DOCSLIB.ORG

Rescue Screens with Secreted Proteins Reveal Compensatory Potential of Receptor Tyrosine Kinases in Driving Cancer Growth

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Rescue Screens with Secreted Proteins Reveal Compensatory Potential of Receptor Tyrosine Kinases in Driving Cancer Growth Published OnlineFirst August 8, 2012; DOI: 10.1158/2159-8290.CD-12-0237 RESEARCH ARTICLE Rescue Screens with Secreted Proteins Reveal Compensatory Potential of Receptor Tyrosine Kinases in Driving Cancer Growth Fred Harbinski 1 , Vanessa J. Craig 3 , Sneha Sanghavi 1 , Douglas Jeffery 1 , Lijuan Liu 1 , Kelly Ann Sheppard 1 , Sabrina Wagner 3 , Christelle Stamm 3 , Andreas Buness 3 , Christian Chatenay-Rivauday3 , Ya o Ya o 5 , Feng He 6 , Chris X. Lu 7 , Vito Guagnano 4 , Thomas Metz 8 , Peter M. Finan 1 , Francesco Hofmann 3 , William R. Sellers 2 , Jeffrey A. Porter 1 , Vic E. Myer1 , Diana Graus-Porta 3 , Christopher J. Wilson1 , Alan Buckler 1 , and Ralph Tiedt 3 Downloaded from cancerdiscovery.aacrjournals.org on September 28, 2021. © 2012 American Association for Cancer Research. Published OnlineFirst August 8, 2012; DOI: 10.1158/2159-8290.CD-12-0237 ABSTRACT The overall power of kinase inhibitors is substantially overshadowed by the acqui- sition of drug resistance. To address this issue, we systematically assessed the potential of secreted proteins to induce resistance to kinase inhibitors. To this end, we developed a high-throughput platform for screening a cDNA library encoding 3,432 secreted proteins in cellular assays. Using cancer cells originally dependent on either MET , FGFR2 , or FGFR3 , we observed a bypass of dependence through ligand-mediated activation of alternative receptor tyrosine kinases (RTK). Our fi ndings indicate a broad and versatile potential for RTKs from the HER and FGFR families as well as MET to compensate for loss of each other. We further provide evidence that combined inhibition of simultaneously active RTKs can lead to an added anticancer effect. SIGNIFICANCE: Although initial tumor responses to kinase inhibitors can be signifi cant, therapeutic ben- efi t is often limited by the emergence of resistance (e.g., as a consequence of mutations in the drug target or through activation of alternative pathways to bypass dependence on the original target). Because the activation of alternative growth-promoting kinases by stimulation with their cognate ligands can consti- tute such a bypass mechanism, the identifi cation of growth factors as possible mediators of resistance to kinase inhibitors is of clinical interest. Cancer Discov; 2(10); 1–12. ©2012 AACR. INTRODUCTION A well-characterized example representing both types of mechanisms is acquired resistance to the EGF receptor Genetic alterations, including point mutations, gene (EGFR) inhibitors gefi tinib and erlotinib in non–small cell amplifi cations, and chromosomal translocations, can render lung cancers carrying activating EGFR mutations ( 5, 6 ). In the kinases oncogenic ( 1, 2 ). The term “oncogene addiction” has majority of patients, this occurs either by emergence of the been used to describe the phenomenon in which growth and secondary resistance mutation T790M in EGFR or by loss of survival of cancer cells becomes dependent on an aberrantly EGFR dependence through activation of the receptor tyro- activated protein, for example, a kinase ( 3, 4 ). Oncogene sine kinase (RTK) MET ( 7–11 ). Although the EGFR-T790M addiction has been validated for several oncogenic “driver” mutation interferes with binding of EGFR inhibitors like kinases in preclinical models and in cancer patients using gefi tinib or erlotinib ( 12 ), MET activation can compensate selective kinase inhibitors. Despite striking initial clinical for loss of EGFR activity by activating an overlapping set activity, the emergence of resistance is becoming a common of downstream signaling molecules including HER3 that problem. One mechanism of resistance involves mutation continue to provide growth-promoting signals ( 8 ). Inter- of the target, thereby compromising binding and activity estingly, MET-driven resistance can occur because of MET of the therapeutic agent. Efforts to understand the specifi c gene amplifi cation ( 8 ) as well as activation by its ligand mechanisms of resistance to imatinib in chronic myelog- hepatocyte growth factor (HGF; refs. 10 , 13 ). On the other enous leukemia have led to second-generation inhibitors hand, it has been shown that MET-amplifi ed cancer cell (e.g., nilotinib) that treat and prevent resistance through lines that are sensitive to MET inhibitors can be rescued increased potency. However, the discovery of parallel or by ligand-induced EGFR activation ( 14 ). EGFR and MET downstream bypass mechanisms of resistance are motivat- are, therefore, considered to be alternative driver kinases ing novel combination therapies as a means to prevent such in the same cancer cells because dependence on one can be bypass events. compensated by activation of the other. In such a setting, simultaneous inhibition of both kinases or intervention at Authors’ Affi liations: 1 Developmental and Molecular Pathways and 2 Dis- a common downstream node is thus required for effective ease Area Oncology, Novartis Institutes for BioMedical Research, Cam- anticancer therapy. bridge, Massachusetts; 3 Disease Area Oncology and 4 Global Discovery Preclinical studies of resistance mechanisms have been Chemistry, Novartis Institutes for BioMedical Research, Basel, Switzerland; conducted using several experimental approaches, including 5 Oncology Translational Medicine, 6 Global Discovery Chemistry, and 7 Devel- opmental and Molecular Pathways, China Novartis Institutes for BioMedical random mutagenesis ( 15–17 ), prolonged drug exposure of sen- Research, Shanghai, PR China; and 8 Oncotest GmbH, Freiburg, Germany sitive cancer models to evoke outgrowth of resistant subclones Note: Supplementary data for this article are available at Cancer Discovery ( 18, 19 ), and expression of potential compensatory proteins in Online (http://www.cancerdiscovery.aacrjournals.org/). drug-exposed cancer cells using cDNA libraries ( 20 ). Prolonged F. Harbinski and V.J. Craig contributed equally to this work. drug-exposure experiments have successfully been used to Corresponding Author: Ralph Tiedt, Novartis Institutes for BioMedical recapitulate autocrine ligand–mediated resistance mechanisms Research, Klybeckstrasse 141, 4057 Basel, Switzerland. Phone: 41-61- that were previously found by hypothesis-driven testing ( 19 , 21 ). 696-1981; Fax: 41-61-696-5511; E-mail: [email protected] However, the potential role of the tumor microenvironment in doi: 10.1158/2159-8290.CD-12-0237 providing paracrine stimuli is not adequately modeled in such © 2012 American Association for Cancer Research. an experimental setup. The rationale for expressing cDNA OCTOBER 2012CANCER DISCOVERY | OF2 Downloaded from cancerdiscovery.aacrjournals.org on September 28, 2021. © 2012 American Association for Cancer Research. Published OnlineFirst August 8, 2012; DOI: 10.1158/2159-8290.CD-12-0237 RESEARCH ARTICLE Harbinski et al. libraries in drug-exposed cancer cells is that resistance to RTK growth compared to a neutral control (vector only). However, inhibitors can occur because of ligand-mediated activation of a small subset of supernatants was able to partially rescue the compensatory kinases. Such rescue ligands can be produced effect of MET inhibition and promote growth of MKN-45 by cancer cells themselves, leading to autocrine stimulation, in the presence of JAA120 ( Fig. 1B ). In line with previously or by stromal cells in the cancer microenvironment, leading to described concepts, members of the EGF family were among paracrine stimulation. the ligands that mediated MET inhibitor rescue ( Fig. 1B and To systematically assess the capability of secreted proteins Supplementary Table S2). In addition, we found that several to drive resistance to kinase inhibitors, we established a members of the fi broblast growth factor (FGF) family could high-throughput “secretome” screening platform. Secreted rescue MKN-45 cells. Among this family, FGF7-mediated proteins are produced in cell media supernatants by transfec- rescue was most pronounced ( Fig. 1B ). tion of a library encompassing 3,432 cDNAs. The resulting Similarly, secretome resistance screening was then carried supernatants are then screened for their potential to rescue out in 4 additional cancer cell lines in which MET activity kinase-dependent cancer cells that are simultaneously treated was known to be a major driver of cell growth. Also in these with a relevant kinase inhibitor. Specifi cally, supernatants additional lines, we observed that members of the EGF and/ leading to the reversal of inhibitor-mediated proliferation in or FGF families could suppress the growth effects of JAA120 MET-dependent and FGFR-dependent cancer cell lines were (Supplementary Table S2). These data suggest that down- discovered. Although the specifi c ligands of each inhibited stream signals emerging from either HER or FGFR family RTK were not capable of reversing growth inhibition, a sur- members that promote cell growth and survival are to some prisingly high degree of fl exibility was observed in bypassing extent qualitatively redundant with signals triggered by MET. oncogene addiction through activation of alternative RTKs. In particular, activation of HER family members could com- Secretome Screening Reveals Broad pensate for inhibition of either MET or FGFRs and activation Compensatory Potential of HER, FGFR, of FGFR or MET could cross-rescue HER inhibition. These and MET RTKs results suggest a specifi c and complementary role of these 3 Next, cancer cell lines that are dependent on a member growth factor pathways and suggest at least partial overlap of of the FGFR family
Load more

Recommended publications
  • ARTICLES Fibroblast Growth Factors 1, 2, 17, and 19 Are The
    0031-3998/07/6103-0267 PEDIATRIC RESEARCH Vol. 61, No. 3, 2007 Copyright © 2007 International Pediatric Research Foundation, Inc. Printed in U.S.A. ARTICLES Fibroblast Growth Factors 1, 2, 17, and 19 Are the Predominant FGF Ligands Expressed in Human Fetal Growth Plate Cartilage PAVEL KREJCI, DEBORAH KRAKOW, PERTCHOUI B. MEKIKIAN, AND WILLIAM R. WILCOX Medical Genetics Institute [P.K., D.K., P.B.M., W.R.W.], Cedars-Sinai Medical Center, Los Angeles, California 90048; Department of Obstetrics and Gynecology [D.K.] and Department of Pediatrics [W.R.W.], UCLA School of Medicine, Los Angeles, California 90095 ABSTRACT: Fibroblast growth factors (FGF) regulate bone growth, (G380R) or TD (K650E) mutations (4–6). When expressed at but their expression in human cartilage is unclear. Here, we deter- physiologic levels, FGFR3-G380R required, like its wild-type mined the expression of entire FGF family in human fetal growth counterpart, ligand for activation (7). Similarly, in vitro cul- plate cartilage. Using reverse transcriptase PCR, the transcripts for tivated human TD chondrocytes as well as chondrocytes FGF1, 2, 5, 8–14, 16–19, and 21 were found. However, only FGF1, isolated from Fgfr3-K644M mice had an identical time course 2, 17, and 19 were detectable at the protein level. By immunohisto- of Fgfr3 activation compared with wild-type chondrocytes and chemistry, FGF17 and 19 were uniformly expressed within the showed no receptor activation in the absence of ligand (8,9). growth plate. In contrast, FGF1 was found only in proliferating and hypertrophic chondrocytes whereas FGF2 localized predominantly to Despite the importance of the FGF ligand for activation of the resting and proliferating cartilage.
    [Show full text]
  • Ectodysplasin Target Gene Fgf20 Regulates Mammary Bud Growth and Ductal Invasion and Branching During Puberty
    View metadata, citation and similar papersbroughtCORE at to core.ac.uk you by provided by Institute of Cancer Research Repository Ectodysplasin target gene Fgf20 regulates mammary bud growth and ductal invasion and branching during puberty Teresa Elo1,#, Päivi H. Lindfors1,#, Qiang Lan1 , Maria Voutilainen1, Ewelina Trela1, Claes Ohlsson2, Sung-Ho Huh3,^, David M. Ornitz3, Matti Poutanen4, Beatrice A. Howard5, Marja L. Mikkola1,* 1 Developmental Biology Program, Institute of Biotechnology, University of Helsinki, Finland 2 Center for Bone and Arthritis Research, Department of Internal Medicine, Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Sweden 3 Department of Developmental Biology, Washington University School of Medicine, St. Louis, Missouri, USA 4 Department of Physiology and Turku Center for Disease Modeling, Institute of Biomedicine, University of Turku, Turku, Finland 5 The Breast Cancer Now Toby Robins Research Centre, Division of Breast Cancer, the Institute of Cancer Research, London, United Kingdom #) these authors contributed equally ^) current address: Holland Regenerative Medicine Program and Department of Developmental Neuroscience, Munroe-Meyer Institute, University of Nebraska Medical Center, Omaha, Nebraska, USA *) Author for correspondence: Marja L. Mikkola Developmental Biology Program Institute of Biotechnology University of Helsinki P.O.B. 56 00014 Helsinki Finland e-mail: [email protected] phone: +358-2-941 59344 fax: +358-2-941 59366 1 Abstract Mammary gland development begins with the appearance of epithelial placodes that invaginate, sprout, and branch to form small arborized trees by birth. The second phase of ductal growth and branching is driven by the highly invasive structures called terminal end buds (TEBs) that form at ductal tips at the onset of puberty.
    [Show full text]
  • FGF Signaling Network in the Gastrointestinal Tract (Review)
    163-168 1/6/06 16:12 Page 163 INTERNATIONAL JOURNAL OF ONCOLOGY 29: 163-168, 2006 163 FGF signaling network in the gastrointestinal tract (Review) MASUKO KATOH1 and MASARU KATOH2 1M&M Medical BioInformatics, Hongo 113-0033; 2Genetics and Cell Biology Section, National Cancer Center Research Institute, Tokyo 104-0045, Japan Received March 29, 2006; Accepted May 2, 2006 Abstract. Fibroblast growth factor (FGF) signals are trans- Contents duced through FGF receptors (FGFRs) and FRS2/FRS3- SHP2 (PTPN11)-GRB2 docking protein complex to SOS- 1. Introduction RAS-RAF-MAPKK-MAPK signaling cascade and GAB1/ 2. FGF family GAB2-PI3K-PDK-AKT/aPKC signaling cascade. The RAS~ 3. Regulation of FGF signaling by WNT MAPK signaling cascade is implicated in cell growth and 4. FGF signaling network in the stomach differentiation, the PI3K~AKT signaling cascade in cell 5. FGF signaling network in the colon survival and cell fate determination, and the PI3K~aPKC 6. Clinical application of FGF signaling cascade in cell polarity control. FGF18, FGF20 and 7. Clinical application of FGF signaling inhibitors SPRY4 are potent targets of the canonical WNT signaling 8. Perspectives pathway in the gastrointestinal tract. SPRY4 is the FGF signaling inhibitor functioning as negative feedback apparatus for the WNT/FGF-dependent epithelial proliferation. 1. Introduction Recombinant FGF7 and FGF20 proteins are applicable for treatment of chemotherapy/radiation-induced mucosal injury, Fibroblast growth factor (FGF) family proteins play key roles while recombinant FGF2 protein and FGF4 expression vector in growth and survival of stem cells during embryogenesis, are applicable for therapeutic angiogenesis. Helicobacter tissues regeneration, and carcinogenesis (1-4).
    [Show full text]
  • FGF20 and DKK1 Are Transcriptional Targets of Catenin and FGF20 Is
    The EMBO Journal (2005) 24, 73–84 | & 2005 European Molecular Biology Organization | All Rights Reserved 0261-4189/05 www.embojournal.org THE EMBO JOJOURNALUR NAL FGF-20 and DKK1 are transcriptional targets of b-catenin and FGF-20 is implicated in cancer and development Mario N Chamorro1,4, Introduction Donald R Schwartz2,5, Alin Vonica3,5, Wnt proteins are secreted glycoproteins that bind and acti- Ali H Brivanlou3, Kathleen R Cho2 1, vate two classes of co-receptors, LDL-related proteins (LRPs) and Harold E Varmus * and members of the Frizzled protein family. Signaling in- 1Cancer Biology and Genetics Program, Sloan-Kettering Institute, itiated by Wnts and their receptors controls a wide variety of Varmus Laboratory, Memorial Sloan-Kettering Cancer Center, New York, cell processes, including cell fate specification, differentia- NY, USA, 2Department of Pathology, The University of Michigan Medical 3 tion, migration, and polarity (reviewed in Peifer and Polakis, School, Ann Arbor, MI, USA, The Laboratory of Vertebrate Embryology, b The Rockefeller University, New York, NY, USA and 4Cell Biology 2000). -Catenin is the major effector of the canonical Wnt Program, Cornell University, Weill Graduate School of Medical Sciences, signaling pathway. In the absence of Wnt, cytosolic b-catenin New York, NY, USA forms a complex with Axin and adenomatous polyposis coli (APC) proteins, and is rapidly degraded by the ubiquitina- b-catenin is the major effector of the canonical Wnt signal- tion–proteosome system. Wnt signaling inactivates the ing pathway. Mutations in components of the pathway that b-catenin destruction complex, so that b-catenin is stabilized, stabilize b-catenin result in augmented gene transcription accumulates in the cytoplasm and nucleus, and forms hetero- and play a major role in many human cancers.
    [Show full text]
  • Ectodysplasin Target Gene Fgf20 Regulates Mammary Bud Growth and Ductal Invasion and Branching During Puberty Teresa Elo University of Helsinki
    Washington University School of Medicine Digital Commons@Becker Open Access Publications 2017 Ectodysplasin target gene Fgf20 regulates mammary bud growth and ductal invasion and branching during puberty Teresa Elo University of Helsinki Päivi H. Lindfors University of Helsinki Qiang Lan University of Helsinki Maria Voutilainen University of Helsinki Ewelina Trela University of Helsinki See next page for additional authors Follow this and additional works at: https://digitalcommons.wustl.edu/open_access_pubs Recommended Citation Elo, Teresa; Lindfors, Päivi H.; Lan, Qiang; Voutilainen, Maria; Trela, Ewelina; Ohlsson, Claes; Huh, Sung-Ho; Ornitz, David M.; Poutanen, Matti; Howard, Beatrice A.; and Mikkola, Marja L., ,"Ectodysplasin target gene Fgf20 regulates mammary bud growth and ductal invasion and branching during puberty." Scientific Reports.7,. (2017). https://digitalcommons.wustl.edu/open_access_pubs/6000 This Open Access Publication is brought to you for free and open access by Digital Commons@Becker. It has been accepted for inclusion in Open Access Publications by an authorized administrator of Digital Commons@Becker. For more information, please contact [email protected]. Authors Teresa Elo, Päivi H. Lindfors, Qiang Lan, Maria Voutilainen, Ewelina Trela, Claes Ohlsson, Sung-Ho Huh, David M. Ornitz, Matti Poutanen, Beatrice A. Howard, and Marja L. Mikkola This open access publication is available at Digital Commons@Becker: https://digitalcommons.wustl.edu/open_access_pubs/6000 www.nature.com/scientificreports OPEN Ectodysplasin target gene Fgf20 regulates mammary bud growth and ductal invasion and branching Received: 28 December 2016 Accepted: 18 May 2017 during puberty Published: xx xx xxxx Teresa Elo1, Päivi H. Lindfors1, Qiang Lan1, Maria Voutilainen1, Ewelina Trela1, Claes Ohlsson2, Sung-Ho Huh3,6, David M.
    [Show full text]
  • Expression of Fgfs During Early Mouse Tongue Development
    Gene Expression Patterns 20 (2016) 81e87 Contents lists available at ScienceDirect Gene Expression Patterns journal homepage: http://www.elsevier.com/locate/gep Expression of FGFs during early mouse tongue development * Wen Du a, b, Jan Prochazka b, c, Michaela Prochazkova b, c, Ophir D. Klein b, d, a State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, 610041, China b Department of Orofacial Sciences and Program in Craniofacial Biology, University of California San Francisco, San Francisco, CA 94143, USA c Laboratory of Transgenic Models of Diseases, Institute of Molecular Genetics of the ASCR, v.v.i., Prague, Czech Republic d Department of Pediatrics and Institute for Human Genetics, University of California San Francisco, San Francisco, CA 94143, USA article info abstract Article history: The fibroblast growth factors (FGFs) constitute one of the largest growth factor families, and several Received 29 September 2015 ligands and receptors in this family are known to play critical roles during tongue development. In order Received in revised form to provide a comprehensive foundation for research into the role of FGFs during the process of tongue 13 December 2015 formation, we measured the transcript levels by quantitative PCR and mapped the expression patterns by Accepted 29 December 2015 in situ hybridization of all 22 Fgfs during mouse tongue development between embryonic days (E) 11.5 Available online 31 December 2015 and E14.5. During this period, Fgf5, Fgf6, Fgf7, Fgf9, Fgf10, Fgf13, Fgf15, Fgf16 and Fgf18 could all be detected with various intensities in the mesenchyme, whereas Fgf1 and Fgf2 were expressed in both the Keywords: Tongue epithelium and the mesenchyme.
    [Show full text]
  • Growth Factors in Multiple Myeloma
    Mahtouk et al. BMC Cancer 2010, 10:198 http://www.biomedcentral.com/1471-2407/10/198 RESEARCH ARTICLE Open Access GrowthResearch article factors in multiple myeloma: a comprehensive analysis of their expression in tumor cells and bone marrow environment using Affymetrix microarrays Karène Mahtouk1, Jérôme Moreaux1,2, Dirk Hose3,4, Thierry Rème1,2, Tobias Meißner3, Michel Jourdan1, Jean François Rossi1,2,5, Steven T Pals6, Hartmut Goldschmidt3,4 and Bernard Klein*1,2,5 Abstract Background: Multiple myeloma (MM) is characterized by a strong dependence of the tumor cells on their microenvironment, which produces growth factors supporting survival and proliferation of myeloma cells (MMC). In the past few years, many myeloma growth factors (MGF) have been described in the literature. However, their relative importance and the nature of the cells producing MGF remain unidentified for many of them. Methods: We have analysed the expression of 51 MGF and 36 MGF receptors (MGFR) using Affymetrix microarrays throughout normal plasma cell differentiation, in MMC and in cells from the bone marrow (BM) microenvironment (CD14, CD3, polymorphonuclear neutrophils, stromal cells and osteoclasts). Results: 4/51 MGF and 9/36 MGF-receptors genes were significantly overexpressed in plasmablasts (PPC) and BM plasma cell (BMPC) compared to B cells whereas 11 MGF and 11 MGFR genes were overexpressed in BMPC compared to PPC. 3 MGF genes (AREG, NRG3, Wnt5A) and none of the receptors were significantly overexpressed in MMC versus BMPC. Furthermore, 3/51 MGF genes were overexpressed in MMC compared to the the BM microenvironment whereas 22/51 MGF genes were overexpressed in one environment subpopulation compared to MMC.
    [Show full text]
  • Role of Fibroblast Growth Factors in Bone Regeneration Pornkawee Charoenlarp, Arun Kumar Rajendran and Sachiko Iseki*
    Charoenlarp et al. Inflammation and Regeneration (2017) 37:10 Inflammation and Regeneration DOI 10.1186/s41232-017-0043-8 REVIEW Open Access Role of fibroblast growth factors in bone regeneration Pornkawee Charoenlarp, Arun Kumar Rajendran and Sachiko Iseki* Abstract Bone is a metabolically active organ that undergoes continuous remodeling throughout life. However, many complex skeletal defects such as large traumatic bone defects or extensive bone loss after tumor resection may cause failure of bone healing. Effective therapies for these conditions typically employ combinations of cells, scaffolds, and bioactive factors. In this review, we pay attention to one of the three factors required for regeneration of bone, bioactive factors, especially the fibroblast growth factor (FGF) family. This family is composed of 22 members and associated with various biological functions including skeletal formation. Based on the phenotypes of genetically modified mice and spatio- temporal expression levels during bone fracture healing, FGF2, FGF9, and FGF18 are regarded as possible candidates useful for bone regeneration. The role of these candidate FGFs in bone regeneration is also discussed in this review. Keywords: Bone regeneration, FGFs, FGF2, FGF9, FGF18, Osteogenesis, Tissue engineering Background growth factors (FGFs) and their roles in bone Tissue engineering is an interdisciplinary field of research regeneration. and clinical applications, which focuses on restoration of FGF signaling in skeletal formation has been demon- impaired function and morphology of tissues and organs strated by identification of gain-of-function mutations in by repair, replacement, or regeneration. It uses a combin- human FGF receptor (FGFR) genes in craniosynostosis ation of several technological approaches beyond trad- and dwarfism patients and skeletal phenotypes in genetic- itional transplantation and replacement therapies.
    [Show full text]
  • FGF/FGFR Signaling in Health and Disease
    Signal Transduction and Targeted Therapy www.nature.com/sigtrans REVIEW ARTICLE OPEN FGF/FGFR signaling in health and disease Yangli Xie1, Nan Su1, Jing Yang1, Qiaoyan Tan1, Shuo Huang 1, Min Jin1, Zhenhong Ni1, Bin Zhang1, Dali Zhang1, Fengtao Luo1, Hangang Chen1, Xianding Sun1, Jian Q. Feng2, Huabing Qi1 and Lin Chen 1 Growing evidences suggest that the fibroblast growth factor/FGF receptor (FGF/FGFR) signaling has crucial roles in a multitude of processes during embryonic development and adult homeostasis by regulating cellular lineage commitment, differentiation, proliferation, and apoptosis of various types of cells. In this review, we provide a comprehensive overview of the current understanding of FGF signaling and its roles in organ development, injury repair, and the pathophysiology of spectrum of diseases, which is a consequence of FGF signaling dysregulation, including cancers and chronic kidney disease (CKD). In this context, the agonists and antagonists for FGF-FGFRs might have therapeutic benefits in multiple systems. Signal Transduction and Targeted Therapy (2020) 5:181; https://doi.org/10.1038/s41392-020-00222-7 INTRODUCTION OF THE FGF/FGFR SIGNALING The binding of FGFs to the inactive monomeric FGFRs will Fibroblast growth factors (FGFs) are broad-spectrum mitogens and trigger the conformational changes of FGFRs, resulting in 1234567890();,: regulate a wide range of cellular functions, including migration, dimerization and activation of the cytosolic tyrosine kinases by proliferation, differentiation, and survival. It is well documented phosphorylating the tyrosine residues within the cytosolic tail of that FGF signaling plays essential roles in development, metabo- FGFRs.4 Then, the phosphorylated tyrosine residues serve as the lism, and tissue homeostasis.
    [Show full text]
  • Halotag Is an Effective Expression and Solubilisation Fusion Partner for a Range of Fibroblast Growth Factors
    HaloTag is an eVective expression and solubilisation fusion partner for a range of fibroblast growth factors Changye Sun1,3 , Yong Li1,3 , Sarah E. Taylor1, Xianqing Mao2, Mark C. Wilkinson1 and David G. Fernig1 1 Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Liverpool, UK 2 Department of Oncology, Laboratory of Cellular and Molecular Oncology, Luxembourg Institute of Health, Luxembourg 3 These authors contributed equally to this work. ABSTRACT The production of recombinant proteins such as the fibroblast growth factors (FGFs) is the key to establishing their function in cell communication. The production of recombinant FGFs in E. coli is limited, however, due to expression and solubility problems. HaloTag has been used as a fusion protein to introduce a genetically- encoded means for chemical conjugation of probes. We have expressed 11 FGF proteins with an N-terminal HaloTag, followed by a tobacco etch virus (TEV) protease cleavage site to allow release of the FGF protein. These were purified by heparin-aYnity chromatography, and in some instances by further ion-exchange chromatography. It was found that HaloTag did not adversely aVect the expression of FGF1 and FGF10, both of which expressed well as soluble proteins. The N-terminal HaloTag fusion was found to enhance the expression and yield of FGF2, FGF3 and FGF7. Moreover, whereas FGF6, FGF8, FGF16, FGF17, FGF20 and FGF22 were only expressed as insoluble proteins, their N-terminal HaloTag fusion counterparts (Halo-FGFs) were soluble, and could be successfully purified. However, cleavage of Halo-FGF6, -FGF8 and -FGF22 with TEV resulted in aggregation of the FGF protein.
    [Show full text]
  • Dissertation LFS Corrected 08-22-16
    EXPRESSION AND ROLE OF FIBROBLAST GROWTH FACTOR 9 (FGF9) IN BOVINE FOLLICULOGENESIS By LUÍS FERNANDO SCHÜTZ Doctor of Veterinary Medicine Federal University of Rio Grande do Sul Porto Alegre, Brazil 2007 Master of Science in Animal Science Santa Catarina State University Lages, Brazil Submitted to the Faculty of the Graduate College of the Oklahoma State University in partial fulfillment of the requirements for the Degree of DOCTOR OF PHILOSOPHY July, 2016 EXPRESSION AND ROLE OF FIBROBLAST GROWTH FACTOR 9 (FGF9) IN BOVINE FOLLICULOGENESIS Dissertation Approved: Dr. Leon J. Spicer Dissertation Adviser Dr. Robert Wettemann Dr. Glenn Zhang Dr. Peter Hoyt ii ACKNOWLEDGEMENTS To my beloved wife, for her love, support, encouragement, and understanding. She made all the steps of the arduous pathway to achieve this degree much easier. With her by my side, I feel much stronger. To my family in Brazil: my beloved parents, my brother, Lipe, and my sister, Ane, for their love, faith, and support. And to my beautiful niece, Sofia, my sister-in-law, Manuela, and my brother-in-law, Joni. To my advisor and mentor, Dr. Leon Spicer, for accepting me as a student, for giving me priceless lessons and advice whenever I needed, for stimulating me to develop a critical thinking, and for teaching me how to perform sound science in the field of ovarian folliculogenesis. And to his family, Michael, Melissa, Anna, and Richie, for their friendship and support. To the Franceschi family: my mother-in-law, Pierina, and my father-in-law, Adi, for their support and encouragement; to my brother-in-law, Junior, his wife, Janine, and his great boys, Bernardo and Bruno; and to my sister-in-law, Sheila, and her fiancé, Fábio.
    [Show full text]
  • Genetic Variation Infgf20modulates Hippocampal Biology
    5992 • The Journal of Neuroscience, April 28, 2010 • 30(17):5992–5997 Behavioral/Systems/Cognitive Genetic Variation in FGF20 Modulates Hippocampal Biology Herve Lemaitre,1 Venkata S. Mattay,1 Fabio Sambataro,1 Beth Verchinski,1 Richard E. Straub,1 Joseph H. Callicott,1 Raja Kittappa,2 Thomas M. Hyde,1 Barbara K. Lipska,1 Joel E. Kleinman,1 Ronald McKay,2 and Daniel R. Weinberger1 1Clinical Brain Disorder Branch, Genes, Cognition, and Psychosis Program, National Institutes of Health–National Institute of Mental Health, and 2Laboratory of Molecular Biology, National Institutes of Health–National Institute of Neurological Disorders and Stroke, Bethesda, Maryland 20892 Weexploredtheeffectofsingle-nucleotidepolymorphisms(SNPs)inthefibroblastgrowthfactor20gene(FGF20)associatedwithriskfor Parkinson’s disease on brain structure and function in a large sample of healthy young-adult human subjects and also in elderly subjects to look at the interaction between genetic variations and age (N ϭ 237; 116 men; 18–87 years). We analyzed high-resolution anatomical magnetic resonance images using voxel-based morphometry, a quantitative neuroanatomical technique. We also measured FGF20 mRNA expression in postmortem human brain tissue to determine the molecular correlates of these SNPs (N ϭ 108; 72 men; 18–74 years). We found that the T allele carriers of rs12720208 in the 3Ј-untranslated region had relatively larger hippocampal volume ( p ϭ 0.0059) and diminished verbal episodic memory ( p ϭ 0.048) and showed steeper decreases of hippocampal volume with normal aging ( p ϭ 0.026). In postmortem brain, T allele carriers had greater expression of hippocampal FGF20 mRNA ( p ϭ 0.037), consistent with a previously characterized microRNA mechanism.
    [Show full text]