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Master's Dissertation
DOI
Document
Author
Full name
Lidia Hyun Joo Myung
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2019
Supervisor
Committee
Abrão, Mauricio Simões (President)
Coudry, Renata de Almeida
Fernandes, Luiz Flávio Cordeiro
Podgaec, Sérgio
Title in Portuguese
Análise de marcadores protéicos no endométrio de mulheres portadoras de endometriose profunda intestinal
Keywords in Portuguese
Biomarcadores
Endométrio
Endometriose
Espectrometria de massas
Neoplasias
Proteômica
Abstract in Portuguese
INTRODUÇÃO: A endometriose é doença ginecológica prevalente, com intervalo de tempo longo entre o início dos sintomas e o diagnóstico definitivo. A endometriose profunda intestinal se apresenta com sintomas mais severos, e maior morbidade cirúrgica. Um método minimamente invasivo para o diagnóstico precoce se faz necessário, e impediria a progressão para as formas mais profundas da doença. Estudos de biomarcadores com técnicas em proteômica podem trazer melhor entendimento das bases moleculares da doença, possibilidade de diagnóstico precoce, e do desenvolvimento de terapias-alvo. OBJETIVO: Comparar o perfil de expressão de proteínas do endométrio tópico entre pacientes saudáveis e com endometriose profunda intestinal, por técnica de espectrometria de massas shotgun. MÉTODOS: Estudo caso-controle exploratório, com total de 24 pacientes divididas em dois grupos: pacientes com endometriose profunda intestinal com comprovação cirúrgica, e pacientes do grupo controle saúdavel submetidas a cirurgia de laqueadura tubárea. Amostras de endométrio tópico foram obtidas por meio de auxílio de catéter de aspiração. As amostras foram submetidas a extração, digestão, dessalinização peptídica, e analisadas seguindo uma abordagem de espectrometria de massas shotgun, label-free de DDA (aquisição dependente de dados), utilizando o instrumento Orbitrap Elite (Thermo Fisher, EUA). RESULTADOS: a análise proteômica dos resultados foi realizada através do software Progenesis QI (Nonlinear Dynamics, Newcastle upon Tyne, UK), e resultou em um painel de 595 proteínas diferencialmente expressas entre os grupos controle e endometriose. Os resultados revelaram diferentes moléculas de acordo com a fase do ciclo menstrual, e o estádio da doença. As principais proteínas com expressão aumentada no grupo de pacientes com endometriose incluíram SETSIP, SRRM2, CAPS, H2AFV e SAFB, enquanto moléculas reguladas negativamente incluíram BPIFB1, FAM184A, SLPI, PIGR, JCHAÍNA, HLA-A e LTF. As proteínas diferenciais entre os grupos apresentaram o câncer como principal doença associada (p - valor de 4,14E-02 - 3,91E-08), de acordo com as análises realizadas pelo software IPA (Ingenuity Pathway Analysis System; Ingenuity Systems, Redwood City, CA, USA). As principais proteínas com expressão aumentada nos estádios mais avançados da doença foram CHMP4B, DES, EIF3I, CDH13, LIMS1, HSPA4, HSP90AB1, TUBB, NPM1, MYL6, e foram descritas relacionadas a metástases e pior prognóstico de diversos tipos de cânceres. CONCLUSÃO: CHMP4B, DES, EIF3I, CDH13, LIMS1, HSPA4, HSP90AB1, TUBB, NPM1, MYL6 são proteínas com expressão aumentada no endométrio das pacientes portadoras de endometriose profunda intestinal em estádios avançados. O presente estudo revelou proteínas diferencialmente expressas na endometriose em associação com diversos tipos de cânceres. Os achados constituem possíveis marcadores para o desenvolvimento de técnicas para o diagnóstico minimamente invasivo, e potenciais alvos terapêuticos para a doença
Title in English
Analysis of protein markers in the endometrium of women with deep intestinal endometriosis
Keywords in English
Biomarkers
Endometriosis
Endometrium
Mass Spectrometry
Neoplasms
Proteomics
Abstract in English
INTRODUCTION: Endometriosis is a prevalent gynecological disease, with a long time elapsed from onset of symptoms to definitive diagnosis. Deep intestinal endometriosis presents with more severe symptoms, and greater surgical morbidity. A minimally invasive method for early diagnosis becomes necessary and would prevent progression to the deeper presentations of the disease. Studies of biomarkers with proteomics techniques can provide a better understanding of the molecular basis of the disease, the possibility of early diagnosis, and the development of targeted therapies. OBJECTIVE: To compare the expression of eutopic endometrial proteins between healthy and deep intestinal endometriosis patients, using a shotgun mass spectrometry technique. METHODS: An exploratory case-control study with a total of 24 patients divided in two groups: patients with intestinal deep endometriosis with surgical confirmation, and patients in the healthy control group submitted to tubal ligation surgery. Eutopic endometrial samples were obtained by aspiration catheter. The samples were submitted to extraction, digestion, peptide desalting and analyzed using a DDA (data-dependent acquisition) label-free mass spectrometry approach by the Orbitrap Elite instrument (Thermo Fisher, USA). RESULTS: Proteomic analysis was performed using the Progenesis QI software (Nonlinear Dynamics, Newcastle upon Tyne, UK) to profile 595 differentially expressed proteins between the control and endometriosis groups. The results revealed different molecules according to the phase of the menstrual cycle and stage of disease. Top upregulated molecules included SETSIP, SRRM2, CAPS, H2AFV, and SAFB, whereas downregulated molecules included BPIFB1, FAM184A, SLPI, PIGR, JCHAIN, HLA-A, and LTF. Among top diseases associated molecules between groups were related to cancer (p - value of 4.14E-02 - 3.91E-08), according to the analysis performed by IPA software (Ingenuity Pathway Analysis System, Ingenuity Systems, Redwood City, CA, USA). Top upregulated proteins among patientes with advanced stage of endomeriosis were CHMP4B, DES, EIF3I, CDH13, LIMS1, HSPA4, HSP90AB1, TUBB, NPM1, MYL6. Those proteins were related to metastasis and poor survival of several cancer types. CONCLUSION: CHMP4B, DES, EIF3I, CDH13, LIMS1, HSPA4, HSP90AB1, TUBB, NPM1, MYL6 are among the top upregulated proteins in patients with advanced stage of deep intestinal endomeriosis. The present study revealed proteins differentially expressed in endometriosis as potential biomarkers for the development of techniques for minimally invasive diagnosis, and potential therapeutic targets for the disease
 
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Publishing Date
2019-09-04
 
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