Expressing humanized bacterial luciferase in stem cells: Moving beyond fireflyluciferase to expand the informational capacity of animal models for regenerativemedicine - Expressing humanized bacterial luciferase in stem cells: Moving beyond firefly luciferase to
expand the informational capacity of animal models for regenerative medicine
Project Summary
This Small Business Technology Transfer (STTR) Phase II project proposes to develop complementary
autonomously bioluminescent (autobioluminescent) in vitro stem cell lines and in vivo small animal model
systems that enable the continuous, reagent-free, and real-time bioimaging of mesenchymal stem cell (MSC)
localization, differentiation into adipocyte, chondrocyte, and osteocyte lineages, and persistence post-
differentiation at the site of activation. These models will specifically address the National Institutes of Health's
request for new techniques for non-invasive, long-term tracking of stem cell survivability, engraftment, and
migration following in vivo implantation. By addressing this critical need for new methods capable of elucidating
the mechanisms underlying how stem cells identify areas of dysfunction within the body, differentiate into the
relevant tissues required to correct the malady, and persist in synergy with existing tissue to enable long term
functionality, these tools will significantly improve the transition of regenerative medicine studies towards
translational and clinical practice outcomes. The autobioluminescent MSCs developed by 490 BioTech under
our Phase I effort demonstrated the ability to track MSC localization in vitro and in vivo similarly to existing
optical imaging approaches, but with significantly reduced cost and personnel effort. Furthermore, these models
also negated the need for sample destruction or the stressful and potentially influential injection of an activating
chemical concurrent with imaging while simultaneously providing an uninterrupted stream of visual data over
the lifetime of the reporter cell as it interacts with its environment and undergoes differentiation. In partnership
with the University of Tennessee Medical Center, this proposal will expand upon these accomplishments to
develop fully self-contained autobioluminescent MSC-based cellular models capable of specifically reporting on
their differentiation into adipocyte, chondrocyte, and osteocyte lineages, and complementary small animal
models harboring native MSCs genetically programmed to autonomously enact their reporter functionality only
following differentiation into adipocyte, chondrocyte, or osteocyte lineages in response to wounding or
exogenous stimulation. These models will overcome the primary technical hurdles encountered with all existing
bioluminescent and fluorescent stem cells currently on the market from companies such as PerkinElmer,
ThermoFisher/Life Technologies, Promega, and ~30 smaller specialized business entities in the U.S. alone,
which comprise an estimated market value of at least $2B, with a predicated annual growth rate of 16-40%. We
believe that the products developed in this effort will be capable of significantly improving the throughput and
effectiveness of regenerative medicine studies and advancing our understanding of stem cell-based treatment
efficiency and efficacy to improve both public health and consumer safety. The functional demonstrations and
data gathered in this effort will position these models to thrive within this market and produce an immediate
and significant impact on the field of regenerative medicine that will benefit the population at large.
