title: Chemically modified RNA induces osteogenesis of stem cells and human tissue explants as well as accelerates bone healing in rats.
abstract: Limitations associated to the use of growth factors represent a major hurdle to musculoskeletal regeneration. On the one hand, they are needed to induce neo-tissue formation for the substitution of a necrotic or missing tissue. On the other hand, these factors are used in supraphysiological concentrations, are short lived and expensive and result in many side effects. Here we develop a gene transfer strategy based on the use of chemically modified mRNA (cmRNA) coding for human bone morphogenetic protein 2 (hBMP-2) that is non-immunogenic and highly stable when compared to unmodified mRNA. Transfected stem cells secrete hBMP-2, show elevated alkaline phosphatase levels and upregulated expression of RunX2, ALP, Osterix, Osteocalcin, Osteopontin and Collagen Type I genes. Mineralization was induced as seen by positive Alizarin red staining. hBMP-2 cmRNA transfected human fat tissue also yielded an osteogenic response in vitro as indicated by expression of hBMP-2, RunX2, ALP and Collagen Type I. Delivering hBMP-2 cmRNA to a femur defect in a rat model results in new bone tissue formation as early as 2 weeks after application of very low doses. Overall, our studies demonstrate the feasibility and therapeutic potential of a new cmRNA-based gene therapy strategy that is safe and efficient. When applied clinically, this approach could overcome BMP-2 growth factor associated limitations in bone regeneration.
authors.affiliation: Institute of Molecular Immunology and Experimental Oncology, Technical University Munich, Ismaninger Str. 22, 81675 Munich, Germany; Ethris GmbH, Semmelweisstr. 3, 82152 Planegg, Germany; Experimental Trauma Surgery, Klinikum rechts der Isar, Technical University Munich, Ismaninger Str. 22, 81675 Munich, Germany. Electronic address: Elizabeth.rosado-balmayor@tum.de.
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Balmayor
authors.affiliation: Ethris GmbH, Semmelweisstr. 3, 82152 Planegg, Germany.
authors.forename: Johannes P
authors.initials: JP
authors.lastname: Geiger
authors.affiliation: Ethris GmbH, Semmelweisstr. 3, 82152 Planegg, Germany.
authors.forename: Manish K
authors.initials: MK
authors.lastname: Aneja
authors.affiliation: Institute of Molecular Immunology and Experimental Oncology, Technical University Munich, Ismaninger Str. 22, 81675 Munich, Germany; Ethris GmbH, Semmelweisstr. 3, 82152 Planegg, Germany.
authors.forename: Taras
authors.initials: T
authors.lastname: Berezhanskyy
authors.affiliation: Ethris GmbH, Semmelweisstr. 3, 82152 Planegg, Germany.
authors.forename: Maximilian
authors.initials: M
authors.lastname: Utzinger
authors.affiliation: Institute of Molecular Immunology and Experimental Oncology, Technical University Munich, Ismaninger Str. 22, 81675 Munich, Germany; Ethris GmbH, Semmelweisstr. 3, 82152 Planegg, Germany.
authors.forename: Olga
authors.initials: O
authors.lastname: Mykhaylyk
authors.affiliation: Ethris GmbH, Semmelweisstr. 3, 82152 Planegg, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Rudolph
authors.affiliation: Institute of Molecular Immunology and Experimental Oncology, Technical University Munich, Ismaninger Str. 22, 81675 Munich, Germany; Ethris GmbH, Semmelweisstr. 3, 82152 Planegg, Germany. Electronic address: plank@ethris.com.
authors.forename: Christian
authors.initials: C
authors.lastname: Plank
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000378:metabolism; D001861:Bone Regeneration; D002478:Cells, Cultured; D005269:Femur / Q000293:injuries* / Q000378:metabolism / Q000473:pathology / Q000502:physiology; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D008297:Male; D010012:Osteogenesis; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000627:therapeutic use*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014162:Transfection
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D012333:RNA, Messenger
keywords: Bone healing; Bone morphogenetic protein; Chemically modified mRNA; Genetically modified fat graft; In vitro osteogenesis; mRNA delivery