Reproductive Health Adverse Outcomes Modeling Platform enables in vivo and in vitro phenotype modeling and molecular mechanism exploration of reproductive health adverse outcomes. In vivo modeling includes whole-animal toxication and genetic engineering editing, while in vitro modeling involves chemical induction and gene intervention at the cellular and organoid levels. The platform facilities comprise basic experimental platforms, cell/embryo culture rooms, and molecular biology laboratories equipped with instruments such as inverted fluorescence microscopes, protein electrophoresis and blotting equipment, gel imaging systems, and real-time fluorescence quantitative PCR instruments, supporting animal model construction, cell and organoid culture, DNA/RNA/protein analysis, and molecular biomarker detection. Additionally, the platform integrates multi-omics analyses to decipher the genetic, epigenetic, and environmental factors' roles in reproductive health adverse outcomes across human populations, animal models, and cell systems.

The laboratory has conducted several applied case studies using in vivo and in vitro models, yielding a series of achievements in exploring the genetic-environmental interactions, birth defects, and related molecular mechanisms. Representative works include: (1) Establishment of in vivo models of cleft lip and palate and neural tube defects caused by pollutants. Through previous exploration and practice, the laboratory has successfully constructed teratogenic models of various environmental pollutants, including exposure to a variety of heavy metals and organic pollutants, which can lead to cleft lip and palate and neural tube defects in fetal rats. Related research papers have been published in SCI journals such as Clinical Epigenetics, Science of the Total Environment, Free Radical Biology and Medicine. (2) In vitro whole embryo culture technology was used to establish a model of neural tube defects induced by pollutants. Combined with the population-based methylation microarray data, an increased incidence of NTD, hypomethylation, up-regulation of Casp8 and excessive apoptosis were found in the benzo (a) pyrene exposed mouse whole embryo culture model in vitro. The findings were published in the journal Clinical Epigenetics. (3) CRISPR/Cas9 technology was used to construct a neural tube defect disease model caused by gene mutation. A somatic variant of MED12 gene p.Arg1782Cys was found in a previous population study. Using CRISPR/Cas9 technology, functional studies in the mouse model showed that functional variant of MED12 gene could lead to neural tube defects. The findings were published in the journal Human Genetics. (4) neural tube organoid models based on embryonic stem cells or induced pluripotent stem cells and using gene editing technology to study gene function at the cellular level. In combination with population-based whole-genome sequencing and targeted sequencing, we found significant enrichment of rare deleterious RAD9B variants in spina bifida cases. We then found that knockdown of RAD9B in human embryonic stem cells (HESCs) affected PAX6 and OCT4 expression, profoundly affected early differentiation and hindered neural tube organoid formation in vitro. The findings were published in the journal Human Mutation.

Integrating the data results of population results, animal experiments and cell models, and aiming at the pain points in the research of disease gene-environment interaction mechanism and opening up the analysis path of multi-level and multi-omics data, the proposer took the lead in building a "people-animal-cell" multi-level research platform, including model construction, sample collection, imaging observation, molecular marker detection and data analysis. At present, it has effectively supported a number of research works [Sci Total Environ. 2021;764:144245; Free Radic Biol Med. 2018;116:149-158; Front Cell Dev Biol. 9:641831; Epigenetics, 222,17(2):133-146; Human Genetics, 2020,139(10):1299-1314; Human mutation, 41(4), 786-799; Epigenetics, 2020, 17(2):133-146; Clinical Epigenetics, 2019,11(1):13].

Principal Investigator: Linlin Wang

Office: Center Laboratory Building 125

Email: linlinwang@pku.edu.cn