Department of Chemistry, University of Bari Aldo Moro, via Orabona 4, 70126, Bari, Italy.
Department of Pharmacy-Drug Sciences, University of Bari Aldo Moro, via Orabona 4, 70126, Bari, Italy.
Anal Bioanal Chem. 2020 Oct;412(25):6859-6874. doi: 10.1007/s00216-020-02817-z. Epub 2020 Aug 1.
Autism spectrum disorder (ASD) is a broad and heterogeneous group of neurological developmental disorders characterized by impaired social interaction and communication, restricted and repetitive behavioural patterns, and altered sensory processing. Currently, no reliable ASD molecular biomarkers are available. Since immune dysregulation has been supposed to be related with ASD onset and dyslipidaemia has been recognized as an early symptom of biological perturbation, lipid extracts from peripheral blood mononuclear cells (PBMCs), consisting primarily of lymphocytes (T cells, B cells, and NK cells) and monocytes, of 38 children with ASD and their non-autistic siblings were investigated by hydrophilic interaction liquid chromatography (HILIC) coupled with electrospray ionization and Fourier-transform mass spectrometry (ESI-FTMS). Performances of two freeware software for data extraction and processing were compared with acquired reliable data regardless of the used informatics. A reduction of variables from 1460 by the untargeted XCMS to 324 by the semi-untargeted Alex software was attained. All-ion fragmentation (AIF) MS scans along with Alex software were successfully applied to obtain information related to fatty acyl chain composition of six glycerophospholipid classes occurring in PBMC. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were explored to verify the occurrence of significant differences in the lipid pool composition of ASD children compared with 36 healthy siblings. After rigorous statistical validation, we conclude that phospholipids extracted from PBMC of children affected by ASD do not exhibit diagnostic biomarkers. Yet interindividual variability comes forth from this study as the dominant effect in keeping with the existing phenotypic and etiological heterogeneity among ASD individuals. Graphical abstract.
自闭症谱系障碍（ASD）是一组广泛且异质的神经发育障碍，其特征为社交互动和沟通受损、行为模式受限和重复、感官处理改变。目前，尚无可靠的 ASD 分子生物标志物。由于免疫失调被认为与 ASD 的发病有关，而血脂异常已被认为是生物扰动的早期症状，因此研究了 38 名 ASD 儿童及其非自闭症兄弟姐妹的外周血单个核细胞（PBMC）的脂提取物，主要由淋巴细胞（T 细胞、B 细胞和 NK 细胞）和单核细胞组成。通过亲水相互作用液相色谱（HILIC）与电喷雾电离和傅里叶变换质谱（ESI-FTMS）联用进行了研究。比较了两种免费软件的数据提取和处理性能，无论使用哪种信息学方法，都与可靠的已获取数据一致。通过无靶向 XCMS 将变量从 1460 个减少到半靶向 Alex 软件的 324 个。全离子碎裂（AIF）MS 扫描以及 Alex 软件成功用于获得与 PBMC 中六种甘油磷脂类的脂肪酸链组成相关的信息。探索了主成分分析（PCA）和偏最小二乘判别分析（PLS-DA），以验证与 36 名健康兄弟姐妹相比，ASD 儿童的脂质池组成是否存在显著差异。经过严格的统计验证，我们得出结论，从受 ASD 影响的儿童的 PBMC 中提取的磷脂没有表现出诊断生物标志物。然而，从这项研究中可以看出个体间的变异性是主要影响因素，这与 ASD 个体之间存在的现有表型和病因异质性一致。
