H2AJ as a regulator of placental senescence and genome organization

H2AJ 作为胎盘衰老和基因组组织的调节剂

• 批准号: 10677156
• 负责人: Tyler Barry Jensen
• 金额: $ 3.26万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-16 至 2027-04-15
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677156
• 关键词: 3-Dimensional; Affect; Architecture; BRCA1 gene; BRCT Domain; Cell Aging; Cell Cycle Arrest; Cells; Characteristics; Chromatin; Complex; DNA Damage; DNA Repair; Data Set; Deposition; Development; Disease; Down-Regulation; Drug Design; Drug Targeting; Eclampsia; Endometrium; Enhancers; Euchromatin; Fibroblasts; First Pregnancy Trimester; Fluorescent in Situ Hybridization; Foundations; Functional disorder; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Genome; Genome Mappings; Genome Stability; Goals; Heterochromatin; Hi-C; Histone H2A; Histones; Hormones; Human; Human Development; Immunoprecipitation; Inflammatory; Invaded; Knowledge; Life; Link; Maintenance; Mass Spectrum Analysis; Mentorship; Modality; Modeling; Molecular Conformation; Morbidity - disease rate; Nuclear; Organ; Outcome; Pathogenesis; Patients; Pattern; Phenotype; Physicians; Placenta; Placentation; Pre-Eclampsia; Pregnancy; Pregnant Women; Production; Proteins; Research; Role; Sampling; Scientist; Signal Pathway; Specific qualifier value; Stains; Syncytiotrophoblast; System; Tail; Techniques; Tertiary Protein Structure; Therapeutic; Therapeutic Intervention; Third Pregnancy Trimester; Time; Training Activity; Transcriptional Regulation; Up-Regulation; Variant; Villous; cell type; chromosome conformation capture; cohesin; drug development; embryonic stem cell; experience; fetal; human disease; human embryonic stem cell; insight; knock-down; learning strategy; mortality; normotensive; novel therapeutics; nuclease; placental mammal; pregnancy disorder; pregnant; promoter; response; segregation; senescence; skills; therapeutic target; transcriptome sequencing; transcriptomics; trophoblast; trophoblast stem cell

PROJECT SUMMARY New mechanistic insights into placentation and the signaling pathways that maintain proper trophoblast differentiation and invasion of the endometrium will provide the foundation for new therapeutics against pre- eclampsia. Senescent cells are characteristic of the developing and mature placenta, and changes in the senescence-associated secretory phenotype (SASP) have been observed in placental dysfunction. In patients with pre-eclampsia, increased SASP factors are associated with their disease status. Senolytics and senomorphics have recently been proposed as therapeutic interventions for patients with pre-eclampsia, reducing the intensity of SASP. Histone variants organize the senescent genome, and a newly characterized histone variant, H2AJ, is enriched in the placenta and is known to control SASP. This proposal aims to perturb H2AJ in human embryonic stem cells and establish foundational knowledge about H2AJ and its role in trophoblast development and genome architecture. Our human trophoblast cells derived from human embryonic stem cells recapitulate many aspects of fetal placental development and upregulate H2AJ (Figure 5,6,7). This study will serve the long-term goal of identifying new regulators of trophoblast development by identifying protein partners of H2AJ that may serve as therapeutic drug targets. Differentiating human embryonic stem cells with and without H2AJ, we will examine transcriptional, chromatin, and secretory changes in the generated trophoblast lineages. Using protein immunoprecipitation, we will identify unknown interactors of H2AJ, providing new insight into the role of H2AJ loading in the genome. It has been shown that during the transition to senescence, cells undergo dramatic chromatin changes, segregating their heterochromatin into large aggregates in the nuclear interior while maintaining SASP genes in highly expressed euchromatin (Figure 8). The mobilization of heterochromatin away from the nuclear periphery and the maintenance of euchromatic boundaries are not well understood in trophoblast cells. While it is known that cells become senescent in the placenta, there are few studies describing their chromatin architecture. As H2AJ appears critical for the upregulation of transcription at specific SASP loci, loss of H2AJ may be associated with demarcation errors between the strict domains of euchromatin and heterochromatin. In this proposal, we will identify H2AJ's deposition in the genome and its role in transcription, characterizing the three-dimensional chromatin architecture of H2AJ depleted and control trophoblast cells. Furthermore, we will perform this system's first Hi-C and chromatin tracing studies. To date, this will be the first study to characterize H2AJ in human development and its role in trophoblast cells. This knowledge will be crucial for discovering new treatment modalities for patients experiencing pre-eclampsia.

项目摘要 对胎盘和信号通路的新机械洞察力，以维持适当的滋养细胞 子宫内膜的分化和入侵将为预先抗衡的新疗法提供基础 子痫。衰老细胞是发育和成熟胎盘的特征，并且变化 在胎盘功能障碍中已经观察到与衰老相关的分泌表型（SASP）。在患者中 在先兆子痫之前，SASP因素增加与其疾病状况有关。 Senolotics和 最近提出了鼻象作为治疗前宾夕法尼亚患者的治疗干预措施， 降低SASP的强度。组蛋白变体组织衰老基因组，并有一个新的特征 组蛋白变体H2AJ富集在胎盘中，已知可以控制SASP。该建议旨在扰动 H2AJ在人类胚胎干细胞中，并建立了有关H2AJ及其在其中的作用的基础知识 滋养细胞的发展和基因组建筑。我们的人类滋养细胞细胞来自人类胚胎 干细胞概括了胎儿胎盘发育的许多方面并上调H2AJ（图5,6,7）。这 研究将实现长期目标，即通过识别蛋白质来识别滋养细胞开发的新调节剂 H2AJ的合作伙伴可以用作治疗性药物靶标。将人类干细胞与 没有H2AJ，我们将检查生成的转录，染色质和分泌变化 滋养细胞谱系。使用蛋白质免疫沉淀，我们将确定H2AJ的未知相互作用者，提供 对H2AJ载荷在基因组中的作用的新见解。已经表明，在过渡到 衰老，细胞发生急剧的染色质变化，将其异染色质隔离到大型 核内部的聚集体，同时在高度表达的圣木素中保持SASP基因（图8）。 异染色质的动员从核外围远离和维持正念 在滋养细胞细胞中，边界没有很好地理解。虽然知道细胞在 胎盘，很少有研究描述其染色质结构。由于H2AJ对于 特定SASP基因座的转录上调，H2AJ的损失可能与界定误差有关 在斑塑素和异染色质的严格结构之间。在此提案中，我们将确定H2AJ的 基因组中的沉积及其在转录中的作用，表征三维染色质结构 H2AJ耗尽并控制滋养细胞的细胞。此外，我们将执行该系统的第一个Hi-C和 染色质追踪研究。迄今为止，这将是第一个描述H2AJ在人类发展和 它在滋养细胞中的作用。这些知识对于发现患者的新治疗方式至关重要 体验前宾夕法尼亚。