Nanotransfection-based vasculogenic cell reprogramming drives functional recovery in a mouse model of ischemic stroke

Abstract

Ischemic stroke causes vascular and neuronal tissue deficiencies that could lead to substantial functional impairment and/or death. Although progenitor-based vasculogenic cell therapies have shown promise as a potential rescue strategy following ischemic stroke, current approaches face major hurdles. Here, we used fibroblasts nanotransfected with Etv2, Foxc2, and Fli1 (EFF) to drive reprogramming-based vasculogenesis, intracranially, as a potential therapy for ischemic stroke. Perfusion analyses suggest that intracranial delivery of EFF-nanotransfected fibroblasts led to a dose-dependent increase in perfusion 14 days after injection. MRI and behavioral tests revealed ~70% infarct resolution and up to ~90% motor recovery for mice treated with EFF-nanotransfected fibroblasts. Immunohistological analysis confirmed increases in vascularity and neuronal cellularity, as well as reduced glial scar formation in response to treatment with EFF-nanotransfected fibroblasts. Together, our results suggest that vasculogenic cell therapies based on nanotransfection-driven (i.e., nonviral) cellular reprogramming represent a promising strategy for the treatment of ischemic stroke.

Modeling the gene delivery process of the needle array-based tissue nanotransfection

“…For determination of mRNA expression of Ascl1, quantified by real-time or quantitative (Q)PCR assay was performed using the double-stranded DNA binding dye SYBR Green-I [2,54].…”

“…Tissue nanotransfection (TNT) is an emerging approach in the field of regenerative medicine that enables non-viral direct reprogramming of terminally differentiated cells to another type in vivo [1][2][3][4]. During the TNT procedure, by applying electric pulses between the nanofabricated chip and the target tissue or skin, reprogramming factors can be delivered to the target tissue cells through the mechanism of electroporation and electrophoresis.…”

“…In 2017, nanoporous chip-based TNT has successfully demonstrated that skin can be reprogrammed into vascular cells and neuron cells in the mouse model [1]. Later, the hollow needle array-based TNT chip was fabricated and tested on mouse skin to investigate the cutaneous wound healing because of its improved gene delivery efficiency [2].…”

“…The needle array-based electrode for electroporation was also proposed for drug delivery and gene vaccination for skin tissue, but the needle arrays mainly acted as the electrode [43,44]. Nanofabricated chip-based TNT approach has been proved to be an efficient, fast, and determined gene delivery approach based on non-virus mediated vector [1][2][3][4]. However, prior to clinical applications for human, TNT parameters such as needle design, voltage amplitude, and pulse duration must be optimized for a safe operation and high efficiency.…”

“…Compared to our previous works [1,2], this work aims to theoretically establish a physical model to describe the gene delivery process on the complex skin structure via hollow needle array-based TNT chips. Our prior works did not consider the coexisting influences of the electroporation and electrophoresis contributions in the gene delivery process.…”