Penn integrated Human Pancreas procurement and Analysis Program

宾夕法尼亚大学人类胰腺采购与分析计划 摘要 利用我们现有的基础设施和科学协作，我们已经组装了6个核心， 从胰腺采购和胰岛隔离到数据集成的专业知识 全面和综合的人类胰腺采购和分析计划，基于 宾夕法尼亚大学。核心A将获取一系列人类胰腺和 详细的捐献者病史；由下一代进行高分辨率的HLA配型 测序；分离胰岛；并将胰岛和组织分配到其他岩心进行进一步分析 或处理过程中。核心B将对孤立的胰岛进行生理表型鉴定。核心C将 应用流式细胞仪和单细胞定量聚合酶链式反应检测记忆性T细胞亚群 分析；表征Tregs的抑制活性和相关效应细胞的能力 抑制；B细胞表型；并生成增强子的染色质可及性图 致病细胞类型。核心D将为分子执行多种高级模式 分离的胰岛的概况，包括分类的胰岛细胞群体的RNAseq和microRNAseq； 20多种细胞表面和细胞内单细胞定量的质量细胞术 标记和单细胞RNAseq.Core E将使用多种方式处理组织，这将 允许使用多路免疫荧光等先进技术进行分析 染色、组合条形码FISH(组合FISH)、整个幻灯片成像和定量图像 蛋白质标志物和免疫细胞渗入的分析。这一核心将适应2维 使用多路离子束成像(MIBI)技术对胰腺切片进行质量细胞术。 这个核心还将存档组织以及DNA和血液，并促进样本分发 给HPPAP批准的研究人员。最后，核心F将组装、注释和维护一个开放的 访问该计划及其成员-研究人员的数据库，并在 来自两个程序的数据共享。整个程序将由一个 由绩效指标组成的行政核心，在执行委员会的协助下 由核心领导人组成。行政核心将与一个外部委员会建立联系 审查HPPAP生物样本应用的申请，并将与HIRN合作。这个 计划还将与HPPAP成员/PANC DB用户社区互动，以提供 以组织为基础的生理学、基因组和免疫学数据的丰富注释来源 主宰T1D的景观。

Penn Human Pancreas Procurement and Analysis Program Abstract Utilizing our existing infrastructure and scientific collaborations, we have assembled 6 cores with expertise ranging from pancreas procurement and islet isolation to data integration for a comprehensive and integrated Human Pancreas Procurement and Analysis Program based at the University of Pennsylvania. Core A will procure a spectrum of human pancreata and detailed donor medical history; perform high resolution HLA typing by next generation sequencing; isolate islets; and distribute islets and tissues to the other Cores for further analysis or processing. Core B will perform physiological phenotyping on the isolated islets. Core C will quantify and characterize memory T cell subsets by flow cytometry and single cell qPCR analysis; characterize suppressive activity of Tregs and the ability of related effector cells to be suppressed; B cell phenotyping; and generate chromatin accessibility maps of enhancers in pathogenic cell types. Core D will perform multiple advanced modalities for the molecular profiling of isolated islets including RNAseq and microRNAseq of sorted islet cell populations; mass cytometry for single cell quantification of more than 20 cell surface and intracellular markers; and single cell RNAseq. Core E will process tissues using multiple modalities that will allow for analysis using advanced technologies such as multiplexed immunoflourescent staining, combinatorial barcoded FISH (combFISH), whole slide imaging, and quantitative image analysis of protein markers and immune cell infiltrates. This Core will adapt 2-dimensional mass cytometry to pancreatic sections utilizing multiplexed ion beam imaging (MIBI) technology. This Core will also archive tissues as well as DNA and blood, and facilitate sample distribution to HPPAP approved researchers. Finally, Core F will assemble, annotate and maintain an open access database for the Program and its member-researchers, and collaborate with the HIRN in the sharing of data from both programs. The entire Program will be executed by an Administrative Core consisting of the PIs, with assistance from an Executive Committee consisting of the core leaders. The Administrative Core will interface with an external committee to review applications for HPPAP biosample use, and will collaborate with the HIRN. The Program will also interact with the HPPAP member/PANC DB user community to provide a richly annotated source of physiologic, genomic and immunologic data on the tissue-based landscape governing T1D.