Development of adenovirus immobilization strategies for <i>in situ</i> gene therapy

Hu, Wei Wen; Lang, M.; Krebsbach, Paul H.

doi:10.1002/jgm.1233

Cited by 23 publications

(41 citation statements)

References 36 publications

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“…Our findings extend previous work showing biomaterialmediated gene delivery using viral vectors (20,(40)(41)(42)(43)(44)(45)(46)(47)(48)(49). The majority of this work demonstrated the ability to specify the spatial location of delivered vectors, rather than the induction of tissuespecific differentiation.…”

Section: Discussionsupporting

confidence: 86%

“…An important feature of the current study includes the use of self-inactivating lentiviral vectors in place of γ-retrovirus (43), nonintegrating viral vectors (16,(40)(41)(42)55), or naked plasmid DNA (15). Although considerable progress has been made with regard to spatial control of gene delivery to cells, transient gene expression and poor transfection/transduction efficiencies may not allow sufficient control of cell phenotype and tissue development.…”

Section: Discussionmentioning

confidence: 99%

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Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Brunger

Huynh

Guenther

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

117

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The ability to develop tissue constructs with matrix composition and biomechanical properties that promote rapid tissue repair or regeneration remains an enduring challenge in musculoskeletal engineering. Current approaches require extensive cell manipulation ex vivo, using exogenous growth factors to drive tissue-specific differentiation, matrix accumulation, and mechanical properties, thus limiting their potential clinical utility. The ability to induce and maintain differentiation of stem cells in situ could bypass these steps and enhance the success of engineering approaches for tissue regeneration. The goal of this study was to generate a self-contained bioactive scaffold capable of mediating stem cell differentiation and formation of a cartilaginous extracellular matrix (ECM) using a lentivirus-based method. We first showed that poly-L-lysine could immobilize lentivirus to poly(e-caprolactone) films and facilitate human mesenchymal stem cell (hMSC) transduction. We then demonstrated that scaffoldmediated gene delivery of transforming growth factor β3 (TGF-β3), using a 3D woven poly(e-caprolactone) scaffold, induced robust cartilaginous ECM formation by hMSCs. Chondrogenesis induced by scaffold-mediated gene delivery was as effective as traditional differentiation protocols involving medium supplementation with TGF-β3, as assessed by gene expression, biochemical, and biomechanical analyses. Using lentiviral vectors immobilized on a biomechanically functional scaffold, we have developed a system to achieve sustained transgene expression and ECM formation by hMSCs. This method opens new avenues in the development of bioactive implants that circumvent the need for ex vivo tissue generation by enabling the long-term goal of in situ tissue engineering.biomaterials | regenerative medicine | genetic engineering | gene therapy | chondrocyte

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Section: Discussionsupporting

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Brunger

Huynh

Guenther

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

117

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“…Surface tethering of gene vectors on implantable biomaterials (also termed as substrate-mediated gene delivery or solid phase gene delivery) has been implemented in cell culture and animal experiments using both specific (antigen-antibody [18][19][20] , avidin-biotin 21,22 ) and non-specific [23][24][25][26] (charge, van der Waals) interactions. The covalent attachment of vectors to the surface of the implanted device has been previously considered as non-functional since excessively strong bonds with the surface preclude vector internalization by target cells.…”

Section: Introductionmentioning

confidence: 99%

Vascular Gene Transfer from Metallic Stent Surfaces Using Adenoviral Vectors Tethered through Hydrolysable Cross-linkers

Fishbein

Forbes

Adamo

et al. 2014

JoVE

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In-stent restenosis presents a major complication of stent-based revascularization procedures widely used to re-establish blood flow through critically narrowed segments of coronary and peripheral arteries. Endovascular stents capable of tunable release of genes with anti-restenotic activity may present an alternative strategy to presently used drug-eluting stents. In order to attain clinical translation, gene-eluting stents must exhibit predictable kinetics of stent-immobilized gene vector release and site-specific transduction of vasculature, while avoiding an excessive inflammatory response typically associated with the polymer coatings used for physical entrapment of the vector. This paper describes a detailed methodology for coatless tethering of adenoviral gene vectors to stents based on a reversible binding of the adenoviral particles to polyallylamine bisphosphonate (PABT)-modified stainless steel surface via hydrolysable cross-linkers (HC). A family of bifunctional (amine- and thiol-reactive) HC with an average t1/2 of the in-chain ester hydrolysis ranging between 5 and 50 days were used to link the vector with the stent. The vector immobilization procedure is typically carried out within 9 hr and consists of several steps: 1) incubation of the metal samples in an aqueous solution of PABT (4 hr); 2) deprotection of thiol groups installed in PABT with tris(2-carboxyethyl) phosphine (20 min); 3) expansion of thiol reactive capacity of the metal surface by reacting the samples with polyethyleneimine derivatized with pyridyldithio (PDT) groups (2 hr); 4) conversion of PDT groups to thiols with dithiothreitol (10 min); 5) modification of adenoviruses with HC (1 hr); 6) purification of modified adenoviral particles by size-exclusion column chromatography (15 min) and 7) immobilization of thiol-reactive adenoviral particles on the thiolated steel surface (1 hr). This technique has wide potential applicability beyond stents, by facilitating surface engineering of bioprosthetic devices to enhance their biocompatibility through the substrate-mediated gene delivery to the cells interfacing the implanted foreign material.

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“…Viral capsid proteins have also been modified to facilitate conjugation to material surfaces. Amine groups on chitosan surfaces were used for bioconjugation to bind virus via avidin-biotin [12] and antibody-antigen interactions [13]. Viral surfaces were covalently modified by biotin or digoxigenin while the infectivity was preserved.…”

Section: Introductionmentioning

confidence: 99%

“…Viral surfaces were covalently modified by biotin or digoxigenin while the infectivity was preserved. Compared to randomly conjugating avidin to biomaterials, immobilization of avidin onto previously biotinylated materials increases avidin orientation and therefore may result in enhanced binding efficiency [12]. The potential shortcoming of these methods, however, is that they may be limited to materials having inherent functional groups on the surface.…”

Section: Introductionmentioning

confidence: 99%

The effects of Runx2 immobilization on poly (ɛ-caprolactone) on osteoblast differentiation of bone marrow stromal cells in vitro

et al. 2010

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In vivo regenerative gene therapy is a promising approach for bone regeneration and can help to address cell-source limitations through surgical implantation of osteoinductive materials and subsequent recruitment of host-derived cells. Localized viral delivery may reduce risk of virus dispersion, enhance transduction efficiency, and reduce administration/injection dosing, which subsequently increases patient safety. In this manuscript, we present a custom-tailored strategy to immobilize adenovirus expressing runt-related transcription factor 2 (AdRunx2) by using reactive polymer coatings to enhance in vitro osteoblast differentiation of bone marrow stromal cells (BMSCs). A thin polymer film of poly[p-xylylene carboxylic acid pentafluorophenol ester-co-pxylylene] equipped with amine-reactive active ester groups was deposited on the surface of poly (ε-caprolactone) (PCL) using the CVD polymerization technique and then anti-adenovirus antibody was conjugated on the material with an amide chemical bond. Following antibody conjugation, AdRunx2 was conjugated to the PCL surface through antibody-antigen interaction. Osteoblast differentiation of BMSCs was induced by incubation in osteogenic medium. Alkaline phosphatase (ALP) activity, calcium deposition, and matrix mineralization were confirmed as markers of osteoblast formation. Incubation of the BMSCs in the presence of AdRunx2 modified PCL resulted

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Development of adenovirus immobilization strategies for in situ gene therapy

Cited by 23 publications

References 36 publications

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Vascular Gene Transfer from Metallic Stent Surfaces Using Adenoviral Vectors Tethered through Hydrolysable Cross-linkers

The effects of Runx2 immobilization on poly (ɛ-caprolactone) on osteoblast differentiation of bone marrow stromal cells in vitro

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