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Updated: Sep 1, 2025
Forward Genetic Approach to Uncover Stress Resistance Genes in Mice — A High-throughput Screen in ES Cells
Published on: November 11, 2015
Using high throughput screens to predict miscarriages with placental stem cells and long-term stress effects with
Elizabeth E Puscheck1,2,3, Ximena Ruden1, Aditi Singh1,4
1CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, Detroit, Michigan, USA.
Abstract:
A problem in developmental toxicology is the massive loss of life from fertilization through gastrulation, and the surprising lack of knowledge of causes of miscarriage. Half to two-thirds of embryos are lost, and environmental and genetic causes are nearly equal. Simply put, it can be inferred that this is a difficult period for normal embryos, but that environmental stresses may cause homeostatic responses that move from adaptive to maladaptive with increasing exposures. At the lower 50% estimate, miscarriage causes greater loss-of-life than all cancers combined or of all cardio- and cerebral-vascular accidents combined. Surprisingly, we do not know if miscarriage rates are increasing or decreasing. Overshadowed by the magnitude of miscarriages, are insufficient data on teratogenic or epigenetic imbalances in surviving embryos and their stem cells. Superimposed on the difficult normal trajectory for peri-gastrulation embryos are added malnutrition, hormonal, and environmental stresses. An overarching hypothesis is that high throughput screens (HTS) using cultured viable reporter embryonic and placental stem cells (e.g., embryonic stem cells [ESC] and trophoblast stem cells [TSC] that report status using fluorescent reporters in living cells) from the pre-gastrulation embryo will most rapidly test a range of hormonal, environmental, nutritional, drug, and diet supplement stresses that decrease stem cell proliferation and imbalance stemness/differentiation. A second hypothesis is that TSC respond with greater sensitivity in magnitude to stress that would cause miscarriage, but ESC are stress-resistant to irreversible stemness loss and are best used to predict long-term health defects. DevTox testing needs more ESC and TSC HTS to model environmental stresses leading to miscarriage or teratogenesis and more research on epidemiology of stress and miscarriage. This endeavor also requires a shift in emphasis on pre- and early gastrulation events during the difficult period of maximum loss by miscarriage.
Insights
Developmental toxicology faces a critical knowledge gap regarding miscarriage causes. High-throughput screening of embryonic and placental stem cells offers a novel approach to understanding environmental impacts on early embryonic loss.
Area of Science:
- Developmental toxicology
- Reproductive biology
- Stem cell research
Background:
- Massive embryonic loss occurs from fertilization through gastrulation, with limited understanding of miscarriage causes.
- Environmental and genetic factors contribute nearly equally to early embryonic loss, a significant public health concern.
- Existing data on teratogenic or epigenetic imbalances in surviving embryos and their stem cells are insufficient.
Purpose of the Study:
- To propose high-throughput screening (HTS) using embryonic stem cells (ESC) and trophoblast stem cells (TSC) to rapidly assess environmental stresses.
- To investigate the differential responses of ESC and TSC to various stressors, predicting miscarriage or long-term health defects.
- To advocate for increased research in developmental toxicology (DevTox) focusing on pre-gastrulation events and miscarriage epidemiology.
Main Methods:
- Utilizing cultured viable reporter ESC and TSC with fluorescent reporters to monitor cellular responses.
- Exposing stem cells to a range of hormonal, environmental, nutritional, drug, and diet supplement stresses.
- Analyzing changes in stem cell proliferation, stemness, and differentiation as indicators of stress impact.
Main Results:
- Hypothesized that ESC and TSC HTS can rapidly test a wide array of environmental stresses.
- Predicted that TSC are more sensitive to miscarriage-inducing stress, while ESC predict long-term health defects.
- Emphasized the need for more ESC and TSC HTS to model miscarriage and teratogenesis.
Conclusions:
- DevTox testing requires enhanced HTS models using ESC and TSC to accurately predict developmental toxicities.
- Further research into the epidemiology of stress and miscarriage is crucial.
- A shift in focus towards pre- and early gastrulation events is necessary to address maximum embryonic loss.
