(4lg) Engineering Targeted Delivery Systems for Gene Therapy and Gene Editing

Millions of people worldwide suffer from genetic disorders which could benefit greatly from gene supplementation and, more recently, gene editing therapies. Successful clinical translation of these technologies is limited by the need to deliver gene supplementation therapies and gene editing agents to target cell types and subcellular compartments. Although major advances have been made in the development of both viral and non-viral delivery technologies for the delivery of gene therapies with RNA or DNA cargoes, no single technology will likely be most optimal for all potential applications and opportunities remain to further develop synthetic nanoparticles, viruses, and more recently virus-like particles, for the delivery of DNA and RNA gene therapies. Therefore, my research interest focuses on engineering non-viral, viral, and virus-like particle systems to enable targeted delivery of therapeutic gene supplementation and gene editing therapies for the treatment of genetic diseases, with a particular focus on hematological and immunological genetic diseases.

My research aims are therefore spread across three major thrusts: 1) engineer strategies to target specific intracellular transport pathways for enabling effective delivery of macromolecular cargoes to desired subcellular compartments, 2) develop high-throughput screening methodologies with a focus on functional, gene editing readouts in target cells and tissues, and 3) apply technologies identified in the first two aims in disease-relevant ex vivo and in vivo models to elucidate and overcome disease-specific determinants of and bottlenecks to delivery and gene editing. My independent research group will combine interdisciplinary approaches from biomolecular engineering, immunology, material science, and chemistry, all rooted in chemical engineering principles, to achieve these goals. Taken together, this work aims to elucidate complex interactions between delivery vehicles and biological systems at the tissue, cellular, and subcellular levels with immediate implications for the development of next-generation gene supplementation and gene editing therapies.

Research Experience

My doctoral and post-doctoral training has prepared me to carry out this research vision. During my PhD with Professors Robert Langer and Daniel Anderson at MIT, I focused on engineering delivery vehicles to improve the safety and efficacy of inhaled mRNA therapies and mRNA vaccines. To this end, I worked on systematically engineering each of the mRNA-LNP components, including the lipids, the mRNA, and the buffer, using a combination of high-throughput screening and rational design approaches to overcome physical and biological barriers to in vivo mRNA delivery. Based on my PhD thesis, I received the Controlled Release Society’s Best Doctoral Thesis in Drug Delivery Research Award.

For inhaled mRNA therapies, I developed nebulizer-stable LNPs for effective delivery of mRNA to the lung epithelium through 1) high-throughput screening of novel ionizable lipids in ex vivo lung models to improve mRNA delivery to the lung epithelium; and 2) rational design of the nebulization buffer based on DLVO theory to stabilize LNPs during nebulization and prevent LNP aggregation (Nature Nanotechnology, 2024). The combinatorially optimized, nebulized LNPs demonstrate state-of-the-art delivery of mRNA to the lung epithelium in both healthy and muco-obstructive disease lung models. Based on this work and our greater understanding of LNP stability, I further rationally designed nebulized LNPs by developing PEG-alternative polymer modifications to the LNPs to enhance nebulizer stability and improve mRNA delivery to the lungs.

In response to the COVID-19 pandemic, I engineered safer and more effective mRNA vaccines by developing a multiply adjuvanted mRNA-LNP system. I achieved this by combining an immunostimulatory ionizable lipid, identified through batch-based in vivo screening of a large lipid library, with a rationally designed mRNA transcript genetically fusing the encoded antigen to an immune-activating protein of the complement pathway (Nature Biomedical Engineering, 2024). Vaccination with the lead LNP formulation and fusion mRNA vaccine resulted in synergistically improved immune responses following either intramuscular or intranasal administration in mice. The multiply adjuvanted mRNA vaccines I developed have the potential to improve not only the safety and efficacy of mRNA vaccines but also the ease of administration.

As a postdoctoral researcher with Professor David Liu at the Broad Institute of Harvard and MIT, I am developing new approaches towards the in vivo delivery of therapeutic gene editors, including prime editors and derivative technologies for gene-sized integration, to diverse tissues and cell types. Working on these projects has helped to elucidate the limitations of current technologies for delivering more complex cargoes required for in vivo gene editing as well as identify bottlenecks of gene editing efficiency unique to in vivo applications.

Collectively, my work has deepened our understanding of the complex interactions between delivery vehicles, macromolecular cargoes, and biological systems, enabling the development of new technologies for overcoming physical and biological barriers to intracellular delivery in vivo.

Teaching Interests

As a chemical engineer by training, I am eager to teach core undergraduate and graduate courses. Throughout my academic career, I have sought out opportunities to help students master core chemical engineering concepts while also developing my skills as a teacher. As an undergraduate at Cornell University, I served as the head teaching assistant for undergraduate fluid mechanics. Additionally, as president of the undergraduate chapter of AIChE at Cornell, I founded a weekend program for high-school students to participate in experiments designed to expose students early on to engineering principles. During my PhD studies, I was a teaching assistant for senior undergraduate electrochemical engineering and kinetics courses. I also served as a laboratory instructor for an undergraduate laboratory course on microfluidics, designing a set of experiments that provided students with the opportunity to synthesize microfluidic chips and use the chips to formulate nanoparticles. To further develop my skills as a teacher, I participated in the Kauffman Teaching Certificate program at MIT, equipping myself with the tools needed to build a diverse, equitable, and inclusive teaching environment.

Given my background combined with my teaching and research experiences, I am prepared to teach any chemical engineering course with a particular interest in transport and thermodynamics, which are subjects that have been integral to my research. I am also excited to teach bioengineering-related subjects and aim to develop a course focused on applying chemical engineering principles to the rational design of drug delivery systems.

Selected Awards

• Controlled Release Society Best Doctoral Thesis in Drug Delivery Research Award, 2024
• AIChE Division 15 Oral Presentation Award, 2022
• Koch Institute Peter Karches Mentorship Prize, MIT, 2022
• Marble Center for Cancer Nanomedicine Symposium Outstanding Poster Award, MIT, 2022
• Robert F. Smith School of Chemical and Biomolecular Engineering Outstanding Research by an Undergraduate Award, Cornell University, 2018
• Barry M. Goldwater Scholarship, 2017
• Genentech Scheele Outstanding Junior Award, Cornell University, 2016

Selected Publications

• Jiang, A.Y.*, Witten, J.*, Raji, I.O.*, Eweje, F., MacIsaac, C., Meng, S., Oladimeji, F.A., Langer, R., Anderson, D.G. “Combinatorial development of nebulized mRNA delivery formulations for the lungs”. Nature Nanotechnology, 19: 364-375 (2024)
• Li, B.*, Jiang, A.Y.*, Raji, I.*, Atyeo, C., Raimondo, T.R., Gordon, A.G.R, Rhym, L.H., Samad, T., MacIsaac, C., Witten, J., Mughal, H., Chicz, T., Xu, Y., McNamara, R., Bhatia, S., Alter, G., Langer, R., Anderson, D.G. “Enhancing the immunogenicity of lipid-nanoparticle mRNA vaccines by adjuvanting the ionizable lipid and the mRNA”. Nature Biomedical Engineering, 1-18 (2023)
• Barbier, A.J., Jiang, A.Y., Zhang, P., Wooster, R., Anderson, D.G., “The clinical progress of mRNA vaccines and immunotherapies”. Nature Biotechnology, 40: 840–854 (2022)
• Jiang, A.Y.*, Lathwal, S.*, Meng, S., Witten, J., Raji, I.O., Manan, R., Langer, R., Anderson, D.G. “Zwitterionic polymer-modified lipid nanoparticles for nebulized mRNA delivery to the lungs”. Submitted

*denotes equal contribution