Microsampling Assays for Immunosuppressive Drugs in Children

项目摘要 免疫抑制治疗是实体器官和骨移植后长期预后的基础。 骨髓/干细胞移植和作为各种自身炎症性疾病的治疗，包括类风湿性关节炎 关节炎、湿疹、牛皮癣、克罗恩病和成人和儿童的肾病综合征。最佳血液 这些药物的浓度对于使毒性最小化和防止移植受者的排斥是关键的。 这些药物需要频繁且通常是终身的治疗药物监测（TDM），以确保给药 维持最佳治疗浓度。用于给药指导的当前TDM（临床标准） 环孢霉素A（CYA）、他克莫司（TAC）和西罗莫司（SIR）中的至少一种衍生自全血免疫测定或 液相色谱-串联质谱（LC-MS/MS）测定。这需要免疫抑制 个人去门诊实验室进行静脉切开术，可能会增加他们获得感染的风险。 开发一种易于使用、体积小的血液采样方法可以促进家庭采样， 减轻儿科患者及其家庭的负担。目前的TDM需要0.5至1.0 mL全血， 通常通过静脉切开术。使用Mitra装置或Tasso-M20装置进行体积吸收微量取样（VAMS） 允许收集固定的少量血液（例如，20 µ L），而无需 静脉切开术最近在成人中进行了使用Mitra器械的CYA、TAC和SIR的临床确认研究， 报道虽然一项研究显示TAC分析具有良好的相关性，但另一项研究显示不一致 使用Mitra设备进行样本采集。因此，Mitra器械用于常规TDM的潜在效用是 不确定FDA批准的Tasso-M20设备允许准确，精确，无痛和一致的收集 一个固定的小体积的血液。然而，Tasso-M20器械在临床环境中的应用， TDM需要分析和临床验证。拟定微量取样试验的目的是 儿童免疫抑制药物EN（MAIDEN）研究旨在：1）验证Tasso-M20 VAMS LC-MS/MS分析 对于CYA、TAC和SIR，2）在以下条件下评价Tasso-M20器械中CYA、TAC和SIR的体外稳定性： 运输和储存条件，3）临床验证CYA、TAC的Tasso-M20 VAMS LC-MS/MS测定， 和SIR对目前儿科患者静脉全血TDM免疫测定，和4）证实稳定性 在运输和储存条件下，Tasso-M20 VAMS临床样本中的CYA、TAC和SIR。如果 成功的，这项研究将使微量采样方法用于TDM的临床设置 （避免了侵入性静脉切开术的需要），以及在家里（减轻家庭带来的负担）。 免疫抑制的儿童走出家门进行常规TDM），并彻底改变远程采样的TDM 儿童和成人的免疫抑制剂。

PROJECT SUMMARY Immunosuppressive therapy is the foundation of successful long-term outcomes after solid organ and bone marrow/stem cell transplants and as a treatment for various auto-inflammatory conditions, including rheumatoid arthritis, eczema, psoriasis, Crohn's disease, and nephrotic syndrome in adults and children. The optimal blood concentrations of these drugs are critical to minimize toxicity and to prevent rejection in transplant recipients. These drugs require frequent and often life-long therapeutic drug monitoring (TDM) to ensure that dosing maintains the optimal therapeutic concentrations. Current TDM (clinical standard) used for the dosing guidance of cyclosporine A (CYA), tacrolimus (TAC), and sirolimus (SIR) are derived from whole blood immunoassays or liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays. This requires immunosuppressed individuals to go to outpatient laboratories for phlebotomy, potentially increasing their risk of acquiring infections. The development of an easy-to-use, small volume, blood sampling approach can facilitate home sampling and reduce the burden on pediatric patients and their families. Current TDM requires 0.5 to 1.0 mL of whole blood, typically via phlebotomy. Volumetric absorptive microsampling (VAMS) with Mitra devices or Tasso-M20 devices allows for the collection of a fixed small volume of blood (e.g., 20 µL) from a capillary needle without the need for phlebotomy. Clinical validation studies for CYA, TAC, and SIR with Mitra devices in adults were recently reported. While one study showed a good correlation for TAC analysis, the other study showed inconsistency with sample collection using Mitra devices. Therefore, the potential utility of Mitra devices for routine TDM is uncertain. FDA-approved Tasso-M20 devices allow for an accurate, precise, painless, and consistent collection of a fixed small volume of blood. However, the implementation of Tasso-M20 devices into the clinical setting for TDM requires both analytical and clinical validation. The objectives of the proposed Microsampling Assays for Immunosuppressive Drugs in childrEN (MAIDEN) study is to: 1) validate Tasso-M20 VAMS LC-MS/MS assays for CYA, TAC, and SIR, 2) evaluate in vitro stability of CYA, TAC, and SIR in Tasso-M20 devices under the conditions of shipping and storage, 3) clinically validate Tasso-M20 VAMS LC-MS/MS assays for CYA, TAC, and SIR against current venous whole blood TDM immunoassays in pediatric patients, and 4) confirm the stability of CYA, TAC, and SIR in Tasso-M20 VAMS clinical samples under the conditions of shipping and storage. If successful, this research will enable the use of the microsampling method for TDM in the clinical setting (obviating the need for invasive phlebotomy), and at home (reducing the burden on families to bring the immunosuppressed children out of the home for routine TDM), and revolutionize remote sampling for TDM of immunosuppressant drugs in children and adults.