Dissecting the role of miR-9 in normal and malignant mast cell biology

剖析 miR-9 在正常和恶性肥大细胞生物学中的作用

• 批准号: 8805547
• 负责人: Joelle M Fenger
• 金额: $ 13.09万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-16 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8805547
• 关键词: 3' Untranslated Regions; Adoptive Transfer; Allergic; Anaphylaxis; Angiogenic Factor; Antigens; Area; Arthritis; Award; Behavior; Benign; Bioinformatics; Biological; Biological Assay; Biological Process; Biology; Bone Marrow; Canis familiaris; Carboxypeptidase; Cardiovascular Diseases; Cell Count; Cell Degranulation; Cell Line; Cells; Cellular biology; Chemotactic Factors; Chemotaxis; Chronic Disease; Chymase; Cytoplasmic Granules; Development; Dinoprostone; Disease; Distant; Doctor of Philosophy; EMSA; Ectopic Expression; Effector Cell; Environment; Evans blue stain; Exhibits; Extracellular Matrix; Extracellular Matrix Degradation; Extravasation; Fibroblast Growth Factor 2; Foundations; Gene Expression; Genetic; Genetic Transcription; Hematologic Neoplasms; Histologic; IgE; Immune response; Immunity; In Vitro; Infiltration; Inflammation; Inflammatory; Internal Medicine; Knockout Mice; Laboratories; Lentivirus Vector; Lesion; Licensing; London; MMP2 gene; Malignant - descriptor; Malignant Neoplasms; Mast Cell Neoplasm; Mediating; Medical Oncologist; Medicine; Mentors; Mentorship; MicroRNAs; Modeling; Molecular; Mus; Mutation; Neoplasm Metastasis; Ohio; Passive Cutaneous Anaphylaxis; Pathologic; Pathologic Processes; Peptide Hydrolases; Phenotype; Physiological; Physiological Processes; Platelet-Derived Growth Factor; Principal Investigator; Production; Property; Proto-Oncogenes; Reporter; Research; Research Personnel; Residencies; Role; Scientist; Small Interfering RNA; Staining method; Stains; Techniques; Testing; Therapeutic Intervention; Time; Tissues; Tolonium chloride; Training; Transgenic Mice; Transgenic Model; Transgenic Organisms; Transmission Electron Microscopy; Tumor Biology; Universities; Untranslated RNA; Vascular Endothelial Growth Factors; Veterinarians; Veterinary Medicine; Work; angiogenesis; career; cell behavior; cell motility; college; comparative; crosslink; density; experience; in vivo; mast cell; mastocytosis; migration; mouse model; mutant; oncology; pre-clinical; professor; programs; promoter; public health relevance; research study; response; skills; therapeutic target; tumor; tumor progression

 DESCRIPTION (provided by applicant): Mast cells are key effector cells in a wide variety of physiological and pathological processes, including innate immune responses and allergic disorders, chronic inflammatory diseases such as cardiovascular disease and arthritis, and tumor progression. Activated mast cells secrete a diverse array of factors that mediate their roles in inflammation, immunity, and tissue remodeling. However the exact mechanisms through which various genetic factors influence multiple aspects of mast cell biology have yet to be fully defined. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and their dysregulation is implicated in numerous pathologic conditions. Recent work by our laboratory found that miR-9 over-expression was associated with aggressive, metastatic canine mast cell tumors (MCT), a well-established model of spontaneous malignant mast cell disease. Furthermore, enforced high expression of miR-9 in normal and malignant mouse mast cells with low basal levels of this miR using lentiviral vectors promoted invasion and enhanced the expression of CMA1, a mast cell-specific protease involved in tissue remodeling. To better study the role of miR9 in mast cell biology, we have generated a transgenic model of tissue specific induced miR-9 expression, have crossed these mice with the carboxypeptidase-3-Cre transgenic mice generated by our collaborator Dr. Stephen Galli (Stanford University), and confirmed the functionality of this model including enhanced invasive properties of mast cells from the CPA3-Cre/miR-9 double transgenics. We therefore hypothesize that miR-9 enhances CMA1 expression and promotes invasion in mast cells through the modulation of factors responsible for tissue remodeling and angiogenesis. We further hypothesize that in normal mast cells, miR-9 alters cell motility and enhances sensitivity to antigen stimulation in vivo, and thatin mouse models of mastocytosis, miR-9 contributes to enhanced cell invasion and angiogenesis and is associated with a more aggressive biological behavior. To test these hypotheses, we will complete the following specific aims: 1) Identify the molecular mechanisms responsible for miR-9-induced CMA1 expression and enhanced invasion in normal mast cells; 2) Assess the effects of miR-9 on normal mast cell biology in vivo utilizing our transgenic mouse model of tissue specific miR-9 expression; and 3) Investigate the contribution of miR-9 in promoting mast cell invasion, metastasis, and tumor progression in mouse models of malignant mast cell disease. In summary, the studies outlined in this K01 proposal will provide a more complete understanding of the molecular mechanisms through which miR-9 regulates mast cell behavior both in vitro and in vivo, particularly as it relates to induction of the metastatic phenotype. The Candidate: Dr Joelle Fenger, a licensed veterinarian and Board Certified Veterinary Medical Oncologist, is currently completing her PhD in the Graduate Studies Program in Comparative and Veterinary Medicine at The Ohio State University in the context of a dual Residency/PhD program. During the K01 SERCA award period, Dr. Fenger's efforts will first be dedicated to completing her PhD thesis studies and completing her dissertation. Over the final years, she will be appointed as a Research Assistant Professor during which time she will gain further skill sets and develop a refined research focus that will serve as the foundation for her transition to an independent academic scientist in the final year of the award. The Environment: Dr. Fenger's co-mentors, Dr. Cheryl London and Dr. Guido Marcucci are both Professors and Principal Investigators at OSU; Dr. London in the Department of Veterinary Biosciences, College of Veterinary Medicine, and Dr. Marcucci in the Department of Internal Medicine, College of Medicine. Dr. London has extensive experience in the biology of malignant mast cell disease and translational oncology, and Dr. Marcucci has substantial expertise in the role of miRNA in hematologic malignancies and preclinical therapeutic targeting of miRNAs. As part of the overall development and training plan during the period of the award, Dr. Fenger will receiving mentorship from a diverse, inter-disciplinary group of advisors and collaborators. She will develop skill sets in the broader areas of miRNA dysregulation in disease, receive formal laboratory training in mast-cell specific techniques, and advanced training in mouse pathobiology and mouse models of disease. Additionally, throughout the duration of the award, Dr. Fenger will be mentored in executing hypothesis-driven research and establishing collaborative research partnerships that will prepare her as she transitions to a career as an independent researcher.

 描述（由适用提供）：肥大细胞是多种物理和病理过程中的关键效应细胞，包括先天免疫调查和过敏性疾病，慢性炎症性疾病，例如心血管疾病和关节炎以及肿瘤的进展。活化的肥大细胞秘密介导了一系列因素，这些因素介导了其在炎症，免疫学和组织重塑中的作用。然而，通过microRNA（miRNA）的确切机制是调节基因表达的小非编码RNA，其失调在许多病理条件下隐含。我们的实验室最近的工作发现，miR-9的过表达与侵略性转移性犬类肥大细胞肿瘤（MCT）有关，这是一个良好的赞助恶性肥大细胞病模型。此外，使用慢病毒载体在正常和恶性小鼠肥大细胞中强制执行MiR-9的高表达，促进了侵袭，并增强了CMA1的表达，CMA1的表达是一种参与组织重塑的肥大细胞特异性蛋白。为了更好地研究miR9在肥大细胞生物学中的作用，我们已经生成了组织特异性诱导的miR-9表达的转基因模型，已与我们的合作者斯蒂芬·加利（Stephen Galli）博士（斯坦福大学）（斯坦福大学）生成的羧肽酶-3-cre转基因小鼠（斯坦福大学）产生了这些小鼠，并证实了该模型的功能，包括增强的MIRS/MIRS/MIR的功能。因此，我们假设miR-9通过调节负责组织重塑和血管生成的因素来增强CMA1表达并促进肥大细胞的侵袭。我们进一步假设，在正常肥大细胞中，miR-9改变了细胞的运动，并增强了对体内抗原刺激的敏感性，而肥大性小鼠模型miR-9有助于增强细胞侵袭和血管生成，并与更具侵略性的生物学行为相关。为了检验这些假设，我们将完成以下特定目的：1）确定负责miR-9诱导的CMA1表达的分子机制，并增强正常肥大细胞中的浸润； 2）使用组织特异性miR-9表达的转基因小鼠模型评估miR-9对体内正常肥大细胞生物学的影响； 3）研究miR-9在恶性肥大细胞疾病的小鼠模型中促进肥大细胞侵袭，转移和肿瘤进展的贡献。总而言之，该K01提案中概述的研究将提供对MiR-9在体外和体内调节肥大细胞行为的分子机制的更完整理解，尤其是与转移性表型的诱导有关。候选人：乔伊尔·芬格（Joelle Fenger）博士是一名有执照的兽医和董事会认证的兽医医学肿瘤学家，目前正在俄亥俄州立大学的比较和兽医医学研究生研究计划中，在双重居住/博士学位课程的背景下。在K01 SERCA奖期间，Fenger博士的努力将首先致力于完成她的博士学位论文研究并完成她的论文。在最后几年中，她将被任命为研究助理教授，在此期间，她将获得进一步的技能，并建立精致的研究重点，这将是她在奖项的最后一年向独立学术科学家过渡的基础。环境：Fenger博士的联合官员Cheryl London博士和Guido Marcucci博士是OSU的教授兼首席研究员；伦敦博士在兽医学院兽医生物科学系和医学院内科系的Marcucci博士。伦敦博士在恶性肥大细胞疾病和转化肿瘤学的生物学方面拥有丰富的经验，而Marcucci博士在miRNA在血液学恶性肿瘤和miRNA的临床前疗法中的作用方面具有丰富的专业知识。作为奖励期间整体发展和培训计划的一部分，Fenger博士将从多样性，跨学科的顾问和合作者中获得心态。她将在疾病中miRNA失调的更广泛领域开发技能，接受桅杆特定技术的正式实验室培训以及小鼠病理学和疾病小鼠模型的高级培训。此外，在整个奖项期间，Fenger博士将在执行以假设为驱动的研究和建立合作研究合作伙伴关系方面进行指导，这将使她在过渡到独立研究人员的职业时做好准备。