CORE--ANALYSIS

核心--分析

• 批准号: 6668362
• 负责人: Neil R. Hackett
• 金额: $ 19.94万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6668362
• 关键词: bioimaging /biomedical imaging; biomedical facility; computer data analysis; cooperative study; gene therapy; immunology

The Analysis Core provides the Weill Cornell PEGT investigators access to advanced technologies in the general areas of biophysical analysis, quantitative assessment of DNA and RNA levels, biological imaging and host response to gene transfer vectors. To relieve PEGT investigators from the necessary to purchase expensive equipment and learn complex techniques, the group has been assembled under the direction by Neil Hackett Ph.D. and Philip Leopold Ph.D. Located primarily in the ninth floor of the Belfer Gene Therapy Core Facility, adjacent to the DNA Vector Core and Experimental Animal Core, the Analysis Core will interact with all Weill Cornell PEGT preclinical and clinical projects. In the area of new vector (RNA and DNA) development and characterization of clinical (GMP) vector preparations, the Analysis Core will perform biophysical methods including spectroscopy (NMR, UV/visible, fluorescent), transmission electron microscopy of negatively stained and frozen specimens, particle size and surface charge measurements and analytical HPLC. In support of PEGT projects the Analysis Core will enable pharmacokinetics and biodistribution studies by quantitative PCR, genotyping PCR, genotyping of hosts and analysis of gene expression profiles by array methods. The biological imaging facilities include photographic and quantitative digital data collection of visible and fluorescent micrographs, scanning and transmission EM, flow cytometry and a range of methods of sample preparation (immunohistochemistry, immunofluorescent, in situ hybridization). The host responses section will assist in assessment of humoral (antigen-0specific antibody by class, neutralizing antibody), cellular (T cell proliferation, cytotoxic T lymphocyte) and cytokine (ELISPOT, serum cytokine) responses to in vivo gene therapy with a particular focus on interpreting human response in the two clinical projects.

分析核心使威尔康奈尔PEGT研究人员能够获得生物物理分析、DNA和RNA水平的定量评估、生物成像和宿主对基因转移载体的反应等一般领域的先进技术。为了免除PEGT研究人员购买昂贵设备和学习复杂技术的需要，该小组在Neil Hackett博士和Philip Leopold博士的指导下组建。分析核心主要位于Belfer基因治疗核心设施的九楼，毗邻DNA载体核心和实验动物核心，将与所有威尔·康奈尔PEGT临床前和临床项目互动。在新载体(RNA和DNA)的开发和临床(GMP)载体制剂的表征方面，分析核心将执行生物物理方法，包括光谱学(核磁共振、UV/可见光、荧光)、负染和冷冻标本的透射电子显微镜、颗粒尺寸和表面电荷测量以及分析性高效液相。为了支持PEGT项目，分析核心将通过定量聚合酶链式反应、基因分型聚合酶链式反应、宿主基因分型和通过阵列方法分析基因表达谱来实现药代动力学和生物分布研究。生物成像设施包括可见和荧光显微照片的摄影和定量数字数据收集、扫描和透射电子显微镜、流式细胞术和一系列样品制备方法(免疫组织化学、免疫荧光、原位杂交)。宿主反应部分将协助评估体液(按类别划分的抗原0特异性抗体，中和抗体)、细胞(T细胞增殖，细胞毒性T淋巴细胞)和细胞因子(ELISPOT，血清细胞因子)对体内基因治疗的反应，特别侧重于解释这两个临床项目中的人类反应。