G-quadruplex aptamer-functionalized liposomes for lung cancer therapy

Detalhes bibliográficos
Autor(a) principal: Leitão, Daniela Ramos Vaz
Data de Publicação: 2023
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.6/14186
Resumo: Lung cancer (LC) has the greatest impact on mortality rates worldwide, particularly nonsmall cell lung cancer (NSCLC). The treatment of NSCLC depends on the stage of the disease and may involve a combination of surgery, chemotherapy, radiotherapy or immunotherapy. However, these therapies have some limitations, since that most patients develop resistance to treatments and also have side effects. In this sense, the aptamers have emerged as a promising strategy in delivery systems for NSCLC therapies. Aptamers are known as chemical antibodies since they can bind to proteins with high specificity. In addition, they have several advantages over antibodies, such as great stability and lower immunogenicity. AT11-L2 is a derivative of the AS1411 aptamer that contain a guanine-rich sequence capable of adopting a G-quadruplex (G4) structure and therefore targets nucleolin (NCL), a protein overexpressed in the cell membrane of NSCLC cells. Thus, the aim of this study was to develop two liposomal systems containing encapsulated doxorubicin and BRACO-19 and AT11-L2 on the surface to target NCL. To this end, the structure of AT11-L2 was first characterized by biophysical studies, and then the interaction of the aptamer with the ligands was studied through molecular dynamics simulations. In addition, the potential of the ligands to stabilize AT11-L2 was also assessed by biophysical studies. Liposomes were then synthesized and characterized. Finally, the selectivity and cellular uptake of the functionalized liposomes containing the compounds were tested on human alveolar basal epithelial adenocarcinoma cells (A549) and human fetal lung fibroblast cells (MRC-5). The results showed that AT11-L2 adopts a G4 structure in the presence of K+ and when associated with PhenDC3 through p-p stacking interactions, increases its stability. In addition, the AT11-L2/NCL complex was shown to have a high affinity with a KD in the nanomolar range. In vitro studies showed that functionalized liposomes containing BRACO-19 and doxorubicin were selectively internalized by cancer cells, with greater selectivity being recorded for liposomes with doxorubicin. These results show the therapeutic potential of liposomes functionalized with AT11-L2 against NSCLC.
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spelling G-quadruplex aptamer-functionalized liposomes for lung cancer therapyAptameros de G-QuadruplexCancro do Pulmão de Não Pequenas CélulasLipossomasNucleolinaDomínio/Área Científica::Engenharia e Tecnologia::BiotecnologiaLung cancer (LC) has the greatest impact on mortality rates worldwide, particularly nonsmall cell lung cancer (NSCLC). The treatment of NSCLC depends on the stage of the disease and may involve a combination of surgery, chemotherapy, radiotherapy or immunotherapy. However, these therapies have some limitations, since that most patients develop resistance to treatments and also have side effects. In this sense, the aptamers have emerged as a promising strategy in delivery systems for NSCLC therapies. Aptamers are known as chemical antibodies since they can bind to proteins with high specificity. In addition, they have several advantages over antibodies, such as great stability and lower immunogenicity. AT11-L2 is a derivative of the AS1411 aptamer that contain a guanine-rich sequence capable of adopting a G-quadruplex (G4) structure and therefore targets nucleolin (NCL), a protein overexpressed in the cell membrane of NSCLC cells. Thus, the aim of this study was to develop two liposomal systems containing encapsulated doxorubicin and BRACO-19 and AT11-L2 on the surface to target NCL. To this end, the structure of AT11-L2 was first characterized by biophysical studies, and then the interaction of the aptamer with the ligands was studied through molecular dynamics simulations. In addition, the potential of the ligands to stabilize AT11-L2 was also assessed by biophysical studies. Liposomes were then synthesized and characterized. Finally, the selectivity and cellular uptake of the functionalized liposomes containing the compounds were tested on human alveolar basal epithelial adenocarcinoma cells (A549) and human fetal lung fibroblast cells (MRC-5). The results showed that AT11-L2 adopts a G4 structure in the presence of K+ and when associated with PhenDC3 through p-p stacking interactions, increases its stability. In addition, the AT11-L2/NCL complex was shown to have a high affinity with a KD in the nanomolar range. In vitro studies showed that functionalized liposomes containing BRACO-19 and doxorubicin were selectively internalized by cancer cells, with greater selectivity being recorded for liposomes with doxorubicin. These results show the therapeutic potential of liposomes functionalized with AT11-L2 against NSCLC.O cancro do pulmão (CP) é uma das doenças oncológicas com maior impacto na taxa de mortalidade em todo o mundo, particularmente o cancro do pulmão de não pequenas células (CPNPC). O tratamento do CPNPC depende do estadio da doença e pode envolver uma combinação de terapias, tais como cirurgia, quimioterapia, radioterapia ou imunoterapia. No entanto, algumas abordagens apresentam limitações, uma vez que a maioria dos doentes desenvolvem resistência aos tratamentos e também apresentam efeitos secundários. Neste sentido, os aptameros surgiram como uma ferramenta promissora em sistemas de entrega de fármacos para terapias direcionadas ao CPNPC. Os aptameros são conhecidos como anticorpos químicos, uma vez que se podem ligar a proteínas com elevada especificidade. Além disso, têm várias vantagens sobre os anticorpos apresentando maior estabilidade e menor imunogenicidade. O AT11-L2 é um derivado do aptamero AS1411, formado por uma sequência rica em guaninas capaz de adotar uma estrutura G-quadruplex (G4) e tem como alvo a nucleolina (NCL), uma proteína sobre expressa na membrana celular das células cancerigenas nomeadamente em CPNPC. Assim, o objetivo deste estudo foi desenvolver dois sistemas lipídicos contendo doxorrubicina e BRACO-19 encapsulados e AT11-L2 na superfície. A estrutura do AT11-L2 foi primeiramente caracterizada por estudos biofísicos, e seguidamente a interação do aptamero com os ligandos foi estudada através de simulações de dinâmica molecular. O potencial dos ligandos para estabilizar o AT11-L2 foi também avaliado por diferentes estudos biofísicos. Seguidamente, os lipossomas foram sintetizados e caracterizados. Finalmente, a seletividade e a absorção celular dos lipossomas funcionalizados contendo os compostos em estudo foram determinadas em células de adenocarcinoma epitelial basal alveolar humano (A549) e em células de fibroblastos pulmonares fetais humanos (MRC-5). Os resultados mostraram que o AT11-L2 adopta uma estrutura G4 na presença de K+ e a sua estabilidade aumenta na presença do ligando de G4, PhenDC3, que interage com a estrutura de G4 do aptamero através de interacções de empilhamento p-p. Além disso, o complexo AT11-L2/NCL demonstrou ter uma elevada afinidade com um valor de KD da ordem do nanomolar. Os estudos in vitro mostraram que os lipossomas funcionalizados contendo BRACO-19 e doxorrubicina são seletivos e capazes de internalizar em células cancerígenas, tendo sido observada uma maior seletividade para os lipossomas com doxorrubicina. Estes resultados mostram o potencial terapêutico dos lipossomas funcionalizados com AT11-L2.Cruz, Carla Patricia Alves Freire Madeira daFigueiredo, Joana Patrícia RodriguesuBibliorumLeitão, Daniela Ramos Vaz2023-11-222023-10-092026-10-08T00:00:00Z2023-11-22T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.6/14186TID:203483642enginfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-01-31T02:32:20Zoai:ubibliorum.ubi.pt:10400.6/14186Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T01:59:19.097650Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv G-quadruplex aptamer-functionalized liposomes for lung cancer therapy
title G-quadruplex aptamer-functionalized liposomes for lung cancer therapy
spellingShingle G-quadruplex aptamer-functionalized liposomes for lung cancer therapy
Leitão, Daniela Ramos Vaz
Aptameros de G-Quadruplex
Cancro do Pulmão de Não Pequenas Células
Lipossomas
Nucleolina
Domínio/Área Científica::Engenharia e Tecnologia::Biotecnologia
title_short G-quadruplex aptamer-functionalized liposomes for lung cancer therapy
title_full G-quadruplex aptamer-functionalized liposomes for lung cancer therapy
title_fullStr G-quadruplex aptamer-functionalized liposomes for lung cancer therapy
title_full_unstemmed G-quadruplex aptamer-functionalized liposomes for lung cancer therapy
title_sort G-quadruplex aptamer-functionalized liposomes for lung cancer therapy
author Leitão, Daniela Ramos Vaz
author_facet Leitão, Daniela Ramos Vaz
author_role author
dc.contributor.none.fl_str_mv Cruz, Carla Patricia Alves Freire Madeira da
Figueiredo, Joana Patrícia Rodrigues
uBibliorum
dc.contributor.author.fl_str_mv Leitão, Daniela Ramos Vaz
dc.subject.por.fl_str_mv Aptameros de G-Quadruplex
Cancro do Pulmão de Não Pequenas Células
Lipossomas
Nucleolina
Domínio/Área Científica::Engenharia e Tecnologia::Biotecnologia
topic Aptameros de G-Quadruplex
Cancro do Pulmão de Não Pequenas Células
Lipossomas
Nucleolina
Domínio/Área Científica::Engenharia e Tecnologia::Biotecnologia
description Lung cancer (LC) has the greatest impact on mortality rates worldwide, particularly nonsmall cell lung cancer (NSCLC). The treatment of NSCLC depends on the stage of the disease and may involve a combination of surgery, chemotherapy, radiotherapy or immunotherapy. However, these therapies have some limitations, since that most patients develop resistance to treatments and also have side effects. In this sense, the aptamers have emerged as a promising strategy in delivery systems for NSCLC therapies. Aptamers are known as chemical antibodies since they can bind to proteins with high specificity. In addition, they have several advantages over antibodies, such as great stability and lower immunogenicity. AT11-L2 is a derivative of the AS1411 aptamer that contain a guanine-rich sequence capable of adopting a G-quadruplex (G4) structure and therefore targets nucleolin (NCL), a protein overexpressed in the cell membrane of NSCLC cells. Thus, the aim of this study was to develop two liposomal systems containing encapsulated doxorubicin and BRACO-19 and AT11-L2 on the surface to target NCL. To this end, the structure of AT11-L2 was first characterized by biophysical studies, and then the interaction of the aptamer with the ligands was studied through molecular dynamics simulations. In addition, the potential of the ligands to stabilize AT11-L2 was also assessed by biophysical studies. Liposomes were then synthesized and characterized. Finally, the selectivity and cellular uptake of the functionalized liposomes containing the compounds were tested on human alveolar basal epithelial adenocarcinoma cells (A549) and human fetal lung fibroblast cells (MRC-5). The results showed that AT11-L2 adopts a G4 structure in the presence of K+ and when associated with PhenDC3 through p-p stacking interactions, increases its stability. In addition, the AT11-L2/NCL complex was shown to have a high affinity with a KD in the nanomolar range. In vitro studies showed that functionalized liposomes containing BRACO-19 and doxorubicin were selectively internalized by cancer cells, with greater selectivity being recorded for liposomes with doxorubicin. These results show the therapeutic potential of liposomes functionalized with AT11-L2 against NSCLC.
publishDate 2023
dc.date.none.fl_str_mv 2023-11-22
2023-10-09
2023-11-22T00:00:00Z
2026-10-08T00:00:00Z
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