Collaborating Investigator: Heidi Feldser, PhD 
Affiliation: GSK  

Funding Source: Internal
Project Period: 03/01/22-02/28/25 

Significance

Pharmaceutical companies such as GSK are increasingly interested in next-generation biological therapeutics, including gene therapy and immunotherapy. CP #2 , which builds on a project with GSK during the first-round funding of the P41, represents a collaborative interest in TR&D2 technologies for translation, with a focus on RNA delivery, and in particular siRNA delivery. While siRNA delivery is not a specific focus of TR&D2, the NIMs used for mRNA and oligonucleotide delivery in TR&D2 Specific Aim 2 can be adapted to encapsulate other genetic cargos such as siRNA and deliver them to immune cells. The goal of CP #2 is safe and efficient siRNA delivery to macrophages to enable new treatments for human diseases. NIM materials, including particles with tunable chemistry and physical size, can enable improved intracellularly delivery to macrophages to knock down multiple target genes with siRNA.

Approach

Aim 1: Evaluate NIM nanoparticle library in macrophages to determine cell uptake and transfection.  NIMs will be evaluated in high-throughput and high-content in vitro assays to understand how chemical structure and physical size modulates uptake and transfection of siRNA-containing nanoparticles. Lead NIMs for siRNA delivery to macrophages will be selected and characterized.

Aim 2: Utilize leading NIMs from Aim 1 to deliver siRNA combinations against GSK identified targets. Knockdown of specific disease-associated genes expressed in macrophages will be targeted with siRNA nanoparticles in vitro. Cells will be evaluated for efficacy, cytotoxicity, and durability.

Push/Pull Interactions

Push: JH-TIE is developing NIMs useful for nucleic acid delivery and is interested in extending the applications. There is an interest in tuning encapsulation conditions of the cationic polymers with RNA to enable efficient encapsulation of siRNA and fabrication of LEAQ/siRNA particles. This project enables this discovery and exploration on NIMs for small oligonucleotide cargos and for new clinical applications.

Pull: GSK has a need for development of novel drugs, specifically siRNA-based therapies and delivery technologies to address immune disorders with high unmet need. Specifically, GSK is exploring new siRNA molecules against new hard-to-drug targets to treat immune disorders. As the siRNA cargos are discovered, there is an immediate need for a method to deliver these cargos intracellularly with high efficacy and safety.