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 胚胎中的非整倍体（染色体异常）被认为是主要的限制之一 人类成功繁殖和妊娠失败的重要原因，约占50% 早期流产在体外受精的人类胚胎中，非整倍体率非常高，约60% 这些胚胎是嵌合体，包含非整倍体和正常整倍体细胞。的频繁发生 镶嵌现象存在于自然受孕和IVF妊娠中。然而，尽管发病率很高， 人类胚胎中的非整倍体，我们对这些非整倍体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，细胞的研究受到限制。 我以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， 人胚胎干细胞（hESC）模型。为了进一步表征非整倍体的细胞生理学， 植入时，需要体内动物模型。普通绒猴表现出自然发生的非整倍体， 使它们成为比啮齿动物更有代表性的人类模型。因此，我提出了一个绒猴模型 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 初步数据显示，非整倍体绒猴胚胎干细胞（cj-ESCs） 进入滋养外胚层谱系响应BMP 4刺激，类似于我观察到的hESC的行为， 我以前的工作，提出了一个保守的作用，非整倍体限制干细胞胚胎外的命运。 在培训期间，我将使用一种独特的绒猴干细胞模型（gastruloid）， 谱系特化和原肠胚形成研究BMP 4信号传导在表型表现中的作用 非整倍体（Aim 1）。为了进一步研究非整倍体的消除和分配， 绒猴胚胎在植入前和植入后胚胎中探索非整倍体细胞的命运和行为 体外开发（目标2）。由于我的初步数据表明，在非整倍体的耐受性较高， 胚胎外谱系，在奖励期的独立阶段，我建议分析基因 研究绒猴胎盘中非整倍体的表达谱，以了解非整倍体对细胞增殖的影响。 胚外组织生理学此外，在此阶段，我将构建一个胎盘/滋养层 从cj-ESCs的类器官中进一步剖析不同胎盘谱系中的非整倍体行为（目的3）。 总之，拟议的研究将提出一个全面的模型，用于研究以前没有特征的 在胚胎发生过程中消除非整倍体的潜在机制，为翻译铺平了道路。 辅助生殖技术的应用。该项目也将作为一个平台， 获得分子和发育生物学，动物生殖科学， 和计算基因组学，这将大大有助于我的职业发展，作为一个独立的 生殖生物学领域的研究者。