Single cell RNA sequencing after moderate traumatic brain injury: effects of therapeutic hypothermia
Abstract Traumatic brain injury (TBI) initiates a cascade of cellular and molecular events that promote acute and long-term patterns of neuronal, glial, vascular, and synaptic vulnerability leading to lasting neurological deficits. These complex responses lead to patterns of programmed cell death, d...
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| Format: | Article |
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BMC
2025-04-01
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| Series: | Journal of Neuroinflammation |
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| Online Access: | https://doi.org/10.1186/s12974-025-03430-6 |
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| author | Nadine A. Kerr James Choi Simone Y. Mohite Praveen Kumar Singh Helen M. Bramlett Jae K. Lee W. Dalton Dietrich |
| author_facet | Nadine A. Kerr James Choi Simone Y. Mohite Praveen Kumar Singh Helen M. Bramlett Jae K. Lee W. Dalton Dietrich |
| author_sort | Nadine A. Kerr |
| collection | DOAJ |
| description | Abstract Traumatic brain injury (TBI) initiates a cascade of cellular and molecular events that promote acute and long-term patterns of neuronal, glial, vascular, and synaptic vulnerability leading to lasting neurological deficits. These complex responses lead to patterns of programmed cell death, diffuse axonal injury, increased blood-brain barrier disruption, neuroinflammation, and reactive gliosis, each a potential target for therapeutic interventions. Posttraumatic therapeutic hypothermia (TH) has been reported to be highly protective after brain and spinal cord injury and studies have investigated molecular mechanisms underlying mild hypothermic protection while commonly assessing heterogenous cell populations. In this study we conducted single-cell RNA sequencing (scRNA-seq) on cerebral cortical tissues after experimental TBI followed by a period of normothermia or hypothermia to comprehensively assess multiple cell type-specific transcriptional responses. C57BL/6 mice underwent moderate controlled cortical impact (CCI) injury or sham surgery and then placed under sustained normothermia (37⁰C) or hypothermia (33⁰C) for 2 h. After 24 h, cortical tissues including peri-contused regions were processed for scRNA-seq. Unbiased clustering revealed cellular heterogeneity among glial and immune cells at this subacute posttraumatic time point. The analysis also revealed vascular and immune subtypes associated with neovascularization and debris clearance, respectively. Compared to normothermic conditions, TH treatment altered the abundance of specific cell subtypes and induced reactive astrocyte-specific modulation of neurotropic factor gene expression. In addition, an increase in the proportion of endothelial tip cells in the hypothermic TBI group was documented compared to normothermia. These data emphasize the importance of early temperature-sensitive glial and vascular cell processes in producing potentially neuroprotective downstream signaling cascades in a cell-type-dependent manner. The use of scRNA-seq to address cell type-specific mechanisms underlying therapeutic treatments provides a valuable resource for identifying targetable biological pathways for the development of neuroprotective and reparative interventions. |
| format | Article |
| id | doaj-art-49e22b80d01b43c7bc0d7b511e28a5db |
| institution | DOAJ |
| issn | 1742-2094 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj-art-49e22b80d01b43c7bc0d7b511e28a5db2025-08-20T03:18:39ZengBMCJournal of Neuroinflammation1742-20942025-04-0122112010.1186/s12974-025-03430-6Single cell RNA sequencing after moderate traumatic brain injury: effects of therapeutic hypothermiaNadine A. Kerr0James Choi1Simone Y. Mohite2Praveen Kumar Singh3Helen M. Bramlett4Jae K. Lee5W. Dalton Dietrich6The Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami Miller School of MedicineThe Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami Miller School of MedicineThe Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami Miller School of MedicineThe Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami Miller School of MedicineThe Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami Miller School of MedicineThe Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami Miller School of MedicineThe Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami Miller School of MedicineAbstract Traumatic brain injury (TBI) initiates a cascade of cellular and molecular events that promote acute and long-term patterns of neuronal, glial, vascular, and synaptic vulnerability leading to lasting neurological deficits. These complex responses lead to patterns of programmed cell death, diffuse axonal injury, increased blood-brain barrier disruption, neuroinflammation, and reactive gliosis, each a potential target for therapeutic interventions. Posttraumatic therapeutic hypothermia (TH) has been reported to be highly protective after brain and spinal cord injury and studies have investigated molecular mechanisms underlying mild hypothermic protection while commonly assessing heterogenous cell populations. In this study we conducted single-cell RNA sequencing (scRNA-seq) on cerebral cortical tissues after experimental TBI followed by a period of normothermia or hypothermia to comprehensively assess multiple cell type-specific transcriptional responses. C57BL/6 mice underwent moderate controlled cortical impact (CCI) injury or sham surgery and then placed under sustained normothermia (37⁰C) or hypothermia (33⁰C) for 2 h. After 24 h, cortical tissues including peri-contused regions were processed for scRNA-seq. Unbiased clustering revealed cellular heterogeneity among glial and immune cells at this subacute posttraumatic time point. The analysis also revealed vascular and immune subtypes associated with neovascularization and debris clearance, respectively. Compared to normothermic conditions, TH treatment altered the abundance of specific cell subtypes and induced reactive astrocyte-specific modulation of neurotropic factor gene expression. In addition, an increase in the proportion of endothelial tip cells in the hypothermic TBI group was documented compared to normothermia. These data emphasize the importance of early temperature-sensitive glial and vascular cell processes in producing potentially neuroprotective downstream signaling cascades in a cell-type-dependent manner. The use of scRNA-seq to address cell type-specific mechanisms underlying therapeutic treatments provides a valuable resource for identifying targetable biological pathways for the development of neuroprotective and reparative interventions.https://doi.org/10.1186/s12974-025-03430-6Traumatic brain injuryHypothermiaSingle-cell RNA sequencingAstrocytes |
| spellingShingle | Nadine A. Kerr James Choi Simone Y. Mohite Praveen Kumar Singh Helen M. Bramlett Jae K. Lee W. Dalton Dietrich Single cell RNA sequencing after moderate traumatic brain injury: effects of therapeutic hypothermia Journal of Neuroinflammation Traumatic brain injury Hypothermia Single-cell RNA sequencing Astrocytes |
| title | Single cell RNA sequencing after moderate traumatic brain injury: effects of therapeutic hypothermia |
| title_full | Single cell RNA sequencing after moderate traumatic brain injury: effects of therapeutic hypothermia |
| title_fullStr | Single cell RNA sequencing after moderate traumatic brain injury: effects of therapeutic hypothermia |
| title_full_unstemmed | Single cell RNA sequencing after moderate traumatic brain injury: effects of therapeutic hypothermia |
| title_short | Single cell RNA sequencing after moderate traumatic brain injury: effects of therapeutic hypothermia |
| title_sort | single cell rna sequencing after moderate traumatic brain injury effects of therapeutic hypothermia |
| topic | Traumatic brain injury Hypothermia Single-cell RNA sequencing Astrocytes |
| url | https://doi.org/10.1186/s12974-025-03430-6 |
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