Project description:Primary Objective:
* To determine whether celecoxib downregulates GATA-6 expression to upregulate 15-LOX-1 expression and induce apoptosis in human rectal tumors, researchers will measure GATA-6 and 15-LOX-1 expression, 13-S-HODE levels, and apoptosis rates in normal and colorectal polyp epithelial tissues before and after 6 months of celecoxib treatment of patients with familial adenomatous polyposis (FAP).
Project description:Despite morphological similarities, relatively little is known about conserved developmental processes in human and mouse pre-implantation embryos. Here we provide the first comprehensive single-cell RNA-sequencing comparison of human and mouse embryos from the zygotic to the blastocyst stage. We establish a robust computational pipeline allowing us to elucidate human-specific transcriptional Programs. Importantly, we validate our RNA-sequencing findings by Immunofluorescence analysis, which further reveals differences in human and mouseembryo gene expression. For example, although key trophectoderm factors Id2, Elf5, Eomes and Tcfap2c/Ap2? are exclusively localized to this lineage in the mouse, the human orthologues are either absent or expressed in alternative lineages. Importantly, we identify several genes exclusively expressed in the human pluripotent epiblast including the transcription factor KLF17 and key components of the TGF-? signaling pathway LEFTY1, LEFTY2, NODAL and ACVRL1/ALK1 whose expression is absent from the mouse inner cell mass. Conversely, we also identify genes with conserved expression dynamics including Foxa2/FOXA2, which we show for the first time is restricted to the primitive endoderm in both human and mouse embryos. Our analysis highlights significant differences in human pre-implantation development compared to mouse and provides a molecular blueprint to understand human embryogenesis. Single-Cell RNA-seq
Project description:STRT-N is a newly optimized single-cell RNA sequencing method for studies of early genome activation in mammalian preimplantation development. Here, single embryos from the oocyte, 2-cell, 4-cell, 8-cell, blastocyst, and morula stages were sampled for experiments and were sequenced using STRT-N method.
Project description:Human naïve pluripotent cells can differentiate to extraembryonic trophoblast and hypoblast cells. Here we report formation of human blastocyst models by self-organization solely of naïve pluripotent stem cells. The embryo models comprise the three founding lineages, epiblast, trophoblast and hypoblast, arranged to mimic the natural blastocyst. Single-cell RNA sequencing validated the identity of each cell type.
Project description:Transcriptional profiling of common marmoset embryo stages spanning zygote to late preimplantation blastocyst was performed by single-cell RNA-seq.
Project description:Many mammals can temporally uncouple conception from parturition by pacing down their development around the blastocyst stage. In mice, this dormant state is achieved by decreasing the activity of the growth-regulating mTOR signaling pathway1. It is unknown whether this ability is conserved in mammals in general and in humans in particular. Here we show that decreasing the activity of the mTOR signaling pathway induces human pluripotent stem cells (hPSCs) and blastoids to enter a dormant state with limited proliferation, developmental progression, and capacity to attach to endometrial cells. These in vitro assays show that, similar to other species, the ability to enter dormancy is active in human cells around the blastocyst stage and is reversible at both functional and molecular levels. The pacing of human blastocyst development has potential implications for reproductive therapies.
Project description:Many mammals can temporally uncouple conception from parturition by pacing down their development around the blastocyst stage. In mice, this dormant state is achieved by decreasing the activity of the growth-regulating mTOR signaling pathway1. It is unknown whether this ability is conserved in mammals in general and in humans in particular. Here we show that decreasing the activity of the mTOR signaling pathway induces human pluripotent stem cells (hPSCs) and blastoids to enter a dormant state with limited proliferation, developmental progression, and capacity to attach to endometrial cells. These in vitro assays show that, similar to other species, the ability to enter dormancy is active in human cells around the blastocyst stage and is reversible at both functional and molecular levels. The pacing of human blastocyst development has potential implications for reproductive therapies.