DOCSLIB.ORG

Autocrine BMP-4 Signaling Is a Therapeutic Target in Colorectal

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Autocrine BMP-4 Signaling Is a Therapeutic Target in Colorectal Published OnlineFirst June 13, 2017; DOI: 10.1158/0008-5472.CAN-17-0112 Cancer Molecular and Cellular Pathobiology Research Autocrine BMP-4 Signaling Is a Therapeutic Target in Colorectal Cancer Yuichiro Yokoyama1,2, Toshiaki Watanabe2, Yusuke Tamura1, Yoshinobu Hashizume3, Kohei Miyazono1, and Shogo Ehata1 Abstract Poor prognoses for colorectal cancer patients with meta- LDN-193189 elevated expression of the phosphatase DUSP5 static lesions have driven demand for the development of in colorectal cancer cells, inducing apoptosis via dephosphor- novel targeted therapies. Here, we demonstrate that expres- ylation of Erk MAPK. Administering LDN-193189 to mice sion of bone morphogenetic protein 4 (BMP-4) is universally diminished tumor formation of colorectal cancer cells. Our upregulated in human colorectal cancer cells and tissues, findings suggest inhibition of autocrine BMP-4 as a candidate resulting in activated BMP signaling. Inhibition of endoge- treatment strategy for colorectal cancer. Cancer Res; 77(15); nous BMP signaling by the BMP type I receptor inhibitor 4026–38. Ó2017 AACR. Introduction their ability to bind certain type I receptors. Upon binding to type I and type II receptors, BMPs form heterotetrameric com- Colorectal cancer is the third most common cancer and the plexes; the protein kinase of the type II receptor activates the fourth most common cause of cancer-related death worldwide protein kinase of the type I receptor, which in turn phosphor- (1). Although surgical resection can cure early-stage colorectal ylates the BMP-specific receptor-regulated Smads (R-Smads), cancer, a combination of surgery and chemotherapeutic agents is Smad1 and Smad5. Phosphorylated R-Smads induce a hetero- recommended in advanced stages of colorectal cancer (1). In meric assembly with common-partner Smad (Co-Smad; addition to conventional cytotoxic agents, new agents targeting Smad4) and translocate into the nucleus, regulating the tran- VEGF signaling and EGFR signaling have been introduced during scription of target genes. BMPs can also activate non-Smad the last decade (2). Although colorectal cancer prognoses have signaling pathways, including the MAPK pathway (3). steadily improved, the 5-year survival rate remains low, especially Divergent roles of BMPs have been reported during cancer in patients with metastatic lesions (1). Thus, the development of progression (4–6). BMPs inhibit proliferation of gastric cancer, new molecular targets for treatment of advanced colorectal cancer breast cancer, and prostate cancer cells, induce differentiation of is critical for improving patient outcomes. glioma-initiating cells, and inhibit glioblastoma tumor formation Bone morphogenetic proteins (BMP) are members of the (5), indicating a tumor-suppressive role of BMPs. In contrast, TGF-b family and are multifunctional cytokines (3–5). BMPs BMPs have been reported to enhance the motility and invasive- recognize two distinct receptors, termed type I and type II ness of various types of cancer cells, such as breast cancer, prostate receptors, with serine/threonine and tyrosine kinase activities. cancer, and malignant melanoma cells, suggesting that BMPs also Type I BMP receptors include activin receptor-like kinase (ALK)- function as tumor-promoting factors (4). 1, -2, -3, and -6, and type II receptors include BMP type II In this study, the role of BMP-4 produced by colorectal cancer receptor (BMPR-II), activin type II receptor (ActR-II), and cells in cancer progression was investigated. We reveal for the first activin type IIB receptor (ActR-IIB). BMPs are classified into time that inhibition of BMP-4 induces the apoptosis of colorectal several subgroups, including the BMP-2/4 group, BMP-5/6/7/8 cancer cells through the attenuation of MAPK activity in culture group, BMP-9/10 group, and growth and differentiation factor and that the small-molecule BMP inhibitor LDN-193189 (GDF)-5/6/7 group, according to structural similarities and diminishes colorectal cancer formation in vivo. 1Department of Molecular Pathology, Graduate School of Medicine, The Univer- Materials and Methods sity of Tokyo, Tokyo, Japan. 2Department of Surgical Oncology, The University of Cell culture and reagents 3 Tokyo, Bunkyo-ku, Tokyo, Japan. RIKEN Program for Drug Discovery and Human colon adenocarcinoma cells HT29 and DLD-1 (Japa- Medical Technology Platforms, Wako, Saitama, Japan. nese Cancer Research Resource Bank) were cultured in RPMI Note: Supplementary data for this article are available at Cancer Research containing 10% FBS, penicillin, and streptomycin. Human colon Online (http://cancerres.aacrjournals.org/). adenocarcinoma SW480 cells (ATCC) were cultured in DMEM Corresponding Authors: Shogo Ehata, Graduate School of Medicine, The containing 10% FBS, penicillin, and streptomycin. Routine Myco- University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. Phone: plasma testing was performed by PCR regularly on these cells. The 813-5841-3356; Fax: 813-5841-3354; E-mail: [email protected]; and Kohei cells were bought in 2002 and have been stocked as cryopreserved Miyazono, [email protected] aliquots in liquid N2. The cells were used within 8 passages after doi: 10.1158/0008-5472.CAN-17-0112 thawing and reauthenticated by short tandem repeat profiling in Ó2017 American Association for Cancer Research. 2017. LDN-193189 was obtained from Wako or RIKEN. 4026 Cancer Res; 77(15) August 1, 2017 Downloaded from cancerres.aacrjournals.org on October 3, 2021. © 2017 American Association for Cancer Research. Published OnlineFirst June 13, 2017; DOI: 10.1158/0008-5472.CAN-17-0112 BMP-4 as a New Therapeutic Target in Colorectal Cancer Quantitative real-time RT-PCR analyses and chromatin NCI-60 cell line panel indicated that, among various BMPs, immunoprecipitation–qRT-PCR analyses expression of BMP4 was commonly elevated in colon cancer cells Quantitative real-time RT-PCR (qRT-PCR) analysis and chro- (Fig. 1A). NCBI GEO database GSE14258 revealed that expression matin immunoprecipitation (ChIP)–qRT-PCR analysis were of BMP4, but not of other BMPs, was significantly higher in performed as previously described (7, 8). Primer sequences colon cancer tissues than in normal colon tissues (Fig. 1B). Next, are described in Supplementary Table S1. Anti-TCF4 antibody the correlation between BMP4 expression and colorectal cancer (sc-8631) was purchased from Santa Cruz Biotechnology. patient prognosis was examined. GSE14333 showed that elevated expression of BMP4 was associated with poor prognosis in Immunoblotting patients with stage II colorectal cancer (Fig. 1C). Furthermore, Immunoblotting was performed as previously described (7). multivariate analysis demonstrated that BMP4 expression was an Antibodies are described in Supplementary Materials and Meth- independent prognostic factor in stage II colorectal cancer (Sup- ods. ImageJ (NIH) was used to quantify blot band intensities. plementary Table S2). Expression of BMP-4 and activation of Smad-dependent BMP signaling was then examined using human Apoptosis assay colorectal cancer tissues and cells. IHC analysis revealed that Terminal deoxynucleotidyl transferase-mediated dUTP expression levels of BMP-4 and phosphorylated Smad1/5 were nick end labeling (TUNEL) assay was performed as previously upregulated in colorectal cancer tissues compared with those in described (7). Fluorescence was examined using a Leica corresponding normal tissues (Fig. 1D). ELISAs demonstrated DMI6000 B. that these colorectal cancer cells produced BMP-4, whereas, with siRNA the exception of SUIT-2 pancreatic cancer cells, other cancer cells fi Stealth RNAi Pre-Designed siRNAs targeting CTNNB1, BMP4 examined did not (Fig. 1E). These ndings suggest that colorectal and DUSP5 were synthesized by Thermo Fisher Scientific. Cells cancer cells produce BMP-4, which may act in an autocrine were transfected in the presence of 30 nmol/L siRNA or control manner, and that BMP-4 expression may be related to colorectal siRNA in a 500 mL volume with 3 mL RNAiMAX reagent (Thermo cancer progression. BMP4 Fisher Scientific) per well in a 6-well plate. Next, we sought to clarify the mechanism by which mRNA was elevated in colorectal cancer. Mutations in the APC Lentiviral production and infection gene occur in the early phase of colorectal cancer progression, We used a lentiviral vector system to induce specific gene which in turn increases the stability of b-catenin (15). Because introduction and knockdown as previously described (9, 10). BMP4 expression is reported to be regulated by the Wnt/b-catenin The target sequences for shRNA are described in the Supplemen- pathway (16), the involvement of Wnt/b-catenin in the induction tary Materials and Methods. of BMP4 expression in colorectal cancer cells was assessed. b-Cate- nin protein levels were elevated in colorectal cancer cells but not in RNA-sequence analyses non-colorectal cancer cells, such as pancreatic cancer cells (SUIT- RNA-sequence (RNA-seq) analysis was performed as described 2) and breast cancer cells (MDA-231-D; Fig. 2A). Knockdown of previously (11). Raw and processed data are available at GEO the CTNNB1 gene (encoding b-catenin) in colorectal cancer cells (GSE96914). Gene Ontology analysis was performed using CLC by siRNAs suppressed BMP4 mRNA expression and BMP-4 pro- Genomics Workbench (Qiagen Bioinformatics). tein production, as well as expression of a direct downstream target of the Wnt/b-catenin pathway, AXIN2 (Fig. 2B and C). Subcutaneous xenograft model Similar to colorectal cancer cells, introduction
Load more

Recommended publications
  • Characterization of Pulmonary Arteriovenous Malformations in ACVRL1 Versus ENG Mutation Carriers in Hereditary Hemorrhagic Telangiectasia
    © American College of Medical Genetics and Genomics ORIGINAL RESEARCH ARTICLE Characterization of pulmonary arteriovenous malformations in ACVRL1 versus ENG mutation carriers in hereditary hemorrhagic telangiectasia Weiyi Mu, ScM1, Zachary A. Cordner, MD, PhD2, Kevin Yuqi Wang, MD3, Kate Reed, MPH, ScM4, Gina Robinson, RN5, Sally Mitchell, MD5 and Doris Lin, MD, PhD5 Purpose: Pulmonary arteriovenous malformations (pAVMs) are mutation carriers to have pAVMs (P o 0.001) or multiple lesions major contributors to morbidity and mortality in hereditary (P = 0.03), and to undergo procedural intervention (P = 0.02). hemorrhagic telangiectasia (HHT). Mutations in ENG and ACVRL1 Additionally, pAVMs in ENG carriers were more likely to exhibit underlie the vast majority of clinically diagnosed cases. The aims of bilateral lung involvement and growth over time, although this did this study were to characterize and compare the clinical and not reach statistical significance. The HHT severity score was morphologic features of pAVMs between these two genotype significantly higher in ENG than in ACVRL1 (P = 0.02). groups. Conclusion: The propensity and multiplicity of ENG-associated Methods: Sixty-six patients with HHT and affected family pAVMs may contribute to the higher disease severity in this members were included. Genotype, phenotypic data, and imaging genotype, as reflected by the HHT severity score and the frequency were obtained from medical records. Morphologic features of of interventional procedures. pAVMs were analyzed using computed tomography angiography. Genet Med HHT symptoms, pAVM imaging characteristics, frequency of advance online publication 19 October 2017 procedural intervention, and HHT severity scores were compared Key Words: ACVRL1; ENG; genotype-phenotype correlation; between ENG and ACVRL1 genotype groups.
    [Show full text]
  • Hereditary Hemorrhagic Telangiectasia: Diagnosis and Management From
    REVIEW ARTICLE Hereditary hemorrhagic telangiectasia: Ferrata Storti diagnosis and management from Foundation the hematologist’s perspective Athena Kritharis,1 Hanny Al-Samkari2 and David J Kuter2 1Division of Blood Disorders, Rutgers Cancer Institute of New Jersey, New Brunswick, NJ and 2Hematology Division, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA ABSTRACT Haematologica 2018 Volume 103(9):1433-1443 ereditary hemorrhagic telangiectasia (HHT), also known as Osler- Weber-Rendu syndrome, is an autosomal dominant disorder that Hcauses abnormal blood vessel formation. The diagnosis of hered- itary hemorrhagic telangiectasia is clinical, based on the Curaçao criteria. Genetic mutations that have been identified include ENG, ACVRL1/ALK1, and MADH4/SMAD4, among others. Patients with HHT may have telangiectasias and arteriovenous malformations in various organs and suffer from many complications including bleeding, anemia, iron deficiency, and high-output heart failure. Families with the same mutation exhibit considerable phenotypic variation. Optimal treatment is best delivered via a multidisciplinary approach with appropriate diag- nosis, screening and local and/or systemic management of lesions. Antiangiogenic agents such as bevacizumab have emerged as a promis- ing systemic therapy in reducing bleeding complications but are not cur- ative. Other pharmacological agents include iron supplementation, antifibrinolytics and hormonal treatment. This review discusses the biol- ogy of HHT, management issues that face
    [Show full text]
  • Sequence Variations of ACVRL1 Play a Critical Role in Hepatic Vascular
    Giraud et al. Orphanet Journal of Rare Diseases (2020) 15:254 https://doi.org/10.1186/s13023-020-01533-2 RESEARCH Open Access Sequence variations of ACVRL1 play a critical role in hepatic vascular malformations in hereditary hemorrhagic telangiectasia Sophie Giraud1, Claire Bardel2,3,4, Sophie Dupuis-Girod1,5, Marie-France Carette6, Brigitte Gilbert-Dussardier7,8, Sophie Riviere9, Jean-Christophe Saurin3,10, Mélanie Eyries11, Sylvie Patri12, Evelyne Decullier13, Alain Calender1,3,14 and Gaëtan Lesca1,3* Abstract Background: Hereditary Hemorrhagic Telangiectasia (HHT) is an autosomal dominant disorder characterized by multiple telangiectases and caused by germline disease-causing variants in the ENG (HHT1), ACVRL1 (HHT2) and, to a lesser extent MADH4 and GDF2, which encode proteins involved in the TGF-β/BMP9 signaling pathway. Common visceral complications of HHT are caused by pulmonary, cerebral, or hepatic arteriovenous malformations (HAVMs). There is large intrafamilial variability in the severity of visceral involvement, suggesting a role for modifier genes. The objective of the present study was to investigate the potential role of ENG, ACVRL1, and of other candidate genes belonging to the same biological pathway in the development of HAVMs. Methods: We selected 354 patients from the French HHT patient database who had one disease causing variant in either ENG or ACVRL1 and who underwent hepatic exploration. We first compared the distribution of the different types of variants with the occurrence of HAVMs. Then, we genotyped 51 Tag-SNPs from the Hap Map database located in 8 genes that encode proteins belonging to the TGF-β/BMP9 pathway (ACVRL1, ENG, GDF2, MADH4, SMAD1, SMAD5, TGFB1, TGFBR1), as well as in two additional candidate genes (PTPN14 and ADAM17).
    [Show full text]
  • Activin Receptor Type IA (ACVR1) Antibody (Center N153) Purified Rabbit Polyclonal Antibody (Pab) Catalog # AP7101A
    10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 Activin Receptor Type IA (ACVR1) Antibody (Center N153) Purified Rabbit Polyclonal Antibody (Pab) Catalog # AP7101A Specification Activin Receptor Type IA (ACVR1) Antibody (Center N153) - Product Information Application WB, IHC-P,E Primary Accession Q04771 Other Accession P80201, P37172, Q28041 Reactivity Human Predicted Bovine, Mouse, Rat Host Rabbit Clonality Polyclonal Isotype Rabbit Ig Antigen Region 138-170 Activin Receptor Type IA (ACVR1) Antibody Western blot analysis of ACVR1 (arrow) using (Center N153) - Additional Information rabbit polyclonal ACVR1 Antibody (Center N153) (Cat.#AP7101a). 293 cell lysates (2 Gene ID 90 ug/lane) either nontransfected (Lane 1) or transiently transfected with the ACVR1 gene Other Names (Lane 2) (Origene Technologies). Activin receptor type-1, Activin receptor type I, ACTR-I, Activin receptor-like kinase 2, ALK-2, Serine/threonine-protein kinase receptor R1, SKR1, TGF-B superfamily receptor type I, TSR-I, ACVR1, ACVRLK2 Target/Specificity This Activin Receptor Type IA (ACVR1) antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 138-170 amino acids from the Central region of human Activin Receptor Type IA (ACVR1). Dilution WB~~1:1000 ACVR1 Antibody (Cat. #AP7101a) western IHC-P~~1:50~100 blot analysis in HL-60 cell line lysates (35ug/lane).This demonstrates the ACVR1 Format antibody detected the ACVR1 protein (arrow). Purified polyclonal antibody supplied in PBS with 0.09% (W/V) sodium azide. This antibody is prepared by Saturated Ammonium Sulfate (SAS) precipitation followed by dialysis against PBS. Storage Page 1/3 10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 Maintain refrigerated at 2-8°C for up to 2 weeks.
    [Show full text]
  • ALK2 / ACVR1 Antibody (Internal) Goat Polyclonal Antibody Catalog # ALS12616
    10320 Camino Santa Fe, Suite G San Diego, CA 92121 Tel: 858.875.1900 Fax: 858.622.0609 ALK2 / ACVR1 Antibody (Internal) Goat Polyclonal Antibody Catalog # ALS12616 Specification ALK2 / ACVR1 Antibody (Internal) - Product Information Application WB Primary Accession Q04771 Reactivity Human, Mouse, Rat, Rabbit, Monkey, Pig, Horse, Bovine, Dog Host Goat Clonality Polyclonal Calculated MW 57kDa KDa ALK2 / ACVR1 Antibody (Internal) - Additional Antibody (0.3 ug/ml) staining of Human Information Umbilical Cord lysate (35 ug protein in RIPA buffer). Gene ID 90 Other Names ALK2 / ACVR1 Antibody (Internal) - Activin receptor type-1, 2.7.11.30, Activin Background receptor type I, ACTR-I, Activin receptor-like kinase 2, ALK-2, Serine/threonine-protein On ligand binding, forms a receptor complex kinase receptor R1, SKR1, TGF-B consisting of two type II and two type I superfamily receptor type I, TSR-I, ACVR1, transmembrane serine/threonine kinases. Type ACVRLK2 II receptors phosphorylate and activate type I receptors which autophosphorylate, then bind Target/Specificity and activate SMAD transcriptional regulators. Human ACVR1 / ALK2. Receptor for activin. May be involved for left-right pattern formation during Reconstitution & Storage embryogenesis (By similarity). Store at -20°C. Minimize freezing and thawing. ALK2 / ACVR1 Antibody (Internal) - References Precautions ALK2 / ACVR1 Antibody (Internal) is for Matsuzaki K.,et al.J. Biol. Chem. research use only and not for use in diagnostic or therapeutic procedures. 268:12719-12723(1993). ten Dijke P.,et al.Oncogene 8:2879-2887(1993). Oppermann F.S.,et al.Mol. Cell. Proteomics ALK2 / ACVR1 Antibody (Internal) - Protein Information 8:1751-1764(2009). Umasankar P.K.,et al.Nat.
    [Show full text]
  • Context-Dependent Roles in Cell and Tissue Physiology
    Downloaded from http://cshperspectives.cshlp.org/ on September 24, 2021 - Published by Cold Spring Harbor Laboratory Press TGF-b and the TGF-b Family: Context-Dependent Roles in Cell and Tissue Physiology Masato Morikawa,1 Rik Derynck,2 and Kohei Miyazono3 1Ludwig Cancer Research, Science for Life Laboratory, Uppsala University, Biomedical Center, SE-751 24 Uppsala, Sweden 2Department of Cell and Tissue Biology, University of California at San Francisco, San Francisco, California 94143 3Department of Molecular Pathology, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan Correspondence: [email protected] The transforming growth factor-b (TGF-b) is the prototype of the TGF-b family of growth and differentiation factors, which is encoded by 33 genes in mammals and comprises homo- and heterodimers. This review introduces the reader to the TGF-b family with its complexity of names and biological activities. It also introduces TGF-b as the best-studied factor among the TGF-b family proteins, with its diversity of roles in the control of cell proliferation and differentiation, wound healing and immune system, and its key roles in pathology, for exam- ple, skeletal diseases, fibrosis, and cancer. lthough initially thought to stimulate cell TGF-b has been well documented in most cell Aproliferation, just like many growth factors, types, and has been best characterized in epithe- it became rapidly accepted that transforming lial cells. The bifunctional and context-depen- growth factor b (TGF-b) is a bifunctional reg- dent nature of TGF-b activities was further con- ulator that either inhibits or stimulates cell pro- firmed in a large variety of cell systems and liferation.
    [Show full text]
  • Primary Chicken and Duck Endothelial Cells Display a Differential Response to Infection with Highly Pathogenic Avian Influenza Virus
    G C A T T A C G G C A T genes Article Primary Chicken and Duck Endothelial Cells Display a Differential Response to Infection with Highly Pathogenic Avian Influenza Virus Zhen Wei Marcus Tong 1,†, Anjana C. Karawita 1,2,† , Colin Kern 3, Huaijun Zhou 3 , Jane E. Sinclair 1 , Limin Yan 1, Keng Yih Chew 1, Sue Lowther 2, Lee Trinidad 2, Arjun Challagulla 2 , Karel A. Schat 4 , Michelle L. Baker 2 and Kirsty R. Short 1,5,* 1 School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane 4072, Australia; [email protected] (Z.W.M.T.); [email protected] (A.C.K.); [email protected] (J.E.S.); [email protected] (L.Y.); [email protected] (K.Y.C.) 2 CSIRO, Australian Centre for Disease Preparedness, Health, and Biosecurity Business Unit, Geelong 3219, Australia; [email protected] (S.L.); [email protected] (L.T.); [email protected] (A.C.); [email protected] (M.L.B.) 3 Department of Animal Science, University of California, Davis, CA 95616, USA; [email protected] (C.K.); [email protected] (H.Z.) 4 Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA; [email protected] 5 Australian Infectious Diseases Research Centre, The University of Queensland, Brisbane 4072, Australia * Correspondence: [email protected] † These authors contributed equally to this work. Citation: Tong, Z.W.M.; Karawita, A.C.; Kern, C.; Zhou, H.; Sinclair, J.E.; Abstract: Highly pathogenic avian influenza viruses (HPAIVs) in gallinaceous poultry are associated Yan, L.; Chew, K.Y.; Lowther, S.; Trinidad, L.; Challagulla, A.; et al.
    [Show full text]
  • Tgfβ/BMP Signaling Pathway in Cartilage Homeostasis
    cells Review TGFβ/BMP Signaling Pathway in Cartilage Homeostasis Nathalie G.M. Thielen , Peter M. van der Kraan and Arjan P.M. van Caam * Experimental Rheumatology, Radboud University Medical Center, Geert Grooteplein 28, 6525 GA Nijmegen, The Netherlands * Correspondence: [email protected]; Tel.: +31-24-10513; Fax: +31-24-3540403 Received: 2 July 2019; Accepted: 19 August 2019; Published: 24 August 2019 Abstract: Cartilage homeostasis is governed by articular chondrocytes via their ability to modulate extracellular matrix production and degradation. In turn, chondrocyte activity is regulated by growth factors such as those of the transforming growth factor β (TGFβ) family. Members of this family include the TGFβs, bone morphogenetic proteins (BMPs), and growth and differentiation factors (GDFs). Signaling by this protein family uniquely activates SMAD-dependent signaling and transcription but also activates SMAD-independent signaling via MAPKs such as ERK and TAK1. This review will address the pivotal role of the TGFβ family in cartilage biology by listing several TGFβ family members and describing their signaling and importance for cartilage maintenance. In addition, it is discussed how (pathological) processes such as aging, mechanical stress, and inflammation contribute to altered TGFβ family signaling, leading to disturbed cartilage metabolism and disease. Keywords: transforming growth factor β; bone morphogenetic proteins; osteoarthritis; cartilage; SMADs; aging; joint loading; inflammation; linker modifications 1. Introduction The transforming growth factor β (TGFβ) family of polypeptide growth factors controls development and homeostasis of many tissues, including articular cartilage. Articular cartilage is the connective tissue covering joint surfaces and is a type of hyaline cartilage. This tissue is key in facilitating movement with its smooth lubricated surface, and it functions as a shock absorber to disperse forces acting upon movement with its physical properties.
    [Show full text]
  • Signalling Pathways Regulating Organ-Specific Endothelial
    REVIEWS The quiescent endothelium: signalling pathways regulating organ- specific endothelial normalcy Nicolas Ricard 1, Sabine Bailly2, Christophe Guignabert 3,4 and Michael Simons 1,5 ✉ Abstract | Endothelial cells are at the interface between circulating blood and tissues. This position confers on them a crucial role in controlling oxygen and nutrient exchange and cellular trafficking between blood and the perfused organs. The endothelium adopts a structure that is specific to the needs and function of each tissue and organ and is subject to tissue-specific signalling input. In adults, endothelial cells are quiescent, meaning that they are not proliferating. Quiescence was considered to be a state in which endothelial cells are not stimulated but are instead slumbering and awaiting activating signals. However, new evidence shows that quiescent endothelium is fully awake, that it constantly receives and initiates functionally important signalling inputs and that this state is actively regulated. Signalling pathways involved in the maintenance of functionally quiescent endothelia are starting to be identified and are a combination of endocrine, autocrine, paracrine and mechanical inputs. The paracrine pathways confer a microenvironment on the endothelial cells that is specific to the perfused organs and tissues. In this Review, we present the current knowledge of organ-specific signalling pathways involved in the maintenance of endothelial quiescence and the pathologies associated with their disruption. Linking organ- specific pathways
    [Show full text]
  • TGF-Β Prodomain Alignments Reveal Unexpected Cysteine Conservation Consistent with Phylogenetic Predictions of Cross-Subfamily
    | INVESTIGATION TGF-b Prodomain Alignments Reveal Unexpected Cysteine Conservation Consistent with Phylogenetic Predictions of Cross-Subfamily Heterodimerization Robert G. Wisotzkey* and Stuart J. Newfeld†,1 *Sema4 Genomics, Stamford, Connecticut 06902 and †School of Life Sciences, Arizona State University, Tempe, Arizona 85287-4501 ORCID ID: 0000-0003-1400-7978 (S.J.N.) ABSTRACT Evolutionary relationships between prodomains in the TGF-b family have gone unanalyzed due to a perceived lack of conservation. We developed a novel approach, identified these relationships, and suggest hypotheses for new regulatory mechanisms in TGF-b signaling. First, a quantitative analysis placed each family member from flies, mice, and nematodes into the Activin, BMP, or TGF-b subfamily. Second, we defined the prodomain and ligand via the consensus cleavage site. Third, we generated alignments and trees from the prodomain, ligand, and full-length sequences independently for each subfamily. Prodomain alignments revealed that six structural features of 17 are well conserved: three in the straitjacket and three in the arm. Alignments also revealed unexpected cysteine conservation in the “LTBP-Association region” upstream of the straitjacket and in b8 of the bowtie in 14 proteins from all three subfamilies. In prodomain trees, eight clusters across all three subfamilies were present that were not seen in the ligand or full- length trees, suggesting prodomain-mediated cross-subfamily heterodimerization. Consistency between cysteine conservation and prodomain clustering provides support for heterodimerization predictions. Overall, our analysis suggests that cross-subfamily interac- tions are more common than currently appreciated and our predictions generate numerous testable hypotheses about TGF-b function and evolution. KEYWORDS Activin; alignments/trees; arm/bowtie/straitjacket; BMP; cleavage site; heterodimer ECRETED TGF-b family members perform a myriad of to cell surface receptors.
    [Show full text]
  • Supplementary Data
    Gene Expressional Differences in MCF-7 vs. MCF-10A cells, Venn Groups e k g b Smad target genes (48) l o n i a h m j d Transcription Factors, And Genes Regulating Developmental Processes (44) f c Adhesion and TGF Superfamily Extracellular Molecules Cytokines (27) (44) Minimum Fold expression values ->640, -320, -160, -80, -40, -20, -10, -5, -2.5, 0, 2.5, 5, 10, 20, 40, 80, 160, 320, 640< + Supplemental Fig. 1 Supplemental Table 1. Change of Basal Gene Expressional values in MCF-7 as compared to MCF-10A cell line. Fold T-test Gene # GeneBank Symbol Up/Down p-Value Description /Position Regulation Venn Group a 01 /A01 NM_001105 ACVR1 -4.96 0.0000 Activin A receptor, type I 02 /A02 NM_001616 ACVR2A -2.62 0.0007 Activin A receptor, type IIA 03 /A03 NM_000020 ACVRL1 -1.25 0.7013 Activin A receptor type II-like 1 05 /A05 NM_020547 AMHR2 1.08 0.8043 Anti-Mullerian hormone receptor, type II 16 /B04 NM_004329 BMPR1A -2.40 0.0011 Bone morphogenetic protein receptor, type IA 17 /B05 NM_001203 BMPR1B -7.37 0.0000 Bone morphogenetic protein receptor, type IB 36 /C12 NM_000557 GDF5 -1.38 0.4911 Growth differentiation factor 5 (cartilage-derived morphogenetic protein-1) 37 /D01 NM_001001557 GDF6 1.12 0.9002 Growth differentiation factor 6 38 /D02 NM_182828 GDF7 -1.52 0.4995 Growth differentiation factor 7 53 /E05 NM_020997 LEFTY1 -1.76 0.0529 Left-right determination factor 1 59 /E11 NM_018055 NODAL -3.62 0.1290 Nodal homolog (mouse) 77 /G05 NM_003238 TGFB2 -4.52 0.0566 Transforming growth factor, beta 2 78 /G06 NM_003239 TGFB3 -1.12 0.2902 Transforming
    [Show full text]
  • CER1 Is a Common Target of WNT and NODAL Signaling Pathways in Human Embryonic Stem Cells
    795-799 24/3/06 13:04 Page 795 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 17: 795-799, 2006 795 CER1 is a common target of WNT and NODAL signaling pathways in human embryonic stem cells 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 January 3, 2006; Accepted February 7, 2006 Abstract. Nodal and BMP signaling pathways network with embryogenesis, underwent protein evolution as well as WNT signaling pathway during embryogenesis and carcino- promoter evolution. These facts indicate that molecular genesis. CER1 (Cerberus 1) and GREM3 (CKTSF1B3 or evolution of CER1 orthologs contributes to the significantly CER2) inhibit NODAL signaling through ACVR1B (ALK4) divergent scenarios of early embryogenesis in primates and or ACVR1C (ALK7) to SMAD2 or SMAD3. GREM1 rodents. (CKTSF1B1) inhibits BMP signaling through BMPR1A (ALK3), BMPR1B (ALK6) or ACVR1 (ALK2) to SMAD1, Introduction SMAD5 or SMAD8. CER1, GREM1 and GREM3 are DAN domain (DAND) family members; however, transcriptional TGFB1, TGFB2, TGFB3, NODAL, LEFTY1, LEFTY2, INHA, regulation of DAND family members by canonical WNT INHBA, INHBB, INHBC, INHBE, AMH, BMP2, BMP3, signaling pathway remains unclear. We searched for the BMP4, BMP5, BMP6, BMP7, BMP8A, BMP8B, BMP10, TCF/LEF-binding site within the promoter region of DAND BMP15, GDF1, GDF2, GDF3, GDF5, GDF6, GDF7, GDF8, family genes, including CER1, GREM1, GREM2, GREM3 and GDF9, GDF10, GDF11, and GDF15 are TGFß superfamily NBL1. Because triple TCF/LEF-binding sites were identified genes within the human genome (http://www.gene.ucl.ac.uk). within human CER1 promoter by using bioinformatics and TGFß signals are transduced through type I receptor TGFBR1 human intelligence, comparative genomics analyses on CER1 and type II receptor TGFBR2 to phosphorylate R-SMAD orthologs were further performed.
    [Show full text]