Mitigation of preanalytic factors influencing brain tumor protein phosphorylation

项目总结 弥漫性胶质瘤，包括少突胶质瘤、星形细胞瘤和高度恶性的胶质母细胞瘤。 最常见的原发恶性脑瘤类型。磷脂酰肌醇3的活性增加 (PI3)-激酶/AKT/mTOR信号通路在GBM中普遍存在，可能是GBM的机制之一。 肿瘤从低级别弥漫性胶质瘤向基底膜的恶变。鉴于这件事的重要性 在疾病中，针对它的几种治疗策略正在早期临床试验中进行测试。至 最大限度地提高这些疗法的益处并将其毒性降至最低，然而，为患者提供准确和可靠的生物标志物 需要对治疗的疾病反应进行分层和评估。蛋白质磷酸化的分析 提供有关信号通路活动的集成、功能信息。这项提案的目标是使用 生物科学，以确定和减轻分析前因素，包括肿瘤内的异质性， 组织获取和混淆的非肿瘤细胞，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，准确的评估需要小规模的、保存完好的 以及生物靶向的肿瘤活检。为了研究这一假设，我们建议成立一个多学科团队， 在获取和分析多个和图像引导的活检、脑肿瘤方面具有专业知识的调查人员 生物学、生物银行、神经病理学、成像、生物统计学和神经肿瘤学。首先，我们调查 术前将肿瘤活检定位于具有生物学侵袭性的肿瘤区域的创新方法 5-氨基乙酰丙酸(ALA)为基础的磁共振波谱成像与术中肿瘤代谢 成像。其次，我们使用临床验证的分析和最先进的单细胞质量细胞术来评估 PI3K/AKT/mTOR信号通路活性及非肿瘤细胞对 磷蛋白水平。第三，使用我们开发的循证方法和方法，我们将 蛋白决定PI3K/AKT/mTOR信号通路活性的预后意义 弥漫性胶质瘤中的磷酸化和mTOR抑制剂结合我们的时相治疗的患者 2针对PI3K/AKT/mTOR信号通路的临床试验(NCT02023905)。

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).