A Practical Approach to Tumor-Specific Aptamers for B-Cell Hematologic Malignancies

B 细胞血液恶性肿瘤肿瘤特异性适体的实用方法

• 批准号: 10611461
• 负责人: Qiao Lin
• 金额: $ 18.84万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10611461
• 关键词: Affinity; Antibodies; B lymphoid malignancy; B-Cell Antigen Receptor; B-Cell Lymphomas; B-Cell Neoplasm; B-Lymphocytes; Behavior; Binding; Biochemical; Biological; Blood specimen; COVID-19; Cancer Diagnostics; Cell Line; Cell surface; Cells; Chronic Lymphocytic Leukemia; Classification; Clinical; Consumption; DNA; Detection of Minimal Residual Disease; Development; Diagnostic; Disease; Disease remission; Effectiveness; Evolution; Generations; Hematologic Neoplasms; Hour; Immunoglobulin Idiotypes; In Vitro; Libraries; Ligands; Magic; Magnetic nanoparticles; Malignant - descriptor; Malignant Neoplasms; Mantle Cell Lymphoma; Methods; Microfluidics; Monitor; Monoclonal Antibodies; Multiple Myeloma; Nucleic Acids; Oligonucleotides; Patient Care; Patients; Precision therapeutics; Process; Proteins; Protocols documentation; Randomized; Reagent; Residual Neoplasm; Sampling; Specificity; Surface Immunoglobulins; Technology; Therapeutic; Time; Translations; aptamer; cancer care; cancer cell; clinical decision-making; cohort; cost effective; cross reactivity; improved; innovation; neoplastic cell; novel; peripheral blood; personalized medicine; personalized therapeutic; protein biomarkers; response; targeted treatment; treatment strategy; tumor

Personalized medicine will greatly improve the effectiveness of cancer care; however, the development of practically attainable patient-specific strategies has remained challenging. One unique opportunity exists with B cell-derived malignancies, which often express surface immunoglobulins (sIgs) with variable regions (idiotypes, Ids) within their B-cell receptors (BCRs). As malignant cells originate from a single clone, such sIg-Id molecules are specific to the tumor and unique to each patient. Targeting sIg-Ids can hence enable personalized disease identification and treatment strategies. Early studies using patient-specific anti-sIg-Id antibodies yielded promising results but were deemed unsustainable. A technology to generate personalized ligands in a time-efficient and cost-effective manner remains an unmet need in sIg-Id-based diagnostics and therapeutics. Aptamers, i.e., single-stranded oligonucleotides that specifically bind to biological targets, offer an attractive solution to this unmet need. Aptamers are isolated from a randomized oligonucleotide library via an in vitro process known as SELEX, which is traditionally labor-intensive, time-consuming (up to a month), and impractical for personalized aptamer generation. In contrast, we have developed a microfluidic platform, called microSELEX (μSELEX), which has been used to isolate aptamers for protein biomarkers, including Id regions of monoclonal antibodies from patients with multiple myeloma and COVID-19. Given a monoclonal protein from a patient sample, the platform is capable of rapidly isolating personalized anti-Id aptamers within ~10 hours. We propose to explore time-efficient and cost-effective μSELEX isolation of patient-specific DNA aptamers targeting sIg-Ids of tumor B cells for B-cell hematologic malignancies. We will first establish an optimal μSELEX protocol using B cell-derived cell lines, then isolate anti-sIg aptamers against tumor B cells obtained from peripheral blood samples of B cell lymphoma patients, and finally demonstrate noninvasive peripheral blood-based monitoring of minimal residual disease by using the aptamers to detect circulating tumor B cells. In addition to enabling timely identification of minimal residual disease for more precise clinical decision making, anti-sIg-Id aptamers can also be used as therapeutic ligands to enable personalized and precisely targeted therapy for more effective disease treatment. Such personalized aptamers can hence potentially lead to transformative changes in the care of patients with B-cell hematologic malignancies.

个性化医学将大大提高癌症护理的有效性；但是，实际上可达到的患者特定策略的发展仍然是挑战。一个独特的机会是存在B细胞衍生的恶性肿瘤，通常在其B细胞受体（BCR）中表达具有可变区域（白痴，ID）的表面免疫球蛋白（SIG）。由于恶性细胞起源于单个克隆，因此这种SIG-ID分子特有肿瘤，并且每个患者独有。靶向信号可以实现个性化的疾病鉴定和治疗策略。使用患者特异性抗SIG-ID抗体的早期研究产生了希望的结果，但被认为是不可持续的。在基于SIG-ID的诊断和疗法中，以时间效率且具有成本效益的方式生成个性化配体的技术仍然是未满足的需求。 特别是与生物学靶标结合的单链寡核苷酸，即单链寡核苷酸，为这种未满足的需求提供了有吸引力的解决方案。通过称为SELEX的体外过程，从随机的寡核苷酸库中分离出蜂窝，该过程传统上是实验室密集型的，耗时的（长达一个月），并且对于个性化的摄影仪的产生而言是不切实际的。相比之下，我们开发了一个称为Microselex（μSelex）的微流体平台，该平台已被用来分离用于蛋白质生物标志物的适体，包括来自多发性骨髓瘤和共同率的患者的单克隆抗体的ID区域。鉴于患者样品的单克隆蛋白，该平台能够在约10小时内快速隔离个性化的抗ID适体。 我们将首先使用B细胞衍生的细胞系建立最佳的μSELEX方案，然后将抗SIG适体分离出从B细胞淋巴瘤患者的外周血血液样本中获得的肿瘤B细胞，并最终证明了使用适中探测循环肿瘤B细胞的最小残留疾病的非侵袭性外周血液基于残留的残留疾病。 除了及时确定最小残留疾病以进行更精确的临床决策之外，抗SIG​​-ID适体也可以用作治疗配体，以实现个性化和精确靶向靶向治疗以进行更有效的疾病治疗。因此，这种个性化的适体可能会导致B细胞血液学恶性肿瘤患者的护理变化。