In vivo assessment of granulin dependent myeloid cell formation

颗粒蛋白依赖性骨髓细胞形成的体内评估

• 批准号: 10043081
• 负责人: Raquel Espín Palazón
• 金额: $ 11.48万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10043081
• 关键词: Acute Myelocytic Leukemia; Adult; Affect; Biological Process; Biology; Blood; Cell Count; Cell Differentiation process; Cell Lineage; Cell Proliferation; Data; Defect; Development; Disease; Embryo; Family member; Frontotemporal Dementia; GRN gene; Genes; Goals; Growth; Guide RNA; Hand; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic Stem Cell Specification; Hematopoietic stem cells; Histological Techniques; Human; Human Cell Line; Inflammation; Injections; Knowledge; Lead; Lysosomes; Malignant Neoplasms; Messenger RNA; Microglia; Microscopy; Molecular; Myelogenous; Myeloid Cells; Myelopoiesis; Myeloproliferative disease; National Cancer Institute; Nerve Degeneration; Pathogenesis; Pathway interactions; Patients; Pharmacology; Phosphorylation; Population; Process; Proteins; Public Health; Publishing; RNA; Regulation; Research; Role; STAT protein; Signal Transduction; Survival Rate; Testing; Therapeutic; Tissues; Transgenic Organisms; Translating; Vertebrates; Visualization; Zebrafish; base; genetic manipulation; granulin; granulin 3; granulin 4; granulocyte; improved; in vivo; innovation; macrophage; member; monocyte; mutant; myeloid cell development; neutrophil; novel strategies; novel therapeutics; outcome forecast; segregation; tool; wound healing; zebrafish genome

There is a fundamental gap in understanding how Granulin (GRN) regulates myeloid cell differentiation. Continued existence of this gap represents an important problem because, until it is filled, understanding of how Granulin contributes to the development of acute myeloid leukemia would be unknown, and therefore the manipulation of the Granulin pathway to treat these hematopoietic malignances will remain unreachable. The long-term goal is to improve the prognosis of patients suffering from acute myeloid leukemia by expanding their therapeutic options. The overall objective is to define in vivo the myeloid cell populations that require Granulin for proper development as well as the molecular pathway activated by Granulin in myeloid cells. The central hypothesis is that Granulin is essential for proper myeloid lineage differentiation of granulocytes, neutrophils and macrophages through the activation of signal transducers and activators of transcription (STAT) family members. This hypothesis has been formulated on the basis of preliminary data produced by the applicant. The rationale for the proposed research is that understanding the fundamental molecular mechanisms of hematopoietic regulation by Granulin has the potential to translate into better understanding of the pathogenesis of acute myeloid leukemia, blood malignancies with a low cure rate of 24%. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Identify in vivo the myeloid cell populations that require Grna for proper development; and 2) Determine which STAT family members are activated through Granulin for proper myeloid differentiation. Under the first aim, a published grna mutant which has been described by the applicant to have decreased myeloid cell numbers, will be used to identify in vivo the myeloid cell populations whose development is affected by the absence of Grna, both during embryonic and adult hematopoiesis. Powerful in vivo microscopy tools in the zebrafish embryo, histological techniques, and RNA-hybridization probes that are already on hand will be used. Under the second aim, quantitative PCR to pinpoint the STAT family members activated by Grna, and injection of mRNA for each STAT candidate in grna mutant embryos will be performed to rescue myeloid defects. An innovative approach is proposed by taking advantage of the zebrafish genome duplication that resulted in two copies of the ancestral Granulin gene (grna and grnb) to understand in an unprecedented manner the hematopoietic function of Granulin due to the specialization of grna in hematopoietic processes. Since grna is specific to hematopoietic processes, this avoids disruption of other tissues. In addition, the power of the zebrafish for visualization of in vivo development of hematopoiesis is exploited. The proposed research is significant, since it is expected to vertically advance understanding of how Granulin could be contributing to acute myeloid leukemia and how this protein could be manipulated to treat these disorders.

了解颗粒蛋白（GRN）如何调节髓样细胞分化存在根本的差距。 持续存在此差距是一个重要的问题，因为在填补之前，了解如何了解 颗粒蛋白有助于急性髓样白血病的发展，因此 处理这些造血恶性肿瘤的颗粒素途径的操纵将无法达到无法达到。这 长期目标是通过扩大其急性髓样白血病患者的预后 治疗选择。总体目的是在体内定义需要颗粒蛋白的髓样细胞群体 为了适当发育以及髓样细胞中颗粒蛋白激活的分子途径。中央 假设是颗粒蛋白对于粒细胞，中性粒细胞和 巨噬细胞通过激活信号换能器和转录激活因子（STAT）家族成员的激活。 该假设是根据申请人产生的初步数据提出的。理由 对于拟议的研究是，了解造血的基本分子机制 颗粒蛋白的调节有潜力转化为对急性发病机理的更好理解 髓样白血病，血液恶性肿瘤低24％。在强大的初步数据的指导下， 假设将通过追求两个具体目标来检验：1）在体内识别需要 grna以进行适当的发展； 2）确定通过颗粒蛋白激活哪些统计家庭成员的 适当的髓样分化。在第一个目标下，已发表的GRNA突变体已被描述 将髓样细胞数量减少的申请人将用于识别体内髓样细胞群体 在胚胎和成人造血期间，其发育受到缺乏GRNA的影响。 斑马鱼胚胎，组织学技术和RNA杂交探针中强大的体内显微镜工具 将使用的那些将被使用。在第二个目标下，定量PCR以查明统计系列 由GRNA激活的成员，在GRNA突变胚胎中注射每个统计候选者的mRNA将是 表演以挽救髓样缺陷。通过利用斑马鱼来提出一种创新的方法 基因组重复，导致两份祖先颗粒蛋白基因（GRNA和GRNB）的副本理解 空前的方式是GRNA在GRNA中的专业化而导致的 造血过程。由于GRNA特定于造血过程，因此避免了其他 组织。此外，斑马鱼在可视化造血的体内发展的力量是 被利用。拟议的研究很重要，因为有望垂直提高对如何的理解 颗粒蛋白可能导致急性髓样白血病，以及如何操纵该蛋白以治疗 这些疾病。