Deep sequencing for minimal residual disease detection in Acute Lymphoblastic Leu

深度测序用于急性淋巴细胞白血病微小残留病检测

• 批准号: 8632909
• 负责人: Harlan S. Robins
• 金额: $ 66.01万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-19 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8632909
• 关键词: Acute; Acute Lymphocytic Leukemia; Adoption; Aftercare; Biological Assay; Bone Marrow; Caring; Children' s Oncology Group; Clinical; Clinical Sensitivity; Clinical Trials; Clonal Evolution; Complex; Data; Detection; Detection of Minimal Residual Disease; Development; Diagnosis; Diagnostic; Disease; Disease Outcome; Europe; Evaluation; Flow Cytometry; Gene Rearrangement; Goals; Gold; Hematopathology; High-Throughput Nucleotide Sequencing; Housing; Immunoglobulins; Individual; Laboratories; Manuscripts; Measurement; Measures; Methods; Molecular; Neoadjuvant Therapy; Outcome; Patients; Preparation; Receptor Gene; Receptors, Antigen, B-Cell; Relapse; Research Infrastructure; Residual Neoplasm; Running; Sampling; Sensitivity and Specificity; Specificity; T-Cell Receptor Genes; T-Lymphocyte; Technology; Testing; Time; United States; Universities; Validation; Washington; Wood material; base; chemotherapy; clinically significant; cohort; cost; deep sequencing; design; immunoglobulin receptor; improved; lymphoid neoplasm; next generation sequencing; public health relevance; standard of care

PROJECT SUMMARY/ABSTRACT Acute lymphoblastic leukemia (ALL) is an aggressive immature lymphoid neoplasm. Over the last couple of decades, there has been substantial improvement in disease outcomes for these patients [3]. In part, this improvement is due to enhanced identification of patients requiring additional therapy based on the assessment of minimal residual disease (MRD). Currently, assessment of MRD is achieved through the use of either patient- specific molecular assays (predominantly in Europe), and/or multi-parametric flow cytometry (mpFC), (commonly in the US). At present, implementation of these approaches in a uniform manner is complex and challenging. Individualized molecular assessment of MRD is limited by requirements for a large institutional infrastructure, as molecular assays for each patient must be individually-designed and validated. By contrast, multi-parametric flow cytometry is difficult to standardize, resulting in disparate quality. While it is clear that MRD is important for guiding patient-specific care, current approaches are not robust to achieve this goal consistently. In a preliminary project involving 43 paired pre- and post-treatment samples from an ongoing T-ALL Children's Oncology Group (COG) trial AALL0434 [1], we demonstrate that high-throughput sequencing of T-cell receptor gene rearrangements enhanced detection of very low-level MRD, improving upon the sensitivity and specificity of multi-parametric flow cytometry, without requiring the complex institutional infrastructure required to individualize high-sensitivity molecular evaluation for minimal residual disease. We found that high-throughput sequencing could identify MRD in all patients in whom multi-parametric flow cytometry identified disease. However, sequencing also permitted detection of MRD at a higher sensitivity of approximately 10-fold. We have since completed analysis of another cohort of 99 patients with B-lineage acute lymphoblastic leukemia derived from COG trial AALL0932 (manuscript in preparation). In this study, we also found that next-generation sequencing of immunoglobulin receptor gene rearrangements could provide enhanced detection of MRD. Several questions are raised by these data. First, what is the clinical specificity and sensitivity of these findings? Second, to what extent is next-generation sequencing suitable for routine implementation in the clinical laboratory? In this proposal, we apply high-throughput sequencing to sensitively and comprehensively assess MRD in patients with B- and T-lineage ALL.

项目摘要/摘要 急性淋巴细胞白血病（ALL）是一种侵袭性的未成熟淋巴肿瘤。在最后一对 数十年来，这些患者的疾病预后有了很大的改善[3]。在某种程度上 改善是由于对需要根据评估进行额外治疗的患者的鉴定增强了 最小残留疾病（MRD）。目前，通过使用任何一名患者来实现MRD的评估 - 特定的分子测定（主要在欧洲）和/或多参数流式细胞仪（MPFC）， （通常在美国）。目前，以统一的方式实施这些方法是复杂的，并且 具有挑战性的。 MRD的个性化分子评估受到大型机构的要求的限制 基础设施，作为每个患者的分子测定法必须单独设计和验证。相比之下 多参数流式细胞术很难标准化，从而导致不同的质量。虽然很明显MRD 对于指导患者特定的护理很重要，目前的方法无法始终如一地实现这一目标。 在一个初步项目中，涉及来自正在进行的T-All的43个配对前后的处理前和处理后样品 儿童肿瘤学组（COG）试验AALL0434 [1]，我们证明了T细胞的高通量测序 受体基因重排增强了对非常低水平的MRD的检测，从而提高了灵敏度和 多参数流式细胞术的特异性，而无需复杂的机构基础设施 个性化高敏性分子评估，以最小残留疾病。我们发现高通量 测序可以鉴定所有多参数流式细胞术鉴定疾病的患者中的MRD。 但是，测序还允许在较高灵敏度的大约10倍的敏感性下检测MRD。我们有 自从完成了另一组99例B-linege急性淋巴细胞白血病患者的队列以来 来自COG试验AALL0932（准备中的手稿）。在这项研究中，我们还发现下一代 免疫球蛋白受体基因重排的测序可提供MRD的增强检测。 这些数据提出了几个问题。首先，这些的临床特异性和灵敏度是什么 发现？其次，下一代测序在多大程度上适合于常规实施 临床实验室？在此提案中，我们将高通量测序应用于敏感和全面 评估B-和T-Linege患者的MRD。