New approaches in single cell biology - linking stem cell function with molecular profiles in heterogeneous populations

单细胞生物学的新方法——将干细胞功能与异质群体中的分子谱联系起来

• 批准号: 1800757
• 金额: --
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1800757
• 关键词: New; approaches; single; cell; biology

This proposal focuses on understanding the complex biology of normal and mutant stem cells by linking single cell function with single cell molecular profiles. It will combine single cell functional and molecular assays with flow cytometric index-sorting and mathematical modelling in an iterative manner across both normal and mutant stem cell populations. This project requires partners with diverse expertise in single cell functional biology (Kent group), single cell molecular biology (Gottgens group) and computational modelling (GSK Systems Modeling and Translational Biology Group). Mathematicalmodels will be developed by the industrial partner (GSK) to understand the scale and speed of clonal expansion in vivo and this will be compared to indexsorted single cell proliferation and differentiation data collected in vitro. These data will be integrated with single cell RNA-sequencing data of stem/progenitor cells generated for this project and also other projects in the Kent/Gottgens labs to provide a comprehensive molecular understanding of distinct fate choices in single stem cells. Overall this project has the following three objectives: Objective 1:To understand the extent of blood stem cell heterogeneity in vitro and how it relates to the distinct subtypes observed in single stem cell transplantations. Objective 2: To compare cellular dynamics observed in Objective 1 to mutant stem cells with increased (TET2 loss-of-function) or ecreased (JAK2 gain-of-function) self-renewal activity to determine which properties are associated with durable self-renewal. Objective 3:To determine regulators of stem cell self-renewal and functional heterogeneity at the protein level by index-sorting and tandem flow cytometry / mass spectrometry (CyToF).

该提案的重点是通过将单细胞功能与单细胞分子谱联系起来，了解正常和突变干细胞的复杂生物学。它将联合收割机单细胞功能和分子检测与流式细胞仪指数分选和数学建模相结合，以迭代方式跨越正常和突变干细胞群体。该项目需要在单细胞功能生物学（肯特小组）、单细胞分子生物学（Gottgens小组）和计算建模（GSK系统建模和转化生物学小组）方面具有不同专业知识的合作伙伴。数学模型将由工业合作伙伴（GSK）开发，以了解体内克隆扩增的规模和速度，并将其与体外收集的索引分类的单细胞增殖和分化数据进行比较。这些数据将与该项目以及肯特/戈特金斯实验室的其他项目产生的干/祖细胞的单细胞RNA测序数据相结合，以提供对单个干细胞中不同命运选择的全面分子理解。总体而言，该项目有以下三个目标：目标1：了解体外造血干细胞异质性的程度，以及它如何与单次干细胞移植中观察到的不同亚型相关。目标二：将目标1中观察到的细胞动力学与具有增加的（TET 2功能丧失）或增加的（JAK 2功能获得）自我更新活性的突变干细胞进行比较，以确定哪些特性与持久的自我更新相关。目的3：通过指数分选和串联流式细胞术/质谱（CyToF）在蛋白质水平上确定干细胞自我更新和功能异质性的调节因子。

项目成果

Population dynamics of normal human blood inferred from somatic mutations.

从体细胞突变推断出正常人血的种群动力学。

• DOI: 10.1038/s41586-018-0497-0
• 发表时间: 2018-09
• 期刊: Nature
• 影响因子: 64.8
• 作者: Lee-Six H;Øbro NF;Shepherd MS;Grossmann S;Dawson K;Belmonte M;Osborne RJ;Huntly BJP;Martincorena I;Anderson E;O'Neill L;Stratton MR;Laurenti E;Green AR;Kent DG;Campbell PJ
• 通讯作者: Campbell PJ

Identification of novel regulators of developmental hematopoiesis using Endoglin regulatory elements as molecular probes.

使用内皮糖蛋白调节元件作为分子探针鉴定发育造血的新型调节因子。

• DOI: 10.1182/blood-2016-02-697870
• 发表时间: 2016
• 期刊: Blood
• 影响因子: 20.3
• 作者: Nasrallah,Rabab;Fast,EvaM;Solaimani,Parham;Knezevic,Kathy;Eliades,Alexia;Patel,Rahima;Thambyrajah,Roshana;Unnikrishnan,Ashwin;Thoms,Julie;Beck,Dominik;Vink,ChrisS;Smith,Aileen;Wong,Jason;Shepherd,Mairi;Kent,David;Roychoudhu
• 通讯作者: Roychoudhu

Emerging single-cell tools are primed to reveal functional and molecular heterogeneity in malignant hematopoietic stem cells.

新兴的单细胞工具有望揭示恶性造血干细胞的功能和分子异质性。

• DOI: 10.1097/moh.0000000000000512
• 发表时间: 2019
• 期刊: Current opinion in hematology
• 影响因子: 3.2
• 作者: Shepherd MS

Single-cell approaches identify the molecular network driving malignant hematopoietic stem cell self-renewal

• DOI: 10.1182/blood-2017-12-821066
• 发表时间: 2018-08-23
• 期刊: BLOOD
• 影响因子: 20.3
• 作者: Shepherd, Mairi S.;Li, Juan;Kent, David G.
• 通讯作者: Kent, David G.