Biallelic mutations in LAMA5 disrupts a skeletal noncanonical focal adhesion pathway and produces a distinct bent bone dysplasia
| Autor: | Ralph S. Lachman, Pavel Krejci, Wenjuan Zhang, S. Paige Taylor, Maya Barad, Deborah Krakow, Deborah A. Nickerson, Jennifer Zieba, Ivan Duran, Michael J. Bamshad, Michaela Bosakova, Jessica X. Chong, Daniel H. Cohn, Fabiana Csukasi, Jorge H. Martin |
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| Přispěvatelé: | [Barad,M, Csukasi,F, Martin,JH, Paige Taylor,S, Zieba,J, Cohn,DH, Krakow,D, Duran,I] Department of Orthopaedic Surgery, University of California-Los Angeles, CA, United States. [Csukasi,F, Duran,I] Laboratory of Bioengineering and Tissue Regeneration-LABRET, Department of Cell Biology, Genetics and Physiology, University of Malaga, IBIMA, Málaga,Spain. [Bosakova,M, Krejci,P] Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic. [Bosakova,M, Krejci,P] International Clinical Research Center, St. Anne’s University Hospital, Brno, Czech Republic. [Zhang,W, Cohn,DH] Department of Molecular, Cell and Developmental Biology, University of California-Los Angeles, CA, United States. [Lachman,RS, Krakow,D] International Skeletal Dysplasia Registry, University of California, Los Angeles, United States. [Bamshad,M, Nickerson,D, Chong,JX] University of Washington Center for Mendelian Genomics, University of Washington, Seattle, WA, United States. [Cohn,DH, Krakow,D] Orthopaedic Institute for Children, University of California Los Angeles, Los Angeles, CA, United States. [Krakow,D] Department of Human Genetics, University of California-Los Angeles, CA, United States. [Durán,I] Networking Biomedical Research Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Andalusian Centre for Nanomedicine and Biotechnology-BIONAND, Málaga, Spain., D.K. and D.H.C are supported by the NIH grants R01 AR066124, R01 DE019567, R01 HD070394. Sequencing was provided by the University of Washington Center for Mendelian Genomics (UWCMG) which is funded by the National Human Genome Research Institute (NHGRI) and the National Heart, Lung and Blood Institute (NHLBI) Award 1U54HG006493. P.K was supported by the Ministry of Education, Youth and Sports of the Czech Republic (Grant INTER-ACTION LTAUSA19030), the Agency for Healthcare Research of the Czech Republic (Grant NV18-08-00567), and the Czech Science Foundation (Grant GA19-20123S). |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Phenomena and Processes::Genetic Phenomena::Phenotype [Medical Subject Headings] DNA Mutational Analysis lcsh:Medicine Anatomy::Cells::Connective Tissue Cells::Chondrocytes [Medical Subject Headings] Diseases::Musculoskeletal Diseases::Bone Diseases::Bone Diseases Developmental [Medical Subject Headings] Analytical Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Genetic Association Studies [Medical Subject Headings] Chemicals and Drugs::Amino Acids Peptides and Proteins::Proteins::Glycoproteins::Membrane Glycoproteins::Laminin [Medical Subject Headings] Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings] Extracellular matrix 0302 clinical medicine Laminin 2.1 Biological and endogenous factors Developmental Analytical Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Sequence Analysis::Sequence Analysis DNA::DNA Mutational Analysis [Medical Subject Headings] Aetiology Wnt Signaling Pathway Anatomy::Musculoskeletal System::Skeleton::Bone and Bones [Medical Subject Headings] Phenomena and Processes::Chemical Phenomena::Chemical Processes::Biochemical Processes::Signal Transduction::Wnt Signaling Pathway [Medical Subject Headings] lcsh:R5-920 Displasia fibrosa ósea Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Adhesion [Medical Subject Headings] Wnt signaling pathway Integrina beta1 General Medicine Vinculin Phenomena and Processes::Genetic Phenomena::Genotype::Genetic Predisposition to Disease [Medical Subject Headings] Cell biology Fibrous dysplasia of bone Phenotype src-Family Kinases Laminin alpha 5 030220 oncology & carcinogenesis Skeletal dysplasia Public Health and Health Services Bone Diseases lcsh:Medicine (General) Research Paper Signal Transduction Cell signaling Phenomena and Processes::Genetic Phenomena::Genetic Variation::Mutation [Medical Subject Headings] Clinical Sciences Biology Bent bone Bone and Bones General Biochemistry Genetics and Molecular Biology Focal adhesion 03 medical and health sciences Chondrocytes Skeletal disorder β1 integrin Genetics Cell Adhesion Humans LAMA5 Genetic Predisposition to Disease Phenomena and Processes::Genetic Phenomena::Genetic Structures::Genome::Genome Components::Genes::Alleles [Medical Subject Headings] Alleles Genetic Association Studies Bone Diseases Developmental Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases::Protein-Tyrosine Kinases::src-Family Kinases [Medical Subject Headings] lcsh:R Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Signal Transduction [Medical Subject Headings] Actin cytoskeleton Focal Adhesion Kinase 2 030104 developmental biology Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases::Protein-Tyrosine Kinases::Focal Adhesion Protein-Tyrosine Kinases::Focal Adhesion Kinase 2 [Medical Subject Headings] Musculoskeletal Mutation biology.protein beta 1 integrin Laminin α5 |
| Zdroj: | EBioMedicine, Vol 62, Iss, Pp 103075-(2020) EBioMedicine |
| Popis: | Background Beyond its structural role in the skeleton, the extracellular matrix (ECM), particularly basement membrane proteins, facilitates communication with intracellular signaling pathways and cell to cell interactions to control differentiation, proliferation, migration and survival. Alterations in extracellular proteins cause a number of skeletal disorders, yet the consequences of an abnormal ECM on cellular communication remains less well understood Methods Clinical and radiographic examinations defined the phenotype in this unappreciated bent bone skeletal disorder. Exome analysis identified the genetic alteration, confirmed by Sanger sequencing. Quantitative PCR, western blot analyses, immunohistochemistry, luciferase assay for WNT signaling were employed to determine RNA, proteins levels and localization, and dissect out the underlying cell signaling abnormalities. Migration and wound healing assays examined cell migration properties. Findings This bent bone dysplasia resulted from biallelic mutations in LAMA5, the gene encoding the alpha-5 laminin basement membrane protein. This finding uncovered a mechanism of disease driven by ECM-cell interactions between alpha-5-containing laminins, and integrin-mediated focal adhesion signaling, particularly in cartilage. Loss of LAMA5 altered β1 integrin signaling through the non-canonical kinase PYK2 and the skeletal enriched SRC kinase, FYN. Loss of LAMA5 negatively impacted the actin cytoskeleton, vinculin localization, and WNT signaling. Interpretation This newly described mechanism revealed a LAMA5-β1 Integrin-PYK2-FYN focal adhesion complex that regulates skeletogenesis, impacted WNT signaling and, when dysregulated, produced a distinct skeletal disorder. Funding Supported by NIH awards R01 AR066124, R01 DE019567, R01 HD070394, and U54HG006493, and Czech Republic grants INTER-ACTION LTAUSA19030, V18-08-00567 and GA19-20123S. |
| Databáze: | OpenAIRE |
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