Enabling NF-kB Signal Transduction Studies in Primary Multiple Myeloma Cells

描述(申请人提供)：与主要评估癌症标本状态的高灵敏度基因组和蛋白质组学方法相比，对原发患者样本进行功能分析以评估其对各种实验条件的生物学反应是困难的，因为可以从患者活检样本中获得的活原代细胞数量通常有限。因此，克服这一技术障碍有可能改变我们显著增加转化型癌症研究方法的能力。通过结合癌细胞信号、生物工程和初级患者护理方面的专业知识，Miyamoto-Beebe-Callander团队建议提高我们在初级患者多发性骨髓瘤(MM)样本中对NF-kB信号转导反应进行功能性分析的能力。具体地说，我们建议开发创新的微通道培养设备并实施功能研究，以探索MM中挑战教条的NF-kB生存途径。我们已经开发了原型培养设备，以提高我们分析原代MM细胞的能力。这些微培养系统还提供了研究肿瘤微环境成分的灵活性，例如支持肿瘤的骨髓基质细胞(BMSCs)。在目标1下，我们将使用第一代微型细胞培养箱(MCCCs)剖析原代MM细胞中诱导耐药的NF-kB信号机制。我们还旨在通过共培养来自相同患者的MM细胞和BMSCs来揭示患者的个性化信息，这是对传统实验设置的一种范式转变，在传统实验设置中，MM和BMSCs的患者来源随机混合。在目标2下，我们将进一步改进功能微量分析，增加额外的功能，每个条件下的细胞数量甚至更少，从而极大地扩大MM的转译研究范围。所提议的微培养技术有可能迅速改变用于研究MM和其他血液类型以及可能的实体癌症类型的信号转导研究的方法，包括核因子-kB。

DESCRIPTION (provided by applicant): In contrast to highly sensitive genomic and proteomic methods that primarily evaluate the state of cancer specimens, functional analyses of primary patient samples to assess their biological responses to various experimental conditions are difficult because of the often limited number of live primary cells that can be obtained from patient biopsy samples. Thus, overcoming this technical barrier has the potential to transform our ability to significantly increase translational cancer research approaches. By combining expertise in, cancer cell signaling, bioengineering, and primary patient care, the Miyamoto-Beebe-Callander team proposes to improve our ability to functionally analyze NF-kB signal transduction responses in primary patient multiple myeloma (MM) samples. Specifically, we propose to develop innovative microchannel culture devices and implement functional studies to investigate a dogma-challenging NF-kB-survival pathway in MM. We have already developed prototype culture devices that increase our ability to analyze primary MM cells. These microculture systems also provide the flexibility to study components of the tumor microenvironment, such as tumor-supporting bone marrow stromal cells (BMSCs). Under Aim 1 we will dissect drug resistance-inducing NF-kB signaling mechanisms in primary MM cells using the first generation microscale cell culture chambers (MCCCs). We also aim to reveal patient individualized information by co- culturing MM cells and BMSCs derived from the same patients, a paradigm shift from the conventional experimental setup where patient sources of MM and BMSCs are randomly mixed. Under Aim 2 we will further improve functional micro-scale assays with additional functionalities and with even smaller cell numbers per condition, thus greatly expanding the scope of translational research in MM. The micro-culture technology proposed has the potential to rapidly change the methods used for investigating signal transduction studies, including NF-kB, in MM and other blood and possibly solid cancer types.