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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 Number: R44ES026269
ORGANIZATION: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 09/01/2015
PERIOD OF PERFORMANCE END DATE: 06/30/2022

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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 - Expanding the throughput of real-time toxicological screening of cardiac differentiation by expressing 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.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2023 ( Subtotal = -$5,571 )
2023	2020	490 BIOTECH INC	2450 EJ CHAPMAN DR	KNOXVILLE	TN	37996	KNOX	USA	Environmental Health	000	3	3/16/2023	NON-COMPETING CONTINUATION	-$5,571
														Subtotal = -$5,571

Issue Date FY: 2020 ( Subtotal = $491,375 )
2020	2020	490 BIOTECH INC	2450 EJ CHAPMAN DR	KNOXVILLE	TN	37996	KNOX	USA	Environmental Health	000	3	6/17/2020	NON-COMPETING CONTINUATION	$491,375
2020	2020	490 BIOTECH INC	2450 EJ CHAPMAN DR	KNOXVILLE	TN	37996	KNOX	USA	Environmental Health	001	3	6/23/2020	NON-COMPETING CONTINUATION	$0
														Subtotal = $491,375

Issue Date FY: 2019 ( Subtotal = $499,634 )
2019	2019	490 BIOTECH INC	2450 EJ CHAPMAN DR	KNOXVILLE	TN	37996	KNOX	USA	Environmental Health	000	2	7/2/2019	COMPETING CONTINUATION	$499,634
														Subtotal = $499,634

Grand Total All Awards = $985,438

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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 Number: R44ES026269

ORGANIZATION: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 09/01/2015

PERIOD OF PERFORMANCE END DATE: 06/30/2022

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