Toxicological High-Throughput Screens Based on Prioritized Differentiation of Mouse Viable Potency Reporter, and Viable First Differentiated Lineage Reporter, Mouse Embryonic Stem Cells
WSU Tech#: 15-1279
Technology Summary:
This technology provides systems and methods to detect stress in stem cells where the detected stressor might affect all stem cell lineages and might not kill the stem cells. Testing for stress before differentiation occurs is of great benefit because stress at this stage can affect all later-developing cell lineages. This technology can detect stressors that acts at sub-morbid runting doses that slow growth and negatively impact and imbalance differentiation of stem cells. Accordingly, the invented technology provides for more sensitive stress detection that does not require cell death to observe stress. Imbalances of differentiation forced on cultured stem cells measure organismal responses not needed for adaptation to culture. Thus stress induced differentiation should reduce or replace the need for slower and more costly gestational animal in vivo reproductive toxicology tests.
This technology is based on the finding that stressed stem cells (i) slow their growth and (ii) differentiate under physiological conditions that would normally maintain their toti- or pluri-potency. That is, they differentiate earlier than they normally would. Early differentiation of stem cells depletes the number of stem cells available to differentiate later in the development process, reducing the number of stem cells available to form organ systems that arise later during development. This type of early, stress-induced differentiation is referred to as “prioritized differentiation.” Prioritized differentiation negatively affects development potential.
The technology provides numerous systems and methods to detect stress in stem cells by measuring potency factors (and markers) and prioritized differentiation factors (and markers). “Potency factors” can be thought of as "brake pedals" for differentiation. That is, their expression and activity is associated with toti- or pluri-potency. “Differentiation factors” can be thought of as "accelerator pedals" for differentiation. That is, their expression and activity is associated with normal differentiation that occurs when potency-maintaining extracellular growth factors are removed. Under stress conditions, however, potency factors can decrease and differentiation factors can increase even in the presence of potency-maintaining extracellular growth factors. This type of differentiation is referred to as “compensatory” or “prioritized” differentiation. That is, under compensatory or prioritized differentiation, stem cells can be induced by stress from a toti- or pluripotent state into a more differentiated/committed cell type despite the presence of growth factors that maintain potency. Stress also slows stem cell growth and induces differentiation in fewer-than-normal stem cells to further deplete stemness to “compensate” for fewer cells. Compensatory or prioritized differentiation of stem cells despite potency-maintaining growth factors also prioritizes differentiation to emphasize creating sufficient first lineage differentiation at the sacrifice of later lineages. Decrease in nuclear potency factors and increased in first lineage determining nuclear differentiation factors provide (early lineage>later lineage) “markers” that are evidence that differentiation is enabled, but their activity does not necessarily lead to or cause the potent or differentiated state. The differentiated state requires sufficient loss of nuclear potency factors and/or gain of nuclear differentiation nuclear factors. But, different stressors may more strongly release the brake pedal (decrease potency) or engage the accelerator pedal.
This technology can be used to assess compounds and other environmental conditions for stress effects on stem cells. The systems and methods can be used to screen compounds and other environmental conditions for stress effects on stem cells before the compounds or other environmental conditions are developed, commercialized or released into the workplace or the atmosphere. The systems and methods include modified stem cells, assays and high throughput screening capability (HTS).
Competitive Advantages
Current in vitro embryonic assays look at embryonic cell death as an endpoint; however, this does not capture non-lethal affects that could lead to issues such as failure to thrive and develop normally. This technology can be used to assess compounds and other environmental conditions for sub-lethal stress effects on stem cells. The systems and methods can be used to screen compounds and other environmental conditions for stress effects on stem cells before the compounds or other environmental conditions are developed, commercialized or released into the workplace or the atmosphere. The systems and methods include modified stem cells, assays and high throughput screening capability (HTS).
Benefit Analysis:
May provide a new assay to be used for the testing of 1) Pharmaceutical compounds undergoing Developmental and Reproductive Toxicity (DART) testing, 2) testing of manufactured chemicals, such as those that must meet the criteria for the Regulation for Registration, Evaluation, Authorization and Restriction of Chemicals (REACH) and 3) testing of cosmetics where animal testing is banned. The DART market alone
Stage of Development: Pre-Clinical
Work is currently focused on the development of a high-throughput screening assay system. Reporters are being generated for key biomarkers of potency, differentiation, lineages and other critical criteria.
Patent Status:
Through the Office of Technology Commercialization at Wayne State University, we have filed an extensive and detailed patent application to the USPTO office. This is a detailed methods and composition of matter application with both specific and fairly broad claims. The filing contains definitions for of the technology discoveries, reporters, assays and uses for the technology; which will provide a strong intellectual property position.
Licensing Opportunity:
WSU is looking for a commercial partners interested in furthering the validation of this technology.
Contact for Further Information:
Frank Urban, MS, CBA, BA. email: frank.urban@wayne.edu Phone (mobile): (734) 355-0730