Human Tissue Culture Bioreactor and Hyperpolarized MR for Biomarker Discovery

用于生物标志物发现的人体组织培养生物反应器和超极化 MR

• 批准号: 8670990
• 负责人: Kayvan R Keshari
• 金额: $ 24.9万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8670990
• 关键词: Affect; Animal Model; Area; Award; Benign; Biochemistry; Bioenergetics; Biological Markers; Biomedical Engineering; Bioreactors; Cancer Patient; Cell Culture Techniques; Cell model; Cells; Clinic; Clinical; Data; Development; Disease; Drug Kinetics; Encapsulated; Engineering; Experimental Models; Faculty; Failure; Gases; Gene Expression; Genetic Markers; Goals; Histopathology; Hour; Human; Image; Imaging Techniques; Individual; Knowledge; Label; Life; Malignant - descriptor; Malignant neoplasm of prostate; Measures; Metabolic; Metabolic Marker; Metabolism; Methods; Modeling; Monitor; Mus; Oncogenic; Pathologic; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacodynamics; Pharmacotherapy; Phase; Physiological Processes; Positioning Attribute; Prostate; Prostate Cancer therapy; Proteins; Pyruvate; Pyruvate Metabolism Pathway; Research; Signal Transduction; Slice; System; Techniques; Therapeutic; Therapeutic Agents; Time; Tissues; Training; Translating; Translations; Work; cellular pathology; human FRAP1 protein; human disease; human tissue; imaging modality; improved; in vivo; inhibitor/antagonist; mTOR Inhibitor; member; molecular imaging; novel; pharmacodynamic model; programs; prostate cancer model; public health relevance; rapid detection; research clinical testing; response; skills; small molecule; tissue culture; tissue/cell culture; treatment planning; tumor metabolism

Project Summary 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 is 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.

项目概要 通过这个独立之路奖，我希望获得获得教师资格所需的技能 拥有一个专注于生物工程和新型 3D 实施的独立研究项目 细胞和组织培养生物反应器，并将该平台与超极化 (HP) 13C MR 结合使用 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， 临床迫切需要开发更灵敏和特异的成像标记物以改善前列腺癌 患者特定的治疗计划和治疗失败的早期评估。非凡的新技术 利用超极化 (HP) 代谢底物有可能提供这些 MR 生物标志物。最近的HP 细胞和动物模型中的 MR 研究表明，HP 代谢标记反映了酶通量，并且可能 提供更准确的前列腺癌存在、进展和治疗反应的测量。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，因此我们提出了一种新的 结合 HP 13C MR 和 NMR 兼容的 3D 组织培养生物反应器来研究实时 活人前列腺组织切片（TSC）的新陈代谢。 这项研究的总体目标是设计一个兼容 NMR 的 3D 组织培养生物反应器 与人类 TSC 一起使用，并用它来识别 HP 分子成像标记物，以改善前列腺癌患者的病情 具体的治疗计划和对靶向治疗的反应的早期评估。实现这些目标将 需要原代细胞和组织培养、前列腺生物化学和病理学领域的额外培训， HP 探针开发、微工程、生物转运和药代动力学。利用这种新的培训， 第一个目标是优化在 NMR 兼容的 3D 组织培养物中维持人类前列腺 TSC 的条件 生物反应器并验证 TSC 随着时间的推移的代谢完整性。连续31P将用于监控 生物反应器中组织切片随时间的进展。动态采购 HP 13C MR 将用于 实时计算与丙酮酸和其他探针代谢相关的通量。这个数据会被比较 在生物反应器中培养之前和之后进行组织病理学评估变化。第二个目标是使用这个新的 比较正常和恶性前列腺组织代谢的实验模型，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。这 第三个目标是利用该平台来识别 PI3K/mTOR 抑制剂治疗反应的 HP 标记物。 该提案的目标是开发一个工程系统，该系统可以克服当前的局限性 小鼠和细胞培养模型并有助于开发相关生物标志物以转化为临床。 虽然该独立之路奖的研究重点是前列腺癌，但结合 与 NMR 兼容的原代组织培养生物反应器平台与高灵敏度 HP MR 探针相结合 将在各种疾病和成像模式中具有广泛的适用性。