transcriptome Sequencing of 18,787 Human Induced Pluripotent Stem Cells Identifies Differentially Primed Subpopulations
    Single-cell transcriptome sequencing of 18,787 human induced pluripotent stem cells identifies differentially primed subpopulations Quan H. Nguyen1*, Samuel W. Lukowski1*, Han Sheng Chiu1, Anne Senabouth1, Timothy J. C. Bruxner1, Angelika N. Christ1, Nathan J. Palpant1*, Joseph E. Powell1,2* 1 Institute for Molecular Bioscience, University of Queensland, Brisbane, Australia 2 Queensland Brain Institute, University of Queensland, Brisbane, Australia * These authors contributed equally Supplementary Figures and Tables Table S1. Summary statistics for sequencing and mapping data of five samples Mean Median Total Median Percent Remaining Number Total number reads per genes genes UMIs per mapped cells post of cells of reads cell per cell detected cell reads filtering Sample 1 2,779 71,256 3,662 20,356 17,769 198,022,303 62.6 2,426 Sample 2 545 318,909 4,547 18,557 27,341 173,805,798 63.4 424 Sample 3 3,103 56,459 2,944 19,261 11,214 175,193,813 71.8 2,906 Sample 4 9,192 38,637 2,016 21,491 5,963 355,158,219 68.9 8,294 Sample 5 4,863 26,471 2,804 20,235 10,150 128,728,889 67.2 4,737 1 Table S2. Summary of the cell and gene filtering process Procedure Count Cells removed by library size (outside 3 x MAD range)a 0 Cells removed by number detected genes (outside 3 x MAD range) 77 Cells removed by reads mapped to mitochondrial genes (outside 3 x MAD range) 1,559 Cells removed by reads mapped to ribosomal genes (outside 3 x MAD range) 102 Cells removed by reads mapped to mitochondrial genes (> 20 % total reads)b 0 Cells removed by reads mapped to ribosomal genes (> 50 % total reads) 0 Genes removed by number of expressed cells (< 1 % total cells)c 16,674 Remaining cells post filtering 18,787 Remaining genes post filtering 16,064 aMAD stands for median absolute deviation.
    [Show full text]
  • PDGFRA in Vascular Adventitial Mscs Promotes Neointima Formation in Arteriovenous Fistula in Chronic Kidney Disease
    RESEARCH ARTICLE PDGFRA in vascular adventitial MSCs promotes neointima formation in arteriovenous fistula in chronic kidney disease Ke Song,1,2 Ying Qing,2 Qunying Guo,2 Eric K. Peden,3 Changyi Chen,4 William E. Mitch,2 Luan Truong,5 and Jizhong Cheng2 1Department of Stomatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. 2Selzman Institute for Kidney Health, Section of Nephrology, Department of Medicine, Baylor College of Medicine, Houston, Texas, USA. 3Department of Vascular Surgery, DeBakey Heart and Vascular Institute, Houston Methodist Hospital, Houston, Texas, USA. 4Michael E. DeBakey Department of Surgery, Baylor College of Medicine, Houston, Texas, USA. 5Department of Pathology and Genomic Medicine, Houston Methodist Hospital, Houston, Texas, USA. Chronic kidney disease (CKD) induces the failure of arteriovenous fistulas (AVFs) and promotes the differentiation of vascular adventitial GLI1-positive mesenchymal stem cells (GMCs). However, the roles of GMCs in forming neointima in AVFs remain unknown. GMCs isolated from CKD mice showed increased potential capacity of differentiation into myofibroblast-like cells. Increased activation of expression of PDGFRA and hedgehog (HH) signaling were detected in adventitial cells of AVFs from patients with end-stage kidney disease and CKD mice. PDGFRA was translocated and accumulated in early endosome when sonic hedgehog was overexpressed. In endosome, PDGFRA-mediated activation of TGFB1/SMAD signaling promoted the differentiation of GMCs into myofibroblasts, extracellular matrix deposition, and vascular fibrosis. These responses resulted in neointima formation and AVF failure. KO of Pdgfra or inhibition of HH signaling in GMCs suppressed the differentiation of GMCs into myofibroblasts. In vivo, specific KO of Pdgfra inhibited GMC activation and vascular fibrosis, resulting in suppression of neointima formation and improvement of AVF patency despite CKD.
    [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]
  • WO 2014/054013 Al 10 April 2014 (10.04.2014) P O P C T
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2014/054013 Al 10 April 2014 (10.04.2014) P O P C T (51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every G01N 33/68 (2006.01) kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, (21) International Application Number: BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, PCT/IB2013/059077 DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (22) International Filing Date: HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KN, KP, KR, 2 October 2013 (02. 10.2013) KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, (25) Filing Language: English OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, (26) Publication Language: English SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, (30) Priority Data: ZW. 61/710,491 5 October 2012 (05. 10.2012) 61/824,959 17 May 2013 (17.05.2013) (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (71) Applicant: NESTEC S.A. [CH/CH]; Ave. Nestle 55, CH- GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, SZ, TZ, 1800 Vevey (CH).
    [Show full text]
  • Fgf10 Dosage Is Critical for the Amplification of Epithelial Cell Progenitors and for the Formation of Multiple Mesenchymal Lineages During Lung Development
    Developmental Biology 307 (2007) 237–247 www.elsevier.com/locate/ydbio Fgf10 dosage is critical for the amplification of epithelial cell progenitors and for the formation of multiple mesenchymal lineages during lung development Suresh K. Ramasamy a,1, Arnaud A. Mailleux b,1, Varsha V. Gupte a, Francisca Mata a, Frédéric G. Sala a, Jacqueline M. Veltmaat c, Pierre M. Del Moral a, Stijn De Langhe a, Sara Parsa a, Lisa K. Kelly d, Robert Kelly e, Wei Shia a, Eli Keshet f, Parviz Minoo g, ⁎ David Warburton a, Savério Bellusci a, a Developmental Biology Program, Saban Research Institute of Childrens Hospital Los Angeles, Los Angeles, CA 90027, USA b Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA c Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore d Division of Pediatrics, Childrens Hospital Los Angeles, Los Angeles, CA 90027, USA e Developmental Biology Institute of Marseille Luminy-UMR6216-CNRS-Université de la Méditerranée, France f Department of Molecular Biology, The Hebrew University–Hadassah Medical School, Jerusalem, Israel g Department of Pediatrics, Women's and Children's Hospital, USC Keck School of Medicine, Los Angeles, CA 90033, USA Received for publication 23 October 2006; revised 24 April 2007; accepted 26 April 2007 Available online 3 May 2007 Abstract The key role played by Fgf10 during early lung development is clearly illustrated in Fgf10 knockout mice, which exhibit lung agenesis. However, Fgf10 is continuously expressed throughout lung development suggesting extended as well as additional roles for FGF10 at later stages of lung organogenesis. We previously reported that the enhancer trap Mlcv1v-nLacZ-24 transgenic mouse strain functions as a reporter for Fgf10 expression and displays decreased endogenous Fgf10 expression.
    [Show full text]
  • Control Mechanisms of Lung Alveolar Development and Their Disorders in Bronchopulmonary Dysplasia
    0031-3998/05/5705-0038R PEDIATRIC RESEARCH Vol. 57, No. 5, Pt 2, 2005 Copyright © 2005 International Pediatric Research Foundation, Inc. Printed in U.S.A. Control Mechanisms of Lung Alveolar Development and Their Disorders in Bronchopulmonary Dysplasia JACQUES BOURBON, OLIVIER BOUCHERAT, BERNADETTE CHAILLEY-HEU, AND CHRISTOPHE DELACOURT Inserm U651-Université Paris XII, Faculté de Médecine, F-94010 Créteil, France ABSTRACT Bronchopulmonary dysplasia (BPD) is a chronic lung disease premature baboon or lamb, neonatal exposure to hyperoxia in that occurs in very premature infants and is characterized by rodents, and maternal-fetal infection. These findings open ther- impaired alveologenesis. This ultimate phase of lung develop- apeutic perspectives to correct imbalanced signaling. Unraveling ment is mostly postnatal and allows growth of gas-exchange the intimate molecular mechanisms of alveolar building appears surface area to meet the needs of the organism. Alveologenesis is as a prerequisite to define new strategies for the prevention and a highly integrated process that implies cooperative interactions care of BPD. (Pediatr Res 57: 38R–46R, 2005) between interstitial, epithelial, and vascular compartments of the lung. Understanding of its underlying mechanisms has consider- Abbreviations ably progressed recently with identification of structural, signal- BPD, bronchopulmonary dysplasia ing, or remodeling molecules that are crucial in the process. ECM, extracellular matrix Thus, the pivotal role of elastin deposition in lung walls has been EMAP II, endothelial-monocyte activating polypeptide II demonstrated, and many key control-molecules have been iden- FGF, fibroblast growth factor tified, including various transcription factors, growth factors such GC, glucocorticoids as platelet-derived growth factor, fibroblast growth factors, and MMP, matrix metalloproteinase vascular endothelial growth factor, matrix-remodeling enzymes, PDGF, platelet-derived growth factor and retinoids.
    [Show full text]
  • Targeting the Platelet-Derived Growth Factor Receptor a with a Neutralizing
    Molecular Cancer Therapeutics 369 Targeting the platelet-derived growth factor receptor A with a neutralizing human monoclonal antibody inhibits the growth of tumor xenografts: Implications as a potential therapeutic target Nick Loizos, Yan Xu, Jim Huber, Meilin Liu, polypeptide chains that bind to two receptors simulta- Dan Lu, Bridget Finnerty, Robin Rolser, neously and induce receptor dimerization, autophosphor- Asra Malikzay, Anita Persaud, Erik Corcoran, ylation, and intracellular signaling. PDGFRa can form Dhanvanthri S. Deevi, Paul Balderes, Rajiv Bassi, homodimers as well as heterodimers with the structurally Xenia Jimenez, Christopher J. Joynes, similar PDGFRh. Given that PDGFRh does not bind the Venkata R.M. Mangalampalli, Philipp Steiner, PDGF-A chain with high affinity, PDGF-AA activates only James R. Tonra, Yan Wu, Daniel S. Pereira, aa receptor dimers, PDGF-AB and PDGF-CC activates aa Zhenping Zhu, Dale L. Ludwig, Daniel J. Hicklin, and ah receptor dimers, and PDGF-BB activates all three Peter Bohlen, Larry Witte, and Paul Kussie combinations of receptor dimers. A critical role for PDGFRa during early development is ImClone Systems Incorporated, New York, New York evident by the fact that mice homozygous for a null mutation die during embryogenesis. The homozygotes exhibit cranial Abstract malformations and a deficiency in myotome formation (3). a Platelet-derived growth factor receptor A (PDGFRA)isa At later stages of development, PDGFR is expressed in type III receptor tyrosine kinase that is expressed on a many mesenchymal structures, whereas adjacent epithelial variety of tumor types. A neutralizing monoclonal antibody cells produce PDGFs (reviewed in ref. 4). In the adult, to human PDGFRA, which did not cross-react with the hh PDGFs function in wound healing by activating mitogenesis, form of the receptor, was generated.
    [Show full text]
  • Post-Transcriptional Tuning of FGF Signaling Mediates Neural Crest Induction
    Post-transcriptional tuning of FGF signaling mediates neural crest induction Jacqueline Copelanda and Marcos Simoes-Costaa,1 aDepartment of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14850 Edited by Janet Rossant, The Gairdner Foundation, Toronto, Canada, and approved November 16, 2020 (received for review May 20, 2020) Ectodermal patterning is required for the establishment of multi- In recent years, growing evidence has supported a role for ple components of the vertebrate body plan. Previous studies have microRNA (miRNA)-mediated gene silencing in neural crest demonstrated that precise combinations of extracellular signals development (18–21). In the developing mouse embryo, the induce distinct ectodermal cell populations, such as the neural tissue-specific deletion of DICER, a key enzyme in miRNA crest and the neural plate. Yet, we still lack understanding of how biogenesis, results in a complete loss or malformation of several the response to inductive signals is modulated to generate the neural crest-derived structures (22–25). Furthermore, several proper transcriptional output in target cells. Here we show that lines of evidence have underscored a decisive role for specific posttranscriptional attenuation of fibroblast growth factor (FGF) miRNAs in modulating neural crest stem cell identity (19, 21, signaling is essential for the establishment of the neural crest 26). In other developmental contexts, miRNAs have been shown territory. We found that neural crest progenitors display elevated to modulate the expression of components of signaling cascades, expression of DICER, which promotes enhanced maturation of a thus influencing developmental decisions (27–30). These results set of cell-type-specific miRNAs. These miRNAs collectively target raise the possibility that posttranscriptional mechanisms may components of the FGF signaling pathway, a central player in the play an important role in the formation of ectodermal deriva- process of neural induction in amniotes.
    [Show full text]
  • Derived Growth Factor
    Human keratinocytes are a major source of cutaneous platelet- derived growth factor. J C Ansel, … , M L Usui, C E Hart J Clin Invest. 1993;92(2):671-678. https://doi.org/10.1172/JCI116636. Research Article PDGF has been implicated as one of the principal mitogens involved in cutaneous wound healing. While it has been previously reported that both platelets and monocytes are a source of PDGF in human dermal wound repair, the production of PDGF by human keratinocytes has not yet been described. In this manuscript, we report the production of PDGF by cultured human keratinocytes. Both PDGF A and B chain mRNA can be detected in cultured cells. While only PDGF-AA polypeptide is found in significant levels in keratinocyte-conditioned culture media, all three PDGF isoforms (AA, AB, and BB) are present in detergent-solubilized cell extracts. No evidence of PDGF receptor expression was observed in cultured keratinocytes when analyzed for either mRNA levels or polypeptide expression, suggesting that PDGF does not play an autocrine role in keratinocyte growth. Analysis of cryosections of human cutaneous wounds by immunostaining for PDGF showed that both PDGF A and B chain is constitutively expressed in normal epidermis, as well as in newly reconstituted wound epidermis. No evidence for PDGF receptor polypeptide expression in the epidermis was detected by immunostaining of cryosections. Find the latest version: https://jci.me/116636/pdf Human Keratinocytes Are a Major Source of Cutaneous Platelet-derived Growth Factor John C. Ansel, Jay P. Tiesman,11 John E. Olerud,5 James G. Krueger, Jeffrey F. Krane,t David C.
    [Show full text]
  • Nerve Growth Factor Neutralization Promotes Oligodendrogenesis by Increasing Mir-219A-5P Levels
    cells Article Nerve Growth Factor Neutralization Promotes Oligodendrogenesis by Increasing miR-219a-5p Levels Rossella Brandi 1,†, Marietta Fabiano 1,†, Corinna Giorgi 1,2, Ivan Arisi 1,3, Federico La Regina 1, Francesca Malerba 1, Sabrina Turturro 1, Andrea Ennio Storti 1, Flavia Ricevuti 1 , Susanna Amadio 4 , Cinzia Volontè 4,5 , Simona Capsoni 6,7, Raffaella Scardigli 1,3,* , Mara D’Onofrio 1,3,* and Antonino Cattaneo 6,* 1 European Brain Research Institute (EBRI) “Rita Levi-Montalcini”, Viale Regina Elena, 295, 00161 Rome, Italy; [email protected] (R.B.); [email protected] (M.F.); [email protected] (C.G.); [email protected] (I.A.); [email protected] (F.L.R.); [email protected] (F.M.); [email protected] (S.T.); [email protected] (A.E.S.); [email protected] (F.R.) 2 CNR, Institute of Molecular Biology and Pathology (IBPM), P.le Aldo Moro, 5, 00185 Rome, Italy 3 CNR, Institute of Translational Pharmacology (IFT), Via del Fosso del Cavaliere 100, 00131 Rome, Italy 4 IRCCS Fondazione Santa Lucia, Preclinical Neuroscience, Via del Fosso di Fiorano 65, 00143 Rome, Italy; [email protected] (S.A.); [email protected] (C.V.) 5 CNR, Institute for Systems Analysis and Computer Science, Via Dei Taurini 19, 00185 Rome, Italy 6 Bio@SNS, Scuola Normale Superiore, 56124 Pisa, Italy; [email protected] 7 Institute of Physiology, Department of Neuroscience and Rehabilitation University of Ferrara, 44121 Ferrara, Italy * Correspondence: [email protected] (R.S.); [email protected] (M.D.); [email protected] (A.C.) † These authors contributed equally to this work.
    [Show full text]