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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要对胎盘形成和维持正常滋养层信号通路的新机制见解子宫内膜的分化和侵袭将为预防子宫内膜异位症的新疗法奠定基础。子痫。衰老细胞是胎盘发育和成熟的特征，并且胎盘的变化在胎盘功能障碍中观察到衰老相关的分泌表型（SASP）。在患者中对于先兆子痫，SASP 因子增加与其疾病状态相关。 Senolytics 和 senomorphics 最近被提议作为先兆子痫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，并且新表征组蛋白变体 H2AJ 在胎盘中富集，已知可控制 SASP。这项提议的目的是扰乱人类胚胎干细胞中的 H2AJ 并建立有关 H2AJ 及其作用的基础知识滋养层发育和基因组结构。我们的人类滋养层细胞源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）。这研究将服务于通过鉴定蛋白质来鉴定滋养层发育的新调节因子的长期目标 H2AJ 的伙伴可以作为治疗药物的靶点。分化人类胚胎干细胞如果没有 H2AJ，我们将检查生成的转录、染色质和分泌变化滋养层谱系。使用蛋白质免疫沉淀，我们将鉴定 H2AJ 的未知相互作用因子，提供关于 H2AJ 负载在基因组中的作用的新见解。事实证明，在转型过程中衰老时，细胞经历剧烈的染色质变化，将其异染色质分离成大的聚集在核内部，同时维持高表达的常染色质中的 SASP 基因（图 8）。异染色质远离核周边并维持常染色质滋养层细胞的边界尚不清楚。虽然众所周知，细胞会衰老胎盘，很少有研究描述其染色质结构。由于 H2AJ 似乎对特定 SASP 位点转录上调，H2AJ 丢失可能与分界错误相关常染色质和异染色质的严格结构域之间。在本提案中，我们将确定 H2AJ 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 耗尽并控制滋养层细胞。此外，我们将执行该系统的第一次 Hi-C 和染色质追踪研究。迄今为止，这将是第一项描述 H2AJ 在人类发展和它在滋养层细胞中的作用。这些知识对于为患者发现新的治疗方式至关重要患有先兆子痫。