Trans-megakaryocyte migration as neutrophil amplifier

作为中性粒细胞放大器的跨巨核细胞迁移

• 批准号: 10599971
• 负责人: Pierre Cunin
• 金额: $ 12.73万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10599971
• 关键词: Achievement; Advisory Committees; Amplifiers; Animals; Appointment; Arthritis; Autoimmune Diseases; Award; Biochemistry; Bioinformatics; Biology; Blocking Antibodies; Blood; Blood Platelets; Bone Marrow; Bone Marrow Cells; Boston; Cell Adhesion Molecules; Cell Communication; Cell Nucleus; Cells; Cellular Immunology; Chemotaxis; Circulation; Confocal Microscopy; Cytoplasm; Data; Deposition; Disease; Electron Microscopy; Emperipolesis; Endothelial Cells; Endothelium; Ensure; Environment; Enzyme-Linked Immunosorbent Assay; Exhibits; Exposure to; Faculty; Flow Cytometry; Frequencies; Funding; Goals; Health; Hemostatic function; Human Biology; Hybrids; Imaging Device; Immune; Immunity; Immunology; In Vitro; Inflammation; Inflammatory; Institution; Interleukin-1 beta; International; K/BxN model; Knowledge; Laboratories; Large Megakaryocyte; Leadership; Learning; Marrow; Mass Spectrum Analysis; Mediating; Mediator; Megakaryocytes; Membrane; Mentored Research Scientist Development Award; Mentors; Mentorship; Methods; Microscopy; Modeling; Molecular; Molecular Biology; Mus; Myelogenous; Neutrophil Activation; Operative Surgical Procedures; Organ; Pathologic; Pediatric Hospitals; Pediatrics; Peritonitis; Photons; Physiological; Plasma; Positioning Attribute; Process; Proteins; Reporting; Research; Research Personnel; Research Training; Role; Route; Serum; Slide; Source; Speed; Sterility; TNF gene; Technical Expertise; Testing; Time; Training; Transgenic Mice; Visualization; Work; cell behavior; eptifibatide; exosome; immune function; immunoregulation; in vivo; inhibitor; instructor; intravital imaging; intravital microscopy; medical schools; member; migration; neutrophil; novel; septic; skills; systemic inflammatory response; therapeutic target; training opportunity; transcriptome sequencing; two photon microscopy; two-photon

Project summary / abstract This K01 award is to provide Dr. Pierre Cunin with the advanced research training, protected time, and mentoring needed to become an independent investigator specialized in intravital 2-photon microscopy and committed to understanding the role of megakaryocytes as a direct modulators of immunity. Candidate: Dr. Cunin is an Instructor in Pediatrics at Harvard Medical School (Boston MA), in the laboratory of Dr. Peter Nigrovic in the Division of Immunology at the Boston Children’s Hospital. Dr Cunin’s previous training has provided him with extensive knowledge in the biology of human and murine immune cells, as well as a technical expertise in cellular immunology, molecular biology, biochemistry, and murine experimentation. Research: Dr. Cunin’s K01 project seeks to understand the function of emperipolesis (EP), a cell-in-cell interaction wherein a neutrophil transits through the cytoplasm of a megakaryocyte. Routinely observed in bone marrow from all species examined, EP had never been investigated in depth experimentally. Dr. Cunin found that EP increases in arthritis and other forms of systemic inflammation, suggesting a role in disease. He previously reported that EP is an active and highly regulated process that mediates bi-directional transfer of proteins and membrane between neutrophils and megakaryocytes. Preliminary data show that this exchange has important physiological consequences for neutrophil function. Dr. Cunin will evaluate how EP modulates the immune function of neutrophils in health and with inflammation. Finally, Dr. Cunin will identify ways to manipulate EP via targets on neutrophil or megakaryocyte, opening up the possibility of therapeutic targeting. Mentoring/Training: The long-term goal of the PI is becoming an independent investigator and tenured faculty member at a leading academic center. The proposed studies and training plan will set the stage for Dr. Cunin to obtain independent R01 funding and become an internationally recognized leader at the cutting-edge of the exploration of megakaryocytes as immune cells, as well as of the pathological role of this lineage in autoimmune and inflammatory diseases. With his appointment as Director of the 2-photon facility in the Division of Immunology at BCH, he will become an expert in intravital imaging to observe immune cells behaviors in their organs, in healthy animals or subjected to inflammation. His research training will encompass in vivo cell visualization and live cell analysis, animal surgery, and bioinformatics for RNA sequencing. Additionally, he will learn laboratory leadership skills to facilitate the transition to his own laboratory. The work will be performed in a superb institutional environment with the mentorship of Dr. Peter Nigrovic, an expert in myeloid biology and inflammatory diseases, Dr. Italiano, an expert in megakaryocyte biology, as well as an Advisory Committee including Drs. Bergmeier and Kim (surgery and in vivo assessment). Guidance on 2- photon technical issues and RNAseq analysis will be provided by collaborators Drs. Chen and Lee.

项目概要/摘要 该K01奖项旨在为Pierre Cunin博士提供先进的研究培训、受保护的时间和 成为一名专门从事活体 2 光子显微镜的独立研究者所需的指导 致力于了解巨核细胞作为免疫直接调节剂的作用。 候选人：Cunin 博士是哈佛医学院（马萨诸塞州波士顿）儿科讲师，实验室 Cunin 博士之前在波士顿儿童医院免疫学科接受的培训。 为他提供了人类和小鼠免疫细胞生物学方面的广泛知识，以及 细胞免疫学、分子生物学、生物化学和小鼠实验方面的技术专长。 研究：Cunin 博士的 K01 项目旨在了解伸入 (EP) 的功能，这是一种细胞内细胞 中性粒细胞穿过巨核细胞细胞质的相互作用。 Cunin 博士从未对所有受检物种的骨髓进行过深入的实验研究。 发现 EP 在关节炎和其他形式的全身炎症中增加，表明其在疾病中发挥作用。 先前报道，EP是一个活跃且高度监管的过程，介导双向转移 初步数据表明，中性粒细胞和巨核细胞之间存在蛋白质和膜的交换。 Cunin 博士将评估 EP 如何调节。 最后，库宁博士将确定中性粒细胞在健康和炎症中的免疫功能。 通过靶向中性粒细胞或巨核细胞来操纵 EP，开启了治疗靶向的可能性。 指导/培训：PI 的长期目标是成为一名独立研究者和终身教授 拟议的研究和培训计划将为 Cunin 博士奠定基础。 获得独立的R01资金并成为国际公认的前沿领导者 探索巨核细胞作为免疫细胞，以及该谱系在免疫细胞中的病理作用 他被任命为 2 光子设施主任。 BCH免疫学部，他将成为观察免疫细胞的活体成像专家 他的研究培训将涵盖其器官、健康动物或遭受炎症的行为。 体内细胞可视化和活细胞分析、动物手术以及 RNA 测序的生物信息学。 此外，他还将学习实验室领导技能，以促进向自己实验室的过渡。 将在一流的机构环境中进行，并在专家 Peter Nigrovic 博士的指导下进行 骨髓生物学和炎症性疾病，巨核细胞生物学专家 Italiano 博士以及 顾问委员会包括 Bergmeier 和 Kim 博士（手术和体内评估）。 光子技术问题和 RNAseq 分析将由合作者 Chen 和 Lee 博士提供。