Heterogeneity and cellular population dynamics in the placenta

胎盘的异质性和细胞群动态

• 批准号: BB/R008590/1
• 负责人: Steve Charnock-Jones
• 金额: $ 83.47万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR008590%2F1
• 关键词: Heterogeneity; cellular; population; dynamics; placenta

ContextPlacenta-related complications of pregnancy are a major challenge for women's health. Stillbirth is a major cause of perinatal death, causing more than 3,000 deaths each year in the UK and ~2.6million worldwide. Fetal growth restriction results in short term morbidity, poor educational outcomes and predisposes to diabetes and cardiovascular disease in later life. Mechanistic understanding and effective therapies for these complications are lacking. The work proposed here will provide a detailed atlas of all the different cell types in the placenta -currently we don't know how many different cell types and sub-types there are. We will also determine which RNAs are present in each of these cells and this is directly relevant to cell function. Knowledge of how the different cell types arise is key to understanding normal placental development. Methods for the analysis of the mRNA within single cells have recently been developed. However, as the barrier between maternal blood and fetal tissue is a multi-nucleated syncytium current single "cell" RNA-sequencing methods are inappropriate and new approaches are needed. This proposal describes such new methods and their application to the placenta.AimsThe overarching aim of this work is to use single cell RNA sequencing to characterise placental cellular heterogeneity in mouse and man and to define regulatory steps in trophoblast differentiation. To address this aim we will answer the following 3 specific questions:1) How many cell types are there in the placenta and what are their distinguishing features? This will necessitate the development of new methods, specifically to allow the analysis of individual nuclei and the analysis of non-coding RNA in many thousands of single nuclei.2) How does the placental cell population change over time? We will evaluate the different cell populations by single nucleus sequencing from tissue collected from 5 human placentas in the first trimester and also at term. We will compare these with data from samples of mouse placenta collected at 5 different times in pregnancy.3) What are the mechanisms that regulate this heterogeneity?Murine trophoblast stems cells can be grown under conditions that induce differentiation. We will sequence single cells from multiple time points during differentiation. Sampling a population of differentiating cells at the single cell level allows identification of the intermediate populations during differentiation. We will identify regulatory factors at the branch points in this process and test whether over expression or knocking out these genes alters the differentiation.Potential applications and benefitsPreventing even a small proportion of still births and other complications of pregnancy would be a significant benefit. Good obstetric care means that many pregnancies complicated by poor placental development end in live births. However, babies whose intrauterine life was compromised are at risk for long-term chronic diseases, such as cardiovascular disease and diabetes. Better understanding of placental growth and function has the potential to improve pregnancy outcome and this will have both short and long term health and economic benefits. These are distant benefits but more immediate gains will be in the fields of placental, developmental, gene regulatory biology. The methods proposed are innovative and require cross-disciplinary work (microfluidics, chemistry, molecular biology and bioinformatics) and so will benefit all these disciplines.The work proposed here is to be carried out with a collaborative industrial partner - Sphere Fluidics Limited. Close collaboration between the University, the Babraham Institute and Sphere Fluidics will further enhance interactions among biotech/pharma and academia around Cambridge. This will be a benefit to these three parties and may attract R&D investment and increase economic activity to the benefit of the UK economy.

与胎盘有关的妊娠并发症是妇女健康面临的一个重大挑战。死产是围产期死亡的主要原因，每年在英国造成3,000多人死亡，全球约有260万人死亡。胎儿生长受限导致短期发病率、不良教育结果，并在以后的生活中易患糖尿病和心血管疾病。对这些并发症的机制缺乏了解和有效的治疗方法。这里提出的工作将提供胎盘中所有不同细胞类型的详细图谱-目前我们不知道有多少不同的细胞类型和亚型。我们还将确定这些细胞中存在哪些RNA，这与细胞功能直接相关。了解不同类型的细胞是如何产生的是理解正常胎盘发育的关键。最近已经开发了用于分析单细胞内的mRNA的方法。然而，由于母体血液和胎儿组织之间的屏障是多核合胞体，目前的单“细胞”RNA测序方法是不合适的，需要新的方法。该提案描述了这样的新方法和它们的应用程序的placenta.AimsThe总体目标这项工作是使用单细胞RNA测序来识别胎盘细胞的异质性在小鼠和人，并确定在滋养层分化的监管步骤。为了达到这个目的，我们将回答以下3个具体问题：1）胎盘中有多少种细胞类型，它们的区别特征是什么？这将需要开发新的方法，特别是允许分析单个细胞核和分析数千个单个细胞核中的非编码RNA。我们将通过从5个人胎盘中收集的组织中的单核测序来评估不同的细胞群，这些胎盘是在妊娠早期和足月时收集的。我们将这些数据与在妊娠的5个不同时间收集的小鼠胎盘样品进行比较。3）调节这种异质性的机制是什么？鼠滋养层干细胞可以在诱导分化的条件下生长。我们将在分化过程中的多个时间点对单个细胞进行测序。在单细胞水平上对分化细胞群体进行取样允许在分化期间鉴定中间群体。我们将确定在这个过程中的分支点的调节因子，并测试是否过度表达或敲除这些基因改变了分化。潜在的应用和好处预防即使是一小部分死产和其他妊娠并发症将是一个显着的好处。良好的产科护理意味着许多因胎盘发育不良而复杂化的怀孕最终都是活产。然而，宫内生命受到损害的婴儿有患长期慢性疾病的风险，如心血管疾病和糖尿病。更好地了解胎盘的生长和功能有可能改善妊娠结局，这将具有短期和长期的健康和经济效益。这些都是遥远的好处，但更直接的收益将在胎盘，发育，基因调控生物学领域。提出的方法是创新的，需要跨学科的工作（微流体，化学，分子生物学和生物信息学），因此将有利于所有这些学科。这里提出的工作是与合作的工业合作伙伴-球体流体有限公司进行。大学，Babraham研究所和Sphere Fluidics之间的密切合作将进一步加强剑桥周围生物技术/制药和学术界之间的互动。这对这三方都是有利的，可能会吸引研发投资，增加经济活动，使英国经济受益。

项目成果

IFPA meeting 2018 workshop report II: Abnormally invasive placenta; inflammation and infection; preeclampsia; gestational trophoblastic disease and drug delivery.

IFPA 2018年会议研讨会报告二：异常侵入性胎盘；

• DOI: 10.17863/cam.38021
• 发表时间: 2019
• 作者: Albrecht C

Evolutionary History of Endogenous Human Herpesvirus 6 Reflects Human Migration out of Africa.

• DOI: 10.1093/molbev/msaa190
• 发表时间: 2021-01-04
• 期刊: Molecular biology and evolution
• 影响因子: 10.7
• 作者: Aswad A;Aimola G;Wight D;Roychoudhury P;Zimmermann C;Hill J;Lassner D;Xie H;Huang ML;Parrish NF;Schultheiss HP;Venturini C;Lager S;Smith GCS;Charnock-Jones DS;Breuer J;Greninger AL;Kaufer BB
• 通讯作者: Kaufer BB

IFPA meeting 2018 workshop report II: Abnormally invasive placenta; inflammation and infection; preeclampsia; gestational trophoblastic disease and drug delivery

• DOI: 10.1016/j.placenta.2019.02.006
• 发表时间: 2019-09-01
• 期刊: PLACENTA
• 影响因子: 3.8
• 作者: Albrecht, Christiane;Chamley, Larry;O'Tierney-Ginn, Perrie
• 通讯作者: O'Tierney-Ginn, Perrie

Single-Cell Analysis Identifies Thymic Maturation Delay in Growth-Restricted Neonatal Mice

单细胞分析确定生长受限的新生小鼠胸腺成熟延迟

• DOI: 10.1101/372862
• 发表时间: 2018
• 作者: Bacon W

Single-Cell Analysis Identifies Thymic Maturation Delay in Growth-Restricted Neonatal Mice.

单细胞分析确定了生长限制的新生儿小鼠的胸腺成熟延迟。

• DOI: 10.3389/fimmu.2018.02523
• 发表时间: 2018
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Bacon WA;Hamilton RS;Yu Z;Kieckbusch J;Hawkes D;Krzak AM;Abell C;Colucci F;Charnock-Jones DS
• 通讯作者: Charnock-Jones DS