Quantitative tissue analysis reveals AK2, COL1A1, & PLG protein signatures: Targeted therapeutics for meningioma.

IF 12.5 2区 医学 Q1 SURGERY International journal of surgery Pub Date : 2024-09-17 DOI:10.1097/JS9.0000000000002054
Swati Sharma, Shamjetsabam Nandibala Devi, Kirti Chauhan, M H Yashavarddhan, Poonam Gautam, Prem Prakash, Priyanka Choudhary, Satnam Singh Chhabra, Rajesh Acharya, Samir Kumar Kalra, Anshul Gupta, Sunila Jain, Nirmal Kumar Ganguly, Rashmi Rana
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Abstract

Background: Meningioma is the most prevalent primary intracranial brain tumor and accounts for one-third of all CNS tumors. Meningioma is known to be the most common yet life-threatening brain tumor with a higher recurrence rate. Globally, there is an increase in the healthcare burden due to meningioma and hence in its research. The present clinical approach includes surgical resection, chemotherapy, and radiation therapies to which the malignancy does not seem to respond efficiently. Targeted therapies and molecular markers provide elite patient treatment and care for individuals suffering from meningiomas as compared to conventional measures. Although there is proteomic data on meningioma the knowledge of potential biomarkers differentiating the grades is scarce. To identify the best set of biomarkers, validation of reported markers in large and independent sample cohorts in the future is necessary.

Methods: A total of 12 samples, 3 each of control (which made pool 1) Meningioma grade I (which made 2 sets: pool 2 and pool 3), and Meningioma grade II (which made pool 4) were taken for LC-MS/MS. After this, the expression of three proteins was checked by immunocytochemistry, flow cytometry, and western blotting.

Results: Protein expression was analyzed using various techniques like mass spectrometry, immunocytochemistry, flow cytometry, and western blotting. Mass spectrometry is the most commonly used standard and reliable technique for identifying and quantifying protein expression. We got three highly upregulated proteins namely AK2, COL1A1, and PLG using this technique. The biomarker potential of these proteins was further checked by ICC, western blotting, and flow cytometry. Three important proteins were found to be upregulated namely AK2 (Adenylate Kinase 2), COL1A1 (Collagen 1A1), and PLG (Plasminogen). The order of increased protein expression was control < MG grade I < MG grade II according to mass spectrometry and western blotting. In immunocytochemistry, we found that COL1A1 expression increases significantly with grades in comparison to control. Similarly, AK2 and PLG also showed little increase but not as much as COL1A1. In flow cytometry, PLG showed higher upregulation in grades than control. While AK2 and COL1A1 showed little increase in expression in grades than control. All techniques, especially mass spectrometry and western blotting presented higher expression of these proteins in grades as compared to control.

Conclusions: In the quest to find a suitable therapeutic marker, this study incorporates quantitative labeling and detection followed by flow cytometry, Immunocytochemistry, and western blotting for early diagnosis and treatment of meningioma. The article further explores the efficacy of some proteins namely AK2, COL1A1 & PLG to be the targeted molecules.

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定量组织分析揭示了 AK2、COL1A1 和 PLG 蛋白特征:脑膜瘤的靶向疗法
背景:脑膜瘤是最常见的原发性颅内脑肿瘤,占所有中枢神经系统肿瘤的三分之一。众所周知,脑膜瘤是最常见但威胁生命的脑肿瘤,复发率较高。在全球范围内,脑膜瘤造成的医疗负担日益加重,因此对脑膜瘤的研究也日益增多。目前的临床方法包括手术切除、化疗和放疗,但恶性肿瘤似乎对这些疗法反应不佳。与传统方法相比,靶向疗法和分子标记为脑膜瘤患者提供了精英治疗和护理。虽然有脑膜瘤的蛋白质组数据,但有关区分等级的潜在生物标志物的知识却很少。为了确定最佳的生物标志物,今后有必要在大型独立样本群中对已报告的标志物进行验证:共采集了 12 份样本,其中对照组(第 1 组)、脑膜瘤 I 级(第 2 组和第 3 组)和脑膜瘤 II 级(第 4 组)各 3 份,进行 LC-MS/MS 分析。之后,通过免疫细胞化学、流式细胞术和 Western 印迹法检测三种蛋白质的表达:结果:使用质谱、免疫细胞化学、流式细胞仪和 Western 印迹等多种技术分析了蛋白质的表达。质谱法是鉴定和量化蛋白质表达的最常用、最可靠的标准技术。我们利用这种技术获得了三个高上调蛋白,即 AK2、COL1A1 和 PLG。我们还通过 ICC、Western 印迹和流式细胞术进一步检测了这些蛋白质的生物标记潜力。结果发现,有三种重要的蛋白质表达上调,即 AK2(腺苷酸激酶 2)、COL1A1(胶原蛋白 1A1)和 PLG(血浆蛋白酶原)。根据质谱和 Western 印迹,蛋白质表达增加的顺序为对照组 < MG I 级 < MG II 级。在免疫细胞化学分析中,我们发现与对照组相比,COL1A1的表达随着等级的升高而显著增加。同样,AK2 和 PLG 也有少量增加,但没有 COL1A1 增加得多。在流式细胞术中,与对照组相比,PLG 在各等级中的上调率更高。而 AK2 和 COL1A1 在各等级中的表达比对照组几乎没有增加。与对照组相比,所有技术,尤其是质谱法和 Western 印迹法,都显示等级中这些蛋白质的表达更高:为了寻找合适的治疗标记物,本研究采用了流式细胞术、免疫细胞化学和 Western 印迹法进行定量标记和检测,用于脑膜瘤的早期诊断和治疗。文章进一步探讨了一些蛋白质(即 AK2、COL1A1 和 PLG)作为靶向分子的功效。
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来源期刊
CiteScore
17.70
自引率
3.30%
发文量
0
审稿时长
6-12 weeks
期刊介绍: The International Journal of Surgery (IJS) has a broad scope, encompassing all surgical specialties. Its primary objective is to facilitate the exchange of crucial ideas and lines of thought between and across these specialties.By doing so, the journal aims to counter the growing trend of increasing sub-specialization, which can result in "tunnel-vision" and the isolation of significant surgical advancements within specific specialties.
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