title: Gene therapy vectors with enhanced transfection based on hydrogels modified with affinity peptides.
abstract: Regenerative strategies for damaged tissue aim to present biochemical cues that recruit and direct progenitor cell migration and differentiation. Hydrogels capable of localized gene delivery are being developed to provide a support for tissue growth, and as a versatile method to induce the expression of inductive proteins; however, the duration, level, and localization of expression is often insufficient for regeneration. We thus investigated the modification of hydrogels with affinity peptides to enhance vector retention and increase transfection within the matrix. PEG hydrogels were modified with lysine-based repeats (K4, K8), which retained approximately 25% more vector than control peptides. Transfection increased 5- to 15-fold with K8 and K4 respectively, over the RDG control peptide. K8- and K4-modified hydrogels bound similar quantities of vector, yet the vector dissociation rate was reduced for K8, suggesting excessive binding that limited transfection. These hydrogels were subsequently applied to an in vitro co-culture model to induce NGF expression and promote neurite outgrowth. K4-modified hydrogels promoted maximal neurite outgrowth, likely due to retention of both the vector and the NGF. Thus, hydrogels modified with affinity peptides enhanced vector retention and increased gene delivery, and these hydrogels may provide a versatile scaffold for numerous regenerative medicine applications.
authors.affiliation: Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Road, Tech E136, Evanston, IL 60208, United States.
authors.forename: Jaclyn A
authors.initials: JA
authors.lastname: Shepard
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Wesson
authors.forename: Christine E
authors.initials: CE
authors.lastname: Wang
authors.forename: Alyson C
authors.initials: AC
authors.lastname: Stevans
authors.forename: Samantha J
authors.initials: SJ
authors.lastname: Holland
authors.forename: Ariella
authors.initials: A
authors.lastname: Shikanov
authors.forename: Bartosz A
authors.initials: BA
authors.lastname: Grzybowski
authors.forename: Lonnie D
authors.initials: LD
authors.lastname: Shea
mesh_terms: D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D008962:Models, Theoretical; D020932:Nerve Growth Factor / Q000378:metabolism; D016501:Neurites / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000235:genetics; D023822:Tissue Engineering; D014162:Transfection
chemical_list: D020100:Hydrogels; D010455:Peptides; D004247:DNA; D020932:Nerve Growth Factor