Image guided profiling of the native HSC niche
原生 HSC 利基的图像引导分析
基本信息
- 批准号:10018892
- 负责人:
- 金额:$ 30.86万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2019
- 资助国家:美国
- 起止时间:2019-09-17 至 2022-06-30
- 项目状态:已结题
- 来源:
- 关键词:AddressAnimalsBiological ProcessBlood CellsBone MarrowBone remodelingCXCL12 geneCalvariaCell CompartmentationCell MaintenanceCellsCrowdingDataDepositionDevelopmentEndothelial CellsGoalsGreen Fluorescent ProteinsHematopoieticHematopoietic Stem Cell subsetsHematopoietic stem cellsImageLabelMethodsMinorMolecularMolecular ProfilingMusOpticsOsteoblastsReporterReticular CellSignal TransductionTechniquesTransplantationVascular Endothelial CellVisualizationWorkbasebonecell typeclinical translationcohortgenetic approachhematopoietic stem cell nicheimage guidedimaging approachimprovedintravital imagingsingle cell analysissingle-cell RNA sequencingstem cell biologystem cell nichestem cellstherapy outcometranscriptometranscriptomicstwo-photon
项目摘要
The concept of the stem cell niche is central both to the fundamental understanding of how stem cells are
regulated by their microenvironment, and to clinical translation that targets the microenvironment for
improving therapeutic outcome. The bone marrow (BM), where hematopoietic stem cells (HSCs) reside, is a
crowded space packed with a diversity of cell types derived from both hematopoietic and nonhematopoietic
precursors. A major challenge in studying the HSC niche has been the difficulty in identifying the rare HSCs
and their neighboring cells in the native BM microenvironment. Elegant cell type-specific deletion of molecules
critical for HSC maintenance has led to the identification of vascular endothelial cells (ECs) and
CXCL12-abundant reticular (CAR) cells as two major cell types of the HSC niche. However, deletion of such
factors impacts all ECs and CAR cells that are present throughout the BM, and are therefore not specific in
terms of their local impact in the HSC niche. Direct imaging has the potential to uncover which cell types are in
close contact with the HSCs, provided that specific markers are available for all cell types involved. As
markers for HSCs are now just beginning to emerge, and visualization of minor cell types remains a
challenge, the direct imaging approach has not progressed beyond resolving whether HSCs are in proximity
to ECs, CAR cells, or bone-lining osteoblasts. Imaging on its own also does not provide the molecular
information essential for understanding how the signals from the niche are communicated to the HSCs. We
propose that two things are needed for the field to move forward. First, development of an HSC-specific
reporter mouse will enable the identification of endogenous stem cells in their native microenvironment
without transplantation. Second, development of a method to selectively isolate the cells in close proximity to
the HSCs will enable unbiased profiling of cell types and their molecular signatures (for example, by
single-cell RNA sequencing) involved in HSC maintenance. We have now taken steps to address both of
these needs. First, we have developed (Camargo Lab) a dual genetic strategy in mice that restricts reporter
labeling near exclusively to the most quiescent long-term subset of the HSC compartment (LT-HSCs). This
reporter line is fully compatible with current intravital imaging approaches in the calvarial BM and enables live
animal tracking of native HSCs (Lin Lab) based on the expression of the green fluorescent protein (GFP)
alone, without the need for additional markers and without transplantation. In addition, we have developed a
technique for micropipette aspiration of single cells and cell clusters directly from the BM under two-photon
image guidance, enabling single cell analysis with high spatial definition. Here, we propose to bring the two
teams together to work on an integrated approach for marking, isolating and profiling the native HSCs
together with their neighboring “niche cells”, whose cell types will be identified retrospectively from the
transcriptome profiles.
干细胞生态位的概念对于理解干细胞是如何
受其微环境的调节,以及靶向微环境的临床翻译,
改善治疗效果。造血干细胞(HSC)所在的骨髓(BM)是造血干细胞的主要来源。
一个拥挤的空间挤满了来自造血和非造血细胞类型的多样性
前体研究HSC生态位的一个主要挑战是难以识别罕见的HSC
以及它们在原生BM微环境中的邻近细胞。优雅细胞类型特异性分子缺失
对HSC维持至关重要的研究导致了对血管内皮细胞(EC)的鉴定,
CXCL 12-丰富的网状(CAR)细胞作为HSC小生境的两种主要细胞类型。但是,删除这些
这些因子影响存在于整个BM中的所有EC和CAR细胞,因此在
他们在HSC利基的本地影响方面。直接成像有可能揭示哪些细胞类型在
与HSC密切接触,前提是所有涉及的细胞类型都有特异性标记。作为
HSC的标记物现在才刚刚开始出现,小细胞类型的可视化仍然是一个挑战。
然而,直接成像方法的进展还没有超出解决HSC是否在邻近区域的范围。
到内皮细胞、CAR细胞或骨衬成骨细胞。成像本身也不能提供分子
这些信息对于理解来自小生境的信号如何传递到HSC至关重要。我们
我认为,要使该领域向前发展,需要做两件事。首先,开发HSC特异性
报告小鼠将能够在其天然微环境中鉴定内源性干细胞
没有移植。第二,开发一种方法来选择性地分离细胞,
HSC将能够无偏地分析细胞类型及其分子特征(例如,通过
单细胞RNA测序)。我们现在已经采取措施,
这些需求。首先,我们已经开发了(Camargo实验室)小鼠的双重遗传策略,
几乎专门标记HSC隔室的最静止的长期亚群(LT-HSC)。这
报告子线与颅骨BM中的当前活体成像方法完全兼容,
基于绿色荧光蛋白(GFP)表达的天然HSC的动物追踪(Lin Lab)
单独使用,不需要额外的标记物,也不需要移植。此外,我们还开发了一个
在双光子作用下直接从骨髓中用微管吸取单个细胞和细胞团的技术
图像引导,能够以高空间分辨率进行单细胞分析。在这里,我们建议将两个
团队共同致力于标记,分离和分析天然HSC的综合方法
以及它们邻近的“小生境细胞”,其细胞类型将从
转录组图谱。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Fernando Camargo其他文献
Fernando Camargo的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Fernando Camargo', 18)}}的其他基金
High resolution lineage tracing of developmental hematopoiesis
发育造血的高分辨率谱系追踪
- 批准号:
10585400 - 财政年份:2023
- 资助金额:
$ 30.86万 - 项目类别:
Generation of a temporal, spatial, and molecular map of in situ hematopoiesis
生成原位造血的时间、空间和分子图
- 批准号:
10415468 - 财政年份:2022
- 资助金额:
$ 30.86万 - 项目类别:
Image guided profiling of the native HSC niche
原生 HSC 利基的图像引导分析
- 批准号:
10212380 - 财政年份:2019
- 资助金额:
$ 30.86万 - 项目类别:
Project 1 - Molecular and cellular determinants of hematopoietic clonal expansion
项目 1 - 造血克隆扩增的分子和细胞决定因素
- 批准号:
10641540 - 财政年份:2017
- 资助金额:
$ 30.86万 - 项目类别:
Clonal analysis of hematopoietic stem and progenitor biology in situ
造血干细胞和祖细胞生物学原位克隆分析
- 批准号:
9225236 - 财政年份:2016
- 资助金额:
$ 30.86万 - 项目类别:
Clonal analysis of hematopoietic stem and progenitor biology in situ
造血干细胞和祖细胞生物学原位克隆分析
- 批准号:
9030319 - 财政年份:2016
- 资助金额:
$ 30.86万 - 项目类别:
Reprogramming of liver cell fate by Hippo signaling
通过 Hippo 信号重新编程肝细胞命运
- 批准号:
8676791 - 财政年份:2013
- 资助金额:
$ 30.86万 - 项目类别:
相似海外基金
The earliest exploration of land by animals: from trace fossils to numerical analyses
动物对陆地的最早探索:从痕迹化石到数值分析
- 批准号:
EP/Z000920/1 - 财政年份:2025
- 资助金额:
$ 30.86万 - 项目类别:
Fellowship
Animals and geopolitics in South Asian borderlands
南亚边境地区的动物和地缘政治
- 批准号:
FT230100276 - 财政年份:2024
- 资助金额:
$ 30.86万 - 项目类别:
ARC Future Fellowships
The function of the RNA methylome in animals
RNA甲基化组在动物中的功能
- 批准号:
MR/X024261/1 - 财政年份:2024
- 资助金额:
$ 30.86万 - 项目类别:
Fellowship
Ecological and phylogenomic insights into infectious diseases in animals
对动物传染病的生态学和系统发育学见解
- 批准号:
DE240100388 - 财政年份:2024
- 资助金额:
$ 30.86万 - 项目类别:
Discovery Early Career Researcher Award
Zootropolis: Multi-species archaeological, ecological and historical approaches to animals in Medieval urban Scotland
Zootropolis:苏格兰中世纪城市动物的多物种考古、生态和历史方法
- 批准号:
2889694 - 财政年份:2023
- 资助金额:
$ 30.86万 - 项目类别:
Studentship
Using novel modelling approaches to investigate the evolution of symmetry in early animals.
使用新颖的建模方法来研究早期动物的对称性进化。
- 批准号:
2842926 - 财政年份:2023
- 资助金额:
$ 30.86万 - 项目类别:
Studentship
Study of human late fetal lung tissue and 3D in vitro organoids to replace and reduce animals in lung developmental research
研究人类晚期胎儿肺组织和 3D 体外类器官在肺发育研究中替代和减少动物
- 批准号:
NC/X001644/1 - 财政年份:2023
- 资助金额:
$ 30.86万 - 项目类别:
Training Grant
RUI: Unilateral Lasing in Underwater Animals
RUI:水下动物的单侧激光攻击
- 批准号:
2337595 - 财政年份:2023
- 资助金额:
$ 30.86万 - 项目类别:
Continuing Grant
RUI:OSIB:The effects of high disease risk on uninfected animals
RUI:OSIB:高疾病风险对未感染动物的影响
- 批准号:
2232190 - 财政年份:2023
- 资助金额:
$ 30.86万 - 项目类别:
Continuing Grant
A method for identifying taxonomy of plants and animals in metagenomic samples
一种识别宏基因组样本中植物和动物分类的方法
- 批准号:
23K17514 - 财政年份:2023
- 资助金额:
$ 30.86万 - 项目类别:
Grant-in-Aid for Challenging Research (Exploratory)