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Principal Investigator: Nicola Pozzi
Organization: SAINT LOUIS UNIVERSITY
Fiscal Year: 2024
Award: $227,250
Funding agency: National Heart Lung and Blood Institute
Project summary
This project aims to investigate a novel approach to mitigate the risk of vascular thromboses, which imposes a
significant economic burden in the US. Our strategy is to develop a technology that, by specifically targeting the
lipid-binding domains of factor X (read factor ten) and prothrombin, two essential coagulation factors, regulates
the rate at which factor Xa and thrombin are produced and, consequently, controls the formation of blood clots.
This approach is unique compared to current pharmacologic methods that either directly (DOACs) or indirectly
(heparins) target the active site of clotting proteases or impair the proper synthesis of a family of proteins to
which multiple coagulation factors belong (warfarin). We propose this approach will offer potential benefits for
patients with prothrombotic antiphospholipid antibodies who experience limited effectiveness with existing
medications. Antiphospholipid antibodies are a defining feature of antiphospholipid syndrome (APS), which is an
acquired autoimmune disorder. Additionally, there is evidence of their involvement in infectious diseases,
including COVID-19. By using an in vitro selection process, our preliminary studies identified a nanobody suitable
for testing this hypothesis, which we called NanoProTEN. Aim 1 of this project aims to investigate the biochemical
characteristics of NanoProTEN, including its affinity and specificity towards coagulation factors and plasma
proteins involved in the blood clotting cascade. We will then assess its anticoagulant potential in human plasma
and with purified coagulation factors, and its ability to interfere with antiphospholipid antibodies. These studies
will define the molecular interactions of NanoProTEN, evaluate its potential off-target effects, and establish its
anticoagulant potential compared to existing therapies. Studies in Aim 2 will define the unique mechanism of
action of NanoProTEN through structural studies using cryo-electron microscopy. By determining the structures
of NanoProTEN bound to prothrombin and factor X, we will uncover the structural elements contributing to its
dual selectivity. These studies will determine how NanoProTEN interacts with these factors, offering mechanistic
insights into its mode of action and providing essential information for enhancing its activity. In conclusion, the
primary outcome of this research project is the development of an innovative technology represented by
NanoProTEN. This technology holds promise as a versatile research tool and a potential candidate for a new
class of anticoagulants for improving outcomes in patients with prothrombotic autoantibodies and potentially
other thrombotic disorders.
Terms: <Activated Blood Coagulation Factor X><Activated Factor X><Active Sites><Affinity><Anti-Phospholipid Antibody Syndrome><Anti-phospholipid Syndrome><Anticoagulant Agents><Anticoagulant Drugs><Anticoagulants><Anticoagulation><Antiphospholipid Antibodies><Autoantibodies><Autoimmune Diseases><Autoprothrombin C><Autoprothrombin III><Binding><Biochemical><Blood Clotting><Blood Coagulation Factor><Blood Coagulation Factor II><Blood Coagulation Factor X><Blood Plasma><Blood Vessels><Blood coagulation><COVID-19><CV-19><Calcium ion><Cell membrane><Characteristics><Charge><Clinical Treatment Moab><Clotting><Coagulation><Coagulation Factor II><Coagulation Factor X><Coagulation Factor Xa><Coagulation Factors><Coagulation Process><Communicable Diseases><Complex><Coronavirus Infectious Disease 2019><Cryo-electron Microscopy><Cryoelectron Microscopy><Cytoplasmic Membrane><Development><Differentiation Reversal Factor><Drugs><Economic Burden><Effectiveness><Electron Cryomicroscopy><Elements><Esteroproteases><Factor II><Factor X><Factor Xa><Familial antiphospholipid syndrome><Generations><Goals><Heparin><Heparinic Acid><Hughes syndrome><Human><Impairment><In Vitro><Individual><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Investigation><Laboratories><Lipid Binding><Mediating><Medication><Methods><Modern Man><Molecular Configuration><Molecular Conformation><Molecular Interaction><Molecular Stereochemistry><Monoclonal Antibodies><Names><Oral><Patients><Peptidases><Peptide Hydrolases><Pharmaceutical Preparations><Pharmacology><Phosphatides><Phospholipids><Plasma><Plasma Membrane><Plasma Proteins><Plasma Serum><Process><Productivity><Property><Protease Gene><Proteases><Protein Family><Proteinases><Proteins><Proteolytic Enzymes><Prothrombin><Prower factor><R-Series Research Projects><R01 Mechanism><R01 Program><Reaction><Reagent><Research><Research Grants><Research Project Grants><Research Projects><Resolution><Reticuloendothelial System, Serum, Plasma><Risk><Series><Specificity><Structure><Stuart Factor><Stuart-Prower Factor><Surface><Technology><Testing><Therapeutic><Thrombase><Thrombin><Thrombokinase><Thrombosis><Warfarin><autoimmune antibody><autoimmune condition><autoimmune disorder><autoimmunity disease><autoreactive antibody><blood thinner><clotting factor><conformation><conformational><conformational state><conformationally><conformations><coronavirus disease 2019><coronavirus disease-19><coronavirus infectious disease-19><cryo-EM><cryoEM><cryogenic electron microscopy><design><designing><developmental><drug/agent><experience><fibrinogenase><high risk><improved><improved outcome><innovative technologies><insight><lipid bound><mAbs><monoclonal Abs><name><named><naming><nano-molar><nanobodies><nanobody><nanomolar><new approaches><novel approaches><novel strategies><novel strategy><pharmacologic><plasmalemma><prevent><preventing><primary outcome><prothrombase><recruit><resolutions><risk mitigation><sdAb><self reactive antibody><single domain antibodies><structural determinants><structural factors><thrombopoiesis inhibitor><thrombotic><thrombotic disease><thrombotic disorder><tool><vascular>