Mechanisms of Synergistic Regulation of Stem/Progenitors

干/祖细胞协同调控机制

• 批准号: 8209131
• 负责人: HAL E. BROXMEYER
• 金额: $ 38.12万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8209131
• 关键词: Acetylation; Adult; Aging; Apoptosis; BFU-E; Biology; Blood; Blood Cells; Bone Marrow; Cell Cycle; Cell Cycle Checkpoint; Cell Lineage; Cell Maintenance; Cell physiology; Cells; Cellular Stress; Chemicals; Data; Deacetylase; Deacetylation; Disease; ES Cell Line; Embryonic Stem Cell Transplantation; Enzymes; Equilibrium; Erythroid; Etoposide; Event; Family; Fetal Liver; Genes; Genotoxic Stress; Genotype; Grant; Growth; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Hypoxia; In Vitro; Knowledge; Laboratories; Life; Light; Link; Longevity; Maintenance; Malignant - descriptor; Mammals; Maps; Marrow; Mediating; Mediator of activation protein; Metabolism; Mitogen-Activated Protein Kinases; Mus; Nocodazole; Non-Malignant; Oxygen; Oxygen measurement, partial pressure, arterial; Paclitaxel; Pathway interactions; Phenotype; Phosphorylation; Phosphotransferases; Play; Population; Post-Translational Protein Processing; Process; Production; Progress Reports; Proliferating; Protein p53; Proteins; Proto-Oncogene Protein c-kit; Publications; Radiation; Reactive Oxygen Species; Recording of previous events; Regulation; Relative (related person); Replacement Therapy; Role; SLAM protein; Saccharomycetales; Signal Pathway; Signal Transduction; Signaling Molecule; Spleen; Stem cells; Stress; Techniques; Testing; Umbilical Cord Blood; Undifferentiated; Western Blotting; Withdrawal; age related; biological adaptation to stress; cell type; chemokine; clinical application; cytokine; embryonic stem cell; granulocyte; human FRAP1 protein; improved; in vivo; insight; irradiation; leukemia inhibitory factor; macrophage; member; mutant; neuronal cell body; peripheral blood; progenitor; public health relevance; reconstitution; response; self-renewal; stem; stress tolerance

DESCRIPTION (provided by applicant): Hematopoiesis is balanced by self-renewal, proliferation, survival and differentiation of hematopoietic stem (HSC) and progenitor (HPC) cells. HSC/HPC are used to treat and cure malignant and non-malignant blood and other disorders. Knowledge of factors regulating HSC/HPC function, and mechanistic insight into this will allow for modulation and more efficacious use of HSC and HPC for therapy. Embryonic stem cells (ESC), are pluripotent, have extensive self-renewal capacity, give rise to all cells of the body including blood cells, and are being considered for cell replacement therapy. We believe that insight into factors mediating HSC or ESC function will provide valuable information on mechanisms applicable to the other stem cell type. Sirt1, a member of the Sirtuin family of deacetylases, has been linked to longevity and stress tolerance in budding yeast and other lower eukaryocytes, and to aging, metabolism and stress tolerance in mammals. We recently identified roles for Sirt1 in maintenance and differentiation of mouse ESC and mouse fetal liver and adult marrow HPC in response to stress. We hypothesize that Sirt1 is an important intracellular regulator of mouse HSC and ESC function, effects especially relevant under various types of cellular stress. The following specific aims are proposed to evaluate this hypothesis: (1) Determine role(s) of Sirt1 on hematopoiesis and on HSC and HPC function in vivo and in vitro in mice under normal conditions, and in the context of stress, and on mouse ESC function. (a) Elucidate effects of Sirt1 in vivo on self-renewal, proliferation, survival, and differentiation of HSC and HPC under unperturbed conditions, and in the context of aging. Evaluate absolute numbers and cycling status of functionally- and phenotypically-defined HSC and HPC in fetal liver, and adult bone marrow and spleen, and on mature blood cell lineages in blood, bone marrow and spleen using Sirt1 -/-, Sirt1 +/-, and littermate control +/+ mice. (b) Elucidate influence in vitro of Sirt1 on different stresses (genotoxic: nocodazole, paclitaxel, etoposide; oxidative: hypoxia and ROS; apoptosis: cytokine withdrawal and irradiation) to Sirt 1 -/-, +/-, and +/+ HSC/HPC from fetal liver and adult bone marrow. (c) Determine role(s) of Sirt1 on self-renewal, proliferation, survival, and differentiation of murine ESC in presence and absence of Leukemia Inhibitory Factor (LIF) and under conditions of varying levels of stresses noted in Aim 1b using Sirt 1 -/- cells, parental (+/+) ESC, and Sirt1 -/- cells in which Sirt1 expression has been restored, and on normal ESC in which expression/activity levels of Sirt 1 are up- or down-modulated. (2) Obtain insight into intracellular signals mediating function in murine ESC lines and primary mouse bone marrow HSC and HPC by focusing on candidate molecules (p53, mTOR and MAP Kinase) in the Sirt 1 pathway. PUBLIC HEALTH RELEVANCE: Blood cell production is regulated by the functional activities of hematopoietic stem and progenitor cells which give rise to blood cells, while embryonic stem cells can give rise to all cells in the body, including blood cells. We have focused our studies on Sirt1, an enzyme previously linked to aging, metabolism, and tolerance to stress in mammals, and we hypothesize that Sirt1 is an important regulator within the cell that mediates functions of hematopoietic and embryonic stem cells, especially under various types of stress including chemicals, oxygen levels, and radiation. The specific aims of our grant are to understand the roles that Sirt1 play in the ability of these mouse stem cells to make more of themselves, and to divide, survive, and become more mature functional cells that the body needs to live, and to gain mechanistic insight into these effects, information that will help us better understand how to modulate stem cells for improved, more efficient treatment.

描述（由申请人提供）：造血通过造血茎（HSC）和祖细胞（HPC）细胞的自我更新，增殖，存活和分化来平衡。 HSC/HPC用于治疗和治愈恶性和非恶性血液以及其他疾病。了解调节HSC/HPC功能的因素的知识以及对此的机械洞察力的了解将允许调节并更有效地使用HSC和HPC进行治疗。胚胎干细胞（ESC）具有多能，具有广泛的自我更新能力，引起人体的所有细胞，包括血细胞，并且是 被考虑用于细胞替代疗法。我们认为，洞悉介导HSC或ESC功能的因素将提供有关适用于其他干细胞类型的机制的宝贵信息。 SIRT1是脱乙酰基酶家族的成员，与出现的酵母和其他下真核生细胞的寿命和胁迫耐受性有关，以及哺乳动物的衰老，代谢和胁迫耐受性。我们最近确定了SIRT1在响应压力的响应时，SIRT1在小鼠ESC和小鼠胎儿肝脏以及成年骨髓HPC的维持和分化中的作用。我们假设SIRT1是小鼠HSC和ESC功能的重要细胞内调节剂，在各种类型的细胞应激下尤其相关。提出了以下具体目的来评估这一假设：（1）在正常条件下以及在压力下以及小鼠的情况下，在小鼠的体内和体外确定SIRT1在造血和HPC功能上的作用，以及小鼠ESC功能。 （a）阐明体内SIRT1对HSC和HPC自我更新，增殖，生存和分化的影响，在不受干扰的条件下以及在衰老的背景下。评估胎儿肝脏功能和表型定义的HSC和HPC的绝对数量和循环状态，成年骨髓和脾脏，以及使用SIRT1 - / - ，SIRT1 +/-，以及静脉疱疹控制 +/ +/ +小鼠在血液，骨髓和脾血液，骨髓和脾脏中成熟的血细胞谱系上。 （b）阐明SIRT1体外影响对不同应力的影响 （遗传毒性：诺科唑，紫杉醇，依托泊剂；氧化：缺氧和ROS；凋亡：细胞因子的戒断和辐照）至SIRT 1 - / - ， +/ - ， +/ +/ +HSC/HPC，来自胎儿肝脏和成人骨髓。 (c) Determine role(s) of Sirt1 on self-renewal, proliferation, survival, and differentiation of murine ESC in presence and absence of Leukemia Inhibitory Factor (LIF) and under conditions of varying levels of stresses noted in Aim 1b using Sirt 1 -/- cells, parental (+/+) ESC, and Sirt1 -/- cells in which Sirt1 expression has been restored, and on normal ESC in which expression/activity levels of Sirt 1是向上或下调的。 （2）通过重点关注SIRT 1途径中的候选分子（p53，mTOR和MAP激酶），从而深入了解鼠类ESC系中的细胞内信号介导功能。 公共卫生相关性：血细胞的产生受造血干细胞和祖细胞的功能活性调节，这些功能会导致血细胞，而胚胎干细胞可以引起人体中的所有细胞，包括血细胞。我们将研究集中在SIRT1上，SIRT1先前与哺乳动物的衰老，代谢和对压力的耐受性相关的酶，我们假设SIRT1是细胞中的重要调节剂，它介导了造血和胚胎干细胞功能，尤其是在各种类型的压力下，尤其是包括化学物质，氧气水平，氧气和辐射水平。我们赠款的具体目的是了解SIRT1在这些小鼠干细胞的能力中发挥更多的作用，使自己更加自身，生存，生存并成为人体需要生存的更成熟的功能细胞，并获得对这些效果的机械洞察力，这些信息，这些信息，这些信息，这些信息将帮助我们更好地了解如何调节干细胞以改善，有效地进行高效，有效的治疗。