In the next few paragraphs, the component research projects are listed and we summarize their relationships to the central theme of the P01, which is ‘the regulation of trophoblast lineage development’. Additionally, the core units are highlighted and their planned support for the execution of the proposed research briefly described.
The theme of this research is to elucidate TEAD4-dependent mechanisms regulating trophoblast lineage development. TEAD4 nuclear localization is required for trophoblast determination and maintenance of the TS cell state, an evolutionarily conserved process dependent upon BMP4 signaling. The proposed research is largely based on experimentation with mouse ES and TS cell models and manipulation of mouse embryos. Rodent and human stem cell models will be the primary experimental systems used in the planned experimentation. The proposed investigation will benefit from access to each of the Core Units.
The theme of this research is to investigate the role of SATB proteins in the regulation of the TS cell state. SATB proteins promote the TS cell stem state and inhibit trophoblast differentiation. We seek to identify the position of SATB proteins within the gene regulatory network controlling development of the trophoblast lineage. The proposed research is based on experimentation with mouse and rat TS cells, mice possessing mutations in SATB genes, and localization of SATB1 within presumptive TS cell populations localized to first trimester human placentation sites. The proposed research will continue to utilize a multi-faceted approach and will benefit from access to each of the Core Units.
The theme of this research is to investigate the impact of histone H3K9 methylation state on development of trophoblast lineages. Histone H3K9 methylation status changes dramatically at specific genetic loci during trophoblast cell lineage development. Furthermore, a key histone H3K9 methyltransferase (SUV39H2) and a histone H3K9 demethylase (KDM3A) are differentially regulated during trophoblast lineage development and impact the developmental fate of TS cells (D. Chakraborty and M.J. Soares, unpublished, see RESEARCH PROJECT III). The proposed research will utilize rodent and human stem cell models, in vivo rat models, and first trimester human placentation sites. The proposed investigation will benefit from access to each of the Core Units.
The Administrative Core Unit will provide support for all phases of the program project, including and especially facilitating efficient and productive interactions among the research project investigators. The impact of the Administrative Core Unit will be improved efficiency and increased productivity. The success of the program project will be directly linked to the communication and coordination of the component projects. Central to the Administrative Core unit’s mission is to promote interactions and research collaborations among members of the research team and with members of the External Scientific Advisory Committee. The Administrative Core Unit will also provide administrative support for the research core unit (Core Unit B).
The Stem Cell and Trophoblast Analysis Core Unit will facilitate all aspects of research with mouse and human ES cells and mouse and rat TS cells. Each of the research projects proposes to use stem cells and trophoblast cells in their research. The Core will contribute to the research effort through the following activities: i) acquisition and preparation of cell culture medium and related reagents; ii) maintaining stem cell stocks and standardization of cell culture protocols; iii) mycoplasma testing of cell cultures; iv) cell manipulation and harvesting for large-scale experimentation; v) oversee and schedule use of shared equipment; vi) coordination of genome-wide sequencing and bioinformatic analyses of stem cells; vii) development of new strategies for studying trophoblast lineage development. Access to the Stem Cell and Trophoblast Analysis Core Unit will expand the capabilities of each research project and represent the most cost effective method to carry out essential aspects of the proposed experimentation.