Effects of Extravasated Serum Proteins on Human Glioblastoma Invasion

描述（由申请人提供）：多形性胶质母细胞瘤（GBM）是最常见和最致命的原发性脑肿瘤。尽管各种疗法取得了进步，但 30 多年来生存率基本保持不变。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 结局缺乏影响。我很早就离开了住院医师实习岗位，因为我相信，通过在理解和阻止 GBM 入侵方面取得科学进展，我可以产生更大的积极影响。通过随后的博士前和博士后研究以及医学遗传学住院医师实习，我逐渐掌握了成像物理学、分子生物学和蛋白质组学方面的高级技能。我相信我已经将自己定位在一条独特的道路上，这将使我能够使用经过临床经验锤炼的新颖的多学科范式来成功地处理 GBM 侵袭。我的长期目标是成为一名独立的医师科学家并改善诊断为 GBM 的患者的治疗结果。我的近期目标是完成博士后培训并过渡到学术教职职位。为了实现这些目标，我在两位高产且成功的医师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，该实验室正在研究神经退行性疾病的生物标志物。我的导师，加上华盛顿大学独特的学术和科学环境，为我提供了丰富而肥沃的环境来进行我的研究，这与他们自己的研究兴趣是互补的，但又是独特的。我的研究计划专门针对 GBM 侵袭。 GBM侵入健康脑组织是疾病难治的主要原因。目前尚不清楚 GBM 肿瘤细胞为何侵袭。然而，有令人信服的证据表明，由于血脑屏障（BBB）破坏而导致的特定血清蛋白在细胞外空间的积累可能控制增殖与侵袭表型。该提案的中心假设是外渗血清蛋白（ESP）对人类 GBM 侵袭有直接影响。 ESP 此前并未被认为是入侵的调节剂。为了确定 ESP 对 GBM 的影响及其作用机制，需要进行多项创新。首先，将开发和优化一种新的蛋白质组学方法，用于在各种 BBB 渗透性条件下识别 ESP。这种蛋白质组技术将与一种用于测量 BBB 渗透性的新型动态磁共振成像 (MRI) 技术相结合，以便可以对收集用于蛋白质组分析的组织进行准确分类。目前还没有一种公正的、大规模的技术来识别 GBM 或任何与完全或部分 BBB 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，以监测体内动物模型中人类 GBM 干细胞 (GSC) 的侵袭。目前可用的成像技术无法捕获人类或 GBM 动物模型中的 GBM 侵袭。识别驱动侵袭性表型的特定 ESP 将加深我们对 GBM 的理解，同时还提供新的治疗靶点，这些靶点可能被证明更强大和/或对传统策略进行补充。此外，拟议的工作将建立新的方法和技术，可应用于与神经病理学和/或 BBB 损害相关的许多领域，例如中风、创伤、感染和各种神经退行性和生化疾病。总之，我独特的背景、知名导师以及华盛顿大学的众多科学资源为我提供了一个特殊的机会，为改善 GBM 的结果做出重大和长期的贡献。霍华德·特明癌症研究独立之路奖 (K99/R00) 将使我能够完成博士后培训，并为过渡到教职职位提供关键支持。 公共卫生相关性： 多形性胶质母细胞瘤（GBM）是最常见和最致命的原发性脑肿瘤。尽管各种疗法取得了进步，但 30 多年来生存率基本保持不变。 GBM侵入健康脑组织是疾病难治的主要原因。目前尚不清楚 GBM 肿瘤细胞为何侵袭。然而，有令人信服的证据表明，由于血脑屏障的破坏，特定血清蛋白在细胞外空间的积累可能会使 GBM 肿瘤细胞更具侵袭性。血清蛋白此前并未被认为是侵袭调节剂。鉴定出促进 GBM 细胞迁移的特定血清蛋白将建立新的治疗靶点，旨在防止 GBM 肿瘤细胞在整个大脑中扩散。通过阻止 GBM 的迁移行为，针对大块肿瘤生长的疗法可能会更加有效，并且这种破坏性疾病的生存率可能会在几十年来首次得到改善。

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. PUBLIC HEALTH RELEVANCE: 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. The invasion of GBM into healthy brain tissues is the main 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 may make GBM tumor cells more invasive. Serum proteins have not previously been recognized as modulators of invasion. The identification of specific serum proteins that actuate GBM cell migration would establish new therapeutic targets directed at preventing the spread of GBM tumor cells throughout the brain. By stopping the migratory behavior of GBM, therapies directed at bulk tumor growth may prove more effective and survival from this devastating disease may improve for the first time in many decades.