Using Cell Priming and Telecommunications Modeling to Enhance Gene Delivery for Stem Cell Therapies - PROJECT SUMMARY
The goal of this New Innovator project is to advance gene delivery applications for stem cell therapies through
innovative new delivery systems designed using information gained from priming of endogenous cellular
targets and telecommunications (mathematical) modeling of nonviral DNA transfer.
Gene delivery requires a
carrier to transport exogenous genes as plasmid DNA (pDNA) to cells to produce an encoded protein (i.e.
transfection). Nonviral delivery systems have been extensively investigated as a technique to accomplish
transfection, however they suffer from low efficiency, thereby limiting their application in functional genomics,
tissue engineering, medical devices, and gene therapy. In particular, gene delivery to adult-derived stem cells
has proven especially difficult. Empirical investigations into enhancing transfection of nonviral systems have
often focused on modifications to the pDNA carrier. However, those studies have had limited successes in
enhancing transfection largely due to the lack of information of the molecular factors that facilitate the delivery
process, as well as an incomplete understanding of the process itself. Recently, two different approaches have
been investigated by the PI to improve our understanding of the gene delivery process: pharmacogenomics
screens (i.e. microarray analysis) and pharmacokinetics models. Microarray studies in the PI's laboratory have
been used to profile transfected and untransfected (but treated) cell populations, providing molecular targets to
enhance nonviral transfection. This project proposes to use those targets for cell priming to both increase our
understanding of the role of the targets and develop simple new protocols to enhance transfection. In addition,
we will identify more candidate priming agents through a screen of clinically approved drugs, which can also be
analyzed for their effect on specific gene pathways, to further our understanding of the process. In parallel with
previous microarray analysis efforts, computational models have been developed, providing insight into barriers
and kinetic parameters of the gene transfer process. However previous models are limited in their ability to output
mechanistic information. Here a novel kinetic model of gene delivery using telecommunications queuing theory
will overcome challenges encountered by previous mathematical models and allow for integration of cell priming
modes of transfection. In this telecommunications model, delivery of DNA to the cell nucleus is considered in the
same way as the delivery of a packet of information (DNA) to a destination computer (nucleus) within a packet-
switched computer network (barriers to DNA transfer). This model will allow for a priori predictions about novel
transfection systems. With information gained from this project, I propose to develop new delivery strategies that
incorporate drug priming to chemically modulate key cellular barriers as a simple and clinically-translatable
approach to improve transfection of adult human stem cells, for use in tissue engineering and regenerative
medicine, the delivery and secretion of therapeutic proteins, organ transplantation, and cancer therapy.
