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Updated: Mar 5, 2026
A Simple Method to Identify Kinases That Regulate Embryonic Stem Cell Pluripotency by High-throughput Inhibitor Screening
Published on: May 12, 2017
High-throughput identification of small molecules that affect human embryonic vascular development
Helena Vazão1, Susana Rosa1, Tânia Barata1,2
1Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal.
Abstract:
Birth defects, which are in part caused by exposure to environmental chemicals and pharmaceutical drugs, affect 1 in every 33 babies born in the United States each year. The current standard to screen drugs that affect embryonic development is based on prenatal animal testing; however, this approach yields low-throughput and limited mechanistic information regarding the biological pathways and potential adverse consequences in humans. To develop a screening platform for molecules that affect human embryonic development based on endothelial cells (ECs) derived from human pluripotent stem cells, we differentiated human pluripotent stem cells into embryonic ECs and induced their maturation under arterial flow conditions. These cells were then used to screen compounds that specifically affect embryonic vasculature. Using this platform, we have identified two compounds that have higher inhibitory effect in embryonic than postnatal ECs. One of them was fluphenazine (an antipsychotic), which inhibits calmodulin kinase II. The other compound was pyrrolopyrimidine (an antiinflammatory agent), which inhibits vascular endothelial growth factor receptor 2 (VEGFR2), decreases EC viability, induces an inflammatory response, and disrupts preformed vascular networks. The vascular effect of the pyrrolopyrimidine was further validated in prenatal vs. adult mouse ECs and in embryonic and adult zebrafish. We developed a platform based on human pluripotent stem cell-derived ECs for drug screening, which may open new avenues of research for the study and modulation of embryonic vasculature.
Insights
Researchers developed a human stem cell-based platform to screen drugs affecting embryonic development. This new method identified compounds impacting embryonic vasculature, offering a more effective alternative to animal testing for drug safety.
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Toxicology
Background:
- Birth defects affect 1 in 33 US infants, often due to environmental chemical and drug exposure.
- Current prenatal drug screening relies on low-throughput animal models, lacking human-specific mechanistic insights.
- Understanding human embryonic development and drug effects is crucial for preventing birth defects.
Purpose of the Study:
- To develop a high-throughput screening platform for molecules impacting human embryonic development.
- To utilize human pluripotent stem cell-derived endothelial cells (ECs) for drug screening.
- To identify compounds affecting embryonic vasculature and their mechanisms of action.
Main Methods:
- Differentiated human pluripotent stem cells into embryonic ECs and matured them under arterial flow.
- Screened compounds for effects on embryonic vasculature using the developed EC platform.
- Validated findings using in vitro models (mouse ECs) and in vivo models (zebrafish).
Main Results:
- Identified two compounds with higher inhibitory effects on embryonic ECs than postnatal ECs.
- Fluphenazine (antipsychotic) was found to inhibit calmodulin kinase II.
- Pyrrolopyrimidine (anti-inflammatory) inhibited VEGFR2, decreased EC viability, induced inflammation, and disrupted vascular networks.
Conclusions:
- A novel screening platform using human pluripotent stem cell-derived ECs was established for drug discovery.
- This platform enables the study and modulation of embryonic vasculature, improving drug safety assessment.
- The findings provide new avenues for understanding and mitigating drug-induced developmental toxicity.
