Análise da expressão e do número de cópias de LINE-1 em PBMC de indivíduos dentro do transtorno do espectro autista e nos espermatozoides de seus pais
Data
2024-10-30
Autores
Sousa, Kathleen da Silva 
Orientadores
Stumpp, Taiza
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Objetivo: Estudos recentes demonstram que alterações epigenéticas em elementos transponíveis (TE) estão correlacionadas ao risco de autismo. Entre os TE, o retrotransposon LINE-1 tem demonstrado estar com a expressão, a metilação e o número de cópias alterados em diversos tecidos de coortes autistas. Assim, a presente pesquisa objetivou analisar a expressão e o número de cópias do LINE-1 no genoma de indivíduos dentro do autismo com grau I e II, como também no genoma dos seus respectivos pais. Métodos: Foram coletadas células mononucleares de sangue periférico (PBMC) dos participantes autistas, bem como de PBMC e espermatozoides de pais de participantes autistas. Participantes controles também tiveram as mesmas amostras coletadas. Após a extração de DNA e RNA e síntese de cDNA, as amostras foram submetidas à qPCR utilizando primers previamente desenhados para a análise do número de cópias e da expressão de LINE-1. Resultados: Nas PBMCS, a expressão de LINE-1 não apresentou diferenças significativas entre os grupos, apesar de um aumento significativo no número de cópias desse retrotransposon em indivíduos autistas. Aumento da expressão de LINE-1 foi observado nos espermatozoides dos pais de autistas, sugerindo uma possível alteração nos mecanismos de controle da expressão desse retrotransposon. Limitações metodológicas e tamanho amostral foram reconhecidos, indicando a necessidade de estudos futuros para confirmar e expandir esses achados. Conclusões: Esse estudo fornece indicações preliminares de que existe variação do número de cópias do LINE-1 em pessoas autistas e a alterações da expressão desse retrotransposon nos espermatozoides dos pais de indivíduos autistas. Os resultados obridos levantam questões importantes sobre o envolvimento da desregulação de LINE-1 nos autismos. Embora esse estudo tenha se concentrado na expressão e na variação do número de cópias do LINE-1, pesquisas futuras de nosso grupo estão se dedicando a investigar fatores adicionais, como a regulação epigenética do LINE-1.
Objective: Recent studies have demonstrated that epigenetic alterations in transposable elements (TEs) are correlated with the risk of autism. Among these TEs, the LINE1 retrotransposon has been shown to exhibit altered expression, methylation, and copy number in various tissues of autistic cohorts. The present research aimed to analyze the expression and copy number of LINE1 in the genome of individuals with level I and II autism, as well as in the genomes of their respective parents. Methods: Peripheral blood mononuclear cells (PBMCs) were collected from autistic participants, along with PBMCs and sperm samples from the fathers of autistic participants. Control participants provided the same types of samples. After DNA and RNA extraction and cDNA synthesis, the samples were subjected to qPCR using predesigned primers to assess LINE1 copy number and expression. Results: In PBMCs, LINE1 expression did not show significant differences between groups, although a significant increase in LINE1 copy number was observed in autistic individuals. An increase in LINE1 expression was detected in the sperm of the fathers of autistic individuals, suggesting a potential disruption in the mechanisms controlling retrotransposon expression. Methodological limitations and sample size were acknowledged, underscoring the need for future studies to confirm and expand these findings. Conclusions: This study provides preliminary evidence of an increased LINE1 copy number in autistic individuals and altered LINE1 expression in the sperm of the fathers of autistic individuals. The findings raise important questions regarding the role of LINE1 dysregulation in autism. While this study focused on LINE1 expression and copy number variation, future research from our group will investigate additional factors, such as the epigenetic regulation of LINE1.
Objective: Recent studies have demonstrated that epigenetic alterations in transposable elements (TEs) are correlated with the risk of autism. Among these TEs, the LINE1 retrotransposon has been shown to exhibit altered expression, methylation, and copy number in various tissues of autistic cohorts. The present research aimed to analyze the expression and copy number of LINE1 in the genome of individuals with level I and II autism, as well as in the genomes of their respective parents. Methods: Peripheral blood mononuclear cells (PBMCs) were collected from autistic participants, along with PBMCs and sperm samples from the fathers of autistic participants. Control participants provided the same types of samples. After DNA and RNA extraction and cDNA synthesis, the samples were subjected to qPCR using predesigned primers to assess LINE1 copy number and expression. Results: In PBMCs, LINE1 expression did not show significant differences between groups, although a significant increase in LINE1 copy number was observed in autistic individuals. An increase in LINE1 expression was detected in the sperm of the fathers of autistic individuals, suggesting a potential disruption in the mechanisms controlling retrotransposon expression. Methodological limitations and sample size were acknowledged, underscoring the need for future studies to confirm and expand these findings. Conclusions: This study provides preliminary evidence of an increased LINE1 copy number in autistic individuals and altered LINE1 expression in the sperm of the fathers of autistic individuals. The findings raise important questions regarding the role of LINE1 dysregulation in autism. While this study focused on LINE1 expression and copy number variation, future research from our group will investigate additional factors, such as the epigenetic regulation of LINE1.
Descrição
Citação
SOUSA, Kathleen da Silva. Análise da expressão e do número de cópias de LINE-1 em PBMC de indivíduos dentro do transtorno do espectro autista e nos espermatozoides de seus pais. 2024. 121 f. Dissertação (Mestrado em Biologia Estrutural e Funcional) - Escola Paulista de Medicina, Universidade Federal de São Paulo (Unifesp), São Paulo, 2024.