Deregulation of desmosomal proteins and extracellular matrix proteases in odontogenic keratocyst

OBJECTIVE : Odontogenic keratocyst (OKC) is a benign lesion that tends to recur after surgical treatment. In an attempt to clarify the molecular basis underlining the OKC pathobiology, we aimed to analyze its proteomic profile. MATERIALS AND METHODS : We compared the proteomic profiles of five OKC and matched normal oral mucosa by using liquid chromatography–tandem mass spectrometry (LC-MS/MS). Then, we performed enrichment analysis and a literature search for the immunoexpression of the proteomics targets. RESULTS : We identified 1,150 proteins and 72 differently expressed proteins (log2 fold change ≥ 1.5; p < .05). Twenty-seven peptides were exclusively detected in the OKC samples. We found 35 enriched pathways related to cell differentiation and tissue architecture, including keratinocyte differentiation, keratinization, desmosome, and extracellular matrix (ECM) organization and degradation. The immunoexpression information of 11 out of 50 proteins identified in the enriched pathways was obtained. We found the downregulation of four desmosomal proteins (JUP, PKP1, PKP3, and PPL) and upregulation of ECM proteases (MMP-2, MMP-9, and cathepsins). CONCLUSIONS : Proteomic analysis strengthened the notion that OKC cells have a similar proteomic profile to oral keratinocytes. Contextual investigation of the differentially expressed proteins revealed the deregulation of desmosome proteins and ECM degradation as important alterations in OKC pathobiology.Conselho Nacional de Desenvolvimento Científico e Tecnológico and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior.http://www.wileyonlinelibrary.com/journal/odihj2022Oral Pathology and Oral Biolog

Caracterização morfológica e bioquímica de uma linhagem mutante de Leishmania braziliensis resistente à tunicamicina

Exportado OPUSMade available in DSpace on 2019-08-13T19:49:05Z (GMT). No. of bitstreams: 1 disserta__o__jessica_gardone_vit_rio.pdf: 1779179 bytes, checksum: 8d006a08e9ad60dddd861cfe059a9105 (MD5) Previous issue date: 21As leishmanioses compõem um grupo de doenças causadas por parasitos do gênero Leishmania e possuem grande importância em saúde pública. O controle dessas doenças ainda é um desafio, devido a crescente resistência às drogas utilizadas no tratamento. Nesse contexto, o estudo de modelos resistentes a drogas é essencial para se explorar o potencial metabólico dos parasitos e melhor compreender os mecanismos envolvidos na aquisição de resistência a fármacos. Neste estudo, foi gerada uma linhagem mutante de Leishmania braziliensis resistente ao antibiótico tunicamicina, fármaco que atua na inibição da glicosilação proteica. Para a geração dos mutantes foi utilizado o método de aumento gradual de concentração de droga (stepwise). Com o intuito de avaliar as adaptações metabólicas induzidas durante o processo de aquisição de resistência à tunicamicina, foram realizadas análises genômicas, ensaios de microscopia, infectividade, citometria e análise do metaboloma da linhagem resistente. As promastigotas de Leishmania braziliensis resistentes à tunicamicina (até 80 vezes o valor da DL50), apresentaram amplificação gênica e um concomitante aumento de virulência. Modificações morfológicas foram observadas na linhagem resistente como, desorganização das cisternas do complexo de Golgi indicando possíveis alterações no processo de glicosilação proteica. Alterações no metabolismo de várias fontes de carbono, relacionados à produção de substrato para a síntese de dolicol, molécula que desempenha papel importante na glicosilação de proteínas, foram detectadas como resposta ao bloqueio do processo induzido pela tunicamicina. Os dados gerados permitiram a concepção de um modelo de estudo dos mecanismos de resistência a drogas, bem como a influencia da glicosilação proteica nas relações parasito-hospedeiro em Leishmania.Leishmaniasis is a group of diseases caused by parasites of the Leishmania type and is of great importance in public health. Disease control is still a challenge due to its resistance to the drugs used in treatment. In this context, the study of potential resistance is essential for the exploration of the potential metabolism of parasitism and improve the structures of the drugs of the stress of drugs In this study, a mutant strain of Leishmania braziliensis resistant to the antibiotic tunicamycin, a drug that inhibits the glycosylation of protein, was generated by the gradual increase in drug concentration (stepwise). In order to evaluate the metabolic adaptations caused during the process of developing resistance to tunicamycin. In order to evaluate the metabolic changes during the acquisition of resistance to tunicamycin, genomic analyzes, microscopy, infectivity, cytometry and resistance metabolism analysis were performed. Promastigotes of Leishmania braziliensis resistant to tunicamycin (up to 80 times the value of LD50), gene amplification and a concomitant increase in virulence. Morphological changes were observed in the lineage such, disorganization of cisterns of the Golgi complex, changes in the process of protein glycosylation. Changes in the metabolism of various carbon sources, related to the production of substrate for a synthesis of fatty acid, molecules that play an important role in the glycosylation of proteins, were detected as responses to the induction blocking process of tunicamycin. The results suggests that main mechanisms of drug resistance, as well as the influence of protein glycosylation on host parasitic infections in Leishmania