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Master's Dissertation
DOI
https://doi.org/10.11606/D.17.2021.tde-06122021-122900
Document
Author
Full name
Álex Cardoso de Souza
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2021
Supervisor
Committee
Hartfelder, Klaus Hartmann (President)
Beloti, Márcio Mateus
Saltoratto, Ana Lucia Fachin
Title in Portuguese
Identificação de fatores de transcrição em células tímicas epiteliais medulares (mTECs) por meio de análise sequenciamento de RNA
Keywords in Portuguese
Aire
Células epiteliais tímicas corticais (cTECs)
Células epiteliais tímicas medulares (mTECs)
Fezf2
Sequenciamento de RNA (RNA-Seq)
Abstract in Portuguese
A não responsividade dos autoantígenos que refere à tolerância imunológica central envolve processos de desenvolvimento e seleção de linfócitos T que caracteriza o crosstalk tímico entre os timócitos e células tímicas epiteliais. Em mTECs, o processo de eliminação de timócitos autorreativos contra a expressão de TRAs ocorre por meio da eliminação por apoptose antes destas células chegarem a periferia. Os principais genes envolvidos no processo da tolerância imunológica são o Aire e o Fezf2, correspondendo a aproximadamente 60% de todos os TRAs expressos nas células mTECs, logo, hipotetizamos que 40% desses TRAs expressos possa ser evidenciado e que existem genes ainda não identificados. Foi realizado um ensaio de adesão das células tímicas epiteliais medulares comparado com células tímicas epiteliais corticais aos timócitos extraídos por timectomia de camundongos, onde, demonstrou-se que houve uma redução no número de fatores de transcrição após o processo de adesão causando uma modulação. A análise de bioinformática dos dados de sequenciamento de RNA (análise de qualidade, trimagem, indexação do genoma e análise estatística) mostrou genes diferencialmente expressos que foram identificados e classificados por enriquecimento funcional. Identificamos os potenciais fatores de transcrição antes da adesão com 73 diferencialmente expressos, enquanto que, após adesão 55 genes diferencialmente expressos. Esse estudo demonstra a predição de novos fatores de transcrição no processo de adesão das células tímicas aos timócitos.
Title in English
Identification of transcription factors in medullary epithelial thymic cells (mTECs) through RNA sequencing analysis
Keywords in English
Aire
Cortical thymic epithelial cells (cTECs)
Fezf2
Medullary thymic epithelial cells (mTECs)
RNA sequencing (RNA-Seq)
Abstract in English
The unresponsiveness of autoantigens that refers to central immunological tolerance involves processes of development and selection of T lymphocytes that characterize the thymic crosstalk between thymocytes and epithelial thymic cells. In mTECs, the process of elimination of autoreactive thymocytes against the expression of TRAs occurs through elimination by apoptosis before these cells reach the periphery. The main genes involved in the process of immunological tolerance are Aire and Fezf2, corresponding to approximately 60% of all TRAs expressed in mTECs cells, so we hypothesize that 40% of these TRAs expressed can be evidenced and that there are still unidentified genes. An adhesion assay of medullary epithelial thymic cells compared with cortical epithelial thymic cells to thymocytes extracted by mouse thymectomy was performed, where it was demonstrated that there was a reduction in the number of transcription factors after the adhesion process causing a modulation. Bioinformatics analysis of RNA sequencing data (quality analysis, trimming, genome indexing and statistical analysis) showed differentially expressed genes that were identified and classified by functional enrichment. We identified potential transcription factors before adhesion with 73 differentially expressed, while after adhesion 55 differentially expressed genes. This study demonstrates the prediction of new transcription factors in the process of thymic cell adhesion to thymocytes.
 
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Publishing Date
2022-01-11
 
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