Expressing humanized bacterial luciferase in stem cells: Moving beyond firefly luciferase to expand the informational capacity of animal models for regenerative medicine

DESCRIPTION provided by applicant This Small Business Technology Transfer STTR Phase I project proposes to develop autonomously bioluminescent human stem cells for continuous reagent free and real time bioimaging to address the National Institutes of Healthandapos s request for new techniques for non invasive long term tracking of stem cell survivability engraftment and migration following in vivo implantation The ability of stem cells to self renew and differentiate into other cell lineages has emerged as a valuable therapeutic approach to functionally heal previously irreparable tissues and organs However for the regenerative medicine field to effectively transition toward translational and clinical practice outcomes a strong dependence on animal models will be required to fully understand the capabilities and complexities of stem cells BioTech proposes to expand the informational capacity of animal models by creating stem cell lines that self generate bioluminescent light via expression of a andapos humanizedandapos bacterial luciferase thereby enabling stem cells to be continuously imaged throughout their lifetime as they physiologically function within their animal host This differs significantly from the current market of bioluminescent imaging technologies that rely on a firefly luciferase gene construct that must be provided with a chemical substrate to activate its light emission response resulting in only marginally informative single time point snapshots of cell function in tandem with repetitive animal injections that invoke unknown and potentially interfering interactions and adversely effects animal welfare In partnership with the University o Tennessee Medical Center the specific objectives of this Randamp D effort are to develop piggyBac transposition and lentiviral transduction methods for streamlined integration of the bioluminescent phenotype into adipose derived mesenchymal stem cell lines followed by performance evaluation in in vitro D scaffolds and in vivo mouse models to demonstrate proficiency toward uninterrupted imaging and enriched data flows that far exceed that of existing firefly luciferase methods With no change in instrumentation or fundamental bioluminescent protocols necessary researchers can seamlessly transition from firefly luciferase to BioTechandapos s humanized bacterial luciferase technology to advance their in vivo experimental Randamp D to more informative endpoints with fewer animals required The contribution of this innovative imaging platform to the field of regenerative medicine will provide more physiologically relevant and representative data critical to predicting the efficacy and safety of treatment strategies as they precede to clinical trials PUBLIC HEALTH RELEVANCE Regenerative medicine using stem cell therapies to replenish and restore tissues and organs has the potential to transform human health by curing and remedying previously unmanageable diseases Animal studies have played significant roles in deciphering the therapeutic capacity of stem cells but there exists a disparity between the results obtained from animal experiments and their transition to human clinical trials that are impeding advancements in the regenerative medicine field To increase the amount of experimental information obtainable from animal models BioTech proposes to create stem cell lines that continuously emit bioluminescent light thereby enabling implanted stem cells to be visualized and tracked throughout their lifetime for improved understanding of their therapeutic potential and limitations directly within living animal subjects