A Label-Free Proteomic Approach for the Identification of Biomarkers in the Exosome of Endometrial Cancer Serum

Abstract

Endometrial cancers (ECs) are mostly adenocarcinomas arising from the inner part of the uterus. The identification of serum biomarkers, either soluble or carried in the exosome, may be useful in making an early diagnosis. We used label-free quantification mass spectrometry (LFQ-MS)-based proteomics to investigate the proteome of exosomes in the albumin-depleted serum from 12 patients with EC, as compared to 12 healthy controls. After quantification and statistical analysis, we found significant changes in the abundance (p < 0.05) of 33 proteins in EC vs. control samples, with a fold change of ≥1.5 or ≤0.6. Validation using Western blotting analysis in 36 patients with EC as compared to 36 healthy individuals confirmed the upregulation of APOA1, HBB, CA1, HBD, LPA, SAA4, PF4V1, and APOE. A multivariate logistic regression model based on the abundance of these proteins was able to separate the controls from the EC patients with excellent sensitivity levels, particularly for stage 1 ECs. The results show that using LFQ-MS to explore the specific proteome of serum exosomes allows for the identification of biomarkers in EC. These observations suggest that PF4V1, CA1, HBD, and APOE represent biomarkers that are able to reach the clinical stage, after a validation phase.

MicroRNA Expression in Endometrial Cancer: Current Knowledge and Therapeutic Implications

“…The risk of EC spreading via lymphatic, hematologic, or contiguous routes may influence treatment options in terms of molecular profile, adjuvant therapy, and personalizing surgery with procedures such as lymphadenectomy, omentectomy, or adnexectomy [ 81 , 92 , 93 , 94 , 95 , 96 ]. Moreover, we call attention to other novel molecular signatures, such as circular RNAs and proteins, that are different among EC patients and healthy control subjects [ 97 , 98 ]. In particular, the proteins apolipoprotein A-I (APOA1), hemoglobin subunit beta (HBB), carbonic anhydrase 1 (CA1), hemoglobin subunit delta (HBD), apolipoprotein(a) (LPA), serum amyloid A-4 protein (SAA4), platelet factor 4 variant (PF4V1), and apolipoprotein E (APOE) were upregulated in the serum of EC patients compared to controls [ 97 ].…”

“…Moreover, we call attention to other novel molecular signatures, such as circular RNAs and proteins, that are different among EC patients and healthy control subjects [ 97 , 98 ]. In particular, the proteins apolipoprotein A-I (APOA1), hemoglobin subunit beta (HBB), carbonic anhydrase 1 (CA1), hemoglobin subunit delta (HBD), apolipoprotein(a) (LPA), serum amyloid A-4 protein (SAA4), platelet factor 4 variant (PF4V1), and apolipoprotein E (APOE) were upregulated in the serum of EC patients compared to controls [ 97 ]. In parallel, the circular RNAs hsa-circ-0109046 and hsa-circ-0002577 were upregulated in the serum of EC-affected women [ 98 ], whereas the circular RNA hsa-miR-200c-3p was slightly upregulated in urine [ 99 ].…”

Multi-omics analysis reveals attenuation of cellular stress by empagliflozin in high glucose-treated human cardiomyocytes

“…Proteomics analysis used a nanoLC coupled to a tribrid Orbitrap Lumos (Thermo Fisher Scientific, Bremen). Methods were performed as described previously [ 27 – 29 ]. Detailed mass spectrometry conditions and raw data alignment, filtering, and annotation parameters are described in (Additional file 1 ).…”

Advances in the application of label‐free quantitative proteomics techniques in malignancy research

“…In addition, they found that TMSB4X might be a novel treatment target for endometrial cancer, particularly in patients with premenopausal endometrial cancer. Sommella et al (2022) performed LFQ‐MS to analyze the exosomal proteomes in serum collected from 12 patients with endometrial cancer versus 12 healthy controls; then they validated some proteins from 36 sufferers with endometrial cancer and 36 healthy subjects using Western blotting and identified PF4V1, CA1, HBD, and APOE as possible diagnostic markers for endometrial cancer.…”