Starodubtseva Natalia L, Chagovets Vitaliy V, Nekrasova Maria E, Nazarova Niso M, Tokareva Alisa O, Bourmenskaya Olga V, Attoeva Djamilja I, Kukaev Eugenii N, Trofimov Dmitriy Y, Frankevich Vladimir E, Sukhikh Gennady T
National Medical Research Center for Obstetrics Gynecology and Perinatology Named after Academician V.I., Kulakov of the Ministry of Healthcare of Russian Federation, 117997 Moscow, Russia.
Moscow Institute of Physics and Technology, 141700 Moscow, Russia.
Metabolites. 2022 May 31;12(6):503. doi: 10.3390/metabo12060503.
A dramatic increase in cervical diseases associated with human papillomaviruses (HPV) in women of reproductive age has been observed over the past decades. An accurate differential diagnosis of the severity of cervical intraepithelial neoplasia and the choice of the optimal treatment requires the search for effective biomarkers with high diagnostic and prognostic value. The objective of this study was to introduce a method for rapid shotgun lipidomics to differentiate stages of HPV-associated cervix epithelium transformation. Tissue samples from 110 HPV-positive women with cervicitis ( = 30), low-grade squamous intraepithelial lesions (LSIL) ( = 30), high-grade squamous intraepithelial lesions (HSIL) ( = 30), and cervical cancers ( = 20) were obtained. The cervical epithelial tissue lipidome at different stages of cervix neoplastic transformation was studied by a shotgun label-free approach. It is based on electrospray ionization mass spectrometry (ESI-MS) data of a tissue extract. Lipidomic data were processed by the orthogonal projections to latent structures discriminant analysis (OPLS-DA) to build statistical models, differentiating stages of cervix transformation. Significant differences in the lipid profile between the lesion and surrounding tissues were revealed in chronic cervicitis, LSIL, HSIL, and cervical cancer. The lipids specific for HPV-induced cervical transformation mainly belong to glycerophospholipids: phosphatidylcholines, and phosphatidylethanolamines. The developed diagnostic OPLS-DA models were based on 23 marker lipids. More than 90% of these marker lipids positively correlated with the degree of cervix transformation. The algorithm was developed for the management of patients with HPV-associated diseases of the cervix, based on the panel of 23 lipids as a result. ESI-MS analysis of a lipid extract by direct injection through a loop, takes about 25 min (including preparation of the lipid extract), which is significantly less than the time required for the HPV test (several hours for hybrid capture and about an hour for PCR). This makes lipid mass spectrometric analysis a promising method for express diagnostics of HPV-associated neoplastic diseases of the cervix.
在过去几十年中，已观察到育龄女性中与人类乳头瘤病毒（HPV）相关的宫颈疾病急剧增加。准确鉴别宫颈上皮内瘤变的严重程度并选择最佳治疗方法，需要寻找具有高诊断和预后价值的有效生物标志物。本研究的目的是引入一种快速鸟枪法脂质组学方法，以区分HPV相关宫颈上皮转化的阶段。获取了110例HPV阳性女性的组织样本，其中宫颈炎患者30例、低级别鳞状上皮内病变（LSIL）患者30例、高级别鳞状上皮内病变（HSIL）患者30例以及宫颈癌患者20例。采用无标记鸟枪法对宫颈肿瘤转化不同阶段的宫颈上皮组织脂质组进行了研究。该方法基于组织提取物的电喷雾电离质谱（ESI-MS）数据。脂质组学数据通过正交投影到潜在结构判别分析（OPLS-DA）进行处理，以建立区分宫颈转化阶段的统计模型。在慢性宫颈炎、LSIL、HSIL和宫颈癌中，病变组织与周围组织的脂质谱存在显著差异。HPV诱导的宫颈转化特异性脂质主要属于甘油磷脂：磷脂酰胆碱和磷脂酰乙醇胺。所建立的诊断OPLS-DA模型基于23种标记脂质。这些标记脂质中超过90%与宫颈转化程度呈正相关。基于这23种脂质组成的检测组，开发了用于管理HPV相关宫颈疾病患者的算法。通过定量环直接进样对脂质提取物进行ESI-MS分析大约需要25分钟（包括脂质提取物的制备），这明显少于HPV检测所需的时间（杂交捕获需要数小时，PCR需要约1小时）。这使得脂质质谱分析成为一种有前景的HPV相关宫颈肿瘤疾病快速诊断方法。
