Single cell approach to uncovering factors regulating HSC division symmetry in vivo

单细胞方法揭示体内调节 HSC 分裂对称性的因素

• 批准号: 10208868
• 负责人: Keisuke Ito
• 金额: $ 55.63万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10208868
• 关键词: Acceleration; Affect; Behavior; Biological Assay; Bone Marrow; Bone Marrow Cells; CD34 gene; Candidate Disease Gene; Cell division; Cells; Clinical; Cues; Data; Engineering; Environment; Equilibrium; Gene Expression Profiling; Genes; Goals; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Research; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cell subsets; Hematopoietic stem cells; Heterogeneity; High Dose Chemotherapy; Homeostasis; Image; Immunocompetent; Immunotoxins; Individual; Lasers; Marrow; Methods; Modeling; Molecular; Molecular Target; Mus; Non-Malignant; Outcome; PTPRC gene; Pathway interactions; Pattern; Physiological; Proto-Oncogene Protein c-kit; Recovery; Regimen; Research; SLAM protein; Science; Severities; Signal Pathway; Signal Transduction; Site; Surface; System; Techniques; Testing; Time; Transplantation; base; bone; candidate validation; cell type; clinical practice; cohort; conditioning; cranium; cytopenia; daughter cell; differential expression; hematopoietic stem cell expansion; hematopoietic stem cell fate; hematopoietic stem cell niche; hematopoietic stem cell self-renewal; improved; in vivo; insight; irradiation; mouse model; multiphoton microscopy; novel therapeutic intervention; post-transplant; preservation; prospective; reconstitution; restoration; self-renewal; single-cell RNA sequencing; stem cell division; stem cell engraftment; stem cell niche; transcriptomics

ABSTRACT The balance between hematopoietic stem cell (HSC) self-renewal and differentiation directly impacts hematopoietic homeostasis. We hypothesize that signals from the bone marrow microenvironment (or “niche”), together with cues from cell-intrinsic networks, contribute to fine-tuning this balance. However, our understanding of the niche has been limited by the current approach relying on sequential deletion of individual regulatory factors from candidate cells in available mouse models, and analysis of individual HSCs and their in vivo interactions with the niche has also been hindered by the heterogeneity of available HSC-enriched fractions and the technical challenges of imaging HSC fate in vivo. To illuminate the behavior of individual HSCs in vivo, we have established a new technical regimen which includes prospective isolation of HSCs with high purity based on Tie2 positivity, a local transplantation technique which delivers a single HSC under multiphoton microscopy guidance into the bone marrow of a live mouse, and micropipette aspiration to extract single cells after division directly from the marrow for transcriptomic assay. Our project will utilize these advances to describe the molecular basis of HSC fate choice in the niche. This in turn will facilitate novel therapeutic strategies for cell- fate manipulation which could accelerate hematopoietic recovery after transplantation, and possibly contribute to improved transplantation efficiency for non-malignant blood diseases. Thus, the goals of this proposal are three-fold: (1) to identify molecular mechanisms which enhance symmetric self-renewing division of HSCs, (2) to understand the niche factors governing HSC division balance, and (3) to assess the HSC niche under non- genotoxic conditioning. If successful, the proposed research will positively impact the HSC field by identifying molecular targets that will improve hematopoietic recovery after transplantation, and enable improvements in the ex vivo engineering of niche models.

抽象的 造血干细胞（HSC）自我更新和分化之间的平衡直接影响 造血稳态。我们假设来自骨髓微环境（或“利基”）的信号， 与细胞固有网络的线索一起，有助于微调这种平衡。然而，我们的理解 目前的方法限制了利基市场的发展，该方法依赖于顺序删除个别监管 现有小鼠模型中候选细胞的因子，以及单个 HSC 及其体内的分析 与生态位的相互作用也受到可用 HSC 富集组分的异质性的阻碍， 体内 HSC 命运成像的技术挑战。为了阐明体内单个 HSC 的行为，我们 建立了一种新技术方案，其中包括基于高纯度的 HSC 的前瞻性分离 Tie2 阳性，一种局部移植技术，可在多光子显微镜下输送单个 HSC 引导进入活体小鼠的骨髓，并在分裂后用微量移液器抽吸提取单细胞 直接从骨髓中进行转录组分析。我们的项目将利用这些进步来描述 HSC 在生态位中命运选择的分子基础。这反过来又将促进新的细胞治疗策略 命运操纵可以加速移植后的造血恢复，并可能有助于 提高非恶性血液疾病的移植效率。因此，该提案的目标是 三重：(1) 确定增强 HSC 对称自我更新分裂的分子机制，(2) 了解控制 HSC 部门平衡的利基因素，以及 (3) 评估非非 HSC 利基 遗传毒性调理。如果成功，拟议的研究将通过确定 HSC 领域产生积极影响 分子靶标将改善移植后的造血恢复，并能够改善 利基模型的离体工程。

项目成果

Quantification of bone marrow interstitial pH and calcium concentration by intravital ratiometric imaging.

• DOI: 10.1038/s41467-022-27973-x
• 发表时间: 2022-01-19
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Yeh SA;Hou J;Wu JW;Yu S;Zhang Y;Belfield KD;Camargo FD;Lin CP
• 通讯作者: Lin CP

Mitochondrial Contributions to Hematopoietic Stem Cell Aging.

• DOI: 10.3390/ijms222011117
• 发表时间: 2021-10-15
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Morganti C;Ito K
• 通讯作者: Ito K

Leukemia Stem Cells as a Potential Target to Achieve Therapy-Free Remission in Chronic Myeloid Leukemia.

• DOI: 10.3390/cancers13225822
• 发表时间: 2021-11-20
• 期刊: Cancers
• 影响因子: 5.2
• 作者: Ito K;Ito K
• 通讯作者: Ito K

Multilayer omics analysis reveals a non-classical retinoic acid signaling axis that regulates hematopoietic stem cell identity.

• DOI: 10.1016/j.stem.2021.10.002
• 发表时间: 2022-01-06
• 期刊: CELL STEM CELL
• 影响因子: 23.9
• 作者: Schoenberger, Katharina;Obier, Nadine;Romero-Mulero, Mari Carmen;Cauchy, Pierre;Mess, Julian;Pavlovich, Polina, V;Zhang, Yu Wei;Mitterer, Michael;Rettkowski, Jasmin;Lalioti, Maria-Eleni;Jaecklein, Karin;Curtis, Jonathan D.;Feret, Betty;Sommerkamp, Pia;Morganti, Claudia;Ito, Keisuke;Ghyselinck, Norbert B.;Trompouki, Eirini;Buescher, Joerg M.;Pearce, Erika L.;Cabezas-Wallscheid, Nina
• 通讯作者: Cabezas-Wallscheid, Nina

PML at mitochondria-associated membranes governs a trimeric complex with NLRP3 and P2X7R that modulates the tumor immune microenvironment.

• DOI: 10.1038/s41418-022-01095-9
• 发表时间: 2023-03
• 期刊: CELL DEATH AND DIFFERENTIATION
• 影响因子: 12.4
• 作者: Missiroli, Sonia;Perrone, Mariasole;Gafa, Roberta;Nicoli, Francesco;Bonora, Massimo;Morciano, Giampaolo;Boncompagni, Caterina;Marchi, Saverio;Lebiedzinska-Arciszewska, Magdalena;Vezzani, Bianca;Lanza, Giovanni;Kricek, Franz;Borghi, Alessandro;Fiorica, Francesco;Ito, Keisuke;Wieckowski, Mariusz R.;Di Virgilio, Francesco;Abelli, Luigi;Pinton, Paolo;Giorgi, Carlotta
• 通讯作者: Giorgi, Carlotta