Single-cell RNA sequencing reveals peripheral blood leukocyte responses to spinal cord injury in mice with humanised immune systems
Abstract Next-generation humanised mouse models and single-cell RNA sequencing (scRNAseq) approaches enable in-depth studies into human immune cell biology. Here we used NSG-SGM3 mice engrafted with human umbilical cord haematopoietic stem cells to investigate how human immune cells respond to and/o...
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | Journal of Neuroinflammation |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12974-024-03048-0 |
| _version_ | 1827326635958861824 |
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| author | Ellen R. Gillespie Laura F. Grice Isabel G. Courtney Hong Wa Lao Woncheol Jung Sonny Ramkomuth Jacky Xie David A. Brown James Walsham Kristen J. Radford Quan H. Nguyen Marc J. Ruitenberg |
| author_facet | Ellen R. Gillespie Laura F. Grice Isabel G. Courtney Hong Wa Lao Woncheol Jung Sonny Ramkomuth Jacky Xie David A. Brown James Walsham Kristen J. Radford Quan H. Nguyen Marc J. Ruitenberg |
| author_sort | Ellen R. Gillespie |
| collection | DOAJ |
| description | Abstract Next-generation humanised mouse models and single-cell RNA sequencing (scRNAseq) approaches enable in-depth studies into human immune cell biology. Here we used NSG-SGM3 mice engrafted with human umbilical cord haematopoietic stem cells to investigate how human immune cells respond to and/or are changed by traumatic spinal cord injury (SCI). We hypothesised that the use of such mice could help advance our understanding of spinal cord injury-induced immune depression syndrome (SCI-IDS), and also how human leukocytes change as they migrate from the circulation into the lesion site. Our scRNAseq experiments, supplemented by flow cytometry, demonstrate the existence of up to 11 human immune cell (sub-) types and/or states across the blood and injured spinal cord (7 days post-SCI) of humanised NSG-SGM3 mice. Further comparisons of human immune cell transcriptomes between naïve, sham-operated and SCI mice identified a total of 579 differentially expressed genes, 190 of which were ‘SCI-specific’ (that is, genes regulated only in response to SCI but not sham surgery). Gene ontology analysis showed a prominent downregulation of immune cell function under SCI conditions, including for T cell receptor signalling and antigen presentation, confirming the presence of SCI-IDS and the transcriptional signature of human leukocytes in association with this phenomenon. We also highlight the activating influence of the local spinal cord lesion microenvironment by comparing the transcriptomes of circulating versus infiltrated human immune cells; those isolated from the lesion site were enriched for genes relating to both immune cell activity and function (e.g., oxidative phosphorylation, T cell proliferation and antigen presentation). We lastly applied an integrated bioinformatics approach to determine where immune responses in humanised NSG-SGM3 mice appear congruent to the native responses of human SCI patients, and where they diverge. Collectively, our study provides a valuable resource and methodological framework for the use of these mice in translational research. |
| first_indexed | 2024-03-07T14:48:29Z |
| format | Article |
| id | doaj.art-40a7cde76523424a9dc3bc7e32ce0e7a |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-03-07T14:48:29Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-40a7cde76523424a9dc3bc7e32ce0e7a2024-03-05T19:49:05ZengBMCJournal of Neuroinflammation1742-20942024-03-0121112010.1186/s12974-024-03048-0Single-cell RNA sequencing reveals peripheral blood leukocyte responses to spinal cord injury in mice with humanised immune systemsEllen R. Gillespie0Laura F. Grice1Isabel G. Courtney2Hong Wa Lao3Woncheol Jung4Sonny Ramkomuth5Jacky Xie6David A. Brown7James Walsham8Kristen J. Radford9Quan H. Nguyen10Marc J. Ruitenberg11School of Biomedical Sciences, Faculty of Medicine, The University of QueenslandSchool of Biomedical Sciences, Faculty of Medicine, The University of QueenslandSchool of Biomedical Sciences, Faculty of Medicine, The University of QueenslandSchool of Biomedical Sciences, Faculty of Medicine, The University of QueenslandSchool of Biomedical Sciences, Faculty of Medicine, The University of QueenslandInstitute for Molecular Bioscience, The University of QueenslandSchool of Biomedical Sciences, Faculty of Medicine, The University of QueenslandNeuroinflammation Research Group, Centre for Immunology and Allergy Research, Westmead Institute for Medical ResearchIntensive Care Unit, Princess Alexandra HospitalMater Research Institute, The University of Queensland, Translational Research InstituteInstitute for Molecular Bioscience, The University of QueenslandSchool of Biomedical Sciences, Faculty of Medicine, The University of QueenslandAbstract Next-generation humanised mouse models and single-cell RNA sequencing (scRNAseq) approaches enable in-depth studies into human immune cell biology. Here we used NSG-SGM3 mice engrafted with human umbilical cord haematopoietic stem cells to investigate how human immune cells respond to and/or are changed by traumatic spinal cord injury (SCI). We hypothesised that the use of such mice could help advance our understanding of spinal cord injury-induced immune depression syndrome (SCI-IDS), and also how human leukocytes change as they migrate from the circulation into the lesion site. Our scRNAseq experiments, supplemented by flow cytometry, demonstrate the existence of up to 11 human immune cell (sub-) types and/or states across the blood and injured spinal cord (7 days post-SCI) of humanised NSG-SGM3 mice. Further comparisons of human immune cell transcriptomes between naïve, sham-operated and SCI mice identified a total of 579 differentially expressed genes, 190 of which were ‘SCI-specific’ (that is, genes regulated only in response to SCI but not sham surgery). Gene ontology analysis showed a prominent downregulation of immune cell function under SCI conditions, including for T cell receptor signalling and antigen presentation, confirming the presence of SCI-IDS and the transcriptional signature of human leukocytes in association with this phenomenon. We also highlight the activating influence of the local spinal cord lesion microenvironment by comparing the transcriptomes of circulating versus infiltrated human immune cells; those isolated from the lesion site were enriched for genes relating to both immune cell activity and function (e.g., oxidative phosphorylation, T cell proliferation and antigen presentation). We lastly applied an integrated bioinformatics approach to determine where immune responses in humanised NSG-SGM3 mice appear congruent to the native responses of human SCI patients, and where they diverge. Collectively, our study provides a valuable resource and methodological framework for the use of these mice in translational research.https://doi.org/10.1186/s12974-024-03048-0NeurotraumaNeuroinflammationNeuroimmunologyImmunosuppressionNeutrophilLymphocyte |
| spellingShingle | Ellen R. Gillespie Laura F. Grice Isabel G. Courtney Hong Wa Lao Woncheol Jung Sonny Ramkomuth Jacky Xie David A. Brown James Walsham Kristen J. Radford Quan H. Nguyen Marc J. Ruitenberg Single-cell RNA sequencing reveals peripheral blood leukocyte responses to spinal cord injury in mice with humanised immune systems Journal of Neuroinflammation Neurotrauma Neuroinflammation Neuroimmunology Immunosuppression Neutrophil Lymphocyte |
| title | Single-cell RNA sequencing reveals peripheral blood leukocyte responses to spinal cord injury in mice with humanised immune systems |
| title_full | Single-cell RNA sequencing reveals peripheral blood leukocyte responses to spinal cord injury in mice with humanised immune systems |
| title_fullStr | Single-cell RNA sequencing reveals peripheral blood leukocyte responses to spinal cord injury in mice with humanised immune systems |
| title_full_unstemmed | Single-cell RNA sequencing reveals peripheral blood leukocyte responses to spinal cord injury in mice with humanised immune systems |
| title_short | Single-cell RNA sequencing reveals peripheral blood leukocyte responses to spinal cord injury in mice with humanised immune systems |
| title_sort | single cell rna sequencing reveals peripheral blood leukocyte responses to spinal cord injury in mice with humanised immune systems |
| topic | Neurotrauma Neuroinflammation Neuroimmunology Immunosuppression Neutrophil Lymphocyte |
| url | https://doi.org/10.1186/s12974-024-03048-0 |
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