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，血液衣服和血液癌，每个人都会影响全球数百万 一年，造成了巨大的经济和社会伯恩斯。美国癌症学会估计有178,520血 2020年，美国的癌症病例在2019年在美国，大约有5,600例患者死于贫血。我们的国家 由于血液相关的疾病，面孔每年至少有450亿美元的经济燃烧。但是，电流 血液相关疾病的治疗仅会抑制症状并暂时防止疾病进展。 该项目的目的是调查专门的支持分解调解员（SPMS）派生的角色 在脊椎动物造血期间的多不饱和脂肪酸（PUFA）N-3 docosapentaenoic酸（DPAN-3） 并在体内评估其血液疾病的治疗潜力。我在斑马鱼中进行了化学屏幕 使用几种生态烷衍生的SPM和整个安装原位杂交（愿望）来识别胚胎以识别 能够增加造血干细胞（HSC）和红细胞（RBC）的化合物。这个屏幕 确定了两个有前途的SMP复合命中：7,17 Dihydroxy DPA（7,17 Dihdpan-3）和19,20 Epoxy DPA （19,20 EPDPA）。在此提案中，我将评估它们对下游造血的增殖的影响 通过将野生型斑马鱼胚胎暴露于1.5 µm 7,17，包括髓样和淋巴样谱系 dihdpan-3和2.5 µm 19,20 EPDPA从受精后11-36小时（HPF）。固定后，我会表演愿望 使用C-MYB，GATA1，L-Plastin，Lyz，MPEG，RAG1和CD79 mRNA，和实时定量PCR（RT-QPCR） 问题和底漆以描述性和定量评估两种SPMS暴露的效果 关于造血干细胞，祖细胞性红细胞，粒细胞，中性粒细胞和 巨噬细胞，T和B细胞。我们的初步数据表明，7,17 Dihdpan-3和19,20 EPDPA 促进HSC增殖，增加RBC祖细胞和RBC数，并增加中性粒细胞数量。 因此，我假设7,17 dihdpan-3和19,20 EPDPA暴露增强HSC增殖， 增加脊椎动物中下游成熟的血细胞类型。为了检验我们的中心假设，我开发了 以下两个具体目的：目标1。测试两个选择的SPM会增加HSC数的假设， 导致髓样衍生细胞和淋巴样细胞数的增加，并瞄准2。 在细菌等疾病模型中，两个选定的SPM可以用作治疗干预措施 介导的感染，造血遗传疾病和灌溉后的成年斑马鱼。这项工作将 表征脊椎动物造血期间这两个SPM的作用，并将测试其治疗潜力 体内。该提案的结果将发现这两个SPM在调节中的未知作用 造血，为开发新型的血液相关疾病治疗疗法开辟了新的道路。