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

项目摘要/摘要 美国国立卫生研究院资助的十个同行评审的合作项目和Leading的八个服务项目 超极化磁共振部位是HMTRC背后的科学推动力，表明显然需要 改进的幽门螺杆菌探针和技术、现实的临床前模型和相关的病理学和生物学 这个研发项目将提供的方法论。TR&D2建立在我们在 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相关的核磁共振兼容的临床前细胞/组织培养 以及相关的新型细胞/组织移植小鼠模型。在当前的资金周期中，我们开发了新的13C 糖酵解、TCA循环和细胞氧化还原以及多极化途径的标记HP探针 提供有关新陈代谢、血流和细胞外pH(Phe)的同步信息。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，重点是提高灵敏度， 健壮性、成本效益、易传播性和可译性。这将通过以下方式实现 使用玻璃剂、化学优化的新自由基和极化方法。我们还将 研究新的幽门螺杆菌Phe、ROS和免疫反应探针，并解决由 CP‘s(目标1)。在这次更新中，我们还将研究新的长寿命超极化探测器和使用LOWER 场强磁共振扫描仪，以延长S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是微工程设计的，坚固耐用，成本低廉。 有效生产，并在细胞和组织培养研究中进行广泛测试和验证 在CP需求的驱动下，我们现在提出未来的生物反应器设计，结合新的生物 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小鼠模型(目标2)。Tr&d2还制作了 在制定和实施程序方面取得实质性进展，以提供相关病理、生物和 成像数据对于了解和验证CP中的HP MR发现至关重要。应EAC的要求 和CP的，这些验证研究将通过添加新的代谢组学、酶活性 染色分析和PET/Hp MR相关研究(目标3)。而惠普MR探头、模型系统和 通过高效的推拉，已经专门为CP开发了相关方法 这些技术的发展也使SP和HP MR受益匪浅 通过广泛的传播和培训，并将继续和扩大通过 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.