Biospecimens and Patient-derived xenografts

核心1--项目摘要 核心1的首要目标是提供银行所需的基础设施和专业知识 原发白血病、结肠癌和胰腺癌标本，建立患者来源的异种移植(PDX)， 有机物和细胞模型，并使这些患者来源的资源可供所有计划调查人员使用， 从而促进CSBC调查人员在以下方面进行的翻译和实验室研究 项目1和项目2。这两个项目都需要获取患者样本和从PDX和有机类化合物中提取的细胞 模特们。该核心支持3个功能：(1)冷冻保存和分配来自慢性粒细胞白血病患者的原代细胞 白血病、结肠癌和胰腺癌；(2)异种移植的建立、特征和分布； 器官和这些疾病的细胞培养模型，以及(3)从原发肿瘤中分离细胞群体 用于单细胞分析的样本和组织模型。该核心的功能包括文件和 追踪活组织、血浆或血清以及基因组的知情同意、分离和冷冻保存 从患者样本中提取DNA，并将样本分发给CSBC调查人员。一大批小学 白血病、结肠癌和胰腺肿瘤样本目前在我们现有的存储库中可供以下人员使用 项目调查员。额外的样本将从我们的常规评估的患者那里获得 胃肠道肿瘤和血液系统恶性肿瘤临床。除了将原代细胞分布到 计划调查员，核心1还将维护和分发一批特征良好的PDX，这些PDX被选择用于 在免疫功能低下的小鼠中通过。白血病细胞和原发上皮性肿瘤的移植方法 免疫缺陷小鼠的组织现在已经很好地建立起来了，我们和其他人已经成功地使用了PDX 用于药物测试的模型。有机培养方法将用于补充PDX的努力，并将 提供另一种增殖和分离患者来源的上皮性肿瘤细胞和间质的方法 细胞。CORE 1将维护PDX模型和有机体的库，并分发给项目调查人员 需要的。CORE 1将由威廉·哈恩博士和大卫·温斯托克博士联合执导。哈恩医生是一名医生 DFCI癌症基因组发现中心(CCGD)的负责人。他还与 DFCI的胃肠道肿瘤科，并帮助建立了有机化合物和PDX存储库 来自原发和转移性胰腺癌和结肠癌样本。与CCGD合作，Dr。 哈恩的实验室已经建立了一条管道，用于对同意的患者来源的基因组进行表征 样本。温斯托克博士是一名内科肿瘤学家，领导着DFCI血液恶性肿瘤PDX存储库， 它已经包含了200个可移植的白血病PDX模型，这些模型已经被冷冻保存和 接受全基因组转录组分析、靶向DNA测序、免疫表型和 额外的分子表征。因此，芯1在加工方面具有丰富的经验， 超低温保存、流式细胞仪分析、纯化、繁殖、银行和基因组特征 原生标本和肿瘤模型，以及它们在临床前药物评价中的应用。

Core 1 – Project Summary The overarching goal of Core 1 is to provide the infrastructure and professional expertise needed to bank primary leukemia, colon, and pancreatic cancer specimens, establish patient-derived xenograft (PDX), organoid, and cell models, and make these patient-derived resources available to all Program investigators, thereby facilitating the translational and laboratory-based research performed by CSBC investigators in Projects 1 and 2. Both projects will require access to patient samples and cells derived from PDX and organoid models. The core supports 3 functions: (1) cryopreservation and distribution of primary cells from patients with leukemia, colon, and pancreatic cancer, (2) establishment, characterization, and distribution of xenograft, organoid, and cell culture models of these diseases, and (3) isolation of cell populations from primary tumor samples and tissue models for single cell analysis. The functions of the core include documentation and tracking of informed consent, isolation and cryopreservation of viable tissue, plasma or serum, and genomic DNA from patient samples and distribution of samples to CSBC investigators. A large number of primary leukemia, colon and pancreatic tumor samples are currently available in our existing repositories for use by Project Investigators. Additional samples will be acquired from patients routinely evaluated in our gastrointestinal oncologic and hematologic malignancies clinics. In addition to distributing primary cells to program investigators, Core 1 will also maintain and distribute a bank of well characterized PDXs selected for passage in immunocompromised mice. Methods for engrafting leukemia cells and primary epithelial tumor tissues in immunodeficient mice are now well established, and we and others have successfully used PDX models for drug testing. Organoid culture approaches will be used to complement the PDX effort, and will provide an additional way of propagating and segregating patient-derived epithelial tumor cells and stromal cells. Core 1 will maintain a bank of PDX models and organoids and distribute to program investigators as needed. Core 1 will be jointly directed by Drs. William Hahn and Dr. David Weinstock. Dr. Hahn is a medical oncologist who directs the DFCI Center for Cancer Genome Discovery (CCGD). He also works closely with the Division of Gastrointestinal Oncology at DFCI and has helped establish an organoid and PDX repository derived from primary and metastatic pancreatic and colon cancer samples. Working with the CCGD, Dr. Hahn's laboratory has created a pipeline for the genomic characterization of consented patient derived samples. Dr. Weinstock is a medical oncologist and directs the DFCI Hematologic Malignancy PDX repository, which already contains >200 transplantable leukemia PDX models that have been cryopreserved and undergone genome-wide transcriptome analysis, targeted DNA sequencing, immunophenotyping, and additional molecular characterization. Thus, Core 1 has extensive experience in the processing, cryopreservation, flow cytometric analysis, purification, propagation, banking, and genomic characterization of primary specimens and tumor models, as well as their use in preclinical drug evaluation.