IN SITU HYBRIDIZATION IN HODGKIN'S DISEASE

霍奇金病的原位杂交

• 批准号: 3459526
• 负责人: LAWRENCE M. WEISS
• 金额: $ 9.38万
• 依托单位: CITY OF HOPE NATIONAL MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1996-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3459526
• 关键词: Epstein Barr virus; Hodgkin's disease; cell type; cytokine; gene expression; genome; human subject; immunoglobulins; in situ hybridization; interferons; interleukin 1; interleukin 6; messenger RNA; neoplastic cell; nucleic acid probes; oncogenes; polymerase chain reaction; tumor necrosis factor alpha; tumor necrosis factor beta; virus genetics

Hodgkin's disease represents one of the major human malignancies, yet investigation of its biology has lagged far behind clinical studies. This is largely due to several unique aspects of Hodgkin's disease, including the fact that the neoplastic element, the Reed-Sternberg cells and variants, comprise a rare component of the cellular infiltrate, the difficulty to grow Reed-Sternberg cells in cell culture, and the phagocytic properties of Reed-Sternberg cells; all three of these features have made the study of Hodgkin's disease through standard molecular and immunologic techniques extremely difficult and relatively fruitless.It is proposed to utilize the technique of in situ hybridization to study Hodgkin's disease.Specific aims include the in situ identification and localization of Epstein-Barr viral genomes to a specific cell type in Hodgkin's disease tissues, in situ identification of cytokine mRNA in Hodgkin's disease tissues, in situ identification of oncogene mRNA in Reed-Sternberg cells of Hodgkin's disease tissues, and the in situ identification of immunoglobulin mRNA in Hodgkin's disease tissues, particularly in the "L&H" cells of the nodular lymphocyte predominant subtype. Protocols involving use of oligonucleotide probes on paraformaldehyde-fixed, frozen sections or formalin-fixed, paraffin-embedded sections will be primarily be utilized. In situ hybridization studies of the identification of Epstein-Barr viral genomes in Hodgkin's disease tissues will be initiated using cases known to be positive by slot blot hybridization studies (approximately 20% of cases of Hodgkin's disease), and will eventually be extended to other cases. In addition, polymerase chain reaction studies for EBV genome will also be performed. In situ hybridization studies of cytokine and oncogene mRNA will be performed on a variety of cases of typical Hodgkin's disease. Cytokine studies will include analysis of interleukin 1-6, tumor necrosis factor alpha and beta, and interferon alpha, beta, and gamma mRNA. The studies of oncogene expression will include a range of oncogenes known to be important in the pathogenesis of hematolymphoid and non-hematolymphoid tumors. Finally, cases of nodular lymphocyte predominant Hodgkin's disease will be studied by tissue in situ hybridization studies employing oligonucleotide probes or by the technique of in situ transcription in an attempt to identify light chain mRNA clonal restriction, evidence that would support the hypothesis of a relationship of nodular lymphocyte predominant Hodgkin's disease to B cell neoplasia.

霍奇金病是人类主要恶性肿瘤之一，但 其生物学研究远远落后于临床研究。 这 很大程度上是由于霍奇金病的几个独特方面，包括 事实上，肿瘤成分、里德-斯滕伯格细胞和 变体，包含细胞浸润的罕见成分， Reed-Sternberg 细胞在细胞培养物中生长困难，并且吞噬细胞 里德-斯腾堡细胞的特性；所有这三个功能都使得 通过标准分子和免疫学研究霍奇金病 技术极其困难且相对无果。建议 利用原位杂交技术研究霍奇金氏病 疾病。具体目标包括原位识别和定位 将 Epstein-Barr 病毒基因组转化为霍奇金病的特定细胞类型 组织，原位鉴定霍奇金病细胞因子 mRNA 组织，原位鉴定 Reed-Sternberg 细胞中的癌基因 mRNA 霍奇金病组织的鉴定和原位鉴定 霍奇金病组织中的免疫球蛋白 mRNA，特别是“L&H”中 结节淋巴细胞为主的亚型细胞。 协议涉及 在多聚甲醛固定的冰冻切片上使用寡核苷酸探针 或福尔马林固定、石蜡包埋的切片将主要是 被利用。 原位杂交鉴定研究 霍奇金病组织中的 Epstein-Barr 病毒基因组将启动 使用通过槽印迹杂交研究已知为阳性的病例 （约占霍奇金病病例的 20%），并且最终将被 延伸至其他情况。 此外，聚合酶链反应研究 还将进行 EBV 基因组检测。 原位杂交研究 将对多种病例进行细胞因子和癌基因 mRNA 检测 典型的霍奇金病。 细胞因子研究将包括分析 白细胞介素 1-6、肿瘤坏死因子 α 和 β 以及干扰素 α、β 和 γ mRNA。 癌基因表达的研究将 包括一系列已知在发病机制中重要的癌基因 血淋巴和非血淋巴肿瘤。 最后，结节性病例 淋巴细胞为主的霍奇金病将通过组织原位研究 使用寡核苷酸探针或通过该技术的杂交研究 原位转录以尝试鉴定轻链 mRNA 克隆 限制，支持关系假设的证据 结节性淋巴细胞为主型霍奇金病转变为 B 细胞肿瘤。