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， 2020年美国约有5，600例癌症病例，2019年美国约有5，600例患者死于贫血。我国 由于血液相关疾病，每年面临至少450亿美元的经济负担。但目前 血液相关疾病的治疗只能减轻症状并暂时阻止疾病进展。 本项目的目标是调查的作用所发挥的专门亲解决调解人（SPM）的衍生 从多不饱和脂肪酸（PUFA）n-3二十二碳五烯酸（DPAn-3）在脊椎动物造血过程中 并在体内评价其对血液疾病的治疗潜力。我对斑马鱼进行了化学筛选 胚胎使用几个ecosanoid衍生的SPM和整体安装原位杂交（WISH），以确定 能够增加造血干细胞（HSC）和红细胞（RBC）的化合物。此屏幕 确定了两种有前途的SMP化合物：7，17二羟基DPA（7，17 diHDPAn-3）和19，20环氧DPA (19，20 EpDPA）。在这个建议中，我将评估它们对下游造血细胞增殖的影响。 谱系，包括骨髓和淋巴谱系，通过将野生型斑马鱼胚胎暴露于1.5 µM 7，17 diHDPAn-3和2.5 µM 19，20 EpDPA，从受精后11-36小时（hpf）。固定后，我将执行WISH 以及使用c-myb、gata 1、l-plastin、lyz、mpeg、rag 1和CD 79 mRNA的实时定量PCR（RT-qPCR） 探针和引物，以连续和定量地评估两种SPM暴露的结果 对造血干细胞、红系祖细胞、粒细胞、中性粒细胞的增殖， 巨噬细胞、T和B细胞。我们的初步数据显示，7，17 diHDPAn-3和19，20 EpDPA两者均具有显著的生物学活性。 促进HSC增殖，增加RBC祖细胞和RBC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。为了验证我们的核心假设，我开发了 以下两个具体目标：目标1。为了检验两种选择的SPM增加HSC数量的假设， 导致骨髓来源的细胞和淋巴来源的细胞数量增加，以及Aim 2。为了检验这一假设 这两种选定的SPM可用作疾病模型的治疗干预，如细菌- 介导的感染、造血遗传疾病和辐照后的成年斑马鱼。这项工作将 描述这两种SPM在脊椎动物造血过程中的作用，并测试其治疗潜力 in vivo.这一提议的结果将发现这两个SPM在调节 造血，为开发血液相关疾病的新治疗方法开辟了新的道路。