Table_1_Comprehending Meningioma Signaling Cascades Using Multipronged Proteomics Approaches & Targeted Validation of Potential Markers.docx

Meningiomas are one of the most prevalent primary brain tumors. Our study aims to obtain mechanistic insights of meningioma pathobiology using mass spectrometry-based label-free quantitative proteome analysis to identifying druggable targets and perturbed pathways for therapeutic intervention. Label-free based proteomics study was done from peptide samples of 21 patients and 8 non-tumor controls which were followed up with Phosphoproteomics to identify the kinases and phosphorylated components of the perturbed pathways. In silico approaches revealed perturbations in extracellular matrix remodeling and associated cascades. To assess the extent of influence of Integrin and PI3K-Akt pathways, we used an Integrin Linked Kinase inhibitor on patient-derived meningioma cell line and performed a transcriptomic analysis of the components. Furthermore, we designed a Targeted proteomics assay which to the best of our knowledge for very first-time enables identification of peptides from 54 meningioma patients via SRM assay to validate the key proteins emerging from our study. This resulted in the identification of peptides from CLIC1, ES8L2, and AHNK many of which are receptors and kinases and are difficult to be characterized using conventional approaches. Furthermore, we were also able to monitor transitions for proteins like NEK9 and CKAP4 which have been reported to be associated with meningioma pathobiology. We believe, this study can aid in designing peptide-based validation assays for meningioma patients as well as IHC studies for clinical applications.

脑膜瘤是原发性脑肿瘤中最常见的类型之一。本研究旨在通过基于质谱技术的无标记定量蛋白质组学分析，揭示脑膜瘤病理生物学机制，以识别可药物治疗的靶点和受干扰的通路。无标记蛋白质组学研究基于21例患者和8例非肿瘤对照的肽样本进行，并随后通过磷酸蛋白质组学分析以识别受干扰通路中的激酶和磷酸化组分。通过计算机模拟方法揭示了细胞外基质重塑及其相关级联反应的干扰。为了评估整合素和PI3K-Akt通路的影响程度，我们使用了一种整合素联激酶抑制剂作用于患者来源的脑膜瘤细胞系，并对相关组分进行了转录组学分析。此外，我们设计了一种靶向蛋白质组学检测方法，据我们所知，这是首次通过SRM检测识别54例脑膜瘤患者的肽段，以验证本研究中出现的核心蛋白。这导致了CLIC1、ES8L2和AHNK等肽段的识别，其中许多是受体和激酶，且难以通过传统方法进行表征。此外，我们还能够监测到NEK9和CKAP4等蛋白质的转换，这些蛋白质已被报道与脑膜瘤的病理生物学相关。我们相信，这项研究有助于为脑膜瘤患者设计基于肽段的验证检测方法，以及用于临床应用的免疫组化研究。