ShEEP Request for Leica Laser Capture Microdissection System (LMD7)

ShEEP 请求徕卡激光捕获显微切割系统 (LMD7)

• 批准号: 9908938
• 负责人: Gianfranco D Alpini
• 金额: --
• 依托单位: RLR VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2020-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9908938
• 关键词: Affect; Alcoholic Liver Diseases; Area; Biology; Biomedical Research; Cancer Biology; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cell Culture Techniques; Cell Separation; Cells; Chromosomes; Chronic; Chronic Disease; Cirrhosis; Cloning; Collaborations; Computer software; Data; Diabetes Mellitus; Discipline; Disease; Dissociation; Equipment; Fluorescence; Functional disorder; Funding; Gastroenterology; Genes; Genomic DNA; Goals; Health Benefit; Healthcare; Healthcare Systems; Hepatitis B; Hepatitis C; Hepatology; Human; Indiana; Intracellular Signaling Proteins; Knowledge; Lasers; Lead; Leadership; Light; Liver; Liver Fibrosis; Liver diseases; Malignant Neoplasms; Malignant neoplasm of liver; Maps; Mechanics; Medical; Medical center; Messenger RNA; Metabolic Bone Diseases; Methods; MicroRNAs; Microdissection; Microscope; Molecular; Molecular Analysis; Morbidity - disease rate; Neurologic; Neurosciences; Organ; Pathogenesis; Pathologic; Pathologic Processes; Pathology; Phase; Play; Population; Prevention therapy; Proteomics; Research; Research Personnel; Resolution; Sampling; Science; Sheep; Specimen; Subcellular structure; System; Techniques; Tissue Sample; Tissue Stains; Tissues; Translational Research; United States; Universities; Veterans; animal tissue; bone metabolism; cell type; cholangiocyte; cholestatic liver disease; chronic liver disease; design; diabetic; hazard; human disease; human model; improved; instrument; interest; laser capture microdissection; medical schools; metabolomics; microsystems; mortality; neuropathology; next generation sequencing; non-alcoholic fatty liver disease; novel; novel therapeutics; operation; problem drinker; research facility; single cell analysis; tool; transcriptomics; translational research program

Laser Microdissection LMD7 System enables users to isolate specific single cells or entire areas of tissue. This system allows rapid and accurate microdissection of both small and large groups of cells and can be used with phase contrast for live-cell applications, with stained tissue samples and transmitted light, or with epi- fluorescence for fluorescence tagged targets. Powered by a unique laser design and dynamic software, Leica LMD7 systems allow users to easily isolate Regions of Interest (ROI) from entire areas of tissue down to single cells or even subcellular structures such as chromosomes. Our VA researchers in hepatology, gastroenterology, cholangiocytes biology and diabetic pathology rely on this method. Furthermore, LMD7 system is a perfect tool for live cell culture (LCC), for cloning and re-cultivation, manipulation or downstream analysis. At the Richard L. Roudebush VA Medical Center (RRVA) at Indianapolis, increased interactions and collaborations have been established among multiple investigators with expertise in liver and cholangiocytes pathobiology, neuropathology, cancer biology, cardiovascular pathophysiology, diabetes and bone metabolism. With the purchase of the shared equipment of the Leica Laser Capture Microdissection system LMD7, which can be used by investigators of different disciplines for the single cell analysis in human healthy and diseased tissues and cells, such collaborations could be further enhanced. Several of the proposed users are focused on chronic liver disorders such as alcoholic liver diseases; Non-alcoholic fatty liver disease (NAFLD), cholestatic liver diseases, hepatitis B and C virus infections are severe medical disorders and will further develop to liver fibrosis, cirrhosis and liver cancer. They are the 10th leading cause of death in the United States that are highly relevant to our veterans. The study shows that the hazard of developing chronic liver diseases and liver cancer cirrhosis in the VA health care system significantly increased recently. The translational research approach of the healthy and diseases human liver at single-cell resolution is critical to understanding the pathogenesis and treatment of human liver disease. This single cell analysis approach has been difficult to carry out, mainly because fresh human liver tissue access is scarce and the tissue is difficult to fractionate without damaging fragile resident cell populations. Cells and tissues from animals and humans that model various pathological processes have played a vital part in medical breakthroughs in the past decade that have benefited the health care of US populations including Veterans. The Leica Laser Capture Microdissection System LMD7 requested in this application will be used to study intracellular signaling and protein interactions in single human cell stage under pathological conditions of human liver and other diseases that lead to significant mortality and morbidity in the Veteran population. The long-term objective of the investigators involved in this application is to obtain a better understanding of molecular underpinnings of diseases with the ultimate goal of improving healthcare for Veterans. The requested Leica Laser Capture Microdissection LMD7 system is critical for progress in these important areas. Two specific aims are proposed in this application: Aim 1: To purchase and maintain a Leica LMD7 system for use by dedicated VA researchers. Aim 2: To enhance the quality of science and promote collaboration among dedicated VA researchers. Results of this proposed research will provide a better understanding of underlying mechanisms critical for designing new therapies for the prevention and treatment of these chronic human diseases. The Leica LMD7 system makes it possible to generate the gene profiling data at specific cell type of intact specimens. It also becomes possible to exclude incorrect results caused by mechanically isolation of the cells from the samples. Having this novel, state-of-the-art laser capture microdissection system will open up new avenues of research and collaboration at RRVA that will not be possible without it.

激光显微切割 LMD7 系统使用户能够分离特定的单个细胞或整个组织区域。这 系统可以快速准确地对小型和大型细胞群进行显微切割，并且可以与 用于活细胞应用的相差，使用染色的组织样本和透射光，或使用 Epi- 荧光标记目标的荧光。徕卡采用独特的激光设计和动态软件 LMD7 系统允许用户轻松地将感兴趣区域 (ROI) 从整个组织区域分离到单个区域 细胞甚至亚细胞结构，例如染色体。我们的 VA 研究人员在肝病学、胃肠病学、 胆管细胞生物学和糖尿病病理学依赖于这种方法。此外，LMD7系统是一个完美的工具 用于活细胞培养 (LCC)、克隆和再培养、操作或下游分析。在理查德 位于印第安纳波利斯的 L. Roudebush VA 医疗中心 (RRVA) 加强了互动和合作 由具有肝脏和胆管细胞病理学专业知识的多名研究人员建立， 神经病理学、癌症生物学、心血管病理生理学、糖尿病和骨代谢。随着 购买徕卡激光捕获显微切割系统LMD7的共享设备，可使用 由不同学科的研究人员对人类健康和患病组织进行单细胞分析， 细胞，可以进一步加强这种合作。一些提议的用户专注于慢性肝病 酒精性肝病等疾病；非酒精性脂肪肝（NAFLD）、胆汁淤积性肝病、 乙型和丙型肝炎病毒感染是严重的疾病，并将进一步发展为肝纤维化， 肝硬化和肝癌。它们是美国第十大死因，且与此高度相关 致我们的退伍军人。研究表明患慢性肝病和肝癌肝硬化的危险 VA 医疗保健系统中的人员数量最近显着增加。转化研究方法 单细胞分辨率下的健康和疾病人类肝脏对于理解发病机制和 治疗人类肝脏疾病。这种单细胞分析方法一直难以实施，主要是 因为新鲜的人类肝脏组织很少，并且很难在不损坏肝脏的情况下对组织进行分级 脆弱的常驻细胞群。来自动物和人类的细胞和组织，模拟各种病理 在过去十年中，流程在有益于健康的医学突破中发挥了至关重要的作用 照顾包括退伍军人在内的美国民众。请求 Leica 激光捕获显微切割系统 LMD7 该应用程序将用于研究单个人类细胞中的细胞内信号传导和蛋白质相互作用 人类肝脏病理条件下的阶段和导致显着死亡率的其他疾病 退伍军人群体的发病率。参与本申请的研究人员的长期目标是 更好地了解疾病的分子基础，最终目标是改善疾病 退伍军人的医疗保健。所需的徕卡激光捕获显微切割 LMD7 系统对于 这些重要领域取得进展。本申请提出了两个具体目标： 目标 1：购买和 维护 Leica LMD7 系统，供专门的 VA 研究人员使用。目标2：提高科学质量 并促进退伍军人管理局专门研究人员之间的合作。这项拟议研究的结果将提供 更好地了解对于设计预防和预防新疗法至关重要的潜在机制 治疗这些人类慢性疾病。 Leica LMD7 系统使生成基因成为可能 完整样本的特定细胞类型的分析数据。还可以排除不正确的结果 由细胞与样品的机械分离引起。拥有这种新颖、最先进的激光捕获 显微切割系统将为 RRVA 开辟新的研究和合作途径，而这些途径不会 没有它也可能。