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: 8691806
• 负责人: Kayvan R Keshari
• 金额: $ 24.15万
• 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/8691806
• 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; non-invasive 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.

项目摘要 通过这一获得独立奖的途径，我希望获得获得教师所需的技能 通过独立研究计划的位置，重点是生物工程和实施新颖的3D 细胞和组织培养生物反应器，以及该平台与超极化（HP）13C MR结合 更好地研究癌症代谢。由于前列腺癌的生物学和病理复杂性，有一个 紧急临床需要开发更敏感和特定的成像标记以改善前列腺癌 特定于患者的治疗计划和治疗衰竭的早期评估。非凡的新技术 利用超极化（HP）代谢底物具有提供这些MR生物标志物的潜力。最近的HP 细胞和动物模型的MR研究表明，HP代谢标记物反映了酶通量，可能 提供更准确的衡量前列腺癌存在，进展和对治疗的反应。然而， 可用的鼠和细胞培养模型无法可靠地模仿人类疾病，因此我们提出了一种新颖的 HP 13C MR和与NMR兼容的3D组织培养物生物反应器的组合研究实时 活人前列腺组织切片（TSC）的代谢。 这项研究的总体目的是设计NMR兼容的3D组织培养物生物反应器 与人类TSC一起使用，并使用它来识别HP分子成像标记，以改善前列腺癌患者 - 具体的治疗计划和对靶向治疗的反应的早期评估。完成这些目标将 需要在原代细胞和组织培养区域，前列腺生物化学和病理学领域进行额外的训练， HP探针开发，微型工程，生物转移和药代动力学。利用这种新培训， 第一个目的是优化在NMR兼容的3D组织培养中维持人前列腺TSC的条件 生物反应器并验证随着时间的推移TSC的代谢完整性。连续31p将用于监视 随着时间的流逝，生物反应器中组织切片的进展。 HP 13C MR的动态采集将用于 计算与丙酮酸代谢和其他探针的代谢相关的通量。这些数据将被比较 在生物反应器中培养前后的组织病理学评估变化。第二个目的是使用这个新的 比较正常和恶性前列腺组织代谢的实验模型，并且重要的是确定 HP代谢物是否与病理级及其与代谢和生物转移的关系相关。这 第三个目的是使用此平台来识别对PI3K/MTOR抑制剂的治疗反应的HP标记。 这项提案的目的是开发一个工程系统，该系统可以克服当前的局限性 鼠类和细胞培养模型，并有助于开发相关的生物标志物，以转化为诊所。 虽然该研究的重点是该独立奖的奖励是前列腺癌，但结合了 NMR兼容的原代组织培养物生物反应器平台与高灵敏度HP MR探针结合 将在各种疾病和成像方式中具有广泛的适用性。