La Rocca Giuseppe La, Simboli Giorgia Antonia, Vincenzoni Federica, Rossetti Diana Valeria, Urbani Andrea, Ius Tamara, Della Pepa Giuseppe Maria, Olivi Alessandro, Sabatino Giovanni, Desiderio Claudia
Department of Neurosurgery, Mater Olbia Hospital, 07026 Olbia, Italy.
Institute of Neurosurgery, Fondazione Policlinico Universitario A. Gemelli IRCCS, Catholic University, 00168 Rome, Italy.
Cancers (Basel). 2020 Dec 23;13(1):30. doi: 10.3390/cancers13010030.
The present investigation aimed to characterize the protein profile of cavitating ultrasound aspirator fluid of newly diagnosed and recurrent glioblastoma comparing diverse zones of collection, i.e., tumor core and tumor periphery, with the aid of 5-aminolevulinic acid fluorescence. The samples were pooled and analyzed in triplicate by LC-MS following the shotgun proteomic approach. The identified proteins were then grouped to disclose elements exclusive and common to the tumor state or tumor zones and submitted to gene ontology classification and pathway overrepresentation analysis. The proteins common to the distinct zones were further investigated by relative quantitation, following a label free approach, to disclose possible differences of expression. Nine proteins, i.e., tubulin 2B chain, CD59, far upstream element-binding, CD44, histone H1.4, caldesmon, osteopontin, tropomyosin chain and metallothionein-2, marked the core of newly diagnosed glioblastoma with respect to tumor periphery. Considering the tumor zone, including the core and the fluorescence positive periphery, the serine glycine biosynthesis, pentose phosphate, 5-hydroxytryptamine degredation, de novo purine biosynthesis and huntington disease pathways resulted statistically significantly overrepresented with respect to the human genome of reference. The fluorescence negative zone shared several protein elements with the tumor zone, possibly indicating the presence of pathological aspects of glioblastoma rather than of normal brain parenchyma. On the other hand, its exclusive protein elements were considered to represent the healthy zone and, accordingly, exhibiting no pathways overrepresentation. On the contrary to newly diagnosed glioblastoma, pathway overrepresentation was recognized only in the healthy zone of recurrent glioblastoma. The TGFβ signaling pathway, exclusively classified in the fluorescence negative periphery in newly diagnosed glioblastoma, was instead the exclusive pathway classified in the tumor core of recurrent glioblastoma. These results, preliminary obtained on sample pools, demonstrated the potential of cavitron ultrasonic sur.
本研究旨在借助5-氨基乙酰丙酸荧光，对新诊断和复发性胶质母细胞瘤的超声空化吸引液的蛋白质谱进行表征，比较不同的采集区域，即肿瘤核心和肿瘤周边。将样本汇集后，采用鸟枪法蛋白质组学方法通过液相色谱-质谱联用（LC-MS）进行一式三份分析。然后将鉴定出的蛋白质分组，以揭示肿瘤状态或肿瘤区域独有的和共有的元素，并进行基因本体分类和通路富集分析。采用无标记方法通过相对定量进一步研究不同区域共有的蛋白质，以揭示可能的表达差异。九种蛋白质，即微管蛋白2B链、CD59、远上游元件结合蛋白、CD44、组蛋白H1.4、钙调蛋白、骨桥蛋白、原肌球蛋白链和金属硫蛋白-2，在新诊断的胶质母细胞瘤核心相对于肿瘤周边有显著特征。考虑到肿瘤区域，包括核心和荧光阳性周边，丝氨酸甘氨酸生物合成、磷酸戊糖、5-羟色胺降解、嘌呤从头合成和亨廷顿病通路相对于参考人类基因组在统计学上有显著富集。荧光阴性区域与肿瘤区域共享几个蛋白质元素，这可能表明存在胶质母细胞瘤的病理特征而非正常脑实质。另一方面，其独有的蛋白质元素被认为代表健康区域，因此未表现出通路富集。与新诊断的胶质母细胞瘤相反，通路富集仅在复发性胶质母细胞瘤的健康区域被识别。在新诊断的胶质母细胞瘤中仅在荧光阴性周边分类的转化生长因子β（TGFβ）信号通路，在复发性胶质母细胞瘤的肿瘤核心中却是唯一分类的通路。这些在样本汇集上初步获得的结果证明了超声空化吸引术的潜力。
