Highly efficient intracellular transduction in three-dimensional gradients for programming cell fate

Eltaher, Hoda M.; Yang, Jing; Shakesheff, Kevin M.; Dixon, James E.

Abstract

Fundamental behaviour such as cell fate, growth and death are mediated through the control of key genetic transcriptional regulators. These regulators are activated or repressed by the integration of multiple signalling molecules in spatio-temporal gradients. Engineering these gradients is complex but considered key in controlling tissue formation in regenerative medicine approaches. Direct programming of cells using exogenously delivered transcription factors can by-pass growth factor complexity but there is still a requirement to deliver such activity spatio-temporally. We previously developed a technology termed GAG-binding enhanced transduction (GET) to efficiently deliver a variety of cargoes intracellularly using GAG-binding domains to promote cell targeting, and cell penetrating peptides (CPPs) to allow cell entry. Herein we demonstrate that GET can be used in a three dimensional (3D) hydrogel matrix to produce gradients of intracellular transduction of mammalian cells. Using a compartmentalised diffusion model with a source-gel-sink (So-G-Si) assembly, we created gradients of reporter proteins (mRFP1-tagged) and a transcription factor (TF, myogenic master regulator MyoD) and showed that GET can be used to deliver molecules into cells spatio-temporally by monitoring intracellular transduction and gene expression programming as a function of location and time. The ability to spatio-temporally control the intracellular delivery of functional proteins will allow the establishment of gradients of cell programming in hydrogels and approaches to direct cellular behaviour for many regenerative medicine applications.

Citation

Eltaher, H. M., Yang, J., Shakesheff, K. M., & Dixon, J. E. (2016). Highly efficient intracellular transduction in three-dimensional gradients for programming cell fate. Acta Biomaterialia, 41, 181-192. https://doi.org/10.1016/j.actbio.2016.06.004

• Journal Article Type: Article
• Acceptance Date: Jun 2, 2016
• Online Publication Date: Jun 3, 2016
• Publication Date: 2016-09
• Deposit Date: Jun 6, 2017
• Publicly Available Date: Jun 6, 2017
• Journal: Acta Biomaterialia
• Print ISSN: 1742-7061
• Electronic ISSN: 1878-7568
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 41
• Pages: 181-192
• DOI: https://doi.org/10.1016/j.actbio.2016.06.004
• Keywords: Intracellular transduction; Gradient; Diffusion; Hydrogel; GAG-binding enhanced transduction (GET); CPP; Myogenesis
• Public URL: https://nottingham-repository.worktribe.com/output/802821
• Publisher URL: http://www.sciencedirect.com/science/article/pii/S1742706116302720?via%3Dihub
• Additional Information: This article is maintained by: Elsevier; Article Title: Highly efficient intracellular transduction in three-dimensional gradients for programming cell fate; Journal Title: Acta Biomaterialia; CrossRef DOI link to publisher maintained version: https://doi.org/10.1016/j.actbio.2016.06.004; Content Type: article; Copyright: © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
• Contract Date: Jun 6, 2017