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)是最常见和最致命的原发脑肿瘤。尽管在各种治疗方法上取得了进展，但30多年来，存活率基本保持不变。作为一名内科医生，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，我深深地意识到传统方法对GBM结果缺乏影响。我很早就离开了我的住院医生生涯，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。通过我随后在医学遗传学的博士后和博士后学习以及实习，我已经稳定地掌握了成像物理、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，这条道路将使我能够使用经过临床经验磨练的新的和多学科的范式来成功地处理GBM的侵袭。我的长期目标是成为一名独立的内科科学家，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，并过渡到学术教职。为了实现这些目标，我在两位高生产率和成功的内科科学家的指导和指导下，设计并启动了一个新的项目。Robert C.Rostomly(Mentor)领导着一个分子生物学实验室，研究基底膜的细胞内侵袭机制。张静，医学博士，博士(共同导师)领导着一个蛋白质组实验室，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，再加上华盛顿大学独特的学术和科学环境，为我提供了一个丰富而肥沃的环境来从事我的研究，这与他们自己的研究兴趣相辅相成，但又是独一无二的。我的研究建议专门针对GBM的入侵。基底膜对健康脑组织的侵袭是疾病难治性的主要原因。目前尚不清楚GBM肿瘤细胞为什么会侵袭。然而，有令人信服的证据表明，由于血脑屏障(BBB)的破坏，特定的血清蛋白在细胞外空间积聚，可能主导了增殖型和侵袭型。这一建议的中心假设是，外渗血清蛋白(ESPs)对人GBM的侵袭有直接影响。ESP以前没有被认为是侵袭的调节器。为了确定ESP对GBM的影响及其作用机制，需要进行几项创新。首先，将开发和优化一种新的蛋白质组学方法来鉴定不同血脑屏障通透性条件下的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，用于测量血脑屏障通透性，以便收集用于蛋白质组分析的组织可以准确地分类。目前还没有一种公正、大规模的技术来识别GBM中的ESP或与完全或部分BBB中断相关的任何病理疾病中的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，以在活体动物模型中监测人类GBM干细胞(GSCs)的侵袭。目前可用的成像技术一直无法捕捉到GBM在人类或GBM动物模型中的侵袭。识别导致侵袭性表型的特定ESP将加深我们对GBM的理解，同时也提供可能被证明更强大和/或补充传统策略的新的治疗靶点。此外，拟议的工作将建立新的方法和技术，这些方法和技术可应用于与神经病理和/或血脑屏障损害相关的一些领域，如中风、创伤、感染以及各种神经退行性和生化疾病。总而言之，我独特的背景、可靠的导师和华盛顿大学的大量科学资源为我提供了一个难得的机会，使我能够为改善GBM的成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，并为过渡到教员职位提供关键支持。 公共卫生相关性： 多形性胶质母细胞瘤(GBM)是最常见、最致命的原发性脑肿瘤。尽管在各种治疗方法上取得了进展，但30多年来，存活率基本保持不变。GBM对健康脑组织的侵袭是疾病难治性的主要原因。目前尚不清楚GBM肿瘤细胞为什么会侵袭。然而，有令人信服的证据表明，由于血脑屏障的破坏，特定的血清蛋白在细胞外空间积聚，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的调节物。识别刺激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，针对实体肿瘤生长的治疗可能会被证明更有效，这种毁灭性疾病的存活率可能会几十年来首次提高。

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.