Identification of Cell-Specific Transcriptional Programs that Drive Cervical Remo

识别驱动宫颈 Remo 的细胞特异性转录程序

• 批准号: 10231426
• 负责人: Mariano Colon-Caraballo
• 金额: $ 7.05万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10231426
• 关键词: 3-Dimensional; 37 weeks gestation; ATAC-seq; Address; Automobile Driving; Birth; Cell Culture System; Cell Nucleus; Cell model; Cell physiology; Cells; Centers for Disease Control and Prevention (U.S.); Cervical; Cervix Uteri; Chromatin; Collagen Fiber; Collagen Fibril; Competence; Complex; Cues; Data; Data Set; Defect; Deposition; Developed Countries; Development; Discipline of obstetrics; Early identification; Elastic Fiber; Enhancers; Estrogens; Etiology; Event; Extracellular Matrix; Extracellular Matrix Proteins; Fetus; Fiber; Foundations; Functional disorder; Future; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Genomics; Goals; Hormonal; Hormones; Human; Incidence; Individual; Infant; K-Series Research Career Programs; Knockout Mice; Knowledge; Lead; Mechanics; Mediating; Mediator of activation protein; Modification; Molecular; Molecular Genetics; Morbidity - disease rate; Mus; Newborn Infant; Outcome; Pathway interactions; Perinatal mortality demographics; Phase; Phenotype; Population; Predictive Factor; Pregnancy; Pregnancy Complications; Pregnancy Maintenance; Premature Birth; Preparation; Prevention; Process; Progesterone; Proteins; Regulator Genes; Regulatory Pathway; Research Personnel; Resolution; Risk; Risk Factors; Role; Signal Transduction; Solid; Stromal Cells; Structure; System; Technology; Testing; Time; Tissues; United States; Validation; Wild Type Mouse; biomarker identification; cell type; cervical remodeling; clinically relevant; conditional knockout; decorin; early pregnancy; in vivo; innovation; mouse model; neonate; novel; perinatal morbidity; post-doctoral training; pregnant; preservation; programs; promoter; reproductive; resilience; single-cell RNA sequencing; steroid hormone; therapeutic target; transcription factor; translational impact

Premature Birth (PTB) is the most common complication during pregnancy leading to a high incidence of perinatal morbidity and mortality rates in developed countries. According to the CDC, in 2018, 1 of every 10 newborns delivered preterm in the United States. An understanding of the molecular processes that determine a successful pregnancy and on time parturition is essential in order to elucidate the diverse mechanisms by which this process can go awry and lead to a premature birth. The transformation of the cervix from a closed rigid structure to one that can open sufficiently for passage of a term infant is one such process. The significance of the proposed studies is the potential to advance understanding of the dynamic molecular events that regulate cervical remodeling at the cellular level. The focus of this application is to utilize innovative single cell genomic technologies that will allow us to define gene regulatory networks that drive cervical remodeling. We will identify regions of active transcription at single cell resolution in the mouse cervix from nonpregnant, time points in pregnancy, and in labor. Utilizing novel computational approaches previously used to study cellular reprogramming events in development, this data will be used to understand how each cell type transitions its phenotype to implement the dynamic process of remodeling and identify the cell specific transcription factors that drive these cell specific molecular changes. Functional validation of the identified transcription factors will be carried out using a human 3D-cervical cell culture system and studies in normal mice and mouse models with targeted deletion of the transcription factors. The proposed studies will provide a solid and broad foundation of understanding of cellular events that drive cervical remodeling which will allow the discovery of risk factors for premature birth that are not yet known and will uncover new pathways to explore for therapeutic targets for prevention of preterm birth. In addition, these studies will provide me the scientific and technical knowledge required for a successful postdoctoral training and importantly, will set the foundation to apply for K awards aiding in my transition to becoming an independent investigator in the reproductive field.

在发达国家，早产是妊娠期最常见的并发症，导致围产儿发病率和死亡率很高。根据疾控中心的数据，2018年，美国每10名新生儿中就有1名早产。了解决定成功妊娠和按时分娩的分子过程是至关重要的，以便阐明这一过程可能出错并导致早产的各种机制。宫颈从封闭的僵硬结构转变为能够充分张开以供足月婴儿通过的宫颈就是这样一个过程。所提出的研究的意义在于有可能在细胞水平上促进对调节宫颈重塑的动态分子事件的理解。这项应用的重点是利用创新的单细胞基因组技术，使我们能够定义驱动宫颈重塑的基因调控网络。我们将确定小鼠宫颈中单细胞分辨率的活跃转录区域，从未怀孕、怀孕和分娩的时间点。利用以前用于研究发育中的细胞重编程事件的新的计算方法，这些数据将被用来理解每种细胞类型如何转换其表型来实施重塑的动态过程，并确定驱动这些细胞特异性分子变化的细胞特异性转录因子。已识别的转录因子的功能验证将使用人类3D-宫颈细胞培养系统进行，并在正常小鼠和靶向删除转录因子的小鼠模型中进行研究。拟议的研究将为了解推动宫颈重塑的细胞事件提供坚实和广泛的基础，这将有助于发现尚不清楚的早产危险因素，并将揭示新的途径，以探索预防早产的治疗靶点。此外，这些研究将为我提供成功的博士后培训所需的科学和技术知识，更重要的是，将为申请K奖奠定基础，帮助我过渡到生殖领域的独立研究人员。