Non-Exosomal and Exosome-Derived miRNAs as Promising Biomarkers in Canine Mammary Cancer

Petroušková, Patrícia; Hudáková, Nikola; Maloveská, Marcela; Humeník, Filip; Čı́žková, Dáša

doi:10.3390/life12040524

Cited by 9 publications

(9 citation statements)

References 317 publications

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“…Several non-exosomal and exosome-derived miRNAs are already known to be dysregulated in canine mammary gland cancer, and noteworthy, high similarities in terms of miRNA dynamics and functions have been found between canine mammary and human breast cancer [ 19 , 27 ]. MiRNAs were investigated in primary and metastatic CMT tissues to find candidate biomarkers in distinguishing between different stages of CMTs [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

RT-qPCR Expression Profiles of Selected Oncogenic and Oncosuppressor miRNAs in Formalin-Fixed, Paraffin-Embedded Canine Mammary Tumors

Abbate

Giannetto

Arfuso

et al. 2022

Animals

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MicroRNAs (miRNAs) can act as oncogenes or oncosuppressor genes, and their involvement in nearly all cancer-associated processes makes these small molecules promising diagnostic and prognostic biomarkers in cancer, as well as specific targets for cancer therapy. This study aimed to investigate the expression of 7 miRNAs (miR-18a, miR-18b, miR-22, miR-124, miR-145, miR-21, miR-146b) in formalin-fixed, paraffin-embedded canine mammary tumors (CMTs) by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Twenty-six mammary samples were selected, including 22 CMTs (7 benign; 15 malignant) and 4 control samples (3 normal mammary gland and 1 case of lobular hyperplasia). Oncogenic miR-18a, miR-18b and miR-21 were significantly upregulated in malignant tumors compared with control tissues (p < 0.05). Conversely, oncosuppressor miR-146b was significantly downregulated in benign and malignant mammary tumors compared with control samples (p < 0.05) while, no group-related differences in the expression levels of miR-22, miR-124 and miR-145 were found (p > 0.05). Upregulated miRNAs found here, may regulate genes involved in receptor-mediated carcinogenesis and proteoglycan remodeling in cancer; while miRNA with reduced expression can regulate genes involved in Toll-like receptor and MAPK signaling pathways. According to the results obtained in the current study, the oncogenic and oncosuppressor miRNAs analyzed here are dysregulated in CMTs and the dysregulation of miRNA targets may lead to specific altered cellular processes and key pathways involved in carcinogenesis. Of note, since oncogenic miRNAs predicted to regulate neoplastic cell proliferation and hormonal activities, they may play an active role in neoplastic transformation and/or progression, having mechanistic and prognostic relevance in CMTs.

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Section: Introductionmentioning

confidence: 99%

RT-qPCR Expression Profiles of Selected Oncogenic and Oncosuppressor miRNAs in Formalin-Fixed, Paraffin-Embedded Canine Mammary Tumors

Abbate

Giannetto

Arfuso

et al. 2022

Animals

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“…In particular, non-coding RNAs participate in carcinogenesis by regulating the expression of protein-coding cancer-associated genes, and it is noteworthy that among the different molecular mechanisms studied on ER downregulation, the progressive loss of ER expression in breast cancer would appear to be more epigenetic than genetic in nature [ 41 ]. Conversely, studies approaching epigenetic mechanisms in canine mammary cancer are still scarce and only few studies investigating miRNA expression have been published, to date [ 12 , 31 , 32 ].…”

Section: Discussionmentioning

confidence: 99%

“…Commonly, miR-18a and miR-18b, and their cluster members, are described as onco-microRNAs, as they show increased expression in more aggressive tumor types [ 12 , 31 , 32 ]. Especially in canine mammary cancer, miR-18a is an oncomiR found to be upregulated in exosomes isolated from malignant mammary epithelial cell lines [ 31 ] and overexpressed in sera from canine patients with metastatic CMTs compared with those with non-metastatic mammary tumors [ 32 ], thus suggesting their role as promising prognostic biomarkers.…”

Section: Discussionmentioning

confidence: 99%

“…It is noteworthy that among several epigenetic mechanisms, miRNAs have been confirmed to play a crucial role in the pathophysiology of breast cancer and several basic and interventional attempts to target epigenomic modifiers in breast cancer have recently been shown to be successful [ 10 , 11 ]. Furthermore, due to their relevance in many cellular pathways, miRNA dysregulation is the most studied epigenetic alteration in canine mammary cancer [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

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Epigenetics in Canine Mammary Tumors: Upregulation of miR-18a and miR-18b Oncogenes Is Associated with Decreased ERS1 Target mRNA Expression and ERα Immunoexpression in Highly Proliferating Carcinomas

Abbate

Arfuso

Riolo³

et al. 2023

Animals

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The expression of miRNAs is one of the main epigenetic mechanisms responsible for the regulation of gene expression in mammals, and in cancer, miRNAs participate by regulating the expression of protein-coding cancer-associated genes. In canine mammary tumors (CMTs), the ESR1 gene encodes for ERα, and represents a major target gene for miR-18a and miR-18b, previously found to be overexpressed in mammary carcinomas. A loss in ERα expression in CMTs is commonly associated with poor prognosis, and it is noteworthy that the downregulation of the ESR1 would appear to be more epigenetic than genetic in nature. In this study, the expression of ESR1 mRNA in formalin-fixed, paraffin-embedded (FFPE) canine mammary tumors (CMTs) was evaluated and compared with the expression levels of miR18a and miR18b, both assessed via RT-qPCR. Furthermore, the possible correlation between the miRNA expression data and the immunohistochemical prognostic factors (ERα immunoexpression; Ki67 proliferative index) was explored. A total of twenty-six FFPE mammary samples were used, including 22 CMTs (7 benign; 15 malignant) and four control samples (three normal mammary glands and one case of lobular hyperplasia). The obtained results demonstrate that miR-18a and miR-18b are upregulated in malignant CMTs, negatively correlating with the expression of target ESR1 mRNA. Of note, the upregulation of miRNAs strictly reflects the progressive loss of ERα immunoexpression and increased tumor cell proliferation as measured using the Ki67 index. The results suggest a central role of miR-18a and miR-18b in the pathophysiology of canine mammary tumors as potential epigenetic mechanisms involved in ERα downregulation. Moreover, as miRNA expression reflects ERα protein status and a high proliferative index, miR-18a and miR-18b may represent promising biomarkers with prognostic value. More detailed investigations on a larger number of cases are needed to better understand the influence of these miRNAs in canine mammary tumors.

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“…No data on plasma circulating miR-21-5p in MCT-affected dogs have been reported so far. Several studies have been conducted in veterinary oncology to investigate the sEV cargo and their role in tumors ( 28 – 32 ), but only one study investigated the concentration of EVs in the blood of MCT-affected dogs with different histological grades ( 33 ). Based on the above considerations, this pilot study aimed at evaluating the expression of miR-21-5p in the plasma of dogs affected by cutaneous MCT to confirm the ability of miR-21-5p to be associated with sEV and investigate whether the stage and consequently the prognosis of the disease are correlated to the sEV-mi-R-21-5p level supporting its potential as a biomarker for canine MCT.…”

Section: Introductionmentioning

confidence: 99%

Plasma small extracellular vesicles from dogs affected by cutaneous mast cell tumors deliver high levels of miR-21-5p

Zamarian¹,

Stefanello²,

Ferrari³

et al. 2023

Front. Vet. Sci.

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Small extracellular vesicles (sEV) are a class of extracellular vesicles (30–150 nm), delivering molecules including proteins, metabolites, and microRNAs (miRNAs), involved in physiological intercellular crosstalk and disease pathogenesis. The present pilot study aims are (I) to develop an easy and fast protocol for the isolation of sEV from plasma of mast cell tumor (MCT)-affected dogs; (II) to evaluate if miR-21-5p (sEV-miR-21-5p), a miRNA overexpressed by MCT, is associated with sEV. Seventeen dogs have been enrolled in the study: 4 healthy and 13 (6 with and 7 without nodal metastasis) MCT-affected dogs. sEV were isolated using size exclusion chromatography (SEC) (IZON column 35nm) and were characterized by Western blot, Nanoparticle tracking analysis, and transmission electron microscopy. sEV-miR-21-5p was quantified using digital PCR. sEV expressed the specific markers CD9 and TSG101, and a marker of mast cell tryptase. The sEV mean concentration and size were 2.68E + 10 particles/ml, and 99.6 nm, 2.89E + 10 particles/ml and 101.7 nm, and 3.21E + 10 particles/ml and 124 nm in non-metastatic, nodal metastatic, and healthy samples, respectively. The comparative analysis demonstrated that the level of sEV-miR-21-5p was significantly higher in dogs with nodal metastasis compared to healthy (P = 0.038) and without nodal metastasis samples (P = 0.007). In conclusion, the present work demonstrated that a pure population of sEV can be isolated from the plasma of MCT-affected dogs using the SEC approach and that the level of sEV-miR-21-5p is higher in nodal metastatic MCT-affected dogs compared with healthy and MCT-affected dogs without nodal involvement.

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Non-Exosomal and Exosome-Derived miRNAs as Promising Biomarkers in Canine Mammary Cancer

Cited by 9 publications

References 317 publications

RT-qPCR Expression Profiles of Selected Oncogenic and Oncosuppressor miRNAs in Formalin-Fixed, Paraffin-Embedded Canine Mammary Tumors

RT-qPCR Expression Profiles of Selected Oncogenic and Oncosuppressor miRNAs in Formalin-Fixed, Paraffin-Embedded Canine Mammary Tumors

Epigenetics in Canine Mammary Tumors: Upregulation of miR-18a and miR-18b Oncogenes Is Associated with Decreased ERS1 Target mRNA Expression and ERα Immunoexpression in Highly Proliferating Carcinomas

Plasma small extracellular vesicles from dogs affected by cutaneous mast cell tumors deliver high levels of miR-21-5p

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