Tania Gutierrez-Riquelme, Isabel Karkossa, Kristin Schubert, Gudrun Liebscher, Eva-Maria Packeiser, Ingo Nolte, Martin von Bergen, Hugo Murua Escobar, Matias Aguilera-Rojas, Ralf Einspanier, Torsten Stein
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We aimed to identify CMT subtype-specific proteome profiles by comparing the proteomes of EVs isolated from epithelial cell lines derived from morphologically normal canine mammary tissue, adenomas, and carcinomas.</p><p><strong>Methods: </strong>Whole-cell protein lysates (WCLs) and EV-lysates were obtained from five canine mammary cell lines: MTH53A (non-neoplastic); ZMTH3 (adenoma); MTH52C (simple carcinoma); 1305, DT1406TB (complex carcinoma); and their proteins identified by LC-MS/MS analyses. Gene Ontology analysis was performed on differentially abundant proteins from each group to identify up- and down-regulated biological processes. To establish CMT subtype-specific proteomic profiles, weighted gene correlation network analysis (WGCNA) was carried out.</p><p><strong>Results: </strong>WCL and EVs displayed distinct protein abundance signatures while still showing the same increase in adhesion, migration, and motility-related proteins in carcinoma-derived cell lines, and of RNA processing and RNA splicing factors in the adenoma cell line. WGCNA identified CMT stage-specific co-abundant EV proteins, allowing the identification of adenoma and carcinoma EV signatures not seen in WCLs.</p><p><strong>Conclusions: </strong>EVs from CMT cell lines exhibit distinct protein profiles reflecting malignancy state, allowing us to identify potential biomarkers for canine mammary carcinomas, such as biglycan. Our dataset could therefore potentially serve as a basis for the development of a less invasive clinical diagnostic tool for the characterisation of CMT.</p>","PeriodicalId":9041,"journal":{"name":"BMC Veterinary Research","volume":"20 1","pages":"488"},"PeriodicalIF":2.3000,"publicationDate":"2024-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11515202/pdf/","citationCount":"0","resultStr":"{\"title\":\"Proteomic analysis of extracellular vesicles derived from canine mammary tumour cell lines identifies protein signatures specific for disease state.\",\"authors\":\"Tania Gutierrez-Riquelme, Isabel Karkossa, Kristin Schubert, Gudrun Liebscher, Eva-Maria Packeiser, Ingo Nolte, Martin von Bergen, Hugo Murua Escobar, Matias Aguilera-Rojas, Ralf Einspanier, Torsten Stein\",\"doi\":\"10.1186/s12917-024-04331-1\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Canine mammary tumours (CMT) are among the most common types of tumours in female dogs. 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引用次数: 0
摘要
背景:犬乳腺肿瘤 (CMT) 是母犬最常见的肿瘤类型之一。目前,诊断需要进行侵入性组织活检和组织学分析。肿瘤细胞脱落的胞外囊泡 (EV) 含有 RNA 和蛋白质,具有液体活检诊断的潜力。我们的目的是通过比较从形态正常的犬乳腺组织、腺瘤和癌的上皮细胞系中分离出的EV的蛋白质组,确定CMT亚型特异性蛋白质组图谱:方法:从五种犬乳腺细胞系中获得全细胞蛋白裂解液(WCL)和EV-裂解液:MTH53A(非肿瘤性);ZMTH3(腺瘤);MTH52C(单纯癌);1305、DT1406TB(复杂癌);并通过 LC-MS/MS 分析鉴定了它们的蛋白质。对各组中含量不同的蛋白质进行了基因本体分析,以确定上调和下调的生物过程。为了建立CMT亚型特异性蛋白质组图谱,进行了加权基因相关网络分析(WGCNA):结果:WCL和EVs显示了不同的蛋白质丰度特征,但在癌源细胞系中,粘附、迁移和运动相关蛋白质以及腺瘤细胞系中的RNA加工和RNA剪接因子仍有相同的增加。WGCNA 鉴定出了 CMT 阶段特异性共富集 EV 蛋白,从而鉴定出了 WCLs 中未见的腺瘤和癌 EV 特征:结论:CMT细胞系的EV表现出不同的蛋白质特征,反映了恶性肿瘤的状态,使我们能够识别犬乳腺癌的潜在生物标记物,如biglycan。因此,我们的数据集有可能成为开发侵袭性较小的临床诊断工具的基础,以确定 CMT 的特征。
Proteomic analysis of extracellular vesicles derived from canine mammary tumour cell lines identifies protein signatures specific for disease state.
Background: Canine mammary tumours (CMT) are among the most common types of tumours in female dogs. Diagnosis currently requires invasive tissue biopsies and histological analysis. Tumour cells shed extracellular vesicles (EVs) containing RNAs and proteins with potential for liquid biopsy diagnostics. We aimed to identify CMT subtype-specific proteome profiles by comparing the proteomes of EVs isolated from epithelial cell lines derived from morphologically normal canine mammary tissue, adenomas, and carcinomas.
Methods: Whole-cell protein lysates (WCLs) and EV-lysates were obtained from five canine mammary cell lines: MTH53A (non-neoplastic); ZMTH3 (adenoma); MTH52C (simple carcinoma); 1305, DT1406TB (complex carcinoma); and their proteins identified by LC-MS/MS analyses. Gene Ontology analysis was performed on differentially abundant proteins from each group to identify up- and down-regulated biological processes. To establish CMT subtype-specific proteomic profiles, weighted gene correlation network analysis (WGCNA) was carried out.
Results: WCL and EVs displayed distinct protein abundance signatures while still showing the same increase in adhesion, migration, and motility-related proteins in carcinoma-derived cell lines, and of RNA processing and RNA splicing factors in the adenoma cell line. WGCNA identified CMT stage-specific co-abundant EV proteins, allowing the identification of adenoma and carcinoma EV signatures not seen in WCLs.
Conclusions: EVs from CMT cell lines exhibit distinct protein profiles reflecting malignancy state, allowing us to identify potential biomarkers for canine mammary carcinomas, such as biglycan. Our dataset could therefore potentially serve as a basis for the development of a less invasive clinical diagnostic tool for the characterisation of CMT.
期刊介绍:
BMC Veterinary Research is an open access, peer-reviewed journal that considers articles on all aspects of veterinary science and medicine, including the epidemiology, diagnosis, prevention and treatment of medical conditions of domestic, companion, farm and wild animals, as well as the biomedical processes that underlie their health.