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微量残留 可检测到的疾病对于治愈来说是必要的。付出了相当大的努力 因此，在过去的十年里，人们开发了敏感的方法 检测患者体内这些最小残留的肿瘤细胞。聚合酶链 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) 并不表现出这种非随机染色体 易位和替代策略对于检测最小限度是必要的 残留病灶（MRD）。 MM 细胞重新排列免疫球蛋白 (Ig) 重链或轻链或两者，及其克隆后代具有相同的 重新排列。这种独特的重新排列提供了一个目标 MRD的扩增和检测。此外，竞争性 PCR 检测可以 用于定量评估患者体内的肿瘤负荷。聚合酶链式反应 Ig 重链和轻链基因座的分析将用于扩增 MM 特异性抗原受体。 PCR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 检测并定量接受新疗法的 MM 患者的白血病负担 治疗策略。该核心将提供测试方法 项目 1,2 和 3 中概述的新治疗策略是否有效 能够消除可检测到的 MM 细胞。此外，由于 Ig 基因 重排是肿瘤细胞所独有的，它们提供了潜在的 可用作拟议免疫治疗靶标的肿瘤抗原 策略。为此，我们提出四个具体目标。第一：PCR 扩增 Ig 重链和轻链重排并对其进行测序。其次，为了 设计源自 Ig 序列的等位基因特异性寡核苷酸 可用于检测和跟踪患者微小残留病 与MM。第三，克隆并表达MM患者的独特型。 第四，开发展示独特型和其他特征的工具 用于临床试验的潜在 MM 特异性抗原。我们的总体目标是 提供必要的技术以允许项目 1,2 和 3 评估 MRD 检测和定量的临床意义以及希望 使我们能够识别后续失败的高风险患者。 此外，我们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 这是否可以用作预测结果的替代终点 提议的临床试验。