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

• 批准号: 10012271
• 负责人: Rolf Muller
• 金额: $ 83.8万
• 依托单位: BIOFLUIDICA, INC
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-18 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10012271
• 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

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（全部）是儿童期最常见的恶性疾病 在18岁之前诊断出的所有癌症中，约有30％（1）。所有人的主要死亡原因 患者是疾病的缓解。因此，监测最小残留疾病（MRD）被认为是最大的 急性白血病中预后的有力预测指标，包括B型急性淋巴细胞白血病（B-ALL）。 临床医生可以在肿瘤细胞迅速扩展为氟化，先发制人之前识别患者的MRD 可以通过更好的患者结局进行疗法。对于小儿B-all，有现有的测试 监测MRD的继电器，包括PCR或多参数流式细胞术，但需要骨髓 抽吸物，这可能是痛苦的，并限制了测试的频率（2,3）。如果可以在B-ALL中检测到MRD 外周血而不是骨髓的患者，相应的测定可以帮助指导治疗 启用精确药物，从而获得更好的患者结局。 在此应用程序中，由微流体组件组成的创新测试和相关硬件将是 发达。该测试可以为MRD测试提供较高的临床灵敏度，并且允许通常微创 使用外周血（1 mL）取样而不是侵入性，尤其是针对儿科患者，骨髓 活检。该测定使用微流体装置来分析外周血并寻找循环白血病 细胞（CLC）。在急性髓样白血病（AML）患者的纵向研究中，使用该微流体测定法 在干细胞移植后，与2个月前通过监测CLC的MRD相比 多参数流式细胞仪（MFC）和PCR都使用了骨髓抽吸；微流体测定 比PCR和MFC更敏感的2个级级。由于微流体测定的能力 检测血液中的CLC，与骨骼相比，可以更频繁地测试患者疾病状态 骨髓活检测试。对于B-ALL，固定在微流体设备中的抗CD19抗体可以亲和力 - 表达CD19表面抗原的精选细胞通常由B-ALL淋巴细胞（即CLC）和 正常的B细胞。 CLC通过表达异常标记（例如末端脱氧核苷酸）的表达来鉴定 转移酶（TDT）和跟踪的CLC数量，以确定救济或救济风险。 在此SBIR I/II阶段快速提案中，CLC微流体测试将扩展和开发 商业化以监测B-ALL小儿患者的MRD和潜在缓解，以提供100％的覆盖率。 鉴于产生的迄今为止的强大数据以及对改进易于实施的紧急诊断需求 MRD测定经常监测，拟议的测试满足了小儿区域未满足的临床需求 肿瘤学。值得注意的是，该测试可以重新编程以搜索其他儿科肿瘤学疾病，例如 T细胞全部（仅需要细胞选择抗体的变化）。