title: Therapeutic efficacy in a hemophilia B model using a biosynthetic mRNA liver depot system.
abstract: DNA-based gene therapy has considerable therapeutic potential, but the challenges associated with delivery continue to limit progress. Messenger RNA (mRNA) has the potential to provide for transient production of therapeutic proteins, without the need for nuclear delivery and without the risk of insertional mutagenesis. Here we describe the sustained delivery of therapeutic proteins in vivo in both rodents and non-human primates via nanoparticle-formulated mRNA. Nanoparticles formulated with lipids and lipid-like materials were developed for delivery of two separate mRNA transcripts encoding either human erythropoietin (hEPO) or factor IX (hFIX) protein. Dose-dependent protein production was observed for each mRNA construct. Upon delivery of hEPO mRNA in mice, serum EPO protein levels reached several orders of magnitude (>125 000-fold) over normal physiological values. Further, an increase in hematocrit (Hct) was established, demonstrating that the exogenous mRNA-derived protein maintained normal activity. The capacity of producing EPO in non-human primates via delivery of formulated mRNA was also demonstrated as elevated EPO protein levels were observed over a 72-h time course. Exemplifying the possible broad utility of mRNA drugs, therapeutically relevant amounts of human FIX (hFIX) protein were achieved upon a single intravenous dose of hFIX mRNA-loaded lipid nanoparticles in mice. In addition, therapeutic value was established within a hemophilia B (FIX knockout (KO)) mouse model by demonstrating a marked reduction in Hct loss following injury (incision) to FIX KO mice.
authors.affiliation: Shire Pharmaceuticals, Lexington, MA, USA.
authors.forename: F
authors.initials: F
authors.lastname: DeRosa
authors.affiliation: Shire Pharmaceuticals, Lexington, MA, USA.
authors.forename: B
authors.initials: B
authors.lastname: Guild
authors.affiliation: Shire Pharmaceuticals, Lexington, MA, USA.
authors.forename: S
authors.initials: S
authors.lastname: Karve
authors.affiliation: Shire Pharmaceuticals, Lexington, MA, USA.
authors.forename: L
authors.initials: L
authors.lastname: Smith
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: K
authors.initials: K
authors.lastname: Love
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.|Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: J R
authors.initials: JR
authors.lastname: Dorkin
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.|Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: K J
authors.initials: KJ
authors.lastname: Kauffman
authors.affiliation: Shire Pharmaceuticals, Lexington, MA, USA.
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Biomedical Research Models, Inc., Worcester, MA, USA.
authors.forename: B
authors.initials: B
authors.lastname: Yahalom
authors.affiliation: David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.|Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.|Harvard-MIT Division of Health Sciences & Technology, Cambridge, MA, USA.|Institute of Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA, USA.
authors.forename: D G
authors.initials: DG
authors.lastname: Anderson
authors.affiliation: Shire Pharmaceuticals, Lexington, MA, USA.
authors.forename: M W
authors.initials: MW
authors.lastname: Heartlein
mesh_terms: D000818:Animals; D004921:Erythropoietin / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D005164:Factor IX / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods*; D006400:Hematocrit; D002836:Hemophilia B / Q000628:therapy*; D008099:Liver / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D053758:Nanoparticles / Q000737:chemistry; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D004921:Erythropoietin; D005164:Factor IX