Human Tissue Culture Bioreactor and Hyperpolarized MR for Biomarker Discovery

描述(申请人提供)：通过这个独立之路奖，我希望获得必要的技能，以获得一个独立研究计划的教员职位，专注于生物工程和新型3D细胞和组织培养生物反应器的实施，并将该平台与超极化(HP)13C磁共振结合使用，以更好地研究癌症新陈代谢。由于前列腺癌生物学和病理的复杂性，临床迫切需要开发更敏感、更特异的影像标志物，以改进前列腺癌患者的治疗计划和早期评估治疗失败。一项利用超极化(HP)代谢底物的非凡新技术有可能提供这些MR生物标记物。最近在细胞和动物模型上的Hp MR研究表明，Hp代谢标记物反映了酶的流动，并可能提供一种更准确的前列腺癌存在、进展和治疗反应的测量方法。然而，现有的小鼠和细胞培养模型并不能可靠地模拟人类疾病，因此我们提出了一种新的组合，即HP 13C MR和核磁共振兼容的3D组织培养生物反应器来研究活体人前列腺组织切片(TSCs)的实时代谢。这项研究的总体目标是设计一种核磁共振兼容的3D组织培养生物反应器，用于人类TSCs，并使用它来识别HP分子成像标记，以改进前列腺癌患者的特定治疗计划和早期评估靶向治疗的反应。要实现这些目标，将需要在原代细胞和组织培养、前列腺生物化学和病理学、幽门螺杆菌探针开发、微工程、生物运输和药代动力学等领域进行额外培训。利用这一新的培训，第一个目标是优化在核磁共振兼容的3D组织培养生物反应器中维持人前列腺TSCs的条件，并验证TSCs随着时间的推移的代谢完整性。连续的31P将用于监测生物反应器中组织切片随时间的进展。HP 13C MR的动态采集将用于实时计算与丙酮酸和其他探针代谢相关的通量。这些数据将与生物反应器培养前后的组织病理学进行比较，以评估变化。第二个目的是利用这个新的实验模型来比较正常和恶性前列腺组织的代谢，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。第三个目的是利用这个平台来确定幽门螺杆菌对PI3K/mTOR抑制剂的治疗反应的标志物。这项建议的目标是开发一种工程系统，该系统可以克服当前小鼠和细胞培养模型的局限性，并有助于开发相关的生物标记物，以便翻译到临床上。虽然这项通向独立奖的研究重点是前列腺癌，但核磁共振兼容的原代组织培养生物反应器平台与高灵敏度的惠普磁共振探头相结合，将在各种疾病和成像方式中具有广泛的适用性。 与公共健康相关：通过这个独立之路奖项目，我将获得必要的知识和培训，成为一名独立研究计划的教员，该计划专注于新型3D细胞和组织培养生物反应器的设计和实施，并将该平台与超极化MR结合使用，以更好地研究癌症新陈代谢。

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.