Cargando…

Key Gene and Functional Pathways Identified in Unexplained Recurrent Spontaneous Abortion Using Targeted RNA Sequencing and Clinical Analysis

Identifying the mechanisms underlying unexplained recurrent spontaneous abortion (URSA) can help develop effective treatments. This study provides novel insights into the biological characteristics and related pathways of differentially expressed genes (DEGs) in URSA. Nineteen patients with URSA and...

Descripción completa

Detalles Bibliográficos
Autores principales: Gu, Heng, Li, Longyu, Du, Mengxuan, Xu, Hang, Gao, Mengge, Liu, Xiaohua, Wei, Xiangcai, Zhong, Xingming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8375436/
https://www.ncbi.nlm.nih.gov/pubmed/34421922
http://dx.doi.org/10.3389/fimmu.2021.717832
_version_ 1783740313887571968
author Gu, Heng
Li, Longyu
Du, Mengxuan
Xu, Hang
Gao, Mengge
Liu, Xiaohua
Wei, Xiangcai
Zhong, Xingming
author_facet Gu, Heng
Li, Longyu
Du, Mengxuan
Xu, Hang
Gao, Mengge
Liu, Xiaohua
Wei, Xiangcai
Zhong, Xingming
author_sort Gu, Heng
collection PubMed
description Identifying the mechanisms underlying unexplained recurrent spontaneous abortion (URSA) can help develop effective treatments. This study provides novel insights into the biological characteristics and related pathways of differentially expressed genes (DEGs) in URSA. Nineteen patients with URSA and three healthy fertile women with regular menstruation (control group) were recruited. RNA was extracted from the two groups to determine the differential expression of immunoregulatory gene sequences. Gene ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analyses were used to identify the biological functions and pathways of the identified DEGs. A protein-protein interaction (PPI) network was constructed using the STRING database. Furthermore, qRT-PCR and ELISA were performed to validate the differential expression of the hub genes. We also explored the regulatory mechanism of Th1/Th2 imbalance. A total of 99 DEGs were identified, comprising 94 upregulated and five downregulated genes. Through GO analysis, nine immune cell function-related clusters were selected, and genes with significant differential expression were primarily enriched in eight immune regulatory functions related to the KEGG signalling pathway. Subsequently, five hub genes (TLR2, CXCL8, IFNG, IL2RA, and ITGAX) were identified using Cytoscape software; qRT-PCR confirmed the differential expression among the hub genes, whereas ELISA revealed a significant difference in extracellular IFN-γ and IL-8 levels. The levels of Th1 (IFN-γ) and the Th1/Th2 ratio were higher in the peripheral blood of URSA patients than in control group patients. These findings suggest that the occurrence of URSA may be associated with the abnormal expression of some specific immunoregulatory genes involved in T-cell activation and differentiation. Among the identified DEGs, IFNG may play a key role in regulating maternal immune response. Although further validation is required, our data provide an important theoretical basis for elucidating the pathogenesis of recurrent spontaneous abortion.
format Online
Article
Text
id pubmed-8375436
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-83754362021-08-20 Key Gene and Functional Pathways Identified in Unexplained Recurrent Spontaneous Abortion Using Targeted RNA Sequencing and Clinical Analysis Gu, Heng Li, Longyu Du, Mengxuan Xu, Hang Gao, Mengge Liu, Xiaohua Wei, Xiangcai Zhong, Xingming Front Immunol Immunology Identifying the mechanisms underlying unexplained recurrent spontaneous abortion (URSA) can help develop effective treatments. This study provides novel insights into the biological characteristics and related pathways of differentially expressed genes (DEGs) in URSA. Nineteen patients with URSA and three healthy fertile women with regular menstruation (control group) were recruited. RNA was extracted from the two groups to determine the differential expression of immunoregulatory gene sequences. Gene ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analyses were used to identify the biological functions and pathways of the identified DEGs. A protein-protein interaction (PPI) network was constructed using the STRING database. Furthermore, qRT-PCR and ELISA were performed to validate the differential expression of the hub genes. We also explored the regulatory mechanism of Th1/Th2 imbalance. A total of 99 DEGs were identified, comprising 94 upregulated and five downregulated genes. Through GO analysis, nine immune cell function-related clusters were selected, and genes with significant differential expression were primarily enriched in eight immune regulatory functions related to the KEGG signalling pathway. Subsequently, five hub genes (TLR2, CXCL8, IFNG, IL2RA, and ITGAX) were identified using Cytoscape software; qRT-PCR confirmed the differential expression among the hub genes, whereas ELISA revealed a significant difference in extracellular IFN-γ and IL-8 levels. The levels of Th1 (IFN-γ) and the Th1/Th2 ratio were higher in the peripheral blood of URSA patients than in control group patients. These findings suggest that the occurrence of URSA may be associated with the abnormal expression of some specific immunoregulatory genes involved in T-cell activation and differentiation. Among the identified DEGs, IFNG may play a key role in regulating maternal immune response. Although further validation is required, our data provide an important theoretical basis for elucidating the pathogenesis of recurrent spontaneous abortion. Frontiers Media S.A. 2021-08-05 /pmc/articles/PMC8375436/ /pubmed/34421922 http://dx.doi.org/10.3389/fimmu.2021.717832 Text en Copyright © 2021 Gu, Li, Du, Xu, Gao, Liu, Wei and Zhong https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Gu, Heng
Li, Longyu
Du, Mengxuan
Xu, Hang
Gao, Mengge
Liu, Xiaohua
Wei, Xiangcai
Zhong, Xingming
Key Gene and Functional Pathways Identified in Unexplained Recurrent Spontaneous Abortion Using Targeted RNA Sequencing and Clinical Analysis
title Key Gene and Functional Pathways Identified in Unexplained Recurrent Spontaneous Abortion Using Targeted RNA Sequencing and Clinical Analysis
title_full Key Gene and Functional Pathways Identified in Unexplained Recurrent Spontaneous Abortion Using Targeted RNA Sequencing and Clinical Analysis
title_fullStr Key Gene and Functional Pathways Identified in Unexplained Recurrent Spontaneous Abortion Using Targeted RNA Sequencing and Clinical Analysis
title_full_unstemmed Key Gene and Functional Pathways Identified in Unexplained Recurrent Spontaneous Abortion Using Targeted RNA Sequencing and Clinical Analysis
title_short Key Gene and Functional Pathways Identified in Unexplained Recurrent Spontaneous Abortion Using Targeted RNA Sequencing and Clinical Analysis
title_sort key gene and functional pathways identified in unexplained recurrent spontaneous abortion using targeted rna sequencing and clinical analysis
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8375436/
https://www.ncbi.nlm.nih.gov/pubmed/34421922
http://dx.doi.org/10.3389/fimmu.2021.717832
work_keys_str_mv AT guheng keygeneandfunctionalpathwaysidentifiedinunexplainedrecurrentspontaneousabortionusingtargetedrnasequencingandclinicalanalysis
AT lilongyu keygeneandfunctionalpathwaysidentifiedinunexplainedrecurrentspontaneousabortionusingtargetedrnasequencingandclinicalanalysis
AT dumengxuan keygeneandfunctionalpathwaysidentifiedinunexplainedrecurrentspontaneousabortionusingtargetedrnasequencingandclinicalanalysis
AT xuhang keygeneandfunctionalpathwaysidentifiedinunexplainedrecurrentspontaneousabortionusingtargetedrnasequencingandclinicalanalysis
AT gaomengge keygeneandfunctionalpathwaysidentifiedinunexplainedrecurrentspontaneousabortionusingtargetedrnasequencingandclinicalanalysis
AT liuxiaohua keygeneandfunctionalpathwaysidentifiedinunexplainedrecurrentspontaneousabortionusingtargetedrnasequencingandclinicalanalysis
AT weixiangcai keygeneandfunctionalpathwaysidentifiedinunexplainedrecurrentspontaneousabortionusingtargetedrnasequencingandclinicalanalysis
AT zhongxingming keygeneandfunctionalpathwaysidentifiedinunexplainedrecurrentspontaneousabortionusingtargetedrnasequencingandclinicalanalysis