Microfluidic Synthesis of Nanoparticles for Cell Reprogramming

Louisiana Tech University, Ruston LA

Investigators

Abstract

During normal life, the cells in different parts of the body use their genetic legacy to perform tasks that create human health. During illnesses certain cells may not properly perform these tasks. As appropriate and optimal therapies may vary among patients due to their own genome difference, it would be very helpful to use the patient's own cells to correctly perform their designated tasks. This could become feasible after the identity of a cell is successfully changed from one type to another by proper genetic programming. Ground-breaking progress was made in this field recently by converting adult skin cells into other desirable cell types. Such cell reprogramming methods are more acceptable because the cells that are modified come from the patient and the changes cannot be transmitted to others or to offspring. Unfortunately, current delivery tools for cell reprogramming are very slow and too inefficient to use in therapy. This award investigates a new microfluidic approach to synthesize and manufacture novel polymeric nanoparticles for efficient gene delivery. With its continuous flow operation, this novel approach produces such gene carriers in large quantities and at low cost. Moreover, the high-quality nanoparticles that are generated could significantly increase the success rate of cell reprogramming to produce desired types of a patient's cells with high yield. Its nanomanufacturing potential could accelerate future clinical and pharmaceutical uses of such patient-matched cells. This research will also train graduate students in multidisciplinary skills in science and engineering. The results will be integrated into engineering courses to inspire, educate, and retain undergraduate and minority students through hands-on opportunities in bio- and nanomanufacturing, cell therapy, chemistry, characterization, and process development. Although sustained cell reprogramming is feasible, the low success rate is tied to its transient transfection behavior, namely, poor delivery efficiency of the key transcription factors and low cell survival to allow for repeated delivery, and the lack of appropriate manufacturing process in the production of desired gene carriers and reprogrammed cells. This research project studies a hybrid field microfluidic process to produce homogeneous polyplex nanoparticles for better protection, condensation, and release of genetic probes by limiting the complexation at the fluid interface. Gold nanoparticle carriers help fix the internalized polymer molecules after the dissociation of the polyplex to reduce cytotoxicity. In addition, electroporation treatment promotes transport of genetic material to the nucleus. The combination ensures increased delivery efficiency and the allowance for repeated transfection to sustain the expression of the needed reprogramming factors. The hybrid microfluidic system?s flow operation allows continuous production of gene carriers and later patient-matched cell lines with high throughput and good quality control. The project's success provides not only a powerful manufacturing route in nanoprobe synthesis, but also an effective tool for potential synergy of diagnosis and therapeutics to advance current gene/drug delivery and cell-based bio-manufacturing processes.