CNS HIV Theranostics Based on Intrinsic CEST Contrasts of Antiretroviral Drugs

项目摘要 我们建议开发人类免疫缺陷病毒(HIV)治疗中枢神经系统(CNS)的药物 基于抗逆转录病毒药物的内在化学交换饱和转移(CEST)对比。 Theranostics是治疗学和诊断学的组合，使生物分布成为可能 用磁性等活体成像技术测量靶器官中的抗逆转录病毒药物 磁共振成像(MRI)。艾滋病毒治疗在三个方面有助于抗逆转录病毒治疗的研究。第一, HIV治疗将成为开发针对中枢神经系统病毒库的抗逆转录病毒药物的有力工具。第二，艾滋病毒 Theranostics将有助于开发将抗逆转录病毒药物的非靶标影响降至最低的策略。第三，龙的活体成像 PK和BD对长效药物的发展至关重要。此外，艾滋病毒治疗学使真正的- 对中枢神经系统药物水平的时间监测使其有可能为个人量身定做个性化治疗 病人。 与用显像剂标记药物的传统治疗技术相比，CEST是一种固有的特性 对于ARV，因此其成像不需要外部化学物质。这消除了限制 与显像剂相关的，包括有限的治疗效果和成像敏感性，以及可能的 毒性。我们假设药物分子中质子的联合CEST效应产生了一个独特的CEST 称为CEST指纹的特征，使体内药物检测具有敏感性和特异性。我们 将首先确定一线HIV抗逆转录病毒药物的CEST指纹，并开发基于该药物的核磁共振方法 用于活体检测的CEST指纹。 目的1：研究抗逆转录病毒药物(3TC、ABC、TDF和DTG)的CEST指纹图谱。我们假设 药物的可交换质子产生独特的CEST指纹。原子核中质子的CEST效应 药物分子将被测量并组合起来，以建立药物的CEST指纹。每个人的CEST指纹 药物将在使用人单核细胞来源的巨噬细胞(MDM)的细胞中进一步验证和确定。 目的：应用CEST磁共振成像技术检测抗逆转录病毒药物在人源化小鼠体内的中枢神经系统生物分布。我们假设核磁共振 利用CEST指纹为体内抗逆转录病毒药物的检测提供了敏感性和特异性。要测试 假说、MRI数据采集和分析方法将基于ARV CEST指纹开发 在目标1中进行了测量，并优化了在感染的人源化小鼠中进行体内检测的优化 抗逆转录病毒治疗(DTG/ABC/3TC或DTG/3TC/TDF)。用液体测量脑组织药物浓度 将使用色谱质谱(LC-MS/MS)来验证CEST MRI结果。 这项研究的一个成功成果是一种测量大脑ARV水平的非侵入性MRI技术。这个 这项技术将为开发减轻艾滋病毒中枢神经系统并发症的治疗方法提供强有力的工具 感染和改善艾滋病毒患者的心理健康结果。

Project Summary We propose to develop human immunodeficiency virus (HIV) theranostics for the central nervous system (CNS) based on the intrinsic chemical exchange saturation transfer (CEST) contrasts of antiretroviral drugs. Theranostics is a combination of the terms therapeutics and diagnostics that enables the biodistribution measurements of antiretrovirals (ARVs) in target organs using in vivo imaging techniques such as magnetic resonance imaging (MRI). HIV theranostics benefits the research of antiretroviral therapy in three ways. First, HIV theranostics will be a powerful tool for the development of ARVs targeting CNS viral reservoirs. Second, HIV theranostics will help develop strategies to minimize the off-target effects of ARVs. Third, In vivo imaging of long term PK and BD is critical for the development of long-acting drugs. Moreover, HIV theranostics enables real- time monitoring of CNS drug levels making it possible to design personalized treatments tailored for individual patients. Compared to traditional theranostic technologies that tag drugs with imaging agents, CEST is an intrinsic property of an ARV, therefore no extrinsic chemical agents are required for its imaging. This eliminates the limitations associated with imaging agents including limited therapy effectiveness and imaging sensitivity, and possible toxicity. We posit that the combined CEST effects of the protons in a drug molecule generate a unique CEST characteristic termed as CEST fingerprint that enables in vivo drug detection with sensitivity and specificity. We will first characterize the CEST fingerprints of first-line HIV ARVs, and develop MRI methods based on the drug CEST fingerprints for in vivo detection. Aim 1: To characterize the CEST fingerprints of ARVs (3TC, ABC, TDF and DTG). We hypothesize that exchangeable protons of the drugs generate unique CEST fingerprints. The CEST effects of the protons in a drug molecule will be measured and combined to build the drug's CEST fingerprint. The CEST fingerprint of each drug will be further validated and determined in cells using human monocyte-derived macrophages (MDMs). Aim 2: To measure in vivo CNS biodistribution of ARVs in humanized mice using CEST MRI. We posit that MRI utilizing CEST fingerprints offer sensitivity and specificity for in vivo measurements of ARVs. To test the hypothesis, MRI data acquisition and analysis methods will be developed based on the ARV CEST fingerprints measured in Aim 1, and optimized for in vivo detection in infected humanized mice that treated with combined antiretroviral therapy (DTG/ABC/3TC or DTG/3TC/TDF). Brain tissue drug levels measured using liquid chromatography mass spectrometry (LC-MS/MS) will be used to validate CEST MRI results. A successful outcome of the study is a noninvasive MRI technique that measures brain ARV levels. The technique will provide a powerful tool for the development of treatments that mitigate CNS complications of HIV infection and ameliorate the mental health outcomes of HIV patients.