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Expanding the throughput of real-time toxicological screening of cardiac differentiation by expressing a synthetic luciferase/luciferin genetic pathway in iPSCs

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 2R44ES026269-02
Agency Tracking Number: R44ES026269
Amount: $991,009.00
Phase: Phase II
Program: SBIR
Solicitation Topic Code: NIEHS
Solicitation Number: PA18-574
Timeline
Solicitation Year: 2018
Award Year: 2019
Award Start Date (Proposal Award Date): 2019-07-15
Award End Date (Contract End Date): 2021-06-30
Small Business Information
2450 EJ CHAPMAN DR
Knoxville, TN 37996-0001
United States
DUNS: 968832498
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 DAN MORRISON
 (865) 604-7713
 dan.morrison@490biotech.com
Business Contact
 STEVEN RIPP
Phone: (865) 974-9605
Email: steven.ripp@490biotech.com
Research Institution
N/A
Abstract

Expanding the throughput of real-time toxicological screening of cardiac differentiation byexpressing a synthetic luciferase/luciferin genetic pathway in iPSCs
Project Summary
This Small Business Innovation Research (SBIR) Phase II project proposes to develop and validate autonomously
bioluminescent induced pluripotent stem cells (iPSCs) for use in continuously data producing, reagent-free, and
real-time toxicological screening assays. These tools will address the National Institute of Environmental Health
Sciences (NIEHS) request for novel high-throughput assays to evaluate the effects of chemical compounds on
the differentiation of pluripotent stem cells, as their ability to differentiate along well-defined lineage pathways
offers a powerful approach to understanding how chemical perturbations disrupt metabolic and regulatory
functions along those pathways. The autobioluminescent iPSC-based assay systems developed here will
significantly contribute towards NIEHS’s mission to ‘discover how the environment affects people in order to
promote healthier lives’ by expanding the knowledgebase of chemical exposure toxicological effects. This is
especially important given that the commercial marketplace maintains an inventory of tens of thousands of
chemicals, the majority of which have poorly understood human health impacts, and currently require animal-
based testing approaches that are expensive, time consuming, and ethically contentious to determine their
effects. As an alternative, stem cell-based assays such as those developed here can mimic human disease states
more reliably than animal models while providing valuable information towards understanding how chemical
exposures influence cancer risks, developmental defects, and other adverse health outcomes. A significant
impetus therefore exists to integrate stem cells into chemical screening programs such as Tox21 and ToxCast,
but under the mandate that they function under high-throughput conditions. While this goal is not obtainable
using existing bioluminescent reporter technologies such as firefly luciferase that must be provided with a
chemical substrate to activate its light emission responses, resulting in only marginally informative single time
point snapshots of potential toxicological interactions, 490 BioTech’s synthetic luciferase technology rather
enables reporter cells to emit light continuously and in real-time, thereby providing an uninterrupted stream of
visual data over the lifetime of the cell as it interacts and reacts to chemical perturbations. The goal of this
research effort is to develop improved assays based upon synthetic-luciferase-expressing iPSCs and validate their
ability to report the impact of chemical exposure to cellular health and development in real time. To accomplish
these goals, our specific aims will focus on assay development and optimization, validation against a chemical
subset of the Tox21 10K library, and benchmark comparisons against existing commercial assay systems.Project Narrative
Humans interact with a vast landscape of chemicals that, despite the efforts of numerous U.S. and international
multiagency sponsored programs to test for their toxic effects and ensure consumer safety, often have poorly
understood impacts on our health. Because the primary limitation preventing these agencies from better
characterizing chemical health effects is a lack of testing methods that can rapidly provide data with sufficient
informational content and direct relevance to human health impacts, 490 BioTech is creating and validating a
next-generation testing system that uses human stem cell lines capable of autonomously emitting
bioluminescent light in response to chemical exposure events. This new testing system will enable chemical
toxicity screening to occur throughout the lifetime of the cell and produce an expanded informational database
of toxicological effects with direct relevance to human health.

* Information listed above is at the time of submission. *

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