Roles of DPA derived specialized pre-resolving mediators during vertebrate hematopoiesis

DPA 衍生的专门预解析介质在脊椎动物造血过程中的作用

• 批准号: 10752549
• 负责人: Kim Uyen Nguyen
• 金额: $ 3.17万
• 依托单位: UNIVERSITY OF MISSISSIPPI MED CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752549
• 关键词: Adult; Affect; Agonist; American; American Cancer Society; Anemia; Animal Model; B-Lymphocytes; Bacteria; Blood; Blood Cell Count; Blood Cells; Blood Transfusion; Blood coagulation; Carbon; Cell Count; Cell Lineage; Cells; Cessation of life; Chemicals; Country; Data; Diagnosis; Dianisidine; Disease; Disease Progression; Disease model; Economic Burden; Economics; Embryo; Erythrocytes; Erythroid; Expenditure; Exposure to; Face; Fatty Acids; Fertilization; Foundations; Genetic; Genetic Diseases; Goals; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hour; Human body; Immune system; Immunosuppression; In Situ Hybridization; Infection; Inflammation; Intervention; Isomerism; L-Plastin; Lymphoid; MYB gene; Macrophage; Malignant - descriptor; Malignant Neoplasms; Mediating; Mediator; Messenger RNA; Microscopy; Modeling; Molecular; Myelogenous; N-3 polyunsaturated fatty acid; Natural Compound; Omega-3 Fatty Acids; Oral; Outcome; Patients; Persons; Pharmaceutical Preparations; Play; Polyunsaturated Fatty Acids; Proliferating; Rag1 Mouse; Resolution; Role; Stains; System; T-Lymphocyte; Testing; Therapeutic; Therapeutic Intervention; Time; Transgenic Organisms; Vertebrates; Visualization; Work; Zebrafish; blood treatment; cancer therapy; cell type; cost; docosapentaenoic acid; fluorescence imaging; granulocyte; hematopoietic differentiation; in vivo; meter; monocyte; neutrophil; novel; novel therapeutics; palliating symptoms; pi bond; prevent; progenitor; receptor; sample fixation; screening; self-renewal; social; stem cell proliferation; stem cells

Blood disease, including anemia, blood clotting, and blood cancers, affects millions of people worldwide each year, causing tremendous economic and social burdens. The American Cancer Society estimated 178,520 blood cancer cases in 2020 in America and approximately 5,600 patients died of anemia in 2019 in the US. Our country faces at least a 45 billion dollars economic burden annually due to blood related diseases. However, the current treatments of blood-related diseases only palliate the symptom and temporarily prevent the disease progression. The goal of this project is to investigate the roles played by specialized pro-resolving mediators (SPMs) derived from polyunsaturated fatty acid (PUFA) n-3 docosapentaenoic acid (DPAn-3) during vertebrate hematopoiesis and evaluate in vivo their therapeutic potentials for blood disorders. I performed chemical screen in zebrafish embryos using several ecosanoid derived SPMs and whole-mount in situ hybridization (WISH) to identify compounds that are able to increase Hematopoietic stem cell (HSC) and red blood cell (RBC). This screen identified two promising SMP compound hits: 7,17 dihydroxy DPA (7,17 diHDPAn-3) and 19,20 epoxy DPA (19,20 EpDPA). In this proposal I will evaluate their effects on the proliferation of downstream hematopoietic lineages, including myeloid and lymphoid lineages by exposing wild-type zebrafish embryos to 1.5 µM 7,17 diHDPAn-3 and 2.5 µM 19,20 EpDPA from 11-36 hours post fertilization (hpf). After fixation, I will perform WISH and real-time quantitative PCR (RT-qPCR) using c-myb, gata1, l-plastin, lyz, mpeg, rag1 and CD79 mRNA probes and primers to descriptively and quantitatively evaluate the resulting effects of the two SPMs exposure on the proliferation of hematopoietic stem cells, progenitor erythroid, granulocytic cells, neutrophils, and macrophages, T and B cells respectively. Our preliminary data show that both 7,17 diHDPAn-3 and 19,20 EpDPA promote HSC proliferation, increase RBC progenitors and RBC number, and increase neutrophil cell number. Therefore, I hypothesize that 7,17 diHDPAn-3 and 19,20 EpDPA exposure enhances HSC proliferation, increasing downstream mature blood cell types in vertebrates. To test our central hypothesis, I developed the following two specific aims: Aim 1. To test the hypothesis that the two selected SPMs increase HSC number, leading to increases in myeloid-derived cell and lymphoid-derived cell numbers and Aim 2. To test the hypothesis that the two selected SPMs can be used as therapeutic interventions in disease models such as bacteria- mediated infection, hematopoiesis genetic diseases, and post-irradiated adult zebrafish. This work will characterize the roles of these two SPMs during vertebrate hematopoiesis and will test their therapeutic potential in vivo. The outcome of this proposal will discover an unknown role for these two SPMs in regulating hematopoiesis, opening a new road to developing a novel therapeutic therapy for blood-related diseases.

血液疾病，包括贫血、凝血和血癌，影响着全世界数百万人 年，造成巨大的经济和社会负担。美国癌症协会估计有 178,520 例血液 2020 年美国癌症病例数，2019 年美国约有 5,600 名患者死于贫血。我们的国家 由于血液相关疾病，每年面临至少 450 亿美元的经济负担。然而，当前 血液相关疾病的治疗只能缓解症状并暂时阻止疾病进展。 该项目的目标是调查由专门的促解决调解员（SPM）衍生的角色所扮演的角色 脊椎动物造血过程中来自多不饱和脂肪酸 (PUFA) n-3 二十二碳五烯酸 (DPAn-3) 并评估其对血液疾病的体内治疗潜力。我对斑马鱼进行了化学筛选 使用多种 Ecosanoid 衍生的 SPM 和整体原位杂交 (WISH) 来鉴定胚胎 能够增加造血干细胞（HSC）和红细胞（RBC）的化合物。这个画面 确定了两种有前景的 SMP 化合物：7,17 二羟基 DPA (7,17 diHDPAn-3) 和 19,20 环氧 DPA (19,20 EpDPA)。在这个提案中，我将评估它们对下游造血细胞增殖的影响 通过将野生型斑马鱼胚胎暴露于 1.5 µM 7,17 来分析谱系，包括骨髓和淋巴谱系 受精后 11-36 小时 (hpf) 的 diHDPAn-3 和 2.5 µM 19,20 EpDPA。固定后，我将执行WISH 使用 c-myb、gata1、l-plastin、lyz、mpeg、rag1 和 CD79 mRNA 进行实时定量 PCR (RT-qPCR) 探针和引物，描述性和定量地评估两种 SPM 暴露所产生的影响 对造血干细胞、祖红系细胞、粒细胞、中性粒细胞和 分别为巨噬细胞、T 细胞和 B 细胞。我们的初步数据显示 7,17 diHDPAn-3 和 19,20 EpDPA 促进HSC增殖，增加红细胞祖细胞和红细胞数量，增加中性粒细胞数量。 因此，我假设 7,17 diHDPAn-3 和 19,20 EpDPA 暴露会增强 HSC 增殖， 增加脊椎动物下游成熟血细胞类型。为了检验我们的中心假设，我开发了 以下两个具体目标： 目标 1. 检验两个选定的 SPM 增加 HSC 数量的假设， 导致骨髓源性细胞和淋巴源性细胞数量增加，目标 2。检验假设 两种选定的 SPM 可用作疾病模型（例如细菌）的治疗干预措施 介导的感染、造血遗传疾病和照射后的成年斑马鱼。这项工作将 描述这两种 SPM 在脊椎动物造血过程中的作用，并将测试它们的治疗潜力 体内。该提案的结果将发现这两个 SPM 在监管方面的未知作用 造血，为开发血液相关疾病的新型治疗方法开辟了新的道路。