Nanotechnology-based non-viral derivation of induced endothelium for ischemic disorders

Ohio State University, Columbus OH

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Abstract

PROJECT SUMMARY 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