Characterization of aneuploidy, cell fate and mosaicism in early development

早期发育中非整倍性、细胞命运和嵌合体的表征

• 批准号: 10877239
• 负责人: Min Yang
• 金额: $ 24.89万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-04 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10877239
• 关键词: Accounting; Aneuploid Cells; Aneuploidy; Animal Model; Animals; Apoptosis; Apoptotic; Area; Assisted Reproductive Technology; Award; BMP4; Behavior; Biological Assay; Birth; CASP3 gene; CDX2 gene; Callithrix; Callithrix jacchus jacchus; Career Mobility; Cattle; Cell Culture System; Cell Line; Cell physiology; Cells; Cellular biology; Chromosomal Instability; Chromosome abnormality; Chromosomes; Clinic; Competence; Data; Defect; Development; Developmental Biology; Diagnostic tests; Embryo; Embryo Research; Embryonic Development; Ethics; Exhibits; Failure; Fertilization in Vitro; Fetal Development; Fetal Research; Fluorescence; GATA3 gene; Gene Expression Profile; Gene Expression Profiling; Genes; Genomics; Geometry; Goals; Human; Image; In Vitro; Incidence; Knowledge; Maintenance; Modeling; Molecular; Molecular Biology; Morula; Mosaicism; Nature; Organoids; Pattern; Phase; Phenotype; Placenta; Placentation; Postnidation Embryo Development; Pregnancy; Primates; Proliferating; Reproduction; Reproductive Biology; Reproductive Sciences; Reproductive Technology; Research; Research Personnel; Rodent; Rodent Model; Role; SOX17 gene; Signal Pathway; Signal Transduction; Specific qualifier value; Spontaneous abortion; Study models; System; TP53 gene; Techniques; Technology; Testing; Tissues; Training; Up-Regulation; Work; aneuploidy analysis; blastocyst; career development; chromosome number abnormality; clinical practice; embryo tissue; embryonic stem cell; experimental study; gastrulation; genetic technology; human embryonic stem cell; implantation; improved; in vivo; matrigel; migration; mosaic; natural Blastocyst Implantation; nonhuman primate; preference; response; single-cell RNA sequencing; stem cell biology; stem cell model; stem cells; success; transcriptome; translational applications; trophoblast

PROJECT SUMMARY The presence of aneuploidy (chromosomal abnormalities) in embryos is considered one of the major limitations to successful human reproduction and a significant cause of gestation failure, accounting for approximately 50% of early miscarriages. Aneuploidy rates are strikingly high in in vitro fertilized human embryos, and around 60% of these embryos are mosaic, containing both aneuploid and normal euploid cells. The frequent occurrence of mosaicism exists in both naturally conceived and IVF pregnancies. However, despite the high incidence of aneuploidy in human embryos, our knowledge of the molecular mechanisms and developmental fate of these cells is restricted due to the considerable ethical limitations associated with human embryo and fetal research. My previous work demonstrated lineage-specific behavior of aneuploidy in early differentiation using an in vitro human embryonic stem cell (hESC) model. To further characterize the cellular physiology of aneuploidy after implantation, an in vivo animal model is required. Common marmosets exhibit naturally occurring aneuploidy, making them a more representative model for humans than rodents are. Therefore, I propose a marmoset model to further dissect aneuploidy cell fate and its molecular and cellular consequences during early development. My preliminary data uncovered that aneuploid marmoset embryonic stem cells (cj-ESCs) preferentially differentiate into trophectoderm lineages in response to BMP4 stimulation, similar to the behavior I observed with hESCs in my previous work, suggesting a conserved role of aneuploidy in restricting stem cells to extraembryonic fates. During the training period, I will use a unique marmoset stem cell model (gastruloid) that recapitulates early lineage specification and gastrulation to investigate the role of BMP4 signaling in the phenotypic manifestation of aneuploidy (Aim 1). To further investigate the elimination and allocation of aneuploidy, I will construct mosaic marmoset embryos to probe aneuploidy cell fate and behaviors during pre-and post-implantation embryonic development in vitro (Aim 2). Since my preliminary data indicates a higher tolerance of aneuploidy in the extraembryonic lineages, during the independent phase of the award period, I propose to analyze the gene expression profile of aneuploidy in the marmoset placenta to understand the effects of aneuploidy on the cellular physiology of extraembryonic tissue. In addition, during this phase, I will construct a placental/trophoblast organoid from cj-ESCs to further dissect the behaviors of aneuploidy in different placental lineages (Aim 3). Together, the proposed research will present a comprehensive model for studying a previously uncharacterized mechanism underlying the elimination of aneuploidy during embryogenesis, paving the way for translational applications to assisted reproductive technologies. The proposed project will also serve as a platform for me to obtain training and scientific expertise in molecular and developmental biology, animal reproductive sciences, and computational genomics which will contribute significantly to my career development as an independent investigator in the field of reproductive biology.

项目摘要 胚胎中非整倍性（染色体异常）的存在被认为是主要局限性之一 成功的人类繁殖和妊娠失败的重大原因，约占50％ 早期流产。非整倍性速率在体外受精的人类胚胎中的高度高，约60％ 这些胚胎是镶嵌的，既包含非整倍体和正常的伯倍型细胞。经常发生 镶嵌物存在于自然构想和IVF妊娠中。但是，尽管发生率很高 人类胚胎中的非整倍性，我们对这些分子机制和发育命运的了解 由于与人类胚胎和胎儿研究相关的相当大的道德局限性，细胞受到限制。 我以前的工作表明了使用体外的早期分化中非整倍性的谱系特异性行为 人类胚胎干细胞（HESC）模型。为了进一步表征非整倍性的细胞生理 植入需要体内动物模型。常见的marmoset表现出天然存在的非整倍性， 使它们成为人类比啮齿动物更具代表性的模型。因此，我提出了一个摩尔马塞特模型 为了进一步剖析早期发育过程中非整倍性细胞命运及其分子和细胞后果。我的 初步数据发现，非整倍型果酱胚胎干细胞（CJ-ESC）优先区分 响应于BMP4刺激的疗程，类似于我在hESC中观察到的行为 我以前的工作表明，非整倍性在将干细胞限制为胚外命运方面的保守作用。 在训练期间，我将使用独特的摩尔果干细胞模型（胃lo脚），该模型早日概括 谱系规范和胃肠道，以研究BMP4信号在表型表现中的作用 非整倍性（AIM 1）。为了进一步研究非整倍性的消除和分配，我将构建马赛克 在植入前和植入后胚胎期间，果冻胚胎探测非整倍性细胞的命运和行为 体外发展（AIM 2）。由于我的初步数据表明对非整倍性的耐受性 在奖励期的独立阶段，我建议分析基因 摩尔莫斯群胎盘中非整倍性的表达曲线，以了解非整倍性对细胞的影响 胚外组织的生理学。另外，在此阶段，我将构建胎盘/滋养细胞 从CJ-ESC进行的类器官，进一步剖析了不同胎盘谱系中非整倍性的行为（AIM 3）。 拟议的研究将共同​​提出一个综合模型，用于研究以前未经表征 在胚胎发生过程中消除非整倍性的基础机制，为翻译铺平道路 辅助生殖技术的应用。拟议的项目还将成为我的平台 获得分子和发育生物学，动物生殖科学的培训和科学专业知识， 和计算基因组学，这将为我作为独立的职业发展做出重大贡献 生殖生物学领域的研究者。