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

描述（由申请人提供）：与主要评价癌症样本状态的高灵敏度基因组和蛋白质组学方法相比，由于可从患者活检样本中获得的活原代细胞数量通常有限，因此难以对原代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。因此，克服这一技术障碍有可能改变我们的能力，显着增加转化癌症研究方法。Miyamoto-Beebe-卡兰德团队结合了癌细胞信号传导、生物工程和初级患者护理方面的专业知识，提出提高我们在原发性多发性骨髓瘤（MM）患者样本中功能性分析NF-κ B信号转导反应的能力。具体而言，我们建议开发创新的微通道培养装置，并实施功能研究，以调查MM中具有教条挑战性的NF-κ B存活途径。我们已经开发出原型培养装置，提高了我们分析原代MM细胞的能力。这些微培养系统还提供了研究肿瘤微环境组分的灵活性，例如肿瘤支持骨髓基质细胞（BMSC）。在目标1下，我们将使用第一代微型细胞培养室（MCCC）在原代MM细胞中剖析耐药性诱导的NF-κ B信号传导机制。我们还旨在通过共培养来自相同患者的MM细胞和BMSC来揭示患者个体化信息，这是从常规实验设置的范式转变，其中MM和BMSC的患者来源被随机混合。根据目标2，我们将进一步改进功能性微型测定，增加额外的功能，并且每种条件下的细胞数量甚至更小，从而大大扩大MM转化研究的范围。提出的微量培养技术有可能快速改变用于研究MM和其他血液以及可能的实体癌类型中信号转导研究（包括NF-kB）的方法。

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.