CORE--MOLECULAR BIOLOGY AND IMMUNE ASSESSMENT

核心--分子生物学和免疫评估

• 批准号: 6103509
• 负责人: JOHN G. GRIBBEN
• 金额: $ 27.53万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-03 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6103509
• 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，消除微量残留 可检测的疾病是治愈的必要条件。大量努力 因此，在过去的十年中，开发了敏感的方法， 检测出病人体内残留的微小肿瘤细胞。聚合酶链 非随机染色体易位的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）患者没有表现出这种非随机染色体 易位和替代策略是必要的，以检测最小的 残留病（MRD）。MM细胞重排免疫球蛋白（IG） 重链或轻链或两者，并且它们的克隆后代携带相同的 重排这种独特的重排提供了一个目标， MRD的扩增和检测。此外，竞争性PCR测定可 用于定量评估患者体内的肿瘤负荷。PCR 在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， 检测并定量接受新治疗的MM患者的白血病负荷 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 能够消除可检测的MM细胞。此外，由于IG基因 重排是肿瘤细胞所特有的，它们提供了一种潜在的 可用作建议的免疫疗法的靶点的肿瘤抗原 战略布局为此，我们提出了四个具体目标。第一：PCR 扩增和测序IG重链和轻链重排。二是 设计来自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的微小残留疾病， 第三，从MM患者中克隆并表达独特型。 第四，开发用于呈现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，使项目1、2和3能够评估 MRD检测和定量的临床意义， 以使我们能够识别出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， 这是否可以作为预测结果的替代终点， 拟进行临床试验。