Validating Rapid Microfluidic Isolation of Personalized Aptamers for Monitoring Minimal Residual Disease in Multiple Myeloma

验证个性化适体的快速微流体分离用于监测多发性骨髓瘤的微小残留疾病

• 批准号: 9532103
• 负责人: Qiao Lin
• 金额: $ 38.61万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-07 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9532103
• 关键词: Address; Antibodies; Binding; Biological Assay; Biopsy Specimen; Blood; Bone marrow biopsy; Caring; Chemistry; Clinical; Clinical Research; DNA; Detection; Detection of Minimal Residual Disease; Devices; Diagnosis; Disease; Doctor of Medicine; Doctor of Philosophy; Electrophoresis; Evaluation; Evolution; Failure; Generations; Goals; Grant; Hematopoietic Neoplasms; Hour; IgG1; Immunoglobulin Idiotypes; Immunoglobulin Variable Region; Immunoglobulins; In Vitro; Individual; Lead; Libraries; Ligands; Medical; Methods; Microfluidic Microchips; Microfluidics; Modeling; Monitor; Monoclonal Antibodies; Multiple Myeloma; New York; Newly Diagnosed; Oligonucleotides; Pathology; Patient Care; Patients; Plasma Cells; Procedures; Process; Proteins; Protocols documentation; Randomized; Reagent; Residual Neoplasm; Sampling; Sensitivity and Specificity; Serum; Serum Proteins; Specimen; Technology; Test Result; Time; Universities; aptamer; base; cost; follow-up; improved; individual patient; innovation; microfluidic technology; peripheral blood; polyclonal antibody; public health relevance; real time monitoring; rituximab; tumor

 DESCRIPTION (provided by applicant): Multiple myeloma (MM) is a plasma cell derived blood cancer that produces monoclonal immunoglobulins (M-Ig). Detection of minimal residual disease (MRD) is of great importance for MM care. All methods used for MRD detection are based on evaluation of plasma cells obtained from bone marrow (BM) biopsies. These methods are susceptible to failure and do not allow frequent monitoring because of their invasiveness. Detection of MRD in peripheral blood is most ideal but remains an unmet medical challenge. Circulating M-Ig proteins are the hallmark of MM, and their detection by currently available serum-based methods, including serum protein electrophoresis (SPEP) and immunofixation (IFE), is used in diagnosis and follow-up, but not MRD monitoring due to insufficient sensitivity. This problem can be addressed by exploiting variable regions (called "idiotypes") of M-Ig proteins, which are tumor-specific and unique to the patient. The ability to monitor the presence and reappearance of the same idiotype in blood after therapy would lead to sensitive MRD detection. This can be enabled by idiotype-targeting aptamers as personalized reagents. Aptamers are single-stranded oligonucleotides that specifically bind to targets, and have been demonstrated to allow sensitive serum-based detection of analytes including immunoglobulins. Through an in vitro process termed systematic evolution of ligands by exponential enrichment (SELEX), aptamers are generated from randomized libraries of oligonucleotides. MRD detection requires generation of aptamers targeting the unique idiotype of the patient. This is impractical with conventional SELEX methods that are labor-intensive and time-consuming, but can be enabled by our innovative microfluidic technology that allows rapid generation of aptamers. Our preliminary microfluidic devices have demonstrated the capability of completing the SELEX process within ~10 hours for successful isolation of immunoglobulin-targeting aptamers. Our long-term objective is to realize a paradigm for sensitive detection MRD in peripheral blood using personalized aptamers targeting patient- and tumor-specific idiotypes of M-Ig proteins of individual MM patients. The goal of this project is to validate the ability of our microfluidic technology to rapidly generate idiotype- targeting aptamers for individual patients for sensitive MRD detection in peripheral blood. We will first optimize a SELEX protocol using readily available IgG1/kappa monoclonal antibodies, and then use this protocol to validate microfluidic isolation of aptamers targeting patient M-Ig proteins for serum-based M-Ig detection. At the end of the three-year grant period, we would be ready to initiate a clinical study, including longitudinal, real-time monitoring of actual patients with intact immunoglobulin multiple myeloma (IIMM) with increased levels of the IgG1/kappa monoclonal antibody, while in parallel we will expand and generalize our procedure to other classes/subclasses, enabling personalized reagents capable of dramatically improving MRD detection for optimal care for nearly all MM patients.

 描述（由适用提供）：多发性骨髓瘤（MM）是一种血浆细胞衍生的血液癌，可产生单克隆免疫球蛋白（M-Ig）。最小残留疾病（MRD）的检测对于MM护理非常重要。用于MRD检测的所有方法均基于对从骨髓（BM）活检获得的浆细胞的评估。这些方法容易发生故障，并且由于其侵入性不允许经常监测。在外周血中检测MRD是最理想的选择，但仍然是未得到的医学挑战。循环的M-Ig蛋白是MM的标志，它们通过当前可用的基于血清的方法（包括血清蛋白电泳（SPEP）和免疫固定（IFE））进行检测，用于诊断和随访，但由于敏感性不足而非MRD监测。可以通过利用M-Ig蛋白的可变区域（称为“白痴类型”）来解决此问题，该蛋白是肿瘤特异性且患者独有的。治疗后同一典型型在血液中同一典型型的存在和重新出现的能力将导致敏感的MRD检测。这可以通过将定型适应器作为个性化试剂来启用。 apatamers是特异性结合靶标的单链寡核苷酸，并已被证明可以允许基于敏感的血清检测包括免疫球蛋白在内的分析物。通过指数富集（SELEX）的体外过程，称为配体的系统演变，适体是由寡核苷酸的随机文库产生的。 MRD检测需要生成针对患者独特异常的适体。这对于劳动密集型且耗时的传统SELEX方法是不切实际的，但是我们的创新微流体技术可以启用，从而可以快速产生适体。我们的初步微流体设备已经证明了成功隔离免疫球蛋白靶向适体的能力。我们的长期目标是使用针对单个MM患者的M-Ig蛋白的患者和肿瘤特异性认同类型的个性化适体，实现外周血中敏感检测MRD的范例。该项目的目的是验证我们的微流体技术快速生成针对个体患者适体的身份类型的能力，以在外周血中检测敏感的MRD检测。我们将首先使用易于可用的IgG1/kappa单克隆抗体优化SELEX方案，然后使用此方案验证针对患者M-Ig蛋白的适体的微流体隔离，以基于血清的M-Ig检测。 At the end of the three-year grant period, we would be ready to initiate a clinical study, including longitudinal, real-time monitoring of actual patients with intact immunoglobulin multiple myeloma (IIMM) with increased levels of the IgG1/kappa monoclonal antibody, while in parallel we will expand and generalize our procedure to other classes/subclasses, enabling personalized reagents capable of dramatically improving MRD detection几乎所有MM患者的最佳护理。

项目成果

A graphene aptasensor for biomarker detection in human serum.

• DOI: 10.1016/j.electacta.2018.08.062
• 发表时间: 2018-11-10
• 期刊: Electrochimica acta
• 影响因子: 6.6
• 作者: Wang X;Zhu Y;Olsen TR;Sun N;Zhang W;Pei R;Lin Q
• 通讯作者: Lin Q

Measurement of cytokine biomarkers using an aptamer-based affinity graphene nanosensor on a flexible substrate toward wearable applications.

• DOI: 10.1039/c8nr04315a
• 发表时间: 2018-11-29
• 期刊: Nanoscale
• 影响因子: 6.7
• 作者: Hao Z ;Wang Z ;Li Y ;Zhu Y ;Wang X ;De Moraes CG ;Pan Y ;Zhao X ;Lin Q
• 通讯作者: Lin Q

Selective detection of water pollutants using a differential aptamer-based graphene biosensor.

• DOI: 10.1016/j.bios.2018.10.047
• 发表时间: 2019-02-01
• 期刊: Biosensors & bioelectronics
• 影响因子: 12.6
• 作者: Li Y;Zhu Y;Wang C;He M;Lin Q
• 通讯作者: Lin Q

Real-Time Monitoring of Insulin Using a Graphene Field-Effect Transistor Aptameric Nanosensor.

• DOI: 10.1021/acsami.7b07684
• 发表时间: 2017-08-23
• 期刊: ACS applied materials & interfaces
• 影响因子: 9.5
• 作者: Hao Z;Zhu Y;Wang X;Rotti PG;DiMarco C;Tyler SR;Zhao X;Engelhardt JF;Hone J;Lin Q
• 通讯作者: Lin Q