CORE--MOLECULAR BIOLOGY AND IMMUNE ASSESSMENT

核心--分子生物学和免疫评估

• 批准号: 6599292
• 负责人: JOHN G. GRIBBEN
• 金额: $ 10.95万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2003-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6599292
• 关键词: biomedical facility; epitope mapping; gene rearrangement; human genetic material tag; human tissue; immunoglobulin genes; minimal residual disease; molecular cloning; molecular oncology; neoplasm /cancer diagnosis; oligonucleotides; polymerase chain reaction; tumor antigens

Increasing evidence suggests that the eradication of minimal residual detectable disease is necessary for cure. Considerable effort has therefore been made over the past decade to develop sensitive methods to detect these minimal residual tumor cells in the patient. Polymerase chain reaction (PCR) amplification of non-random chromosome translocations permits sensitive detection of tumors. However, the majority of patients with multiple myeloma (MM) do not demonstrate such non-random chromosomal translocations, and alternative strategies are necessary to detect minimal residual disease (MRD). MM cells rearrange either the immunoglobulin (Ig) heavy or light chains or both, and their clonal progeny bear the identical rearrangement. This unique rearrangement provides a target for amplification and detection of MRD. Moreover, competitive PCR assays can be used to assess quantitatively the tumor burden within the patient. PCR analysis at both the Ig heavy and light chain loci will be used to amplify the MM specific antigen receptor. Sequence analysis of the PCR product will enable us to design junctional specific oligonucleotide probes to detect and quantitate leukemic burden in patients with MM undergoing novel treatment strategies. This core will provide the methodologies to test whether novel treatment strategies, outlined in Projects 1,2 and 3, are capable of eliminating detectable MM cells. Furthermore, since the Ig gene rearrangements are unique to the tumor cells, they provide a potential tumor antigen that can be used as a target for proposed immunotherapeutic strategies. To this end, we propose four specific aims. First: to PCR amplify and sequence Ig heavy and light chain rearrangement. Second, to design allele specific oligonucleotides derived from the I g sequences that can be used to detect and follow minimal residual disease in patients with MM. Third to clone and express the idiotype from patients with MM. Fourth to develop tools for the presentation of idiotype and other potential MM specific antigens for clinical trials. Our overall goal is to provide the techniques necessary to allow Projects 1,2 and 3 to assess the clinical significance of detection and quantification of MRD and hopefully to enable us to identify patients at high risk for subsequent failure. Furthermore, we can then assess whether any novel treatment approaches are capable of eradicating minimal disease in these patients, to address whether this can be used as a surrogate end-point predicting outcome in proposed clinical trials.

越来越多的证据表明，根除最小残留 可察觉的疾病是治愈疾病的必要条件。相当大的努力已经 因此，在过去的十年里，我们开发了敏感的方法来 检测患者体内这些微小的残留肿瘤细胞。聚合酶链条 非随机染色体易位的反应(PCR)扩增 允许灵敏地检测肿瘤。然而，大多数患者 多发性骨髓瘤(MM)不显示这种非随机染色体 易位，和替代策略是必要的，以检测最小 残留病(MRD)。MM细胞重排免疫球蛋白(Ig) 重链或轻链或两者兼而有之，其克隆后代带有相同的 重新编排。这种独特的重新安排提供了一个目标 MRD的扩增和检测。此外，竞争性的聚合酶链式反应分析可以 用于定量评估患者体内的肿瘤负担。聚合酶链反应 免疫球蛋白重链和轻链基因座的分析将用于扩增 MM特异性抗原受体。扩增产物的序列分析 将使我们能够设计连接特异性寡核苷酸探针来 新型多发性骨髓瘤患者白血病负荷的检测与量化 治疗策略。该核心将提供用于测试的方法 项目1、2和3中概述的新治疗策略是否 能够消除可检测到的MM细胞。此外，由于免疫球蛋白基因 重排是肿瘤细胞独有的，它们提供了潜在的 可用作拟议免疫治疗靶点的肿瘤抗原 战略。为此，我们提出了四个具体目标。第一：到聚合酶链式反应 扩增和测序Ig重链和轻链重排。第二，到 从Ig序列衍生的设计等位基因特异性寡核苷酸 可用于检测和跟踪患者的微小残留疾病 第三，克隆和表达MM患者的独特型。 第四，开发独特型和其他类型的呈现工具 可能用于临床试验的MM特异性抗原。我们的总体目标是 提供必要的技术，使项目1、2和3能够评估 MRD检测和定量的临床意义 以使我们能够识别后续失败的高危患者。 此外，我们还可以评估是否有任何新的治疗方法 能够根除这些患者的微小疾病，以解决 这是否可以作为替代终点来预测 拟议的临床试验。