Molecular Genetic Study of Stem Cells, Cancer Stem Cells, and Stem Cell Aging

干细胞、癌症干细胞和干细胞衰老的分子遗传学研究

• 批准号: 8552797
• 负责人: xianyu s Hou
• 金额: $ 121.67万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8552797
• 关键词: Adult; Animals; Biological; Biological Process; Blood; Cell Adhesion; Cell Aging; Cell Cycle; Cell Proliferation; Cell division; Cell physiology; Cells; Characteristics; Cyclin D1; Development; Disease model; Drosophila genus; E-Cadherin; Embryo; Eye; Fishes; Fruit; Genes; Genetic; Genetic Screening; Hematopoietic stem cells; Homologous Gene; Intestines; Investigation; Kidney; Knock-out; Knowledge; Laboratories; Lead; Life; Location; Malpighian Tubules; Mammals; Mediating; Midgut; Molecular; Molecular Genetics; Mus; Mutation; Natural regeneration; Organ; Pathway interactions; Pattern; Proteins; Ras/Raf; Regenerative Medicine; Regulation; Reporter; Signal Transduction; Signal Transduction Pathway; Stem cells; Stomach; System; Testing; Testis; Therapeutic Uses; Tissues; Tumor Stem Cells; Work; adult stem cell; autocrine; cancer stem cell; cancer therapy; cell age; cell behavior; cell transformation; cell type; daughter cell; fly; genetic manipulation; human disease; in vivo; inhibitor/antagonist; interest; killings; knock-down; male; medical schools; notch protein; novel; ras Oncogene; receptor; self-renewal; smoothened signaling pathway; stem cell differentiation; stem cell fate; stem cell population; tumor

Molecular Genetic Study of Stem Cell and Cancer Stem Cell Regulation in Drosophila and MiceOur laboratory is interested in understanding how intercellular signals regulate development in embryo and regeneration in adult. We have been studying the JAK-Stat signaling in Drosophila. From our work and that of many others, it has emerged that the same signaling controls numerous decisions in development and also regulates stem cells in various adult tissues, including testis, kidney, stomach, and intestine. Current work is directed at understanding how adult stem cells are regulated in Drosophila and mouse in vivo genetic systems.Our current projects have evolved from our earlier studies of the JAK-Stat signal transduction pathway in Drosophila. In 1996, I identified mutations in the Drosophila stat gene (working with Dr. Norbert Perrimon in Harvard Medical School) and opened the field to the use of fly genetics to understand the JAK-Stat signaling mechanisms and functions (Hou et al., Cell, 1996). Over the years, my lab in NCI has made several major discoveries in understanding of the JAK-Stat signaling, including the identification of a receptor for the JAK/STAT signal transduction pathway (Chen et al., Genes Dev., 2002) and the discovery that the JAK/STAT pathway and Cyclin D/Cdk4 cooperatively regulate tumor development in fly blood and eye (Chen et al., Dev. Cell, 2003). During a genetic screen for genes that interact with the JAK-Stat signaling in male germline stem cells (GSCs), we discovered that a RapGEF/Rap signaling regulates stem cell anchoring to the niche by regulating E-cadherin-mediated cell adhesion (Wang et al., Dev. Cell, 2006). We also generated mice carrying a conditional knockout of the RapGEF gene and found that a RapGEF/SCL pathway regulates development of haematopoietic stem cells (Satya et al., Blood, 2010).Using a GFP reporter (Stat-GFP) for the JAK-Stat signaling in Drosophila, we found that the signaling is activated in stem cells in several adult tissues and the Stat-GFP reporter can be used as a stem cell marker. Using the stem cell marker we identified adult kidney multipotent renal and nephric stem cells (RNSCs) in the Drosophila Malpighian tubules (MTs) and demonstrated that an autocrine JAK/STAT signal regulates the kidney stem cell self-renewal (Singh et al., Cell Stem Cell, 2007). The adult fly kidney stem cells are relatively quiescent and only divide once in one week. However, they can become very active and even develop stem cell tumors upon activating the JAK-STAT signal transduction pathway or expressing the activated form of the Ras oncogene (Singh et al., Cell Stem Cell, 2007; Zeng et al., JCP, 2010). The function and anatomical location of adult kidney stem cells are evolutionarily conserved from fly to fish and maybe also to mammals (Zeng and Hou, Cell Stem Cell, 2011).We recently also identified gastric stem cells (GaSCs) in the adult Drosophila gastric and stomach Organs by using the Stat-GFP reporter (Singh et al., Cell Cycle, 2011). We further found that JAK-STAT signaling regulates GaSC proliferation, Wingless signaling regulates GaSC self-renewal, and Hedgehog signaling regulates GaSC differentiation. The differentiation pattern and genetic control of the Drosophila GaSCs are remarkably similar to what are observed in the mouse gastric stem cells.In all stem cell systems that we have checked so far, a common theme emerges: the JAK-Stat pathway is a major signaling that regulates stem cell proliferation and works in combination with other signals to control stem cell fates. The other signals can be differ in different stem cell systems. For example, the JAK-Stat signaling collaborates with the Notch signaling in posterior midgut intestine stem cells (ISCs), with the Ras/Raf signaling in RNSCs, and with the Wingless and Hedgehog signaling in GaSCs. The Stat-GFP is a general stem cell marker; escargot (esg) is a stem cell marker in GSCs, ISCs, and RNSCs; wingless (wg) and patched (ptc) are stem cell markers in GaSCs. Using these stem cell markers, we are performing genetic screens and have so far identified a number of novel regulators of GSCs, ISCs, RNSCs, and GaSCs. Molecular and genetic characterization of several novel regulators is ongoing. Particularly, we found that knocking down several genes selectively killed transformed stem cells in Drosophila. We are currently testing inhibitors of the mammalian homologues of these proteins to selectively kill cancer stem cells in tumors.Reporters of the JAK-Stat signaling and Wg signaling are stem cell markers in several Drosophila tissues. These signalings also regulate stem cells in several mammalian systems. We reasoned that these reporters might be stem cell markers in mice. We are currently developing several reporter mice of the JAK-Stat and Wnt signaling, which will drive GFP and CreER in the target cells. We plan to study adult stem cells (particularly in kidney and stomach) in mice by using the new mice lines. Our investigation and knowledge of stem cells in Drosophila should help analysis of stem cells in mice. In summary, we have already laid the groundwork for both fly and mouse projects, and expect to reap the fruits in the next few years. Using this cross-species approach we expect both to obtain an enhanced understanding of stem-cell regulation and to identify new targets for the treatment of human diseases.

果蝇和小鼠实验室中干细胞和癌症干细胞调节的分子遗传研究有兴趣了解细胞间信号如何调节成人胚胎的发育和再生。我们一直在研究果蝇中的jak-stat信号传导。从我们的工作和许多其他工作中，相同的信号传导控制着许多发展的决定，并调节包括睾丸，肾脏，胃和肠道在内的各种成年组织中的干细胞。当前的工作旨在了解果蝇和小鼠在体内遗传系统中如何调节成年干细胞。我们当前的项目已经从我们对果蝇中Jak-STAT信号转导途径的早期研究中演变而成。 1996年，我确定了果蝇统计基因（与哈佛医学院的诺伯特·佩里蒙（Norbert Perrimon）一起工作）中的突变，并开放了使用蝇遗传学来了解JAK-STAT信号传导机理和功能（Hou等，Cell，Cell，1996）。多年来，我在NCI的实验室在理解JAK-STAT信号传导方面有了一些主要发现，包括鉴定JAK/Stat STAT信号转导途径的受体（Chen等，Genes Dev。，2002）以及JAK/STAT途径和Cyclin D/CDK4在Fly and Eye中的jak/stat途径和Cyclin D/CDK4合作调节tumor and Eye and Eye and and。在与男性种系干细胞（GSC）中与JAK-STAT信号相互作用的基因的遗传筛选期间，我们发现Rapgef/Rap信号传导通过调节E-钙粘着蛋白介导的细胞粘附来调节锚定在细生位的干细胞（Wang等，Dev。Cell，2006）。 We also generated mice carrying a conditional knockout of the RapGEF gene and found that a RapGEF/SCL pathway regulates development of haematopoietic stem cells (Satya et al., Blood, 2010).Using a GFP reporter (Stat-GFP) for the JAK-Stat signaling in Drosophila, we found that the signaling is activated in stem cells in several adult tissues and the Stat-GFP reporter can be used as a干细胞标记。使用干细胞标记，我们确定了果蝇马尔皮亚小管（MTS）中的成年肾脏多能肾细胞和肾脏干细胞（RNSC），并证明自分泌JAK/STAT信号调节肾脏干细胞自我更新（Singh等人，Singh等，Cell Stem pell。，Cell Stem cell，2007）。成年蝇肾干细胞相对静止，仅在一周内进行一次分裂。然而，它们可能会非常活跃，甚至在激活JAK-STAT信号转导途径或表达RAS癌基因激活形式时会发展干细胞肿瘤（Singh等，Cell Stem Cell，2007; Zeng等，JCP，2010）。成年肾脏干细胞的功能和解剖位置在从飞行到鱼类以及哺乳动物的进化上是保守的（Zeng and Hou，hou，Cell Stem Cell，2011年）。我们最近还通过使用Stat-GFP Reporter（Singh et an cell Cycle Cyclecy，2011），在成人果蝇胃和胃中鉴定出胃干细胞（GASCS）。我们进一步发现，JAK-STAT信号传导调节GASC增殖，无翼信号传导调节GASC自我更新，刺猬信号传导调节GASC分化。果蝇GASC的分化模式和遗传控制与小鼠胃干细胞中观察到的相似之处。其他信号在不同的干细胞系统中可能有所不同。例如，JAK-STAT信号传导与后肠里肠道干细胞（ISC）中的Notch信号传导，RNSC中的RAS/RAF信号，以及GASC中的无翼和刺猬信号传导。 Stat-GFP是一般的干细胞标记。 Escargot（ESG）是GSC，ISC和RNSC中的干细胞标记。无翅（WG）和修补（PTC）是GASC中的干细胞标记。使用这些干细胞标记，我们正在执行遗传筛选，到目前为止，我们已经确定了许多新型GSC，ISC，RNSC和GASC的调节剂。几个新型调节剂的分子和遗传表征正在进行中。特别是，我们发现击倒几个基因在果蝇中有选择地杀死了转化的干细胞。 目前，我们正在测试这些蛋白质的哺乳动物同源物的抑制剂，以选择性地杀死肿瘤中的癌症干细胞。JAK-STAT信号传导和WG信号传导的重复蛋白是几种果蝇组织中的干细胞标记物。这些信号还调节了几个哺乳动物系统中的干细胞。我们认为这些记者可能是小鼠的干细胞标记。我们目前正在开发几只JAK-STAT和WNT信号的记者小鼠，这些小鼠将驱动目标细胞中的GFP和Creer。我们计划使用新的小鼠系研究小鼠的成年干细胞（尤其是在肾脏和胃中）。我们对果蝇中干细胞的研究和知识应有助于分析小鼠干细胞。 总而言之，我们已经为Fly和Mouse Projects奠定了基础，并希望在未来几年内获得果实。使用这种跨物种方法，我们希望既可以增强对干细胞调节的理解，又要确定治疗人类疾病的新目标。