Proteômica de vesículas extracelulares de soro de cadelas com neoplasias mamárias
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Data
2023-07-07
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Universidade Estadual Paulista (Unesp)
Resumo
As neoplasias mamárias são o tipo de tumor mais frequente em cadelas, acometendo cerca de 50% das fêmeas. A espécie é considerada modelo experimental para câncer de mama em mulheres. Em vista disso, novas abordagens buscando biomarcadores e alvos terapêuticos têm encontrado respostas promissoras no estudo de vesículas extracelulares (VEs). As VEs são nanopartículas secretadas por diversos tipos celulares, incluindo células cancerígenas, as quais carreiam informações da sua célula de origem e estão envolvidas na manutenção do microambiente tumoral e metástases. O objetivo desse estudo foi investigar a relação da proteômica de VEs séricas de cadelas acometidas por neoplasias mamárias e a malignidade tumoral. Trinta cadelas foram selecionadas e divididas em 3 grupos, controle (GC, n = 10) composto por cadelas sem neoplasia; cadelas com tumores grau I e II (GI, n = 13) e grau III (GII, n = 7). Durante a exérese cirúrgica o sangue foi colhido, o soro separado e preparado para o isolamento das VEs por cromatografia por exclusão de tamanho. As VEs séricas foram caracterizadas por análise de nanopartículas, microscopia eletrônica de transmissão. A seguir as amostras foram digeridas com tripsina para proteômica com abordagem shotgun. A média ± erro padrão para a idade foi (1,95 ± 0,33; 10,1 ± 0,60; 12,1 ± 0,70 anos), e para o peso corporal (16,2 ± 2,5; 17,9 ± 3,6; 10,9 ± 3,4 kg). Não houve diferença entre a concentração de VEs (5,5 x 1012 ± 4,4 x 1012; 1,3 x 1012 ± 0,66 x 1012; 1,1 x 1012 ± 0,88 x 1012 partículas/mL) e tamanho das VEs (136,2 6,0; 129,3 4,0; 125,5 6,1 nm) entre os GC, GI e GII, respectivamente. Os resultados de VEs isoladamente não foram capazes de diferenciar o GC dos grupos GI e GII, entretanto podem ser considerados uma importante contribuição para a caracterização, quantificação e validação de VEs sérica em cadelas saudáveis e com neoplasia mamária. Na proteômica duas proteínas sintetizadas por genes importantes na supressão de tumores em humanos a keratin 18 e a protocadherin 17 apresentaram diferente abundância entre os grupos, podendo assim servir como candidatas em futuros estudos que busquem identificar biomarcadores para as neoplasias mamárias em cadelas por meio das VEs.
Mammary tumors are the most frequent type of neoplasm in bitches, affecting about 50% of females. The canine species is considered an experimental model for breast cancer in women. In view of this, new approaches seeking biomarkers and therapeutical targets have found encouraging answers in the research of extracellular vesicles (EVs). EVs are nanoparticles secreted by several cell types, including cancer cells, which carry information from their cell of origin and are involved in the maintenance of the tumor microenvironment and metastases. This study aimed to investigate the association between serum EVs proteomics from bitches with mammary neoplasms and tumor malignancy. Thirty bitches were selected and divided into three groups, control (GC, n = 10) composed of bitches without tumors; bitches with grade I and II (GI, n = 13), and grade III (GII, n = 7) tumors. Blood was collected during surgical excision, and the serum was separated and prepared for isolation of EVs by size exclusion chromatography. Serum EVs were characterized by nanoparticle analysis, transmission electron microscopy. Then the samples were digested with trypsin for proteomics with a shotgun approach. The mean ± standard error for age (1.95 ± 0.33; 10.1 ± 0.60; 12.1 ± 0.70 years) and body weight (16.2 ± 2.5; 17.9 ± 3.6; 10.9 ± 3.4 kg). There was no difference in the concentration of EVs (5.5 x 1012 ± 4.4 x 1012; 1.3 x 1012 ± 0.66 x 1012; 1.1 x 1012 ± 0.88 x 1012 particles/mL) and EV size (136.2 ± 6.0; 129.3 ± 4.0; 125.5 ± 6.1 nm) between GC, GI, and GII, respectively. Our EVs results were not able to differentiate the GC from the GI and GII, however, are an important contribution for the characterization quantification and validation of serum EVs in bitches. Also, our proteomics results showed two important proteins from genes that are important for tumor suppression in humans’ keratin 18 and protocadherin 17 that showed differences in regulation among the groups and could be a target for future studies that intent to identify and validate biomarkers for the canine mammary neoplasia.
Mammary tumors are the most frequent type of neoplasm in bitches, affecting about 50% of females. The canine species is considered an experimental model for breast cancer in women. In view of this, new approaches seeking biomarkers and therapeutical targets have found encouraging answers in the research of extracellular vesicles (EVs). EVs are nanoparticles secreted by several cell types, including cancer cells, which carry information from their cell of origin and are involved in the maintenance of the tumor microenvironment and metastases. This study aimed to investigate the association between serum EVs proteomics from bitches with mammary neoplasms and tumor malignancy. Thirty bitches were selected and divided into three groups, control (GC, n = 10) composed of bitches without tumors; bitches with grade I and II (GI, n = 13), and grade III (GII, n = 7) tumors. Blood was collected during surgical excision, and the serum was separated and prepared for isolation of EVs by size exclusion chromatography. Serum EVs were characterized by nanoparticle analysis, transmission electron microscopy. Then the samples were digested with trypsin for proteomics with a shotgun approach. The mean ± standard error for age (1.95 ± 0.33; 10.1 ± 0.60; 12.1 ± 0.70 years) and body weight (16.2 ± 2.5; 17.9 ± 3.6; 10.9 ± 3.4 kg). There was no difference in the concentration of EVs (5.5 x 1012 ± 4.4 x 1012; 1.3 x 1012 ± 0.66 x 1012; 1.1 x 1012 ± 0.88 x 1012 particles/mL) and EV size (136.2 ± 6.0; 129.3 ± 4.0; 125.5 ± 6.1 nm) between GC, GI, and GII, respectively. Our EVs results were not able to differentiate the GC from the GI and GII, however, are an important contribution for the characterization quantification and validation of serum EVs in bitches. Also, our proteomics results showed two important proteins from genes that are important for tumor suppression in humans’ keratin 18 and protocadherin 17 that showed differences in regulation among the groups and could be a target for future studies that intent to identify and validate biomarkers for the canine mammary neoplasia.
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Oncologia, Proteína, Exossomo, Tumor de mama, Biopsia liquida, Neoplasias da Mama