title: Proof-of-concept Studies for siRNA-mediated Gene Silencing for Coagulation Factors in Rat and Rabbit.
abstract: The present study aimed at establishing feasibility of delivering short interfering RNA (siRNA) to target the coagulation cascade in rat and rabbit, two commonly used species for studying thrombosis and hemostasis. siRNAs that produced over 90% mRNA knockdown of rat plasma prekallikrein and rabbit Factor X (FX) were identified from in vitro screens. An ionizable amino lipid based lipid nanoparticle (LNP) formulation for siRNA in vivo delivery was characterized as tolerable and exerting no appreciable effect on coagulability at day 7 postdosing in both species. Both prekallikrein siRNA-LNP and FX siRNA-LNP resulted in dose-dependent and selective knockdown of target gene mRNA in the liver with maximum reduction of over 90% on day 7 following a single dose of siRNA-LNP. Knockdown of plasma prekallikrein was associated with modest clot weight reduction in the rat arteriovenous shunt thrombosis model and no increase in the cuticle bleeding time. Knockdown of FX in the rabbit was accompanied with prolongation in ex vivo clotting times. Results fit the expectations with both targets and demonstrate for the first time, the feasibility of targeting coagulation factors in rat, and, more broadly, targeting a gene of interest in rabbit, via systemic delivery of ionizable LNP formulated siRNA.
authors.affiliation: Thrombosis, Cardiometabolic Diseases, Merck Sharp & Dohme Corp, Kenilworth, New Jersey, USA.
authors.forename: Zhu
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Pharmacology (West Point), Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Bin
authors.initials: B
authors.lastname: Luo
authors.affiliation: Pharmacology (Kenilworth), Merck Sharp & Dohme Corp, Kenilworth, New Jersey, USA.
authors.forename: Tian-Quan
authors.initials: TQ
authors.lastname: Cai
authors.affiliation: Discovery Biology, RNA Therapeutics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Anil
authors.initials: A
authors.lastname: Thankappan
authors.affiliation: Thrombosis, Cardiometabolic Diseases, Merck Sharp & Dohme Corp, Kenilworth, New Jersey, USA.
authors.forename: Yiming
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Thrombosis, Cardiometabolic Diseases, Merck Sharp & Dohme Corp, Kenilworth, New Jersey, USA.
authors.forename: Weizhen
authors.initials: W
authors.lastname: Wu
authors.affiliation: Discovery Biology, RNA Therapeutics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Jillian
authors.initials: J
authors.lastname: DiMuzio
authors.affiliation: Pharmacology (West Point), Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Traci
authors.initials: T
authors.lastname: Lifsted
authors.affiliation: Discovery Biology, RNA Therapeutics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Marty
authors.initials: M
authors.lastname: DiPietro
authors.affiliation: Discovery Biology, RNA Therapeutics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Disa
authors.affiliation: Discovery Biology, RNA Therapeutics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Bruce
authors.initials: B
authors.lastname: Ng
authors.affiliation: Discovery Biology, RNA Therapeutics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Leander
authors.affiliation: Non-Clinical Statistics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Seth
authors.initials: S
authors.lastname: Clark
authors.affiliation: Pharmacology (Kenilworth), Merck Sharp & Dohme Corp, Kenilworth, New Jersey, USA.
authors.forename: Lizbeth
authors.initials: L
authors.lastname: Hoos
authors.affiliation: Pharmacology (Kenilworth), Merck Sharp & Dohme Corp, Kenilworth, New Jersey, USA.
authors.forename: Yuchen
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Pharmacology (Kenilworth), Merck Sharp & Dohme Corp, Kenilworth, New Jersey, USA.
authors.forename: Nina
authors.initials: N
authors.lastname: Jochnowitz
authors.affiliation: Thrombosis, Cardiometabolic Diseases, Merck Sharp & Dohme Corp, Kenilworth, New Jersey, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Jachec
authors.affiliation: Clinical Pathology (West Point), Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Szczerba
authors.affiliation: Pharmaceutical Sciences, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Marian E
authors.initials: ME
authors.lastname: Gindy
authors.affiliation: Discovery Biology, RNA Therapeutics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Walter
authors.initials: W
authors.lastname: Strapps
authors.affiliation: Discovery Biology, RNA Therapeutics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Sepp-Lorenzino
authors.affiliation: Thrombosis, Cardiometabolic Diseases, Merck Sharp & Dohme Corp, Kenilworth, New Jersey, USA.
authors.forename: Dietmar A
authors.initials: DA
authors.lastname: Seiffert
authors.affiliation: Pharmacology (West Point), Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Lubbers
authors.affiliation: Discovery Biology, RNA Therapeutics, Merck Sharp & Dohme Corp, West Point, Pennsylvania, USA.
authors.forename: Marija
authors.initials: M
authors.lastname: Tadin-Strapps