Generation of virus-free induced pluripotent stem cells using fusogenic lipid vesicles to deliver reprogramming proteins to adult neural stem cells
Final Report Abstract
We have designed fusogenic lipids that were able to efficiently deliver the model mKate protein and mRNA encoding for nGFP to cells. Among them the thiocholesterol-based lipid showed highest delivery efficiency. Two other libraries of bioresponsive, fusogenic lipids have been synthesized. These will serve to fine-tune delivery of reprogramming factors in vitro and in vivo. To be able to deliver reprogramming factors as proteins and/or nucleic acids in vivo we have developed a method to generate lipid nanoparticles with small sizes (< 100 nm) and high encapsulation efficiency. Particle sizes smaller than 100 nm will allow the lipid nanoparticles to circulate for a longer time period under in vivo conditions, enabling them to accumulate to the site of disease. We have developed and engineered carriers that are able to target the ischemic area of the heart after myocardial infarction. In future studies we plan to combine these tools for in vivo reprogramming the scar tissue of infarcted heart to fully functional cardiomycoytes. This would allow the regeneration of cardiac tissue without the use of viruses and DNA that could cause insertional mutagenesis.
Publications
- Controlled nucleation of lipid nanoparticles. Pharmaceutical Research. 2012 Aug;29(8):2236-48
J. Nguyen, C. Walsh, J.P. Motion, E. Perttu, F. Szoka
- Nucleic acid delivery: the missing pieces of the puzzle. Accounts of Chemical Research. 2012 Jul 17;45(7):1153-62
J. Nguyen and F. Szoka
- Phosphatase-triggered fusogenic liposomes for cytoplasmic delivery of cell-impermeable compounds. Angewandte Chemie International Edition. June 2012
M. Motion, J. Nguyen and F. Szoka
- Synthesis and characterization of novel zwitterionic lipids with pHresponsive biophysical properties. Chemical Communications (Camb). 2012 Jun 7;48(45):5575-7
C. Walsh, J. Nguyen and F. Szoka
- Synthesis, characterization and evaluation of ionizable lysine-based lipids for siRNA delivery. Bioconjugate Chemistry, Nov. 2012
C. Walsh, J. Nguyen, M. Tiffany and F. Szoka
