Elastin-like polypeptide matrices for enhancing adeno-associated virus-mediated gene delivery to human neural stem cells

Abstract

The successful development of efficient and safe gene delivery vectors continues to be a major obstacle to gene delivery in stem cells. In this study, we have developed an elastin-like polypeptide (ELP)-mediated adeno-associated virus (AAV) delivery system for transducing fibroblasts and human neural stem cells (hNSCs). AAVs have significant promise as therapeutic vectors because of their safety and potential for use in gene targeting in stem cell research. ELP has been recently employed as a biologically inspired 'smart' biomaterial that exhibits an inverse temperature phase transition, thereby demonstrating promise as a novel drug carrier. The ELP that was investigated in this study was composed of a repetitive penta-peptide with [Val-Pro-Gly-Val-Gly]. A novel AAV variant, AAV r3.45, which was previously engineered by directed evolution to enhance transduction in rat NSCs, was nonspecifically immobilized onto ELPs that were adsorbed beforehand on a tissue culture polystyrene surface (TCPS). The presence of different ELP quantities on the TCPS led to variations in surface morphology, roughness and wettability, which were ultimately key factors in the modulation of cellular transduction. Importantly, with substantially reduced viral quantities compared with bolus delivery, ELP-mediated AAV delivery significantly enhanced delivery efficiency in fibroblasts and hNSCs, which have great potential for use in tissue engineering applications and neurodegenerative disorder treatments, respectively. The enhancement of cellular transduction in stem cells, as well as the feasibility of ELPs for utilization in three-dimensional scaffolds, will contribute to the advancement of gene therapy for stem cell research and tissue regenerative medicine.

Effective release of a broad spectrum antibiotic from elastin‐like polypeptide‐collagen composite

“…ELPs have been used for tissue engineering and drug delivery applications . Researchers have demonstrated enhanced delivery of ELP‐bound bioactive agents to solid tumors, intraarticular joint space, and chronic wounds .…”

“…ELPs have been used for tissue engineering and drug delivery applications. [25][26][27][28][29][30][31][32][33][34][35] Researchers have demonstrated enhanced delivery of ELP-bound bioactive agents to solid tumors, 28 intraarticular joint space, 29 and chronic wounds. 30 On the other hand, delivery of bioactive agents has been attempted from ELP scaffolds.…”

Comparison of Ectopic Gene Expression Methods in Rat Neural Stem Cells

“…In addition, to improve viral transduction efficiency, magnetically guided AAV delivery system was used and this significantly enhanced cellular transduction in human NSCs (hNSCs) and reduced the virus incubation time [58]. Elastin-like polypeptide-treated polystyrene surfaces are known to enhance AAV-mediated gene delivery efficiency in hNSCs [59]. It is also reported that ultrasound can enhance retrovirus-mediated gene transfer efficiency in various cell types [60,61].…”

Enhanced selective gene delivery to neural stem cells in vivo by an adeno-associated viral variant

“…In addition to improved transduction of NSCs, AAV r3.45 significantly improved homologous recombination-based gene correction: its use resulted in a fivefold increase in targeted gene correction in NSCs compared with wild-type AAV2 and AAV5 (Jang et al, 2011). When AAV r3.45 was immobilized onto elastinlike peptides, delivery to human NSCs was further enhanced (Kim et al, 2012b).…”