Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Tipo de documento: | Trabalho de conclusão de curso |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/15964 |
Resumo: | Chikungunya virus (CHIKV), which causes Chikungunya fever, is a arbovirus transmitted by the Aedes aegypti mosquito, which periodically causes epidemics around the world and can become a serious public health problem. However, there are no approved vaccines or drugs against CHIKV. So it's research to identify antivirals is fundamental, which can be carried out in a quickly and efficiently by high-throughput screening (HTS) assays, such as replicon-based assays. These are subgenomic systems in which genes encoding structural proteins are replaced by a reporter gene, allowing the identification of inhibitors of viral replication by measuring the activity of the reporter protein, in an easier and safer than traditional methods. In this study, the replicon strain was used. CHIKV reporter that contains the Gaussia luciferase (GLuc) gene as a reporter, called BHK-21-Rep-T7-GLuc-nSP-CHIKV-99650, for the screening of three MMV/DNDi small molecule libraries. Through tests based on replicon it was possible to identify 4 compounds, Itraconazole, GSK-983, Rubitecan and MMV676270, as inhibitors of CHIKV replication with low to moderate toxicity. Two of these molecules, the antifungal Itraconazole and the antiviral Rubitecan, were the most promising with selectivity indices >312.5 and >714.3, respectively, values considerably higher than those observed for the too much. In addition, the inhibition of replication was confirmed by the decrease in relative expression of the replicon RNA in cells treated with the concentrations effects of the 4 compounds. To better characterize the activity of these molecules against CHIKV, viral infection assays will be carried out in collaboration with Prof. Ana Carolina Jardim from the Federal University of Uberlândia (UFU). |
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Policastro, Lucca RochaFernandes, Rafaela Sachettohttp://lattes.cnpq.br/2580624711278661Anibal, Fernanda de Freitashttp://lattes.cnpq.br/491826196877280644c91d54-30f5-47d4-8dd9-5274054cb9242022-05-02T20:48:14Z2022-05-02T20:48:14Z2022-04-12POLICASTRO, Lucca Rocha. Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais. 2022. Trabalho de Conclusão de Curso (Graduação em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2022. Disponível em: https://repositorio.ufscar.br/handle/ufscar/15964.https://repositorio.ufscar.br/handle/ufscar/15964Chikungunya virus (CHIKV), which causes Chikungunya fever, is a arbovirus transmitted by the Aedes aegypti mosquito, which periodically causes epidemics around the world and can become a serious public health problem. However, there are no approved vaccines or drugs against CHIKV. So it's research to identify antivirals is fundamental, which can be carried out in a quickly and efficiently by high-throughput screening (HTS) assays, such as replicon-based assays. These are subgenomic systems in which genes encoding structural proteins are replaced by a reporter gene, allowing the identification of inhibitors of viral replication by measuring the activity of the reporter protein, in an easier and safer than traditional methods. In this study, the replicon strain was used. CHIKV reporter that contains the Gaussia luciferase (GLuc) gene as a reporter, called BHK-21-Rep-T7-GLuc-nSP-CHIKV-99650, for the screening of three MMV/DNDi small molecule libraries. Through tests based on replicon it was possible to identify 4 compounds, Itraconazole, GSK-983, Rubitecan and MMV676270, as inhibitors of CHIKV replication with low to moderate toxicity. Two of these molecules, the antifungal Itraconazole and the antiviral Rubitecan, were the most promising with selectivity indices >312.5 and >714.3, respectively, values considerably higher than those observed for the too much. In addition, the inhibition of replication was confirmed by the decrease in relative expression of the replicon RNA in cells treated with the concentrations effects of the 4 compounds. To better characterize the activity of these molecules against CHIKV, viral infection assays will be carried out in collaboration with Prof. Ana Carolina Jardim from the Federal University of Uberlândia (UFU).O vírus Chikungunya (CHIKV), causador da febre Chikungunya, é um arbovírus transmitido pelo mosquito Aedes aegypti, que periodicamente provoca epidemias ao redor do mundo, podendo se tornar um sério problema de saúde pública. Entretanto, não existem vacinas ou fármacos aprovados contra o CHIKV. Portanto, é fundamental a pesquisa para identificação de antivirais, a qual pode ser realizada de maneira rápida e eficiente por ensaios em modo de triagem de alta performance (highthroughput screening, HTS), como os ensaios baseados em replicon. Estes são sistemas subgenômicos nos quais os genes codificadores das proteínas estruturais são substituídos por um gene repórter, permitindo a identificação de inibidores da replicação viral medindo-se a atividade da proteína repórter, de maneira mais fácil e segura do que os métodos tradicionais. Neste estudo, foi utilizada a linhagem replicon repórter de CHIKV que contém o gene da Gaussia luciferase (GLuc) como repórter, denominada BHK-21-Rep-T7-GLuc-nSP-CHIKV-99650, para a triagem de três bibliotecas de pequenas moléculas do MMV/DNDi. Através dos ensaios baseados em replicon foi possível identificar 4 compostos, Itraconazol, GSK-983, Rubitecan e MMV676270, como inibidores da replicação do CHIKV com baixa a moderada toxicidade. Duas destas moléculas, o antifúngico Itraconazol e o antiviral Rubitecan, foram as mais promissoras com índices de seletividade >312,5 e >714,3, respectivamente, valores consideravelmente maiores do que o observado para os demais. Além disso, a inibição da replicação foi confirmada pela diminuição da expressão relativa do RNA replicon em células tratadas com as concentrações efetivas dos 4 compostos. Para melhor caracterizar atividade destas moléculas contra o CHIKV, ensaios de infecção viral serão realizados em colaboração com a Profa. Ana Carolina Jardim da Universidade Federal de Uberlândia (UFU).Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)2021/01706-0porUniversidade Federal de São CarlosCâmpus São CarlosBiotecnologia - BiotecUFSCarAttribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nc-nd/3.0/br/info:eu-repo/semantics/openAccessAlfavírusArbovirosesEnsaios celularesDescoberta de drogasChikungunyaRepliconCIENCIAS BIOLOGICAS::GENETICAEnsaios celulares baseados em replicon de Chikungunya para identificação de antiviraisChikungunya replicon-based cell assays for antiviral identificationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bachelorThesis6006001a6ee24d-c5f4-4aa1-ac4d-cf987c1aa5b8reponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALTCC - Lucca Rocha Policastro.pdfTCC - Lucca Rocha Policastro.pdfTCC originalapplication/pdf578309https://repositorio.ufscar.br/bitstream/ufscar/15964/3/TCC%20-%20Lucca%20Rocha%20Policastro.pdfa52ebfec4df68f7f28d78ac6c574a873MD53CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufscar.br/bitstream/ufscar/15964/4/license_rdfe39d27027a6cc9cb039ad269a5db8e34MD54TEXTTCC - Lucca Rocha Policastro.pdf.txtTCC - Lucca Rocha Policastro.pdf.txtExtracted texttext/plain49392https://repositorio.ufscar.br/bitstream/ufscar/15964/5/TCC%20-%20Lucca%20Rocha%20Policastro.pdf.txt355e436c5c7aeefd26e48b2061b4c8afMD55THUMBNAILTCC - Lucca Rocha Policastro.pdf.jpgTCC - Lucca Rocha Policastro.pdf.jpgIM Thumbnailimage/jpeg5239https://repositorio.ufscar.br/bitstream/ufscar/15964/6/TCC%20-%20Lucca%20Rocha%20Policastro.pdf.jpg9b76789726459cc1787af378d702bbc8MD56ufscar/159642023-09-18 18:32:20.861oai:repositorio.ufscar.br:ufscar/15964Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:32:20Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais |
dc.title.alternative.eng.fl_str_mv |
Chikungunya replicon-based cell assays for antiviral identification |
title |
Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais |
spellingShingle |
Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais Policastro, Lucca Rocha Alfavírus Arboviroses Ensaios celulares Descoberta de drogas Chikungunya Replicon CIENCIAS BIOLOGICAS::GENETICA |
title_short |
Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais |
title_full |
Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais |
title_fullStr |
Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais |
title_full_unstemmed |
Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais |
title_sort |
Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais |
author |
Policastro, Lucca Rocha |
author_facet |
Policastro, Lucca Rocha |
author_role |
author |
dc.contributor.author.fl_str_mv |
Policastro, Lucca Rocha |
dc.contributor.advisor1.fl_str_mv |
Fernandes, Rafaela Sachetto |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/2580624711278661 |
dc.contributor.advisor-co1.fl_str_mv |
Anibal, Fernanda de Freitas |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/4918261968772806 |
dc.contributor.authorID.fl_str_mv |
44c91d54-30f5-47d4-8dd9-5274054cb924 |
contributor_str_mv |
Fernandes, Rafaela Sachetto Anibal, Fernanda de Freitas |
dc.subject.por.fl_str_mv |
Alfavírus Arboviroses Ensaios celulares Descoberta de drogas |
topic |
Alfavírus Arboviroses Ensaios celulares Descoberta de drogas Chikungunya Replicon CIENCIAS BIOLOGICAS::GENETICA |
dc.subject.eng.fl_str_mv |
Chikungunya Replicon |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::GENETICA |
description |
Chikungunya virus (CHIKV), which causes Chikungunya fever, is a arbovirus transmitted by the Aedes aegypti mosquito, which periodically causes epidemics around the world and can become a serious public health problem. However, there are no approved vaccines or drugs against CHIKV. So it's research to identify antivirals is fundamental, which can be carried out in a quickly and efficiently by high-throughput screening (HTS) assays, such as replicon-based assays. These are subgenomic systems in which genes encoding structural proteins are replaced by a reporter gene, allowing the identification of inhibitors of viral replication by measuring the activity of the reporter protein, in an easier and safer than traditional methods. In this study, the replicon strain was used. CHIKV reporter that contains the Gaussia luciferase (GLuc) gene as a reporter, called BHK-21-Rep-T7-GLuc-nSP-CHIKV-99650, for the screening of three MMV/DNDi small molecule libraries. Through tests based on replicon it was possible to identify 4 compounds, Itraconazole, GSK-983, Rubitecan and MMV676270, as inhibitors of CHIKV replication with low to moderate toxicity. Two of these molecules, the antifungal Itraconazole and the antiviral Rubitecan, were the most promising with selectivity indices >312.5 and >714.3, respectively, values considerably higher than those observed for the too much. In addition, the inhibition of replication was confirmed by the decrease in relative expression of the replicon RNA in cells treated with the concentrations effects of the 4 compounds. To better characterize the activity of these molecules against CHIKV, viral infection assays will be carried out in collaboration with Prof. Ana Carolina Jardim from the Federal University of Uberlândia (UFU). |
publishDate |
2022 |
dc.date.accessioned.fl_str_mv |
2022-05-02T20:48:14Z |
dc.date.available.fl_str_mv |
2022-05-02T20:48:14Z |
dc.date.issued.fl_str_mv |
2022-04-12 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
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bachelorThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
POLICASTRO, Lucca Rocha. Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais. 2022. Trabalho de Conclusão de Curso (Graduação em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2022. Disponível em: https://repositorio.ufscar.br/handle/ufscar/15964. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/15964 |
identifier_str_mv |
POLICASTRO, Lucca Rocha. Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais. 2022. Trabalho de Conclusão de Curso (Graduação em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2022. Disponível em: https://repositorio.ufscar.br/handle/ufscar/15964. |
url |
https://repositorio.ufscar.br/handle/ufscar/15964 |
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Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ |
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openAccess |
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Universidade Federal de São Carlos Câmpus São Carlos Biotecnologia - Biotec |
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Universidade Federal de São Carlos Câmpus São Carlos Biotecnologia - Biotec |
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