Constitutive and inducible co-expression systems for non-viral osteoinductive gene therapy

Transcriptional Activation 0301 basic medicine Bone Morphogenetic Protein 7 Genetic Vectors Osteocalcin Bone Morphogenetic Protein 2 Diseases of the musculoskeletal system Cell Line Mice 03 medical and health sciences Osteogenesis Animals Humans Bone Morphogenetic Protein 7/genetics Orthopedic surgery 0303 health sciences Osteoblasts Mesenchymal Stem Cells/cytology Osteoblasts/cytology Mesenchymal Stem Cells Genetic Therapy Bone Morphogenetic Protein 2/genetics 3. Good health Genetic Vectors/genetics RC925-935 Osteocalcin/genetics RD701-811
DOI: 10.22203/ecm.v027a13 Publication Date: 2016-11-28T05:27:46Z
ABSTRACT
Tissue regenerative gene therapy requires expression strategies that deliver therapeutic effective amounts of transgenes.As physiological patterns are more complex than high-level a singular gene, we aimed at constitutive or inducible co-expression 2 transgenes simultaneously.Co-expression human bone morphogenetic protein and 7 (BMP2/7) from constitutively expressing doxycycline plasmids was evaluated in vitro C2C12 cells with osteocalcin reporter assays standard for osteogenic differentiation.The systems were additionally tested an vivo pilot ectopic formation after repeated naked DNA injection to murine muscle tissue.Inductor controlled differentiation demonstrated co-expression.Both systems, constitutive, achieved significantly better single factor expression.The potency the dependent on relative cassette topology.In vivo, 6/13 animals (46 % efficacy) days 14 28 hind limb muscles as proven by µCT histological evaluation.In findings devised vector BMP2/7 strategy mediates superior osteoinduction, can be applied inductor fashion its efficiency is results indicate non-viral transfer effectively without application biomaterials, recombinant growth factors, offering promising alternative current treatment potential clinical translation future.
SUPPLEMENTAL MATERIAL
Coming soon ....
REFERENCES (0)
CITATIONS (14)