Simultaneous gene transfer of bone morphogenetic protein (BMP) -2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression

Kawai, Mariko; Bessho, Kazuhisa; Maruyama, Haruhiko; Miyazaki, Jun‐ichi; Yamamoto, Tetsuro

doi:10.1186/1471-2474-7-62

Cited by 55 publications

(47 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since several studies have reported the possible involvement of CK2 and its effect Akt in bone metabolism (Bragdon et al, 2010;Moon et al, 2012), here we evaluated and found the antiosteoclastogenic and anabolic activity of CX-4945; CX-4945 inhibited the RANKL-induced osteoclast differentiation accompanied with suppression of Akt/NFATc1 signaling axis, but it enhanced the BMP-2-induced osteoblast differentiation via the ERK1/2-Smad signaling axis. Since BMP-2 enhances the expression of other BMP genes during osteoblast differentiation (Chen et al, 1997;Kawai et al, 2006), these reports demonstrate that CX-4945 could enhance the BMP-2-induced increases in ALP and BMP-4 mRNA levels (Fig. 3C).…”

Section: Discussionmentioning

confidence: 87%

The Protein Kinase 2 Inhibitor CX-4945 Regulates Osteoclast and Osteoblast Differentiation In Vitro

Son

Moon

Kim

2013

Molecules and Cells

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 87%

The Protein Kinase 2 Inhibitor CX-4945 Regulates Osteoclast and Osteoblast Differentiation In Vitro

Son

Moon

Kim

2013

Molecules and Cells

View full text Add to dashboard Cite

“…Several multi-gene approaches have already been applied for tissue regenerative therapies. In bone regeneration it has been shown that combinatorial gene therapy using multiple bone morphogenetic protein (BMP) genes -especially the combination of BMP2 and BMP7 -leads to higher osteogenic bioactivity compared to single factor expression (Zhu et al, 2004;Kawai et al, 2006). This BMP2/7 co-expression strategy has been shown to lead to the expression of a heterodimeric growth factor with higher bioactivity (Israel et al, 1996) compared to the respective single gene derived homodimeric variants (Kawai et al, 2009).…”

Section: Constitutive and Inducible Co-expression Systems For Non-virmentioning

confidence: 99%

“…An alternative strategy to increase the therapeutic effectiveness is plasmid modification (Tolmachov, 2009;Tolmachov, 2011) and the selection of an optimal therapeutic gene to compensate for the low transfection efficacy of plasmid vectors in vivo. Furthermore, gene combinations are beneficial in experimental models (Zhu et al, 2004;Kawai et al, 2006;Steinert et al, 2009;Wan et al, 2012). Several multi-gene approaches have already been applied for tissue regenerative therapies.…”

Section: Constitutive and Inducible Co-expression Systems For Non-virmentioning

confidence: 99%

“…Gene therapeutics lead to an inherent sustained release of expressed therapeutic genes by host cells in situ, which is more effective due to higher bioactivity of the hostproduced growth factor (Bonadio et al, 1999;Bleiziffer et al, 2007). Plasmid DNA has been successfully applied in direct in vivo gene transfer for wound healing and angiogenesis (Michlits et al, 2007;Mittermayr et al, 2008), cardiac regeneration (Sundararaman et al, 2011) and musculoskeletal regeneration (Grossin et al, 2003;Kawai et al, 2006;Osawa et al, 2009;Osawa et al, 2010). Taking these advantages into account, non-viral vectors, such as plasmid DNA, are considered the most likely candidates for clinical translation of tissue regenerative gene therapies.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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

Feichtinger

Hacobian²,

Hofmann³

et al. 2014

eCM

View full text Add to dashboard Cite

Tissue regenerative gene therapy requires expression strategies that deliver therapeutic effective amounts of transgenes. As physiological expression patterns are more complex than high-level expression of a singular therapeutic gene, we aimed at constitutive or inducible co-expression of 2 transgenes simultaneously.Co-expression of human bone morphogenetic protein 2 and 7 (BMP2/7) from constitutively expressing and doxycycline inducible plasmids was evaluated in vitro in C2C12 cells with osteocalcin reporter gene assays and standard assays for osteogenic differentiation. The constitutive systems were additionally tested in an in vivo pilot for ectopic bone formation after repeated naked DNA injection to murine muscle tissue.Inductor controlled differentiation was demonstrated in vitro for inducible co-expression. Both co-expression systems, inducible and constitutive, achieved significantly better osteogenic differentiation than single factor expression. The potency of the constitutive co-expression systems was dependent on relative expression cassette topology. In vivo, ectopic bone formation was demonstrated in 6/13 animals (46 % bone formation efficacy) at days 14 and 28 in hind limb muscles as proven by in vivo µCT and histological evaluation.In vitro findings demonstrated that the devised single vector BMP2/7 co-expression strategy mediates superior osteoinduction, can be applied in an inductor controlled fashion and that its efficiency is dependent on expression cassette topology. In vivo results indicate that co-expression of BMP2/7 applied by non-viral naked DNA gene transfer effectively mediates bone formation without the application of biomaterials, cells or recombinant growth factors, offering a promising alternative to current treatment strategies with potential for clinical translation in the future.

show abstract

“…An alternative strategy is the delivery of BMP transgenes instead of recombinant growth factors in order to generate a local sustained release of host-cell produced, highly bioactive growth factors [3][4][5][6]. Recent experimental data showed a synergistic effect of BMP2 and BMP7 therapeutic gene co-delivery [7][8][9]. This promises to increase therapeutic efficiency in upcoming clinical studies.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of fibrin-based gene-activated matrices for BMP2/7 plasmid codelivery in a rat nonunion model

Kaipel

Schützenberger

Hofmann

et al. 2014

International Orthopaedics (SICOT)

View full text Add to dashboard Cite

PURPOSE: Treatment of large segmental bone defects still is a challenge in clinical routine.Application of Gene-activated matrices (GAMs) based on fibrin, BMP 2/7 plasmids and nonviral transfection reagents (cationic polymers) could be an innovative treatment strategy to overcome this problem.The aim of this study was to determine the therapeutic efficacy of fibrin GAMs with or without additional transfection reagents for bone morphogenic protein (BMP)2 and BMP7 plasmid co-delivery in a rat femur non-union model. METHODS:In this experimental study a critical size femoral defect was created in 27 rats. At 4 weeks after the surgery animals were separated into 4 groups and underwent a second operation. Fibrin clots containing BMP2 and BMP7 plasmids with and without cationic polymer were implanted into the femoral defect. Fibrin clots containing recombinant human (rh) BMP2 served as positive and clots without supplement as negative controls.RESULTS: At 8 weeks animals which received GAMs containing the cationic polymer and BMP 2/7 plasmids showed decreased bone volume compared to animals treated with GAMs and BMP2/7 only. Application of BMP2 and BMP7 plasmids in fibrin GAMs without cationic polymer lead to variable results. Animals which received rhBMP 2 protein showed increased bone volume and osseous unions were achieved in 2 out of 6 animals.2 CONCLUSIONS: Cationic polymers decrease therapeutic efficiency of fibrin GAM based BMP2/7 plasmid co-delivery in bone regeneration. Non-viral gene transfer of BMP2/7 plasmids needs alternative promoters (e.g. by sonoporation, electroporation) to promise beneficial clinical effects.

show abstract

Simultaneous gene transfer of bone morphogenetic protein (BMP) -2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression

Cited by 55 publications

References 32 publications

The Protein Kinase 2 Inhibitor CX-4945 Regulates Osteoclast and Osteoblast Differentiation In Vitro

The Protein Kinase 2 Inhibitor CX-4945 Regulates Osteoclast and Osteoblast Differentiation In Vitro

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

Evaluation of fibrin-based gene-activated matrices for BMP2/7 plasmid codelivery in a rat nonunion model

Contact Info

Product

Resources

About