Latent TGF-β2 Structure and Activation

NIH Pandemic-Era Grants

Pandemic Era Grants

2024

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Principal Investigator: TIMOTHY A SPRINGER
Organization: BOSTON CHILDREN'S HOSPITAL
Fiscal Year: 2024
Award: $692,334
Funding agency: National Heart Lung and Blood Institute

Abstract. Transforming growth factor β2 (TGF-β2) is critically important for heart and vascular development
and repair. TGF-β2 dysregulation is seen in patient TGF-β2 mutations, systemic sclerosis, and Kawasaki
disease, which have cardiovascular sequelae such as aortic aneurysms and cardiac fibrosis. TGF-β1, 2 and 3
are synthesized as proproteins that dimerize and associate with milieu molecules that regulate TGF-β tissue
localization, such as the transmembrane protein glycoprotein A repetitions predominant (GARP) and latent
TGF-β binding proteins (LTBPs) in the extracellular matrix (ECM). Proconvertases cleave between the
prodomain and growth factor (GF) domain; however, the prodomain dimer remains non-covalently associated
with the GF in a proTGF-β–milieu molecule complex after secretion. ProTGF-β–milieu molecule complexes are
inactive because the prodomains encircle the GF and prevent binding to TGF-β receptors. ProTGF-β1 and 3
activation is mediated by binding of integrins αVβ6 and αVβ8 to an RGD-motif in the prodomain and requires
proTGF-β association with a milieu molecule. How proTGF-β2, which lacks an RGD-motif, is activated remains
a mystery. Aim 1 will define the structure of proTGF-β2 to understand its mechanism of latency. Aim 2 will
determine proTGF-β2/milieu molecule complex structures by X-ray crystallography and cryo-EM to define how
milieu molecules bind and alter TGF-β2 latency. We will generate antibodies to use as crystallization
chaperones in addition to using already developed nanobodies to proTGF-β2. Complementary unfolding
studies will test the hypothesis that milieu molecule binding stabilizes proTGF-β2. Aim 3 characterizes TGF-β2
activation. Follow-up studies will identify cell-lines that natively activate TGF-β2 and characterize the
physiologically relevant process. The results of this grant will enhance our understanding of TGF-β2 latency
and activation in extracellular milieus and lay the foundation for developing therapeutics that target proTGF-β2
and its physiologically relevant complexes with milieu molecules.

Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><Actins><Affinity><Antibodies><Aortic Aneurysm><Arg-Gly-Asp><Arginine-Glycine-Aspartic Acid Cell Adhesion Domain><BSC-1 Cell Growth Inhibitor><Binding><Biochemical><Biological><Biological Function><Biological Process><Blood Vessels><Body Tissues><Bone-Derived Transforming Growth Factor><CIF-B><COVID-19 virus><COVID19 virus><Cardiovascular><Cardiovascular Body System><Cardiovascular Organ System><Cardiovascular Physiology><Cardiovascular system><Cartilage-Inducing Factor-B><Cell Body><Cell Line><Cell surface><Cell-Extracellular Matrix><CellLine><Cells><Cellular Matrix><Chaperone><Chimera Protein><Chimeric Proteins><CoV-2><CoV2><Complex><Cryo-electron Microscopy><Cryoelectron Microscopy><Crystallization><Crystallographies><Crystallography><Cytoplasmic Domain><Cytoplasmic Tail><Cytoskeletal System><Cytoskeleton><D11S833E><Defect><Detection><Development><Dimerization><Disease><Disorder><ECM><Electron Cryomicroscopy><Esteroproteases><Extracellular Matrix><Family><Fibrosis in the heart><Fibrosis in the myocardium><Fibrosis within the heart><Fibrosis within the myocardium><Fibrotic myocardium><Follow-Up Studies><Followup Studies><Foundations><Frog><Fusion Protein><GARP gene><GARP protein><GSC-1GI><Genetic Alteration><Genetic Change><Genetic defect><Genetics-Mutagenesis><Glioblastoma-Derived T-Cell Suppressor Factor><Glycoprotein A Repetitions Predominant><Grant><Growth Agents><Growth Factor><Growth Substances><Heart><Heart Vascular><Human><Integral Membrane Protein><Integrin Binding><Integrins><Integrins Extracellular Matrix><Intrinsic Membrane Protein><Kawasaki Disease><Knowledge><LRRC32><LRRC32 gene><LTBP protein><Length><Leucine-Rich Repeat-Containing Protein 32><Loeys-Dietz Syndrome><Mediating><Membrane><Mice><Mice Mammals><Milk Growth Factor><Modeling><Modern Man><Molecular Chaperones><Molecular Configuration><Molecular Conformation><Molecular Interaction><Molecular Stereochemistry><Mucocutaneous Lymph Node Syndrome><Murine><Mus><Mutagenesis><Mutagenesis Molecular Biology><Mutate><Mutation><N-terminal><NH2-terminal><Patients><Peptidases><Peptide Hydrolases><Peptides><Physiologic><Physiological><Pilot Projects><Platelet Transforming Growth Factor><Polyergin><Preparation><Process><Protease Gene><Proteases><Protein Dimerization><Protein Secretion><Proteinases><Proteins Growth Factors><Proteolytic Enzymes><RGD (sequence)><RGD Cell Adhesion Domain><RGD Domain><RGD Motif><RGD Tripeptide Sequence><RGD peptide><RGD tripeptide><Rana><Reagent><Receptor Protein><Research><Role><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV2><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Scleroderma><Serine Endopeptidases><Serine Protease><Serine Protein Hydrolases><Serine Proteinases><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome related corona virus 2><Shapes><Single Crystal Diffraction><Site><Strains Cell Lines><Structure><Systemic Scleroderma><Systemic Sclerosis><TGF B><TGF-Beta 2><TGF-Beta2><TGF-b2><TGF-beta><TGF-beta Receptors><TGF-β><TGF-β Receptors><TGF-β2><TGFbeta><TGFβ><Testing><Therapeutic><Time><Tissues><Transforming Growth Factor Beta 2><Transforming Growth Factor beta><Transforming Growth Factor beta Receptors><Transforming Growth Factor β Receptors><Transforming Growth Factor-Beta Family Gene><Transmembrane Protein><Transmembrane Protein Gene><Vertebrate 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