G-Quadruplex Aptamer Beacon for Detection of Prostate Cancer Biomarker
Autor(a) principal: | |
---|---|
Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.6/10540 |
Resumo: | The prostate is the major male reproductive gland involved in male fertility and plays an important role in triggering of molecular pathways relevant to fertility success. Unfortunately, in Portugal prostate cancer is the most common cancer type among men, being asymptomatic in earlier stages. Thus, is important early detection of disease. NCL is a multifunctional protein involved in multiple biological processes under both physiological and pathological processes and can have several cellular localizations. Cell surface protein overexpression was found restricted to cancer cells, namely in prostate cancer cells. Thus, we can consider NCL as a potential biomarker for cancer diagnosis and a target for cancer treatment. The AS1411 is an aptamer capable to recognise and binds specifically NCL and have a therapeutic effect on cancer cells through of induction of antiproliferative activity. Beyond its therapeutic use, AS1411 can be used in imaging and diagnostic, particularly on aptasensors development. One of the most relevant characteristics of this aptamer is the ability to fold in a G4 conformation, a secondary structure of nucleic acids. G4 structure confers stabilization to sequence and availability to bind NCL. Thus, in this work is presented the first approach of use AS1411 aptamer to prostate cancer diagnosis, namely through the design of molecular beacon (MB) designated by AS1411N5. Initially, biophysical characterization of AS1411-N5 was done by circular dichroism, nuclear magnetic resonance or fluorometric spectroscopies. Additionally, it was performed microfluidic experiments, to detect NCL using AS1411-N5 in biological samples. The results demonstrated that the proposed AS1411-N5 adopt a G4 structure and it is capable to bind with specificity and selectivity NCL, even in plasma of human patients with prostate cancer. |
id |
RCAP_c334b3a0ca3517500a04e4c59cc3a2d7 |
---|---|
oai_identifier_str |
oai:ubibliorum.ubi.pt:10400.6/10540 |
network_acronym_str |
RCAP |
network_name_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository_id_str |
7160 |
spelling |
G-Quadruplex Aptamer Beacon for Detection of Prostate Cancer BiomarkerCancro da PróstataEnsaios BiofísicosG-QuadruplexMicrofluídicaNucleolinaSondaDomínio/Área Científica::Engenharia e Tecnologia::BiotecnologiaThe prostate is the major male reproductive gland involved in male fertility and plays an important role in triggering of molecular pathways relevant to fertility success. Unfortunately, in Portugal prostate cancer is the most common cancer type among men, being asymptomatic in earlier stages. Thus, is important early detection of disease. NCL is a multifunctional protein involved in multiple biological processes under both physiological and pathological processes and can have several cellular localizations. Cell surface protein overexpression was found restricted to cancer cells, namely in prostate cancer cells. Thus, we can consider NCL as a potential biomarker for cancer diagnosis and a target for cancer treatment. The AS1411 is an aptamer capable to recognise and binds specifically NCL and have a therapeutic effect on cancer cells through of induction of antiproliferative activity. Beyond its therapeutic use, AS1411 can be used in imaging and diagnostic, particularly on aptasensors development. One of the most relevant characteristics of this aptamer is the ability to fold in a G4 conformation, a secondary structure of nucleic acids. G4 structure confers stabilization to sequence and availability to bind NCL. Thus, in this work is presented the first approach of use AS1411 aptamer to prostate cancer diagnosis, namely through the design of molecular beacon (MB) designated by AS1411N5. Initially, biophysical characterization of AS1411-N5 was done by circular dichroism, nuclear magnetic resonance or fluorometric spectroscopies. Additionally, it was performed microfluidic experiments, to detect NCL using AS1411-N5 in biological samples. The results demonstrated that the proposed AS1411-N5 adopt a G4 structure and it is capable to bind with specificity and selectivity NCL, even in plasma of human patients with prostate cancer.A próstata é a maior glândula reprodutiva masculina e tem um papel importante nas vias moleculares relevantes para o sucesso da fertilização. Infelizmente, em Portugal o cancro da próstata é o cancro mais comum entre os homens, sendo assintomático em estadios iniciais. Assim é imperativo a deteção precoce da doença. A nucleolina (NCL) é uma proteína multifuncional envolvida em múltiplos processos biológicos sob condições fisiológicas e patológicas, podendo ter várias localizações celulares. A sobre-expressão da proteína na superfície das células é apenas encontrada em células cancerosas, nomeadamente as do cancro da próstata. Assim a NCL pode ser considerada como um potencial biomarcador para o diagnóstico e tratamento do cancro da próstata. O AS411 é um aptamero capaz de reconhecer e ligar especificamente a esta proteína, e de ter um efeito terapêutico nas células cancerosas ao induzir atividade antiproliferativa. Além do uso terapêutico, a sequência pode ser utilizada na imagiologia e diagnóstico, particularmente através do desenvolvimento de aptasensores. Uma das características mais relevantes do aptamero AS1411 é a capacidade de adotar a configuração de G-quadruplex (G4), uma estrutura secundária dos ácidos nucleicos. As estruturas G4 conferem estabilização à sequência e capacidade de ligar à NCL quando adota esta estrutura. Assim, neste trabalho é apresentada uma primeira abordagem do uso do AS1411 no diagnóstico do cancro da próstata, nomeadamente através da construção de uma sonda a partir da sequência deste aptamero designado por AS1411N5. Inicialmente foi efetuada a caracterização biofísica do AS1411-N5 a nível da estrutura e interação com o alvo, recorrendo às espectroscopias dicroísmo circular e ressonância magnética nuclear, e ensaios fluorométricos. Adicionalmente foram efetuadas experiências de microfluídica, para o uso do AS1411N5 como sonda de deteção da NCL. Estes resultados demonstraram, que o AS1411-N5adota a estrutura G4 e é capaz de ligar especificamente e com seletividade com a NCL, mesmo em amostras biológicas.Cruz, Carla Patrícia Alves Freire MadeiraConde, João Pedro Estrela RodriguesChu, VirgíniauBibliorumMiranda, André Filipe Rodrigues2023-06-29T00:30:22Z2020-07-232020-06-302020-07-23T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.6/10540TID:202544826enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-12-15T09:52:26Zoai:ubibliorum.ubi.pt:10400.6/10540Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:50:28.362723Repositório 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 Beacon for Detection of Prostate Cancer Biomarker |
title |
G-Quadruplex Aptamer Beacon for Detection of Prostate Cancer Biomarker |
spellingShingle |
G-Quadruplex Aptamer Beacon for Detection of Prostate Cancer Biomarker Miranda, André Filipe Rodrigues Cancro da Próstata Ensaios Biofísicos G-Quadruplex Microfluídica Nucleolina Sonda Domínio/Área Científica::Engenharia e Tecnologia::Biotecnologia |
title_short |
G-Quadruplex Aptamer Beacon for Detection of Prostate Cancer Biomarker |
title_full |
G-Quadruplex Aptamer Beacon for Detection of Prostate Cancer Biomarker |
title_fullStr |
G-Quadruplex Aptamer Beacon for Detection of Prostate Cancer Biomarker |
title_full_unstemmed |
G-Quadruplex Aptamer Beacon for Detection of Prostate Cancer Biomarker |
title_sort |
G-Quadruplex Aptamer Beacon for Detection of Prostate Cancer Biomarker |
author |
Miranda, André Filipe Rodrigues |
author_facet |
Miranda, André Filipe Rodrigues |
author_role |
author |
dc.contributor.none.fl_str_mv |
Cruz, Carla Patrícia Alves Freire Madeira Conde, João Pedro Estrela Rodrigues Chu, Virgínia uBibliorum |
dc.contributor.author.fl_str_mv |
Miranda, André Filipe Rodrigues |
dc.subject.por.fl_str_mv |
Cancro da Próstata Ensaios Biofísicos G-Quadruplex Microfluídica Nucleolina Sonda Domínio/Área Científica::Engenharia e Tecnologia::Biotecnologia |
topic |
Cancro da Próstata Ensaios Biofísicos G-Quadruplex Microfluídica Nucleolina Sonda Domínio/Área Científica::Engenharia e Tecnologia::Biotecnologia |
description |
The prostate is the major male reproductive gland involved in male fertility and plays an important role in triggering of molecular pathways relevant to fertility success. Unfortunately, in Portugal prostate cancer is the most common cancer type among men, being asymptomatic in earlier stages. Thus, is important early detection of disease. NCL is a multifunctional protein involved in multiple biological processes under both physiological and pathological processes and can have several cellular localizations. Cell surface protein overexpression was found restricted to cancer cells, namely in prostate cancer cells. Thus, we can consider NCL as a potential biomarker for cancer diagnosis and a target for cancer treatment. The AS1411 is an aptamer capable to recognise and binds specifically NCL and have a therapeutic effect on cancer cells through of induction of antiproliferative activity. Beyond its therapeutic use, AS1411 can be used in imaging and diagnostic, particularly on aptasensors development. One of the most relevant characteristics of this aptamer is the ability to fold in a G4 conformation, a secondary structure of nucleic acids. G4 structure confers stabilization to sequence and availability to bind NCL. Thus, in this work is presented the first approach of use AS1411 aptamer to prostate cancer diagnosis, namely through the design of molecular beacon (MB) designated by AS1411N5. Initially, biophysical characterization of AS1411-N5 was done by circular dichroism, nuclear magnetic resonance or fluorometric spectroscopies. Additionally, it was performed microfluidic experiments, to detect NCL using AS1411-N5 in biological samples. The results demonstrated that the proposed AS1411-N5 adopt a G4 structure and it is capable to bind with specificity and selectivity NCL, even in plasma of human patients with prostate cancer. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-07-23 2020-06-30 2020-07-23T00:00:00Z 2023-06-29T00:30:22Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.6/10540 TID:202544826 |
url |
http://hdl.handle.net/10400.6/10540 |
identifier_str_mv |
TID:202544826 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
_version_ |
1799136395310137344 |