(340z) Design and Development of Gold Nanoshell-Liposomes Formulations for Scalable, High-Throughput Ex-Vivo mRNA and DNA Delivery

In 2020, almost 2 million new cases of cancer were diagnosed in the United States and over 600,000 people died from the disease. Immune therapy has recently been validated as a new platform that has the potential for complete cancer remission by altering the patient’s immune system to seek out and destroy tumors. T cells and NK cells can be modified by ex vivo delivery of an mRNA that codes for chimeric antigen receptors (CAR) to identify tumor cells and activate the immune response. To date, clinical efforts to develop mRNA delivery systems have been focused on vaccines such as the Moderna and Pfizer Covid-19 vaccines. However, we believe, that an optimized, high throughput, high viability mRNA delivery method could lead to the use of CAR and other therapies for treatment of a broad range of diseases. Hence, development of high throughput and efficient gene delivery systems are imperative for manufacturing protein-modified immune cells ex vivo. Current strategies that have been investigated to improve RNA delivery, such as condensation of mRNA in lipid nanoparticles or electroporation can be toxic and are inefficient, resulting in large cell losses. There are few available viral vectors for mRNA, and all have drawbacks associated with genome integration, and possible host rejection (immunogenicity and cytotoxicity).

To address this problem, my post-doctoral research focused on the design and development of a high viability ex-vivo mRNA delivery system for transient expression of CARs in NK and other primary cells. I encapsulate the mRNA in neutral liposomes that are then chemically tethered to plasmon-resonant hollow gold nanoshells (HGN). I synthesize the HGN by galvanic replacement chemistry from silver nanocrystals. The HGN plasmon resonance can be tuned to absorb physiologically friendly near infra-red light of 650 – 1040 nm wavelength by controlling the reaction conditions. This system has shown great potential with high transfection efficiency on several cell lines.

Our long-term goal is to create the cell manufacturing machinery to produce a cell bank of mRNA coded CAR NK cells. Transient ex-vivo mRNA modification of NK cells represents a hit and run strategy whereby engineered donor NK cells mediate complete and rapid disease eradication with minimal side effects, followed by extinguishing the mRNA-induced alterations, allowing for normal cell recovery. Patients can use donor supplied NK cells rather than patient specific T cells that induce a graft versus host response. The cell bank could provide patients with immediate treatment without the difficulty of T cell harvesting, transfection and expansion, which can take weeks while the cancer progresses.

Gene therapy is possibly one of the most exciting area of bioengineering. We have reached exceptional levels of control over gene delivery, immune system modulation as well as precise manipulation of the human genome as seen through the rapid manufacture of the COVID vaccine. Combining the various experimental as well as instruments techniques can undoubtedly reshape the face of biomedical research in the near future.

Research Interests

Drug discovery, product formulations, Innovative platforms, nanotechnology, nanomedicine, gene/drug therapy, novel vaccine development, research and development