Nanotransfection-driven gene and cell therapies

Gene and engineered cell therapies hold transformative potential for treating a wide range of conditions. However, current approaches face significant practical and translational challenges, including a heavy reliance on viral vectors and high variability, which often result in inefficient or unpredictable outcomes. To address these limitations, we developed a novel nanotechnology-based approach that enables deterministic cell (in vitro) or tissue nano-transfection (TNT) (in vivo) without the need for viral vectors. These platform technologies offer a minimally disruptive and non-viral solution, making them particularly suited for complex disease systems such as
neurodegenerative conditions and metabolic disorders. Nano-transfection chips were manufactured from silicon or polymeric track etched membranes using cleanroom-based approaches, as described previously. These chips were then used to nanotransfect skin cells, in vitro or in vivo, with pro-vasculogenic or pro-adipogenic gene cocktails, which were then evaluated (for their therapeutic potential) in different murine models of neurodegenerative conditions, including ischemic stroke, and Alzheimer’s disease (AD), as well as models of
metabolic dysfunction. Different molecular, histological, and functional outcomes studies were conducted to assess the extent to which these therapies mitigated disease burden in each model. In murine models of ischemic stroke, nanotransfected fibroblasts improved brain vascularization, perfusion, as well as neuroprotection and neuroregeneration in the motor cortex, which led to improved motor function. In murine models of AD, nanotransfected fibroblasts led to improved brain vascularization, perfusion, and reduced amyloid beta load. This also correlated with improved memory and cognitive function. Finally, in murine models of
metabolic dysfunction, TNT-treated skin cells were successfully coopted to partially fulfill brown
adipogenic functions, which led to improved weight control and cardiometabolic function.
Overall, these findings highlight the potential of cell and tissue nano-transfection to drive therapeutic processes for restoring damaged or diseased tissue structure and function, paving the way for innovative treatments across diverse pathological conditions.