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: 10413583
• 负责人: Qiao Lin
• 金额: $ 19.19万
• 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/10413583
• 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; Development; Devices; Diagnostic; Disease; Disease remission; Effectiveness; Evolution; Generations; Hematologic Neoplasms; Hour; Immunoglobulin Idiotypes; Immunoglobulins; In Vitro; Ions; Lead; Libraries; Ligands; Magic; Magnetic nanoparticles; Malignant - descriptor; Malignant Neoplasms; Mantle Cell Lymphoma; Medicine; Methods; Microfluidics; Monitor; Monoclonal Antibodies; Multiple Myeloma; Nucleic Acids; Oligonucleotides; Pathologic; Patient Care; Patient Isolation; Patients; Precision therapeutics; Process; Proteins; Protocols documentation; Randomized; Reagent; Residual Neoplasm; Residual Tumors; Residual state; Sampling; Specificity; Surface Immunoglobulins; Tars; Technology; Therapeutic; Time; Tin; Translations; aptamer; base; cancer care; cancer cell; clinical decision-making; cohort; cost effective; cross reactivity; improved; innovation; nanoparticle; 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.

个性化医疗将大大提高癌症护理的有效性;然而， 实际上可实现的患者特异性策略仍然具有挑战性。B存在一个独特的机会 细胞来源的恶性肿瘤，通常表达具有可变区（独特型， Ids）在其B细胞受体（BCR）内。由于恶性细胞起源于单个克隆，因此这种sIg-Id分子 是肿瘤特有的，每个病人都是独一无二的。因此，靶向sIg-Ids可以使个性化疾病 识别和治疗策略。早期使用患者特异性抗sIg-Id抗体的研究表明， 有希望的结果，但被认为是不可持续的。一种能在短时间内生成个性化配体的技术- 高效且具有成本效益的方式仍然是基于sIg-Id的诊断和治疗中尚未满足的需求。 适体，即，特异性结合生物靶标的单链寡核苷酸提供了一种有吸引力的 解决这一未满足的需求。通过体外扩增从随机化寡核苷酸文库中分离适体。 被称为SELEX的过程，这是传统的劳动密集型，耗时（长达一个月），不切实际 用于个性化适体生成。相比之下，我们已经开发了一个微流体平台，称为microSELEX （μSELEX），其已用于分离蛋白质生物标志物的适体，包括单克隆抗体的Id区域。 多发性骨髓瘤和COVID-19患者的抗体。从病人身上提取的单克隆蛋白质 该平台能够在约10小时内快速分离个性化抗Id适体。 我们建议探索具有时间效率和成本效益的μSELEX分离患者特异性DNA适体 靶向肿瘤B细胞的sIg-Ids用于B细胞恶性血液病。我们将首先建立一个最优的μSELEX 方案使用B细胞衍生的细胞系，然后分离抗肿瘤B细胞的抗sIg适体， 外周血样本的B细胞淋巴瘤患者，并最终证明非侵入性外周血- 通过使用所述适体检测循环肿瘤B细胞来监测微小残留病。 除了能够及时识别微小残留病，以做出更精确的临床决策外， 因此，抗sIg-Id适体也可以用作治疗配体，以实现个性化和精确的治疗。 更有效的疾病治疗的靶向治疗。因此，这种个性化的适体可能导致 B细胞恶性血液病患者护理的变革。