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: 10201594
• 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 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201594
• 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; Prognosis; 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; 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.

对于颗粒蛋白（GRN）如何调节骨髓细胞分化的理解存在根本性的差距。 这一差距的持续存在代表了一个重要的问题，因为在它被填补之前，了解如何 颗粒蛋白对急性髓系白血病发展的贡献尚不清楚，因此 操纵颗粒蛋白途径来治疗这些造血系统恶性肿瘤仍然是不可能的。这 长期目标是通过扩大治疗范围来改善急性髓系白血病患者的预后 治疗选择。总体目标是在体内定义需要颗粒蛋白的骨髓细胞群 促进骨髓细胞的正常发育以及颗粒蛋白激活的分子途径。中央 假设颗粒蛋白对于粒细胞、中性粒细胞和粒细胞的正常骨髓谱系分化至关重要。 巨噬细胞通过信号转导子和转录激活子 (STAT) 家族成员的激活。 该假设是根据申请人提供的初步数据提出的。理由 所提出的研究的目的是了解造血的基本分子机制 颗粒蛋白的调节有可能转化为更好地了解急性疾病的发病机制 骨髓性白血病等血液恶性肿瘤的治愈率较低，仅为24%。在强有力的初步数据的指导下， 该假设将通过追求两个具体目标来检验：1）在体内鉴定需要的骨髓细胞群 Grna 的适当发展； 2) 确定哪些 STAT 家族成员通过 Granulin 被激活 适当的骨髓分化。在第一个目标下，已发表的 grna 突变体已由 骨髓细胞数量减少的申请人将用于鉴定体内骨髓细胞群 在胚胎和成体造血过程中，其发育均受到 Grna 缺失的影响。 斑马鱼胚胎中强大的活体显微镜工具、组织学技术和 RNA 杂交探针 现有的将被使用。第二个目标是通过定量 PCR 查明 STAT 家族 Grna 激活的成员，并在 grna 突变胚胎中注射每个 STAT 候选者的 mRNA 进行以挽救骨髓缺陷。利用斑马鱼的优势提出了一种创新方法 基因组复制导致了祖先颗粒蛋白基因（grna 和 grnb）的两个拷贝，以了解 由于 grna 的专门化，颗粒蛋白的造血功能以前所未有的方式增强 造血过程。由于 grna 是造血过程所特有的，因此可以避免干扰其他造血过程 组织。此外，斑马鱼在体内造血发育可视化方面的能力是 被剥削。拟议的研究意义重大，因为它有望纵向推进对如何 颗粒蛋白可能导致急性髓系白血病，以及如何操纵这种蛋白质来治疗 这些疾病。