TR&D2: Development of Novel Hyperpolarized MR Molecular Imaging Probes, Realistic Pre-clinical Models and Correlative Science Methods

项目总结/摘要 十个同行评审和NIH资助的合作项目和八个服务项目， 作为HMTRC背后的科学驱动力的超极化MR位点，表明了对以下方面的明确需求： 改进的HP探针和技术，逼真的临床前模型以及相关的病理学和生物学 这将是由这个TR&D项目提供的方法。TR&D2基于我们在以下领域的丰富经验： DNP极化/溶解过程，新型超极化分子探针制剂的开发， 以及开发和测试新的生物学相关NMR相容的临床前细胞/组织培养物 以及相关的新型细胞/组织移植鼠模型。在当前的融资周期中，我们开发了新的13 C 糖酵解、TCA循环和细胞氧化还原的标记HP探针以及多极化方法， 同时提供代谢、灌注和细胞外pH（pHe）的信息。通过这次更新， 我们的目标是极大地改进这些HP探针和方法， 健壮性、成本效益、易于传播和可翻译性。这将通过以下方式实现： 使用玻璃化剂、化学优化的新自由基和极化方法。我们还将 研究pHe，ROS和免疫应答的新型HP探针，并解决由 CP（目标1）。在这次更新中，我们还将研究新的长寿命超极化探针和使用较低的 场MRI扫描仪，以延长超极化探头的T1。在目前的项目中，小说10和5 mm MR和PET兼容的3D细胞和组织培养生物反应器经过微工程设计，坚固耐用，成本低， 有效地生产，并在细胞和组织培养研究中进行了广泛的测试和验证， CP的在CP需求的驱动下，我们现在提出了未来的生物反应器设计， 测量能力，添加新的细胞培养结构，开发微型生物反应器系统， 以及将人类细胞和组织生物反应器研究扩展到新的鼠模型（目标2）。TR&D2还 在制定和实施程序以提供相关的病理、生物和 对于理解和验证CP中的HP MR结果至关重要的成像数据。根据选管会的要求 和CP的，这些验证研究将通过增加新的代谢组学，酶活性 染色分析和PET/HP MR相关研究（目的3）。虽然HP MR探头、模型系统和 相关的方法学已经通过一个高生产力的推拉系统，专门为CP开发出来 这些技术的发展也使SP和HP MR受益匪浅 通过广泛的宣传和培训， 这次更新。

PROJECT SUMMARY/ABSTRACT The ten peer-reviewed and NIH funded Collaborative Projects and the eight Service Projects from leading hyperpolarized MR sites, that are the scientific driving force behind the HMTRC, demonstrate a clear need for the improved HP probes and techniques, realistic pre-clinical models and correlative pathology and biology methodologies that will be provided by this TR&D project. TR&D2 builds on our significant experience in the DNP polarization/dissolution process, the development of new hyperpolarized molecular probe preparations, and in the development and testing of novel biologically relevant NMR-compatible pre-clinical cell/tissue culture and associated novel cell/tissue graft murine models. In the current funding cycle, we developed new 13C labeled HP probes of glycolysis, TCA cycle and cellular redox as well as multi-polarization approaches that provide simultaneous information on metabolism, perfusion, and extracellular pH (pHe). Through this renewal, we aim to greatly improve these HP probes and methods with an emphasis on increased sensitivity, robustness, cost-effectiveness, ease of dissemination, and translatability. This will be accomplished through the use of glassing agents, chemically optimized new radicals, and polarization methods. We will also investigate novel HP probes of pHe, ROS and immune response and to address new questions raised by the CP's (aim 1). In this renewal, we will also investigate new long-lived hyperpolarized probes and use of lower field MRI scanners in order to prolong the T1's of hyperpolarized probes. In the current project, novel 10 and 5 mm MR and PET compatible 3D cell and tissue culture bioreactors were micro-engineered, robustly and cost- effectively produced, and extensively tested and validated in cell and tissue culture studies performed in the CP's. Driven by the needs of the CP's, we now propose future bioreactor designs incorporating new biologic measurement capabilities, addition of novel cell culture constructs, development of a micro-bioreactor system, and extension of human cell & tissue bioreactor studies to novel murine models (aim 2). TR&D2 also made substantial progress developing and implementing procedures to provide correlative pathologic, biologic and imaging data critical for understanding and validating HP MR findings in the CP's. As requested by the EAC and CP's, these validation studies will be augmented by the addition of new metabolomics, enzyme activity staining assays, and PET/HP MR correlative studies (aim 3). While the HP MR probes, model systems and correlative methodologies have been specifically developed for the CP's through a highly productive push-pull approach, these technology developments also have significantly benefited the SP's and the HP MR community in general through extensive dissemination and training as will be continued and expanded through this renewal.