Increased sensitivity of minimal residual disease monitoring using peripheral blood in pediatric patients with acute lymphoblastic leukemia

提高急性淋巴细胞白血病儿科患者外周血微小残留病监测的敏感性

基本信息

批准号：
10020337
负责人：
Rolf Muller
金额：
$ 55.67万
依托单位：
BIOFLUIDICA, INC
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-18 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10020337
关键词：
18 year old Acute Lymphocytic Leukemia Acute Myelocytic Leukemia Acute T Cell Leukemia Acute leukemia Affinity Age Allogenic Antibodies Antigens Area Aspirate substance Automation B-Cell Acute Lymphoblastic Leukemia B-Lymphocytes Biological Assay Blood Blood specimen Bone Marrow Bone marrow biopsy CD19 gene CD34 gene Cause of Death Cell Count Cell Line Cells Child Childhood Childhood Acute Lymphocytic Leukemia Classification Clinical Clinical Research Clinical Sensitivity Cytogenetics DNA Nucleotidylexotransferase Data Detection of Minimal Residual Disease Development Devices Diagnostic Diagnostic Procedure Disease Disease remission Early Intervention Flow Cytometry Frequencies Genotype Hematopoietic stem cells Image Immobilization Liquid substance Longitudinal Studies Malignant - descriptor Measurement Measures Methods Microfluidic Microchips Microfluidics Modeling Monitor Monoclonal Antibodies Mutate Neoadjuvant Therapy Outcome Pain Patient risk Patient-Focused Outcomes Patients Pediatric Oncology Phase Phenotype Procedures Process Reagent Recovery Recurrent disease Relapse Reproducibility Residual Neoplasm Residual Tumors Robot Robotics Sampling Secure Sedation procedure Series Small Business Innovation Research Grant Stains Stem cell transplant Surface Surface Antigens System T-Lymphocyte Techniques Testing White Blood Cell Count procedure acute lymphoblastic leukemia cell aged base cancer diagnosis chemotherapy circulating leukemia cell commercialization experience falls fluorescence microscope improved individual patient innovation instrument leukemia lymphoblast minimally invasive monolayer neoplastic cell outcome prediction pediatric patients peripheral blood phase 1 study precision medicine relapse patients relapse risk response standard of care

项目摘要

Abstract Acute lymphoblastic leukemia (ALL) is the most common malignant disease in childhood and accounts for approximately 30% of all cancers diagnosed before the age of 18 years (1). The primary cause of death for ALL patients is disease relapse. Therefore, monitoring for minimal residual disease (MRD) is considered the most powerful predictor of outcome in acute leukemias, including B-type acute lymphoblastic leukemia (B-ALL). If clinicians could identify a patient’s MRD before the tumor cells rapidly expand to florid relapse, preemptive therapies could be undertaken with better patient outcome. For pediatric B-ALL, there are existing tests for monitoring relapse from MRD including PCR or multi-parameter flow cytometry, but require a bone marrow aspirate, which can be painful and limits the frequency of testing (2,3). If MRD could be detected in B-ALL patients from peripheral blood and not bone marrow, the corresponding assay could assist in guiding therapy to enable precision medicine resulting in better patient outcome. In this application, an innovative test that consists of a microfluidic assay and the associated hardware will be developed. The test can provide high clinical sensitivity for MRD testing and permits frequent minimally invasive sampling using peripheral blood (1 mL) as opposed to an invasive, especially for pediatric patients, bone marrow biopsy. The assay uses a microfluidic device to analyze peripheral blood and search for circulating leukemic cells (CLCs). Using this microfluidic assay in a longitudinal study of acute myeloid leukemia (AML) patients following stem cell transplantation, MRD via monitoring of CLCs was detected ~2 months earlier compared to both multi-parameter flow cytometry (MFC) and PCR, which used bone marrow aspirates; the microfluidic assay was 2-orders of magnitude more sensitive than PCR and MFC. Owing to the ability of the microfluidic assay to detect CLCs in blood, more frequent testing of a patients’ disease status was possible when compared to bone marrow biopsy testing. For B-ALL, anti-CD19 antibodies immobilized within a microfluidic device can affinity- select cells expressing CD19 surface antigen commonly expressed by B-ALL lymphoblasts (i.e., CLC) and normal B-cells. CLCs are identified by expression of aberrant markers, such as Terminal deoxynucleotidyl Transferase (TdT) and the number of CLCs tracked to determine the onset of relapse or the risk of relapse. In this SBIR Phase I/II fast track proposal, the CLC microfluidic test will be expanded and developed for commercialization to monitor MRD and potential relapse in B-ALL pediatric patients to provide coverage of 100%. Given the strong data generated to-date and the urgent diagnostic need for an improved easy-to-implement MRD assay for frequent monitoring, the proposed test fills an unmet clinical need in the area of pediatric oncology. As a note, the test can be reprogrammed to search for other pediatric oncological diseases such as T-cell ALL (requires only a change in the cell selection antibody).

抽象的急性淋巴细胞白血病（ALL）是儿童期最常见的恶性疾病，占大约 30% 的癌症是在 18 岁之前诊断出来的 (1)。因此，对微小残留病（MRD）的监测被认为是最重要的。急性白血病（包括 B 型急性淋巴细胞白血病 (B-ALL)）结果的有力预测因子。勇士们可以在肿瘤细胞迅速扩大到严重复发之前识别患者的 MRD，先发制人对于儿童 B-ALL，现有的治疗方法可以使患者获得更好的结果。监测 MRD 复发，包括 PCR 或多参数流式细胞术，但需要骨髓抽吸，这可能会很痛苦，并且会限制 B-ALL 中检测到 MRD 的频率 (2,3)。来自外周血而不是骨髓的患者，相应的检测可以帮助指导治疗实现精准医疗，从而带来更好的患者治疗效果。在此应用中，将进行由微流体测定和相关硬件组成的创新测试该测试可为 MRD 测试提供高临床灵敏度，并允许微创。使用外周血 (1 mL) 取样，而不是侵入式骨髓取样，尤其是对于儿科患者该检测使用微流体装置分析外周血并寻找循环白血病。在急性髓性白血病 (AML) 患者的纵向研究中使用这种微流体测定。干细胞移植后，通过监测 CLC 检测到的 MRD 比之前提前约 2 个月多参数流式细胞术 (MFC) 和 PCR，均使用骨髓抽吸物进行微流控检测；由于微流控检测的能力，其灵敏度比 PCR 和 MFC 高 2 个数量级。检测血液中的 CLC，与骨相比，可以更频繁地检测患者的疾病状态对于 B-ALL，固定在微流体装置内的抗 CD19 抗体可以亲和- 选择表达 B-ALL 淋巴细胞（即 CLC）通常表达的 CD19 表面抗原的细胞，以及正常 B 细胞通过异常标记（如末端脱氧核苷酸）的表达来识别。跟踪转移酶 (TdT) 和 CLC 数量以确定复发的发生或复发的风险。在此 SBIR I/II 期快速通道提案中，CLC 微流体测试将扩展和开发用于商业化以监测 B-ALL 儿科患者的 MRD 和潜在复发，以提供 100% 的覆盖率。鉴于迄今为止生成的强大数据以及对改进的易于实施的诊断的迫切需求用于频繁监测的 MRD 检测，所提出的检测填补了儿科领域未满足的临床需求需要注意的是，该测试可以重新编程以搜索其他儿科肿瘤疾病，例如 T 细胞 ALL（仅需要改变细胞选择抗体）。