Comprehensive transcriptomic analysis integrating bulk and single-cell RNA-seq with machine learning to identify and validate mitochondrial unfolded protein response biomarkers in patients with ischemic stroke
BackgroundIschemic stroke (IS) represents a significant contributor to morbidity and mortality globally. The relationship between IS and mitochondrial unfolded protein response (UPRmt) was presently uncertain. This study endeavors to explore the fundamental mechanism of UPRmt in IS by utilizing bioi...
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Frontiers Media S.A.
2025-04-01
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2025.1582252/full |
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| author | Lu Zhang Lei Yue Peng Jia Peng Jia Ziqi Cheng Jiwen Liu |
| author_facet | Lu Zhang Lei Yue Peng Jia Peng Jia Ziqi Cheng Jiwen Liu |
| author_sort | Lu Zhang |
| collection | DOAJ |
| description | BackgroundIschemic stroke (IS) represents a significant contributor to morbidity and mortality globally. The relationship between IS and mitochondrial unfolded protein response (UPRmt) was presently uncertain. This study endeavors to explore the fundamental mechanism of UPRmt in IS by utilizing bioinformatics methods.MethodsIn GSE58294, differentially expressed genes (DEGs) were obtained, which were overlapped with key module genes of UPRmt-related gene (UPRmt-RGs) for producing candidate genes. The biomarkers were identified from the candidate genes through machine learning, expression validation, and receiver operating characteristic (ROC) curves. In order to verify the biomarkers, reverse transcription-quantitative PCR (RT-qPCR) experiments were performed on human peripheral blood. Subsequently, a predictive nomogram was created to estimate the likelihood of developing IS. Next, the mechanisms and functions related to the biomarkers were explored by enrichment analysis and immune infiltration. In addition, cells enriched with biomarkers were identified, and the biological processes involved in these cells were analyzed through intercellular communication analysis and virtual knockout experiments.ResultsMCEMP1, CACNA1E, and CLEC4D were identified as biomarkers and subsequently validated by RT-qPCR. RT-qPCR revealed that CLEC4D is the most sensitive biomarker. The nomogram analysis revealed that these biomarkers possess strong diagnostic value. Immune infiltration analysis indicated that all three biomarkers are strongly correlated with neutrophils. Additionally, in the single-cell transcriptome data, these biomarkers were predominantly enriched in neutrophils. Compared to the sham group, the middle cerebral artery occlusion (MCAO) group exhibited enhanced immune-inflammatory responses. Virtual knockout experiments provide preliminary evidence that CLEC4D functions as a regulatory molecule in neutrophil-mediated inflammation, rather than serving merely as a passive marker.ConclusionCLEC4D was identified as the most sensitive biomarker for IS related to UPRmt-RGs, offering a new reference for IS diagnosis and treatment. |
| format | Article |
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| issn | 2296-634X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj-art-c1d799e7b97c480595a68690aa26c8cf2025-08-20T02:13:19ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2025-04-011310.3389/fcell.2025.15822521582252Comprehensive transcriptomic analysis integrating bulk and single-cell RNA-seq with machine learning to identify and validate mitochondrial unfolded protein response biomarkers in patients with ischemic strokeLu Zhang0Lei Yue1Peng Jia2Peng Jia3Ziqi Cheng4Jiwen Liu5Institute of Science and Technology for Brain-Inspired Intelligence, Fudan University, Shanghai, ChinaDepartment of Neurology, Shangrao Municipal Hospital, Shangrao, Jiangxi, ChinaInstitute of Longevity and Aging Research, Zhongshan Hospital, Fudan University, Shanghai, ChinaShanghai Key Laboratory of Medical Epigenetics, Institutes of Biomedical Sciences, Fudan University, Shanghai, ChinaDepartment of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan, Hubei, ChinaDepartment of Emergency Medicine, Shanghai Pudong New Area Gongli Hospital, Shanghai, ChinaBackgroundIschemic stroke (IS) represents a significant contributor to morbidity and mortality globally. The relationship between IS and mitochondrial unfolded protein response (UPRmt) was presently uncertain. This study endeavors to explore the fundamental mechanism of UPRmt in IS by utilizing bioinformatics methods.MethodsIn GSE58294, differentially expressed genes (DEGs) were obtained, which were overlapped with key module genes of UPRmt-related gene (UPRmt-RGs) for producing candidate genes. The biomarkers were identified from the candidate genes through machine learning, expression validation, and receiver operating characteristic (ROC) curves. In order to verify the biomarkers, reverse transcription-quantitative PCR (RT-qPCR) experiments were performed on human peripheral blood. Subsequently, a predictive nomogram was created to estimate the likelihood of developing IS. Next, the mechanisms and functions related to the biomarkers were explored by enrichment analysis and immune infiltration. In addition, cells enriched with biomarkers were identified, and the biological processes involved in these cells were analyzed through intercellular communication analysis and virtual knockout experiments.ResultsMCEMP1, CACNA1E, and CLEC4D were identified as biomarkers and subsequently validated by RT-qPCR. RT-qPCR revealed that CLEC4D is the most sensitive biomarker. The nomogram analysis revealed that these biomarkers possess strong diagnostic value. Immune infiltration analysis indicated that all three biomarkers are strongly correlated with neutrophils. Additionally, in the single-cell transcriptome data, these biomarkers were predominantly enriched in neutrophils. Compared to the sham group, the middle cerebral artery occlusion (MCAO) group exhibited enhanced immune-inflammatory responses. Virtual knockout experiments provide preliminary evidence that CLEC4D functions as a regulatory molecule in neutrophil-mediated inflammation, rather than serving merely as a passive marker.ConclusionCLEC4D was identified as the most sensitive biomarker for IS related to UPRmt-RGs, offering a new reference for IS diagnosis and treatment.https://www.frontiersin.org/articles/10.3389/fcell.2025.1582252/fullbioinformationbiomarkerbulk RNA-seqischemic strokesingle cellmitochondrial unfolded protein response |
| spellingShingle | Lu Zhang Lei Yue Peng Jia Peng Jia Ziqi Cheng Jiwen Liu Comprehensive transcriptomic analysis integrating bulk and single-cell RNA-seq with machine learning to identify and validate mitochondrial unfolded protein response biomarkers in patients with ischemic stroke Frontiers in Cell and Developmental Biology bioinformation biomarker bulk RNA-seq ischemic stroke single cell mitochondrial unfolded protein response |
| title | Comprehensive transcriptomic analysis integrating bulk and single-cell RNA-seq with machine learning to identify and validate mitochondrial unfolded protein response biomarkers in patients with ischemic stroke |
| title_full | Comprehensive transcriptomic analysis integrating bulk and single-cell RNA-seq with machine learning to identify and validate mitochondrial unfolded protein response biomarkers in patients with ischemic stroke |
| title_fullStr | Comprehensive transcriptomic analysis integrating bulk and single-cell RNA-seq with machine learning to identify and validate mitochondrial unfolded protein response biomarkers in patients with ischemic stroke |
| title_full_unstemmed | Comprehensive transcriptomic analysis integrating bulk and single-cell RNA-seq with machine learning to identify and validate mitochondrial unfolded protein response biomarkers in patients with ischemic stroke |
| title_short | Comprehensive transcriptomic analysis integrating bulk and single-cell RNA-seq with machine learning to identify and validate mitochondrial unfolded protein response biomarkers in patients with ischemic stroke |
| title_sort | comprehensive transcriptomic analysis integrating bulk and single cell rna seq with machine learning to identify and validate mitochondrial unfolded protein response biomarkers in patients with ischemic stroke |
| topic | bioinformation biomarker bulk RNA-seq ischemic stroke single cell mitochondrial unfolded protein response |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2025.1582252/full |
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