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: 10525693
• 负责人: Min Yang
• 金额: $ 14.33万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10525693
• 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; Fetal Development; Fetal Research; Fluorescence; GATA3 gene; Gene Expression Profile; Genes; Genomics; Geometry; Goals; Human; Image; In Vitro; Incidence; Knowledge; Maintenance; Modeling; Molecular; Molecular Biology; Morula; Mosaicism; Nature; Organoids; Phase; Phenotype; Placenta; Placentation; Pregnancy; Primates; Reproduction; Reproductive Biology; Reproductive Sciences; Reproductive Technology; Research; Research Personnel; Rodent; Rodent Model; Role; SOX17 gene; Signal Pathway; Signal Transduction; Spontaneous abortion; Study models; System; TP53 gene; Techniques; Technology; Testing; Time; Tissues; Training; Up-Regulation; Work; Yang; aneuploidy analysis; base; career development; chromosome number abnormality; clinical practice; embryonic stem cell; experimental study; gastrulation; genetic technology; human embryonic stem cell; implantation; improved; in vivo; matrigel; migration; natural Blastocyst Implantation; nonhuman primate; preference; preimplantation; 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.

项目概要 胚胎中非整倍性（染色体异常）的存在被认为是主要限制之一 是人类成功繁殖和妊娠失败的重要原因，约占50% 早期流产。体外受精人类胚胎的非整倍体率惊人地高，约为 60% 这些胚胎是嵌合体，包含非整倍体和正常整倍体细胞。频繁出现的情况 自然受孕和体外受精妊娠中都存在嵌合现象。然而，尽管发病率很高 人类胚胎的非整倍性，我们对这些胚胎的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伦理限制，细胞受到限制。 我之前的工作证明了非整倍体在早期分化中的谱系特异性行为，使用体外 人类胚胎干细胞（hESC）模型。为了进一步表征非整倍体的细胞生理学 植入，需要体内动物模型。常见的狨猴表现出自然发生的非整倍体， 使它们成为比啮齿类动物更具代表性的人类模型。因此，我提出了狨猴模型 进一步剖析非整倍体细胞的命运及其在早期发育过程中的分子和细胞后果。我的 初步数据显示，非整倍体狨猴胚胎干细胞（cj-ESC）优先分化 响应 BMP4 刺激进入滋养外胚层谱系，类似于我在 hESC 中观察到的行为 我之前的工作表明，非整倍性在限制干细胞胚胎外命运方面发挥着保守作用。 在训练期间，我将使用独特的狨猴干细胞模型（原肠胚）来概括早期 谱系规范和原肠胚形成研究 BMP4 信号传导在表型表现中的作用 非整倍体（目标 1）。为了进一步研究非整倍体的消除和分配，我将构建镶嵌 狨猴胚胎探测植入前和植入后胚胎期间非整倍体细胞的命运和行为 体外开发（目标 2）。因为我的初步数据表明对非整倍体的容忍度更高 胚胎外谱系，在颁奖期间的独立阶段，我建议分析基因 狨猴胎盘中非整倍性的表达谱，以了解非整倍性对细胞的影响 胚胎外组织的生理学。此外，在这个阶段，我将构建一个胎盘/滋养层 来自 cj-ESC 的类器官，以进一步剖析不同胎盘谱系中非整倍性的行为（目标 3）。 总之，拟议的研究将提出一个综合模型，用于研究以前未表征的 胚胎发生过程中消除非整倍性的机制，为转化铺平道路 辅助生殖技术的应用。拟议的项目也将作为一个平台，让我 获得分子和发育生物学、动物生殖科学方面的培训和科学专业知识， 和计算基因组学，这将对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。