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Principal Investigator: Christian Kastrup
Organization: VERSITI BLOOD HEALTH, INC.
Fiscal Year: 2024
Award: $707,172
Funding agency: National Heart Lung and Blood Institute
SUMMARY
BACKGROUND: The balance between clot formation (coagulation) and degradation (fibrinolysis) is disrupted in
patients with bleeding disorders or severe hemorrhage, resulting in increased bleeding. The extent to which
fibrinolysis contributes to bleeding from the onset of acute bleeding in these contexts is unclear, and this gap in
knowledge limits the development of therapies that inhibit fibrinolysis. Current therapeutics are often not sufficient
for patients with bleeding disorders; for example, antifibrinolytic drugs are widely used to manage acute bleeding
in these patients, but are unsuitable for long-term prophylaxis due to short half-lives that decrease efficacy.
RATIONALE: We hypothesize that fibrinolysis contributes to the incidence and severity of bleeding starting from
the onset of bleeding, and that long-acting RNA agents targeting plasminogen, the zymogen precursor of the
main fibrinolytic enzyme plasmin, can decrease fibrinolysis and help manage bleeding long-term for people
across all classes of bleeding disorders, including diagnosed and undiagnosed disorders. This knowledge will
provide insights into the mechanisms that impact outcomes in traumatic hemorrhage. These RNA agents will be
delivered using lipid nanoparticles (LNPs), a clinically approved delivery platform we have unique expertise in.
SPECIFIC AIMS: We will develop an RNA therapeutic targeting plasminogen, using small interfering RNA
(siRNA)-mediated gene silencing (siPlg). This therapy is long-acting over several weeks per dose and highly
specific for the target protein, which decreases the burden of frequent administration. The specific aims of this
proposal are to (1) Develop approaches for knockdown of plasminogen and inhibition of fibrinolysis in mice, pigs,
and dogs, and characterize the safety of this approach; (2) Determine the contribution of fibrinolysis to blood
loss at the onset and in early stages of hemorrhage in a traumatic injury model in swine; and (3) Investigate the
role of fibrinolysis in bleeding disorders by testing the effect of plasminogen knockdown in mouse and canine
models of hemophilia A, and mouse models of hemophilia B and von Willebrand disease.
INNOVATION: This work has both mechanistic and therapeutic innovation. It will immediately determine the
contribution of fibrinolysis to bleeding in early stages of severe hemorrhage and in bleeding disorders. This will
likely be the first development of RNA-LNP therapies targeting circulating proteins in large animal models of
bleeding. These findings will address a fundamental question in trauma and hemostasis research. Separately, it
will aid the development of improved, long-acting therapies for controlling bleeding in bleeding disorders. These
therapies would be particularly beneficial for women with menorrhagia caused by bleeding disorders.
EXPECTED OUTCOMES: We expect to (1) Develop species-specific siPlg that have minimal adverse health
effects; and (2-3) Demonstrate that knocking down plasminogen inhibits fibrinolysis and decreases blood loss in
swine models of traumatic hemorrhage, and in murine and canine models of bleeding disorders.
Terms: <AMCA><AMCHA><Acute><American><Angiohemophilia><Animal Model><Animal Models and Related Studies><Animals><Antifibrinolysins><Antifibrinolytic Agents><Antifibrinolytics><Antiplasmin><Antithrombotic Agents><Antithrombotic Drugs><Biochemistry><Biological Chemistry><Bleeding><Bleeding Time><Bleeding time procedure><Blood Coagulation Disorders><Canine Species><Canis familiaris><Christmas Disease><Clinical><Clotting><Coagulation><Coagulation Disorder><Coagulation Process><Coagulopathy><DNA Therapy><Data><Development><Diagnosis><Disease><Disorder><Dogs><Dogs Mammals><Dose><EC 3.4.21.7><Enzyme Gene><Enzyme Precursors><Enzymes><Equilibrium><FDA approved><Factor IX Deficiency><Factor VIII Deficiency><Family suidae><Fibrinolyses><Fibrinolysin><Fibrinolysis><Fibrinolytic Agents><Fibrinolytic Drugs><Gene Inactivation><Gene Silencing><Gene Transfer Clinical><Genetic><Genetic Intervention><Glu-Plasmin><Half-Life><Health><Hemophilia><Hemophilia A><Hemophilia B><Hemorrhage><Hemostasis><Hemostatic Agents><Hemostatic function><Hemostatics><Hereditary><Hour><Hypermenorrhea><In Vitro><Incidence><Inherited><Injury><Investigation><Investigators><Knowledge><Measures><Mediating><Menorrhagia><Methods><Mice><Mice Mammals><Modeling><Morbidity><Morbidity - disease rate><Murine><Mus><Nerve><Non-Polyadenylated RNA><Operative Procedures><Operative Surgical Procedures><Outcome><PAI-1><PAI1><PLANH1><Patients><Persons><Pigs><Plasmin><Plasmin Antiactivator><Plasminogen><Plasminogen Activator Inhibitor 1><Predisposition><Process><Proenzymes><Profibrinolysin><Prophylactic treatment><Prophylaxis><Protease F><Proteins><Puncture procedure><Punctures><QOL><Quality of life><RNA><RNA Gene Products><RNA based therapeutics><RNA based therapy><RNA targeting drug><RNA targeting therapeutics><RNA therapy><RNA-targeting therapy><Research><Research Personnel><Researchers><Ribonucleic Acid><Role><Rupture><Safety><Saphenous Vein><Serine or Cysteine Proteinase Inhibitor Clade E Member 1><Severities><Short interfering RNA><Small Interfering RNA><Suidae><Surgical><Surgical Interventions><Surgical Procedure><Susceptibility><Swine><Symptoms><Tail><Testing><Therapeutic><Thrombolytic Agents><Thrombolytic Drugs><Toxic effect><Toxicities><Tranexamic Acid><Trauma><Traumatic Hemorrhage><Traumatic injury><Type 1 Plasminogen Activator Inhibitor><Vascular Hemophilia><Von willebrand factor deficiency><Willebrand disease><Woman><Work><Zymogens><alpha 2-Plasmin Inhibitor><alpha-2-Antiplasmin><antithrombotic medication><antithrombotics><balance><balance function><bleeding disorder><blood loss><canine><canine animal model><canine model><clotting disorder><compare to control><comparison control><develop therapy><developmental><disease model><disorder model><dog model><domestic dog><gene repair therapy><gene therapy><gene-based therapy><genetic therapy><genomic therapy><improved><in silico><in vivo><injuries><innovate><innovation><innovative><insight><intervention development><joint damage><joint injury><joint trauma><knock-down><knockdown><lipid based nanoparticle><lipid nanoparticle><model of animal><mortality><mouse model><murine model><nanoparticle therapy><novel><pig model><piglet model><porcine><porcine model><prevent><preventing><prophylactic><siRNA><small molecular inhibitor><small molecule inhibitor><social role><suid><surgery><swine model><t-AMCHA><targeted agent><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic RNA><therapeutic nanoparticles><therapy development><tool><transcriptional silencing><treatment development><von Willebrand Disease>