DNA Sequencing and FACS Core Facilities

DNA 测序和 FACS 核心设施

基本信息

批准号：
8350117
负责人：
Paul Robbins
金额：
$ 175.31万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8350117
关键词：
Address Adoptive Immunotherapy Adoptive Transfer Antigens Autologous Binding Sites Biological Models CD4 Positive T Lymphocytes CTAG1 gene Cell Separation Cell physiology Cell surface Cells Characteristics Clinical Clinical Trials Core Facility Cytotoxic T-Lymphocytes DNA DNA Sequence DNA Sequencing Facility DNA analysis Differentiation Antigens Epitopes Family Genes Genetic Goals Growth Helper-Inducer T-Lymphocyte Human In Vitro Investigation Laboratories Lead Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of ovary Malignant neoplasm of prostate Mediating Melanoma Cell Modification Monophenol Monooxygenase Mus Mutate Nature Neoplasm Metastasis Operative Surgical Procedures Patients Peripheral Blood Mononuclear Cell Phase II Clinical Trials Phenotype Population Postdoctoral Fellow Proteins Publishing RNA Reagent Renal carcinoma Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role Sampling Services Synovial Cell T-Cell Receptor T-Lymphocyte Tumor Antigens Tumor-Infiltrating Lymphocytes base cancer testis antigen clinically significant gene cloning immune function in vitro Assay malignant breast neoplasm melanocyte melanoma member novel patient population peripheral blood research study response sarcoma tumor

项目摘要

Current studies have on the identification of novel antigens expressed in melanoma and renal cancers as well as the characterization of T cells that recognize epitopes of previously described antigens. In a recent studies, several novel point-mutated genes have been identified as the targets of melanoma-reactive TIL that mediated long terms clinical tumor regression following adoptive transfer. Current investigations are being carried out to evaluate the potential role of these mutated gene products in promoting the growth or metastasis of tumor cells. Additional studies have involved the identification of T cell receptors (TCRs) directed against widely shared antigens. Recent studies have focused on the identification of TCRs that recognize cancer/testis (C/T) antigens, molecules that are expressed in melanoma as well as a variety of highly prevalent malignancies such as breast and prostate cancer. A TCR that recognizes the NY-ESO-1 C/T antigen was used to transduce autologous PBMC, and results of a Phase II clinical trial published in January of 2011 indicate that approximately 50% of either melanoma or synovial cell sarcoma patients who received transudced PBMC demonstrated objective clinical responses to adoptively transferred autologous PBMC that were transduced with the NY-ESO-1 TCR. The DNA core facility provides services to many of the members of the Surgery Branch, which includes expertise and reagents used for RT-PCR, quantatitive RT-PCR, gene cloning and RNA/DNA analysis. For some of these studies, sequences were analyzed on an Applied Biosystems 3100-Avant Genetic Analyzer that is a part of the Surgery Branch DNA Sequencing Core. The FACS Core Facility is currently being utilized for the analysis of T cell populations that are administered to patients as a part of the analysis of ongoing clinical cancer adoptive immunotherapy trials. In addition, the FACS Core is utilized on a daily basis for the analysis of the results of experiments to analyze the results of in vitro experiments to examine factors that influence the phenotype and function of tumor reactive T cells as well as to carry out the separation of cells based upon their expression of a wide variety of cell surface markers. For these studies phenotypic analyses were carried out on a BD FACSCalibur, FACSCantoI and FACSCantoII, and cell separations were carried out on a BD FACSAriaIIu.

当前的研究已经对鉴定在黑色素瘤和肾脏癌中表达的新型抗原以及识别先前描述的抗原表位的T细胞的表征。在最近的一项研究中，已经确定了几个新型点突变的基因为黑色素瘤反应性的靶标，直到继产后介导了长期临床肿瘤消退。目前正在进行研究，以评估这些突变基因产物在促进肿瘤细胞的生长或转移中的潜在作用。其他研究涉及针对广泛共享抗原的T细胞受体（TCR）的鉴定。最近的研究集中在识别识别癌症/睾丸（C/T）抗原的TCR，在黑色素瘤中表达的分子以及多种高度普遍的恶性肿瘤，例如乳腺癌和前列腺癌。识别NY-ESO-1 C/T抗原的TCR用于转导自体PBMC，并于2011年1月发表的II期临床试验的结果表明，大约50％的黑色素瘤或滑膜细胞肉瘤患者，这些患者对接收性临床的临床反应表现出对继发性转移pBMC的客观临床反应，该反应与自动pBMC相关。 DNA核心设施为手术分支的许多成员提供服务，其中包括用于RT-PCR，定量RT-PCR，Gene Cloning和RNA/DNA分析的专业知识和试剂。对于其中一些研究，对应用的生物系统3100遗传分析仪进行了分析，该遗传分析仪是手术分支分支DNA测序核心的一部分。目前，FACS核心设施用于分析对患者进行的T细胞群体，作为正在进行的临床癌症收养免疫疗法试验的一部分。此外，每天都使用FACS核心来分析实验结果，以分析体外实验的结果，以检查影响肿瘤反应性T细胞的表型和功能的因素，并基于其表达各种细胞表面标记的表达来进行细胞的分离。对于这些研究，在BD FACSCALIBUR，FACSCANTOI和FACSCANTOII上进行了表型分析，并在BD FACSARIAIIU上进行了细胞分离。