Dissect the mechanisms underlying interspecies pluripotent stem cell competition

剖析种间多能干细胞竞争的机制

• 批准号: 10576347
• 负责人: Jun Wu
• 金额: $ 35.26万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-18 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10576347
• 关键词: Animals; Apoptosis; Attention; Automobile Driving; Cell Death; Cell Survival; Cells; Chemicals; Chimera organism; Chimerism; Coculture Techniques; Competence; Complement; Complex; Data; Development; Disabled Persons; Distant; Embryo; Embryonic Heart; Endogenous Retroviruses; Endothelium; Engraftment; Ensure; Generations; Genes; Genetic; Growth; Homeostasis; Human; In Vitro; Innate Immune Response; Inner Cell Mass; Kidney; Knockout Mice; Macaca mulatta; Medical; Molecular; Mus; Nature; Organ; Organ Donor; Organ Transplantation; Organogenesis; Pancreas; Play; Pluripotent Stem Cells; Process; RELA gene; Rattus; Research; Research Personnel; Rhesus; Role; Shapes; Signal Transduction; Source; System; TP53 gene; Thymus Gland; Tissues; Transmission Electron Microscopy; Work; blastocyst; comparative; embryo culture; fetal; fitness; human pluripotent stem cell; hypoxia inducible factor 1; implantation; improved; in vitro Model; in vivo; innovation; p53 Signaling Pathway; p65; pluripotency; single-cell RNA sequencing; stem cell survival; success; transcriptome; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Shortage of organs for transplantation is one of the largest unmet medical needs. Researchers are currently working on a variety of ways to increase the number of organs available. Recently, the prospect of producing human organs in animals via interspecies blastocyst complementation has raised important attention and may constitute an innovative approach to overcome the worldwide shortage of donor organs. Interspecies blastocyst complementation works by injecting donor pluripotent stem cells (PSCs) from one species into the organogenesis-disabled blastocyst of a different species. The growing host embryo can provide an emptied developmental organ niche for donor cells to occupy. Despite the potential, however, recent work revealed that although donor PSCs can initially engraft to inner cell mass (ICM) of blastocysts from distantly related host species, their chimeric contribution to early post-implantation stages is low. These results suggest the existence of embryonic xenogeneic barriers between evolutionarily distant species. Unlike chimera formation within the same species, a multitude of factors can differ significantly between species, which preclude robust chimerism. Cell competition, a conserved fitness-sensing process during which fitter cells eliminate neighboring less-fit but viable cells, has been proposed as a surveillance mechanism to ensure normal development and tissue homeostasis, and has also been suggested to act as a barrier to interspecies chimerism. During chimera formation, xenogeneic donor PSCs may be treated as unfit or aberrant cells and targeted for elimination. Most recently, we developed an interspecies PSC co-culture strategy and discovered a previously unknown mode of cell competition between species. Interspecies competition between PSCs occurred in primed but not naive pluripotent cells, and between evolutionarily distant species. By comparative transcriptome analysis, we found that genes related to the NF-κB and p53 signaling pathways, among others, were upregulated in less-fit ‘loser’ human cells. Genetic inactivation of TP53, a core component (P65, also known as RELA) and an upstream regulator (MYD88) of the NF-κB complex in human PSCs could overcome their competitive elimination by co- cultured mouse PSCs, thereby improving the survival and chimerism of human cells in early mouse embryos. Based on extensive preliminary results, in this proposal, we will further dissect the mechanisms underlying interspecies primed PSC competition in both loser and winner cells. And, by suppressing interspecies PSC competition, we aim to improve chimerism and provide the proof-of-concept of interspecies blastocyst complementation from donor PSCs of rhesus monkey, our close kin, in distantly related mouse host.

项目摘要/摘要 移植器官短缺是最大的未满足医疗需求之一。研究人员目前是 采用多种方法来增加可用器官的数量。最近，生产的前景 通过种间胚泡完成的动物中的人体器官引起了重要的关注，可能 构成一种创新的方法来克服全球捐助器官的短缺。种间胚泡 互补是通过从一个物种中注入供体多能干细胞（PSC）的作用 不同物种的器官生成的胚泡。成长的寄主胚胎可以提供一个空的 供体细胞占据的发育器官利基市场。尽管有潜力，但最近的工作表明 尽管供体PSC最初可以植入远距离相关宿主的胚泡的内部细胞质量（ICM） 物种，它们对早期植入后阶段的嵌合贡献很低。这些结果表明存在 进化较远的物种之间的胚胎异构屏障。与嵌合在 同一物种之间，多种因素在物种之间可以有所不同，这排除了鲁棒的嵌合。 细胞竞争是一种配置的健身感应过程，在此过程中，拟合细胞消除了相邻的较少拟合，但 已提出可行的细胞作为监视机制，以确保正常发育和组织 体内平衡，还被建议作为种间嵌合体的障碍。在嵌合体期间 形成，异类供体PSC可以被视为不适或异常细胞，并以靶向消除。最多 最近，我们开发了一种种间PSC共培养策略，发现了以前未知的模式 物种之间的细胞竞争。 PSC之间的种间竞争发生在Primed但不天真 多能细胞，以及在进化上不同的物种之间。通过比较转录组分析，我们发现 与NF-κB和p53信号通路相关的基因在较小的“失败者”中被上调 人类细胞。 TP53，核心成分（p65，也称为RELA）和上游的遗传失活 人类PSC中NF-κB复合物的调节剂（MYD88）可以通过共同克服其竞争性消除 培养的小鼠PSC，从而改善了早期小鼠胚胎中人类细胞的生存和嵌合体。 基于广泛的初步结果，在此提案中，我们将进一步剖析基本机制 种间种间启动了失败者和赢家细胞中的PSC竞争。并且，通过抑制种间PSC 竞争，我们旨在改善嵌合体并提供种间胚泡的概念验证 来自遥远相关的小鼠宿主的恒河猴的供体PSC的补充。