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Principal Investigator: Vladimir R Muzykantov
Organization: UNIVERSITY OF PENNSYLVANIA
Fiscal Year: 2024
Award: $653,281
Funding agency: National Heart Lung and Blood Institute
No anti-thrombotic agent (ATA) is safe and effective in the many patients at a combined risk of
acute thrombosis and bleeding, e.g., in the early post-surgery period. To address this unmet need, we
develop drug delivery systems (DDS) executing two main functions: A) Block access of ATA to off-
target sites, e.g., hemostatic plugs formed after surgery, while B) Optimize pharmacokinetics and
deliver ATA into subsequent thrombi, where ATA is activated by thrombin. ATA fused with single-
chain fragments (scFv) targeted to red blood cells (RBC) bind to these carriers that execute dual
blocking/delivering function. Proof-of-concept is emerging in models of pre-existing and nascent clots
in animals. Here we devise humanized scFv/ATA targeted to human RBC and will test them in a
humanized microfluidic system (HMF), in transgenic (TG) mice expressing humanized target epitopes
on blood cells, and in the perfusion of isolated human lungs. We will pursue three aims. Aim 1. RBC
loading. We will characterize scFv/ATA loading onto RBC: A) Binding (copies/cell, on/off kinetics); B)
Effect on RBC functionality, biocompatibility and biomechanics; and, B) Regulation of distribution of
scFv/ATA between RBC in circulation. We also will characterize biomechanical factors modulating
RBC/ATA delivery and effect on clot dynamics and structure, in particular, impact of RBC
rigidification, caused by either drug loading or by intrinsic pathophysiological changes in patient's
blood. Aim 2. Mechanistic insights. We will interrogate previously unrecognized yet critically
aspects of the RBC/ATA workings, in particular their interaction with vascular endothelium and
transfer of the drug cargo to these and other vascular cells. In this Aim we will use standard mouse in
vivo models, microfluidic model and perfusion of isolated human lungs model. Aim 3. Appraisal of
benefit/risk ratio. We are developing TM mice expressing human RBC determinants in mouse EBC,
in order to study scFv/ATA loaded on "human RBC" in vivo: A) PK/BD, complement activation,
phagocyte uptake and vascular adhesion of RBC/ATA in TG mice; B) Define the time window/extent
of anti-thrombotic effect of human RBC/ATA in models of arterial vs venous thrombosis in TG mice;
B) Affirm the safety of RBC/ATA. We will detect adversities of scFv/ATA including abnormalities of
RBC. To defuse potential issues, if necessary, we will use more benign loading regimen. Together,
these studies will advance mechanistic insights and clinical translation of a novel way to mitigate
thrombosis in currently unprotected patients by providing a new and tractable approach to
understanding thrombus development and a rational approach to deliver cell-directed therapeutics
Terms: <Abnormal Erythrocytes><Abnormal Red Blood Cell><Acquired brain injury><Acute><Address><Adhesions><Adhesiveness><Animal Model><Animal Models and Related Studies><Animals><Antigenic Determinants><Antiinflammatory Effect><Antithrombotic Agents><Antithrombotic Drugs><Benign><Binding><Binding Determinants><Biodistribution><Biological Agent><Biological Products><Biomechanics><Bleeding><Blood><Blood Cells><Blood Coagulation Factor III><Blood Platelets><Blood Reticuloendothelial System><Blood Vessels><Blood erythrocyte><Brain Injuries><CD142 Antigens><COVID-19><CV-19><Cell Body><Cells><Circulation><Clinic><Clinical><Clinical Research><Clinical Study><Clotting><Coagulation><Coagulation Factor III><Coagulation Process><Coagulin><Complement Activation><Complex><Coronavirus Infectious Disease 2019><Coupled><Coupling><Cytolysis><Data><Development><Disease><Disorder><Dose><Drug Delivery><Drug Delivery Systems><Drug Kinetics><Drugs><Embolism><Embolus><Endothelium><Endotoxemia><Epitopes><Erythrocytes><Erythrocytic><Factor III><Fibrinolyses><Fibrinolysis><Fibrinolytic Agents><Fibrinolytic Drugs><Glomerular Procoagulant Activity><Glycoproteins><Goals><Hemorrhage><Hemostatic Agents><Hemostatics><Hour><Human><IV Infusion><Immobilization><Impairment><Inflammation><Inflammatory><Injections><Intravenous infusion procedures><Isolated Perfusion><Isolation Perfusion><Isolation Perfusion Therapy><Kinetics><Lead><Leg><Life><Lung><Lung Respiratory System><Lysis><Marrow erythrocyte><Marrow platelet><Mediating><Mediator><Medical><Medication><Mice><Mice Mammals><Microfluidics><Modeling><Modern Man><Modification><Molecular><Molecular Interaction><Murine><Mus><Operative Procedures><Operative Surgical Procedures><Outcome><Pathologic><Patients><Pb element><Perfusion><Peripheral Blood Cell><Phagocytes><Phagocytic Cell><Pharmaceutical Preparations><Pharmacokinetics><Phlebothrombosis><Physicians><Pilot Projects><Plasminogen Activator><Platelets><Post-Operative><Postoperative><Postoperative Period><Primates><Primates Mammals><Prophylactic treatment><Prophylaxis><Prothrombinase><Red Blood Cells><Red Cell><Regimen><Regional Perfusion><Regulation><Rhesus><Risk><Safety><Site><Source><Structure><Surgical><Surgical Hemostasis><Surgical Interventions><Surgical Procedure><System><Testing><Therapeutic><Thrombase><Thrombin><Thrombocytes><Thrombolytic Agents><Thrombolytic Drugs><Thrombomodulin><Thromboplastin><Thrombosis><Thrombus><Time><Tissue Factor><Tissue Factor Procoagulant><Tissue Thromboplastin><Transgenic Mice><Transgenic Organisms><Trauma><U-PA><U-Plasminogen Activator><Urinary Plasminogen Activator><Urokinase><Urokinase Plasminogen Activator><Urokinase-Type Plasminogen Activator><Urothromboplastin><Vascular Diseases><Vascular Disorder><Vascular Endothelium><Venous Thrombosis><Work><activated Protein C><amebocyte><anti-inflammatory effect><antithrombotic medication><antithrombotics><biocompatibility><biologics><biomaterial compatibility><biomechanical><biomechanical analyses><biomechanical analysis><biomechanical assessment><biomechanical characterization><biomechanical evaluation><biomechanical measurement><biomechanical profiling><biomechanical test><biopharmaceutical><biotherapeutic agent><blood corpuscles><blood loss><blood vessel disorder><brain damage><brain-injured><clinical applicability><clinical application><clinical translation><clinically translatable><complement pathway regulation><coronavirus disease 2019><coronavirus disease-19><coronavirus infectious disease-19><design><designing><developmental><drug/agent><fibrinogenase><heavy metal Pb><heavy metal lead><high risk><in vivo><in vivo Model><insight><interest><intravenous infusion><invention><model of animal><mouse model><murine model><mutant><new approaches><novel><novel approaches><novel strategies><novel strategy><off-target site><orthopedic freezing><pilot study><pre-clinical><preclinical><preservation><prevent><preventing><prophylactic><prototype><pulmonary><risk/benefit ratio><surgery><targeted agent><thromboinflammation><thromboinflammatory><thrombotic><thrombotic disease><thrombotic disorder><transgenic><translational study><uptake><vascular><vascular dysfunction><vasculopathy><µfluidic>