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Frederico Marianetti Sorianihttp://lattes.cnpq.br/3671650166497515https://lattes.cnpq.br/6177388935668766Marina Pimenta Braga2023-08-16T15:27:40Z2023-08-16T15:27:40Z2023-07-07http://hdl.handle.net/1843/57842A Hanseníase é uma doença crônica e infecto contagiosa, tendo como agente etiológico a Mycobacterium leprae. Atualmente, é uma doença que, apesar de ter tido umgrande progresso na diminuição de sua transmissão, ainda acomete um número consideravelmente grande de pessoas. É caracterizada por um amplo espectro clínico que varia entre a forma de resistência (tuberculóide -TT) e a forma de susceptibilidade (virchowiana – VV), que se manifestam sob diferentes perfis de resposta imune. O controle da resposta imune envolve uma miríade de processos celulares e teciduais e, dentre eles, o controle da expressão gênica é fator essencial. Resultados prévios demonstram que existe um papel importante de miRNAs, pequenos RNAs não codificantes, nas formas polares da hanseníase. Previamente, identificamos o perfil de expressão de miRNAs diferencial em pacientes com as formas polares da hanseníase (TT e VV) e indivíduos controle não doentes, e estes perfis foram utilizados para identificar, in silico, vias celulares enriquecidas e que pudessem estar relacionadas à fisiopatologia da doença. Dentre essas vias, destacam-se as vias de reconhecimento de patógenos ou de PAMPs, como a sinalização mediada por TLR4 (Toll-like receptor 4). Esta via possui um regulador central, o let-7a-5p, que possui como alvo o próprio receptor TLR4 e apresenta-se diferencialmente expresso nas formas clínicas da doença. Nesse sentido, considerando que let-7a-5p pode ter uma função importante na regulação do reconhecimento do patógeno por TLR4, nas formas clínicas polares da hanseníase, o objetivo deste trabalho foi caracterizar a função do miRNA let-7a-5p no controle da sinalização celular mediada por TLR4 e correlacionar com a fisiopatologia da hanseníase. Para tanto foram padronizados sistemas in vitro, em cultura de células HEK293T e L929. Os resultados demonstram que TLR4 é um alvo do miRNA e pode ter sua expressão controlada, pelo menos em parte, por let-7a-5p. Além disso, a presença do mímico de let-7a-5 é capaz de inibir parcialmente o aumento de CXCL1, após o estímulo de fibroblastos L929 com LPS, e observamos também a alteração da expressão do miRNA após o estímulo das células com LPS e a alteração de mRNA de TLR4 em células tratadas com o mímico do miRNA. Os resultados gerados neste trabalho demonstram que let-7a-5p é um controlador da resposta inflamatória mediada por TLR4 na hanseníase.Leprosy is a chronic and contagious infectious disease, whose etiological agent is Mycobacterium leprae. Currently, it is a disease that, despite having made great progress in reducing its transmission, still affects a considerably large number of people. It is characterized by a wide clinical spectrum that varies between the form of resistance (tuberculoid -TT) and the form of susceptibility (virchowian -VV), which manifest in different profiles of immune response. The control of the immune response involves a myriad of cellular and tissue processes and, among them, the control of gene expression is an essential factor. Previous results demonstrate that there is an important role for miRNAs, small non-coding RNAs, in the polar forms of leprosy. Previously, we identified the differential expression profile of miRNAs in patients with the polar forms of leprosy (TT and VV) and non-ill control subjects, and these profiles were used to identify, in silico, enriched cellular pathways that could be related to the pathophysiology of the disease. Among these pathways, the recognition pathways of pathogens or PAMPs stand out, such as signaling mediated by TLR4 (Toll-like receptor 4). This pathway has a central regulator, let-7a-5p, which targets the TLR4 receptor itself and is differentially expressed in the clinical forms of the disease. In this sense, considering that let-7a-5p may play an important role in regulating the recognition of the pathogen by TLR4, in the polar clinical forms of leprosy, the objective of this work was to characterize the function of the miRNA let- 7a-5p in the signaling control cell mediated by TLR4 and correlate with the pathophysiology of leprosy. For this purpose, in vitro systems were standardized in HEK293T and L929 cell cultures. The results demonstrate that TLR4 is a miRNA target and may have its expression controlled, at least in part, by let-7a-5p. Furthermore, the presence of the let-7a-5 mimic is capable of partially inhibiting the increase in CXCL1 after stimulating L929 fibroblasts with LPS, and we also observed changes in miRNA expression after stimulating cells with LPS and TLR4 mRNA alteration in cells treated with the miRNA mimic. The results generated in this work demonstrate that let-7a-5p is a controller of the TLR4-mediated inflammatory response in leprosy.FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas GeraisporUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em GenéticaUFMGBrasilhttp://creativecommons.org/licenses/by-nc-nd/3.0/pt/info:eu-repo/semantics/openAccessGenéticaHanseníaseMycobacterium lepraeMicroRNAsReceptor 4 Toll-LikeHanseníasemiRNAsMycobacterium lepraeTLR4Caracterização do papel do miRNA let-7a-5p no controle da resposta imune e inflamatória na hanseníaseinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALUFMG-dissertação de mestrado Marina Braga (4).pdfUFMG-dissertação de mestrado Marina Braga (4).pdfapplication/pdf1849736https://repositorio.ufmg.br/bitstream/1843/57842/1/UFMG-disserta%c3%a7%c3%a3o%20de%20mestrado%20Marina%20Braga%20%284%29.pdf43f71a630daaa8d5a86a9dd1dde55561MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufmg.br/bitstream/1843/57842/2/license_rdfcfd6801dba008cb6adbd9838b81582abMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82118https://repositorio.ufmg.br/bitstream/1843/57842/3/license.txtcda590c95a0b51b4d15f60c9642ca272MD531843/578422023-08-16 12:27:40.9oai:repositorio.ufmg.br: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ório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2023-08-16T15:27:40Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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