Effects of Extravasated Serum Proteins on Human Glioblastoma Invasion

描述（申请人提供）：多形性胶质母细胞瘤（GBM）是最常见、最致命的原发性脑肿瘤。尽管各种治疗方法都取得了进展，但30多年来生存率基本没有变化。作为一名医生，我亲眼目睹了与GBM相关的毁灭性的发病率和死亡率。GBM是我进入神经外科的主要原因。然而，在我的临床和手术经验中，我深刻认识到传统方法对GBM结果的影响不足。我很早就离开了实习期，因为我相信我可以通过推动科学进步来理解并阻止GBM的入侵，从而产生更大的积极影响。通过随后在医学遗传学的博士前、博士后学习和实习，我逐渐掌握了影像物理、分子生物学和蛋白质组学方面的先进技能。我相信我已经把自己定位在一条独特的道路上，这条道路将使我能够使用经过临床经验锤炼的新颖多学科范式来成功地接近GBM的侵袭。我的长期目标是成为一名独立的内科医生和科学家，并改善被诊断为GBM的患者的预后。我的近期目标是完成博士后培训并过渡到学术教师职位。为了实现这些目标，我在两位高产且成功的内科科学家的指导和指导下，设计并启动了一个新颖的项目。Robert C. Rostomily，医学博士（导师）领导一个分子生物学实验室，研究GBM的细胞内侵袭机制。张静，医学博士，博士（共同导师）领导一个蛋白质组学实验室，研究神经退行性疾病的生物标志物。我的导师们，加上华盛顿大学独特的学术和科学环境，为我的研究提供了丰富而肥沃的环境，这对他们自己的研究兴趣是互补的，但也是独特的。我的研究计划专门针对GBM的入侵。GBM侵入健康脑组织是该病难治性的主要原因。GBM肿瘤细胞侵袭的原因尚不清楚。然而，有令人信服的证据表明，由于血脑屏障（BBB）的破坏，特异性血清蛋白在细胞外空间的积累可能决定了增殖性与侵袭性表型。该建议的中心假设是外渗血清蛋白（ESPs）对人类GBM侵袭有直接影响。ESPs以前没有被认为是侵袭的调节剂。为了确定ESPs对GBM的影响及其作用机制，需要进行一些创新。首先，将开发和优化一种新的蛋白质组学方法，用于鉴定血脑屏障通透性不同条件下的esp。这种蛋白质组学技术将与一种新的动态磁共振成像（MRI）技术相结合，用于测量血脑屏障的通透性，以便收集用于蛋白质组学分析的组织可以准确分类。目前还没有一种公正的、大规模的技术来识别GBM或任何与血脑屏障完全或部分破坏相关的病理疾病中的esp。最后，一种被称为快速结合池分数成像（FBFI）的最先进的MRI技术将被优化，以监测人类GBM干细胞（GSCs）在体内动物模型中的入侵。目前可用的成像技术还无法捕捉到GBM在人类或动物模型中的侵袭。识别驱动侵袭性表型的特定ESPs将加深我们对GBM的理解，同时也提供新的治疗靶点，可能被证明更强大和/或补充传统策略。此外，这项工作将建立新的方法和技术，应用于与神经病理学和/或血脑屏障损害相关的许多领域，如中风、创伤、感染和各种神经退行性疾病和生化疾病。总之，我独特的背景、成熟的导师和华盛顿大学丰富的科学资源为我提供了一个难得的机会，可以为改善GBM的成果做出重大而长期的贡献。Howard Temin癌症研究独立之路奖（K99/R00）将允许我完成我的博士后培训，并为过渡到教师职位提供关键支持。

DESCRIPTION (provided by applicant): Glioblastoma multiforme (GBM) is the most common and the most lethal primary brain tumor. Despite advances in a variety of therapies, survival has remained largely unchanged for more than 30 years. As a physician, I have personally witnessed the devastating morbidity and mortality associated with GBM. GBM was the principal reason that I entered Neurological Surgery. During my clinical and surgical experiences, however, I gained a deep appreciation for the lack of effect on GBM outcomes by traditional approaches. I departed my residency early with the belief that I could have a greater positive impact by making scientific advances towards understanding and then stopping GBM invasion. Through my subsequent pre- and post-doctoral studies and residency in Medical Genetics, I have steadily equipped myself with advanced skills in imaging physics, molecular biology, and proteomics. I believe I have uniquely positioned myself on a path that will enable me to use novel and multidisciplinary paradigms tempered by clinical experience to successfully approach GBM invasion. My long-term goals are to become an independent physician-scientist and to improve outcomes in patients diagnosed with GBM. My immediate goal is to complete my post-doctoral training and transition to an academic faculty position. To achieve these goals, I have designed and initiated a novel project under the guidance and mentorship of two highly productive and successful physician-scientists. Robert C. Rostomily, M.D. (mentor) leads a molecular biology laboratory that studies intracellular mechanisms of invasion in GBM. Jing Zhang, M.D., Ph.D. (co-mentor) leads a proteomics laboratory that is studying biomarkers of neurodegenerative diseases. My mentors, coupled with the unique academic and scientific environment at the University of Washington, have given me a rich and fertile setting to pursue my research, which is complementary, but unique, to their own research interests. My research proposal specifically addresses GBM invasion. The invasion of GBM into healthy brain tissues is the predominant reason for disease intractability. It is unclear why GBM tumor cells invade. However, there is compelling evidence that suggests the accumulation of specific serum proteins in the extracellular space due to disruption of the blood-brain-barrier (BBB) may govern the proliferative versus invasive phenotype. The central hypothesis of this proposal is that extravasated serum proteins (ESPs) have a direct effect on human GBM invasion. ESPs have not previously been recognized as modulators of invasion. To determine the effect of ESPs on GBM and their mechanism of action, several innovations are required. First, a novel proteomic methodology for identifying ESPs under various conditions of BBB permeability will be developed and optimized. This proteomic technique will be coupled with a novel dynamic magnetic resonance imaging (MRI) technique for measuring BBB permeability so that tissues collected for proteomic analysis can be accurately categorized. An unbiased, wide-scale technique for identifying ESPs in GBM or in any pathologic disease associated with complete or partial BBB disruption is not presently available. And finally, a state-of-the-art MRI technique known as fast bound pool fraction imaging (FBFI) will be optimized to monitor invasion of human GBM stem cells (GSCs) in an in vivo animal model. Currently available imaging techniques have been unable to capture GBM invasion in humans or in animal models of GBM. The identification of specific ESPs that actuate the invasive phenotype would deepen our understanding of GBM, while also providing new therapeutic targets that may prove more robust and/or complementary to conventional strategies. In addition, the proposed work will establish novel methodologies and technologies that have applications in a number of fields associated with neurological pathology and/or compromise of the BBB such as stroke, trauma, infection, and a variety of neurodegenerative and biochemical disorders. In summary, my unique background, established mentors, and the multitude of scientific resources at the University of Washington have provided me with an exceptional opportunity to make a significant and long- term contribution towards the improvement of outcomes in GBM. The Howard Temin Pathway to Independence Award for Cancer Research (K99/R00) will allow me to complete my post-doctoral training and provide critical support for the transition to a faculty position.