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

个性化医疗将极大地提高癌症护理的有效性；然而， 实际可实现的针对患者的策略仍然具有挑战性。B存在一个独特的机会 细胞性恶性肿瘤，通常表达具有可变区域的表面免疫球蛋白(SIGs)(独特型， 在它们的B细胞受体(BCR)内)。由于恶性细胞起源于单个克隆，这样的sig-ID分子 是肿瘤特有的，对每个患者来说都是独一无二的。因此，针对SIG-ID可以实现个性化疾病 识别和治疗策略。使用患者特异性抗SIG-ID抗体的早期研究产生 有希望的结果，但被认为是不可持续的。一种在一段时间内产生个性化配体的技术- 在基于SIG-ID的诊断和治疗中，高效和经济高效的方式仍然是一个未得到满足的需求。 适配子，即与生物靶标特异结合的单链寡核苷酸，提供了一种诱人的 解决这一未得到满足的需求。通过体外实验从随机寡核苷酸文库中分离适体 称为SELEX的过程，传统上是劳动密集型、耗时(长达一个月)且不切实际 用于个性化适配子生成。相比之下，我们开发了一种微流控平台，称为microSELEX (μSELEX)，已被用于分离蛋白质生物标记物的适体，包括单克隆的ID区 多发性骨髓瘤患者和新冠肺炎患者的抗体。从病人那里得到了一种克隆蛋白 该平台能够在~10小时内快速分离出个性化的抗ID适配子。 我们建议探索既省时又经济的DNASELEX分离患者特异性μ适配子 靶向肿瘤B细胞SIG-ID治疗B细胞恶性血液病。我们将首先建立一个最优μSELEX 使用B细胞来源的细胞系，然后分离抗肿瘤B细胞的抗Sig适配子 B细胞淋巴瘤患者的外周血样本，最终证实无创外周血- 通过使用适配子检测循环中的肿瘤B细胞来监测微小残留病。 除了能够及时识别最小残留疾病，以便做出更准确的临床决策 制造、抗SIG-ID适体也可以用作治疗配体，使个性化和精确化 为更有效的疾病治疗提供靶向治疗。因此，这种个性化的适配子可能会导致 到B细胞恶性血液病患者护理的变革性变化。

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.