Gene therapy based on vector VTVAF17
Produced the gene therapy DNA vectors based on the gene therapy DNA vector for treatment of diseases associated with disorders of osteogenesis, formation and regeneration of bone and cartilage tissues, including, in case of bone fractures, increased brittleness of bones, reduction of bones mineralisation, for improvement in osteoinduction of bone implants. The gene therapy DNA vector contains the coding region of the COL1A1, COL1A2, BMP2 or BMP7 therapeutic genes. Methods of producing or use the gene therapy DNA vector carrying therapeutic genes. Escherichia coli strain SCS110-AF/VTvaf17-COL1A1, SCS110-AF/VTvaf17-COL1A2, SCS110-AF/VTvaf17-BMP2 or SCS110-AF/VTvaf17-BMP7 obtains by the method described above carrying gene therapy DNA vector VTvaf17-COL1A1, VTvaf17-COL1A2, VTvaf17-BMP2 or VTvaf17-BMP7. The method of producing the gene therapy DNA vector carrying COL1A1, or COL1A2, or BMP2, or BMP7 therapeutic gene uses on an industrial scale.
1. A DNA vector selected from the group consisting of (i) DNA vector VTvaf17-COL1A1 that has the nucleotide sequence of SEQ ID NO: 1; (ii) DNA vector VTvaf17-COL1A2 that has the nucleotide sequence of SEQ ID NO: 2; (iii) DNA vector VTvaf17-BMP2 that has the nucleotide sequence of SEQ ID NO: 3; and (iv) DNA vector VTvaf17-BMP7 that has the nucleotide sequence of SEQ ID NO: 4.
2. A gene therapy DNA vector selected from the group consisting of (i) DNA vector VTvaf17-BMP2 that has the nucleotide sequence of SEQ ID NO: 3, and (ii) DNA vector VTvaf17-BMP7 that has the nucleotide sequence of SEQ ID NO: 4.
3. A method of producing a DNA vector, comprising cloning a coding region of COL1A1, COL1A2, BMP2, or BMP7 into gene therapy DNA vector VTvaf17 to produce (i) DNA vector VTvaf17-COL1A1 that has the nucleotide sequence of SEQ ID NO: 1, (ii) DNA vector VTvaf17-COL1A2 that has the nucleotide sequence of SEQ ID NO: 2, (iii) DNA vector VTvaf17-BMP2 that has the nucleotide sequence of SEQ ID NO: 3, or (iv) DNA vector VTvaf17-BMP7 that has the nucleotide sequence of SEQ ID NO: 4, respectively.
4. A method of improving osteoinduction or bone formation in a patient, comprising:
(i) injecting, into a bone in need of osteoinduction or bone formation of the patient, autologous dermal fibroblasts transfected with VTvaf17-BMP2 that has the nucleotide sequence of SEQ ID NO: 3, and/or VTvaf17-BMP7 that has the nucleotide sequence of SEQ ID NO: 4; or
(ii) injecting, into a bone in need of osteoinduction or bone formation of the patient, VTvaf17-BMP2 that has the nucleotide sequence of SEQ ID NO: 3, and/or VTvaf17-BMP7 that has the nucleotide sequence of SEQ ID NO: 4.
5. An Escherichia coli strain selected from the group consisting of (i) SCS110-AF/VTvaf17-COL1A1, deposited at the Russian National Collection of Industrial Microorganisms (VKPM) as accession number B-13165, (ii) SCS110-AF/VTvaf17-COL1A2, deposited at the Russian National Collection of Industrial Microorganisms (VKPM) as accession number B-13164, (iii) SCS110-AF/VTvaf17-BMP2, deposited at the Russian National Collection of Industrial Microorganisms (VKPM) as accession number B-13167, and (iv) SCS110-AF/VTvaf17-BMP7, deposited at the Russian National Collection of Industrial Microorganisms (VKPM) as accession number B-13166.