Human Tissue Culture Bioreactor and Hyperpolarized MR for Biomarker Discovery

描述（由申请人提供）：通过这个途径独立奖，我希望获得必要的技能，以获得一个独立的研究计划，专注于生物工程和实施新型3D细胞和组织培养生物反应器的教师职位，并使用这个平台与超极化（HP）13 C MR结合，以更好地研究癌症代谢。由于前列腺癌的生物学和病理学复杂性，临床上迫切需要开发更敏感和特异性的成像标记物，以改善前列腺癌患者特异性治疗计划和治疗失败的早期评估。一种利用超极化（HP）代谢底物的非凡新技术有可能提供这些MR生物标志物。最近在细胞和动物模型中的HP MR研究表明，HP代谢标志物反映了酶通量，并可能提供更准确的测量前列腺癌的存在、进展和对治疗的反应。然而，现有的小鼠和细胞培养模型不能可靠地模拟人类疾病，因此我们提出了HP 13 C MR和NMR兼容的3D组织培养生物反应器的新组合，以研究活体人类前列腺组织切片（TSC）的实时代谢。 本研究的总体目标是设计一种NMR兼容的3D组织培养生物反应器，用于人TSC，并使用其鉴定HP分子成像标记物，以改善前列腺癌患者特异性治疗计划和靶向治疗反应的早期评估。实现这些目标将需要在原代细胞和组织培养、前列腺生物化学和病理学、HP探针开发、微工程、生物运输和药代动力学等领域进行额外的培训。利用这种新的训练，第一个目的是优化在NMR兼容的3D组织培养生物反应器中维持人前列腺TSC的条件，并验证TSC随时间的代谢完整性。将使用连续31 P监测生物反应器中组织切片随时间的进展。HP 13 C MR的动态采集将用于真实的实时计算与丙酮酸和其他探针代谢相关的通量。将该数据与生物反应器中培养前后的组织病理学进行比较，以评估变化。第二个目的是使用这种新的实验模型来比较正常和恶性前列腺组织的代谢，并且重要的是，确定HP代谢物是否与病理分级相关以及它们与代谢和生物转运的关系。第三个目的是使用该平台来鉴定对PI 3 K/mTOR抑制剂的治疗反应的HP标志物。 该提案的目标是开发一种工程系统，该系统可以克服当前鼠和细胞培养模型的局限性，并有助于开发相关生物标志物用于临床。虽然独立之路奖的研究重点是前列腺癌，但NMR兼容的原代组织培养生物反应器平台与高灵敏度HP MR探针的结合将在各种疾病和成像模式中具有广泛的适用性。 公共卫生相关性：通过这个独立之路奖项目，我将获得必要的知识和培训，成为一名独立研究项目的教师，该项目专注于新型3D细胞和组织培养生物反应器的工程和实施，并使用该平台与超极化MR结合使用，以更好地研究癌症代谢。

DESCRIPTION (provided by applicant): Through this Pathway to Independence Award, I hope to acquire the skills necessary to obtain a faculty position with an independent research program focused on the bioengineering and implementation of novel 3D cell and tissue culture bioreactors, and the use this platform in conjunction with hyperpolarized (HP) 13C MR to better study cancer metabolism. Due to the biologic and pathologic complexity of prostate cancer, there is an urgent clinical need to develop more sensitive and specific imaging markers for improved prostate cancer patient-specific treatment planning and early assessment of therapeutic failure. An extraordinary new technique utilizing hyperpolarized (HP) metabolic substrates has the potential to provide these MR biomarkers. Recent HP MR studies in cell and animal models suggest that HP metabolic markers reflect enzymatic fluxes and may provide a more accurate measure of prostate cancer presence, progression and response to therapy. However, available murine and cell culture models don't reliably mimic human disease, thus we propose a novel combination of HP 13C MR and NMR-compatible 3D tissue culture bioreactors to study the real-time metabolism of living human prostate tissue slices (TSCs). The overall objective of this research are to engineer an NMR-compatible, 3D Tissue Culture Bioreactor for use with human TSCs and use it to identify HP molecular imaging markers for improved prostate cancer patient- specific treatment planning and early assessment of response to targeted therapy. Accomplishing these aims will require additional training in the areas of primary cell and tissue cultures, prostate biochemistry and pathology, HP probe development, micro-engineering, biotransport, and pharmacokinetics. Utilizing this new training, the first aim i to optimize conditions for maintaining human prostate TSCs in an NMR-compatible, 3D tissue culture bioreactor and to verify the metabolic integrity of TSCs over time. Continuous 31P will be used to monitor the progression of tissue slices in the bioreactor with time. Dynamic acquisitions of HP 13C MR will be used to calculate fluxes associated with metabolism of pyruvate and other probes in real time. This data will be compared to histopathology before and after culture in the bioreactor to assess changes. The second aim is to use this new experimental model to compare normal and malignant prostate tissues metabolism, and importantly, determine whether HP metabolites correlate with pathologic grade and their relationship to metabolism and biotransport. The third aim is to use this platform to identify HP markers of therapeutic response to PI3K/mTOR inhibitors. It is the goal of this proposal to develop an engineered system, which can overcome the limitations of current murine and cell cultures models and aid in the development of relevant biomarkers for translation to the clinic. While the focus of the research in this Pathway to Independence Award is on prostate cancer, the combination of NMR-compatible primary tissue culture bioreactor platform combined with high sensitivity HP MR probes would have wide applicability across a variety of diseases and imaging modalities. PUBLIC HEALTH RELEVANCE: Through this Pathway to Independence Award project, I will gain the necessary knowledge and training to become a faculty member with an independent research program focused on the engineering and implementation of novel 3D cell and tissue culture bioreactors, and the use this platform in conjunction with hyperpolarized MR to better study cancer metabolism.