Mitigation of preanalytic factors influencing brain tumor protein phosphorylation

项目摘要 弥漫性胶质瘤，包括少突胶质细胞瘤、星形细胞瘤和高度恶性胶质母细胞瘤（GBM）， 最常见的原发性恶性脑肿瘤。磷脂酰肌醇3的活化增加 （PI 3）-激酶/AKT/mTOR信号通路在GBM中是常见的，并且似乎是GBM的一种机制。 肿瘤从低级别弥漫性胶质瘤向GBM的恶性转化。考虑到这件事的重要性 在疾病中，靶向它的几种治疗策略正在早期临床试验中进行测试。到 最大化这些疗法的益处并最小化毒性，然而， 需要对疾病对治疗的反应进行分层和评估。蛋白质磷酸化分析 提供关于信号传导通路活性的整合的功能信息。该提案的目的是利用 生物标本科学，以确定和减轻分析前的因素，包括肿瘤内异质性， 组织采集和混淆非肿瘤细胞，这可能会模糊蛋白质的准确分析 弥漫性胶质瘤中的磷酸化。我们假设，准确的评估将需要小，保存完好的 和生物靶向肿瘤活检。为了调查这一假设，我们提出了一个多学科的团队， 具有采集和分析多个和图像引导活检、脑肿瘤 生物学、生物库、神经病理学、成像、生物统计学和神经肿瘤学。首先我们调查 采用术前靶向肿瘤活检至生物侵袭性肿瘤区域的创新方法 基于5-氨基乙酰丙酸（ALA）的MR光谱成像和术中肿瘤代谢 显像其次，我们使用临床验证的检测方法和最先进的单细胞质谱仪来评估 PI 3 K/AKT/mTOR信号通路的活性，并确定非肿瘤细胞对整体肿瘤的贡献。 磷蛋白水平。第三，使用我们开发的基于证据的方法和途径，我们将 研究由蛋白质测定的PI 3 K/AKT/mTOR信号通路活性的预后意义 在弥漫性胶质瘤和用mTOR抑制剂治疗的患者中，通过整合我们的阶段， 2项靶向PI 3 K/AKT/mTOR信号通路的临床试验（NCT 02023905）。

PROJECT SUMMARY Diffuse gliomas, including oligodendroglioma, astrocytoma, and the highly malignant glioblastoma (GBM), are the most common type of primary malignant brain tumor. Increased activation of the phosphatidylinositol 3 (PI3)-kinase/AKT/mTOR signaling pathway is common in GBM and appears to be one mechanism of malignant transformation of tumors from lower grade diffuse glioma to GBM. Given the importance of this pathway in disease, several therapeutic strategies that target it are being tested in early phase clinical trials. To maximize benefit and minimize toxicity to these therapies, however, accurate and robust biomarkers for patient stratification and assessment of disease response to therapy are needed. Analysis of protein phosphorylation provides integrated, functional information about signaling pathway activity. The goal of this proposal is to use biospecimen sciences to identify and mitigate the preanalytic factors, including intratumoral heterogeneity, tissue acquisition, and confounding non-neoplastic cells, that can obscure accurate analysis of protein phosphorylation in diffuse glioma. We hypothesize that accurate assessment will require small, well-preserved and biologically targeted tumor biopsies. To investigate this hypothesis we propose a multidisciplinary team of investigators with expertise in the acquisition and analysis of multiple and image-guided biopsies, brain tumor biology, biobanking, neuropathology, imaging, biostatistics, and neuro-oncology. First, we investigate innovative approaches to target tumor biopsies to biologically aggressive tumor regions using pre-operative MR spectroscopic imaging and intra-operative tumor metabolism using 5-aminolevulinic acid (ALA)-based imaging. Second, we use clinically validated assays and state-of-the-art single cell mass cytometry to evaluate PI3K/AKT/mTOR signaling pathway activity and determine the contribution of non-neoplastic cells to overall phosphoprotein levels. Third, using the evidence-based methodologies and approaches we develop, we will investigate the prognostic significance of PI3K/AKT/mTOR signaling pathway activity as determined by protein phosphorylation in diffuse glioma and in patients treated with an mTOR inhibitor by integration with our Phase 2 clinical trial targeting the PI3K/AKT/mTOR signaling pathway (NCT02023905).