Navajas Rosana, Ramos-Fernandez Antonio, Herraiz Ignacio, Galindo Alberto, Bartha José Luis, Corrales Fernando, Paradela Alberto
Functional Proteomics Facility, Centro Nacional de Biotecnología (CNB-CSIC), ProteoRed-ISCIII, Madrid, Spain.
Proteobotics, Madrid, Spain.
Clin Proteomics. 2022 Feb 10;19(1):5. doi: 10.1186/s12014-022-09342-4.
The high incidence of pre-eclampsia, which affects 2-7% of all pregnancies, remains a major health concern. Detection of pre-eclampsia before the appearance of clinical symptoms is essential to allow early intervention, and would benefit from identification of plasma/serum biomarkers to help guide diagnosis and treatment. Liquid biopsy has emerged as a promising source of protein biomarkers that circumvents some of the inherent challenges of proteome-wide analysis of plasma/serum. In this respect, purified exosomes have the added benefit of being carriers of intercellular communication both in physiological and pathological conditions.
We compared the protein complement of purified exosomes from three different collections of control and pre-eclamptic serum samples, obtained at the end of the second trimester of pregnancy and at delivery. We employed shotgun label-free proteomics to investigate differential protein expression, which was then validated by targeted proteomics.
We developed a purification method that yielded highly enriched exosome preparations. The presence of specific pregnancy protein markers suggested that a significant proportion of purified exosomes derived from tissues related to pregnancy. Quantitative proteomic analyses allowed us to identify 10, 114 and 98 differentially-regulated proteins in the three sample collections, with a high degree of concordance. Functional analysis suggested that these proteins participate in biological processes related to pre-eclampsia, including angiogenesis, inflammation and cell migration. The differential abundance of 66 proteins was validated by targeted proteomics. Finally, we studied the impact of the pre-eclampsia-associated exosomes in the proteome using an in vitro cellular model.
We have identified and validated differential exosomal proteins in liquid biopsy of pregnant women that open new possibilities for early detection of pre-eclampsia. Additionally, the functional impact of the proteome composition of purified pre-eclamptic exosomes in target cells provides new information to better understand changes in embryo-maternal interactions and, consequently, the pathogenesis of this disease.
子痫前期的发病率较高，影响着2%至7%的所有妊娠，仍然是一个主要的健康问题。在临床症状出现之前检测子痫前期对于早期干预至关重要，并且将受益于血浆/血清生物标志物的鉴定以帮助指导诊断和治疗。液体活检已成为一种有前景的蛋白质生物标志物来源，它规避了血浆/血清蛋白质组全分析的一些固有挑战。在这方面，纯化的外泌体在生理和病理条件下都具有作为细胞间通讯载体的额外优势。
我们比较了来自三个不同对照组和子痫前期血清样本集合的纯化外泌体的蛋白质组成，这些样本在妊娠中期结束时和分娩时获得。我们采用鸟枪法无标记蛋白质组学来研究差异蛋白质表达，然后通过靶向蛋白质组学进行验证。
我们开发了一种纯化方法，可产生高度富集的外泌体制剂。特定妊娠蛋白标志物的存在表明，很大一部分纯化的外泌体源自与妊娠相关的组织。定量蛋白质组分析使我们能够在三个样本集合中鉴定出10种、114种和98种差异调节的蛋白质，具有高度的一致性。功能分析表明，这些蛋白质参与了与子痫前期相关的生物学过程，包括血管生成、炎症和细胞迁移。66种蛋白质的差异丰度通过靶向蛋白质组学得到验证。最后，我们使用体外细胞模型研究了子痫前期相关外泌体对蛋白质组的影响。
我们已经在孕妇的液体活检中鉴定并验证了差异外泌体蛋白质，这为子痫前期的早期检测开辟了新的可能性。此外，纯化的子痫前期外泌体蛋白质组组成在靶细胞中的功能影响提供了新信息，以更好地理解胚胎 - 母体相互作用的变化，从而更好地理解这种疾病的发病机制。
