Direct cell reprogramming has the potential to facilitate the development of safer and more effective patient- specific cell-based therapies. However, current methodologies that induce direct reprogramming face major translational hurdles, including heavy reliance on viral transfection. While the results are promising, biosafety concerns, capsid size constraints and/or the stochastic nature of conventional methods (viral and non-viral) pose significant limitations. We developed and novel 3D nanochannel electroporation (3D NEP) platform technology that overcomes these barriers by enabling deterministic transduction of reprogramming factors with single-cell resolution, and without the need for viral vectors. This nanotechnology-based approach promotes remarkably fast and efficient direct cellular reprogramming, as demonstrated with well-established and newly-developed models of induced neurons and endothelium, respectively. Non-viral direct derivation of induced endothelial cells, in particular, could find applications in the treatment of a number of disorders, including critical limb ischemia and stroke. Ischemic strokes, for example, result in significant cellular deficiencies (e.g., vascular, neuronal) that could lead to death or major morbidity. Nevertheless, currently no study has looked into developing methods for virus-free direct reprograming of endothelial cells. Moreover, the regenerative potential of these cells has not been investigated within the context of stroke. Here we are proposing to develop an optimized method for direct derivation of endothelial cells by 3D NEP, and to study the extent to which these cells induce functional recovery in a middle cerebral artery occlusion (MCAO) mouse stroke model. In the US, strokes occur every ~40 s and have a death rate of ~42%. With an estimated cost of >70 billion dollars, strokes represent a substantial burden to the health care system. 1

Public Health Relevance

Although direct cell reprogramming could enable safer and more effective cell therapies, status quo reprogramming methodologies face major translational hurdles. So as to overcome these barriers, we are proposing to use 3D NEP to develop a novel reprograming model of induced endothelium, and study its regenerative potential in a mouse stroke model. With an estimated death rate of ~42%, and >70 billion dollars in costs, strokes represent a substantial burden to the health care system in the US. 1

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21NS099869-02
Application #
9334334
Study Section
Nanotechnology Study Section (NANO)
Program Officer
Bosetti, Francesca
Project Start
2016-09-01
Project End
2019-08-31
Budget Start
2017-09-01
Budget End
2019-08-31
Support Year
2
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Ohio State University
Department
Surgery
Type
Schools of Medicine
DUNS #
832127323
City
Columbus
State
OH
Country
United States
Zip Code
43210
Shukla, Vasudha C; Kuang, Tai-Rong; Senthilvelan, Abirami et al. (2018) Lab-on-a-Chip Platforms for Biophysical Studies of Cancer with Single-Cell Resolution. Trends Biotechnol 36:549-561
Gallego-Perez, Daniel; Pal, Durba; Ghatak, Subhadip et al. (2017) Topical tissue nano-transfection mediates non-viral stroma reprogramming and rescue. Nat Nanotechnol 12:974-979