Single-Nucleus RNA Sequencing Reveals the Spatiotemporal Dynamics of Disease-Associated Microglia in Amyotrophic Lateral Sclerosis
Disease-associated microglia (DAM) are observed in neurodegenerative diseases, demyelinating disorders, and aging. However, the spatiotemporal dynamics and evolutionary trajectory of DAM during the progression of amyotrophic lateral sclerosis (ALS) remain unclear. Using a mouse model of ALS that exp...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Association for the Advancement of Science (AAAS)
2024-01-01
|
| Series: | Research |
| Online Access: | https://spj.science.org/doi/10.34133/research.0548 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850247267100917760 |
|---|---|
| author | Lu-Xi Chen Mei-Di Zhang Hai-Feng Xu Hai-Qin Ye Dian-Fu Chen Pei-Shan Wang Zhi-Wei Bao Sheng-Mei Zou Yong-Ting Lv Zhi-Ying Wu Hong-Fu Li |
| author_facet | Lu-Xi Chen Mei-Di Zhang Hai-Feng Xu Hai-Qin Ye Dian-Fu Chen Pei-Shan Wang Zhi-Wei Bao Sheng-Mei Zou Yong-Ting Lv Zhi-Ying Wu Hong-Fu Li |
| author_sort | Lu-Xi Chen |
| collection | DOAJ |
| description | Disease-associated microglia (DAM) are observed in neurodegenerative diseases, demyelinating disorders, and aging. However, the spatiotemporal dynamics and evolutionary trajectory of DAM during the progression of amyotrophic lateral sclerosis (ALS) remain unclear. Using a mouse model of ALS that expresses a human SOD1 gene mutation, we found that the microglia subtype DAM begins to appear following motor neuron degeneration, primarily in the brain stem and spinal cord. Using reverse transcription quantitative polymerase chain reaction, RNAscope in situ hybridization, and flow cytometry, we found that DAM increased in number as the disease progressed, reaching their peak in the late disease stage. DAM responded to disease progression in both SOD1G93A mice and sporadic ALS and C9orf72-mutated patients. Motor neuron loss in SOD1G93A mice exhibited 2 accelerated phases: P90 to P110 (early stage) and P130 to P150 (late stage). Some markers were synchronized with the accelerated phase of motor neuron loss, suggesting that these proteins may be particularly responsive to disease progression. Through pseudotime trajectory analysis, we tracked the dynamic transition of homeostatic microglia into DAM and cluster 6 microglia. Interestingly, we used the colony-stimulating factor 1 receptor (CSF1R) inhibitor PLX5622 to deplete microglia in SOD1G93A mice and observed that DAM survival is independent of CSF1R. An in vitro phagocytosis assay directly confirmed that DAM could phagocytose more beads than other microglia subtypes. These findings reveal that the induction of the DAM phenotype is a shared cross-species and cross-subtype characteristic in ALS. Inducing the DAM phenotype and enhancing its function during the early phase of disease progression, or the time window between P130 and P150 where motor neuron loss slows, could serve as a neuroprotective strategy for ALS. |
| format | Article |
| id | doaj-art-91a34361e33a4def9ef94a9c33ca2891 |
| institution | OA Journals |
| issn | 2639-5274 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | American Association for the Advancement of Science (AAAS) |
| record_format | Article |
| series | Research |
| spelling | doaj-art-91a34361e33a4def9ef94a9c33ca28912025-08-20T01:58:59ZengAmerican Association for the Advancement of Science (AAAS)Research2639-52742024-01-01710.34133/research.0548Single-Nucleus RNA Sequencing Reveals the Spatiotemporal Dynamics of Disease-Associated Microglia in Amyotrophic Lateral SclerosisLu-Xi Chen0Mei-Di Zhang1Hai-Feng Xu2Hai-Qin Ye3Dian-Fu Chen4Pei-Shan Wang5Zhi-Wei Bao6Sheng-Mei Zou7Yong-Ting Lv8Zhi-Ying Wu9Hong-Fu Li10Department of Medical Genetics and Center for Rare Diseases, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.Department of Medical Genetics and Center for Rare Diseases, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.Institute of Developmental and Regenerative Biology, Zhejiang Key Laboratory of Organ Development and Regeneration, College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, China.Department of Medical Genetics and Center for Rare Diseases, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.Department of Medical Genetics and Center for Rare Diseases, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.Department of Medical Genetics and Center for Rare Diseases, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.Department of Medical Genetics and Center for Rare Diseases, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.Department of Medical Genetics and Center for Rare Diseases, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.Department of Medical Genetics and Center for Rare Diseases, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.Disease-associated microglia (DAM) are observed in neurodegenerative diseases, demyelinating disorders, and aging. However, the spatiotemporal dynamics and evolutionary trajectory of DAM during the progression of amyotrophic lateral sclerosis (ALS) remain unclear. Using a mouse model of ALS that expresses a human SOD1 gene mutation, we found that the microglia subtype DAM begins to appear following motor neuron degeneration, primarily in the brain stem and spinal cord. Using reverse transcription quantitative polymerase chain reaction, RNAscope in situ hybridization, and flow cytometry, we found that DAM increased in number as the disease progressed, reaching their peak in the late disease stage. DAM responded to disease progression in both SOD1G93A mice and sporadic ALS and C9orf72-mutated patients. Motor neuron loss in SOD1G93A mice exhibited 2 accelerated phases: P90 to P110 (early stage) and P130 to P150 (late stage). Some markers were synchronized with the accelerated phase of motor neuron loss, suggesting that these proteins may be particularly responsive to disease progression. Through pseudotime trajectory analysis, we tracked the dynamic transition of homeostatic microglia into DAM and cluster 6 microglia. Interestingly, we used the colony-stimulating factor 1 receptor (CSF1R) inhibitor PLX5622 to deplete microglia in SOD1G93A mice and observed that DAM survival is independent of CSF1R. An in vitro phagocytosis assay directly confirmed that DAM could phagocytose more beads than other microglia subtypes. These findings reveal that the induction of the DAM phenotype is a shared cross-species and cross-subtype characteristic in ALS. Inducing the DAM phenotype and enhancing its function during the early phase of disease progression, or the time window between P130 and P150 where motor neuron loss slows, could serve as a neuroprotective strategy for ALS.https://spj.science.org/doi/10.34133/research.0548 |
| spellingShingle | Lu-Xi Chen Mei-Di Zhang Hai-Feng Xu Hai-Qin Ye Dian-Fu Chen Pei-Shan Wang Zhi-Wei Bao Sheng-Mei Zou Yong-Ting Lv Zhi-Ying Wu Hong-Fu Li Single-Nucleus RNA Sequencing Reveals the Spatiotemporal Dynamics of Disease-Associated Microglia in Amyotrophic Lateral Sclerosis Research |
| title | Single-Nucleus RNA Sequencing Reveals the Spatiotemporal Dynamics of Disease-Associated Microglia in Amyotrophic Lateral Sclerosis |
| title_full | Single-Nucleus RNA Sequencing Reveals the Spatiotemporal Dynamics of Disease-Associated Microglia in Amyotrophic Lateral Sclerosis |
| title_fullStr | Single-Nucleus RNA Sequencing Reveals the Spatiotemporal Dynamics of Disease-Associated Microglia in Amyotrophic Lateral Sclerosis |
| title_full_unstemmed | Single-Nucleus RNA Sequencing Reveals the Spatiotemporal Dynamics of Disease-Associated Microglia in Amyotrophic Lateral Sclerosis |
| title_short | Single-Nucleus RNA Sequencing Reveals the Spatiotemporal Dynamics of Disease-Associated Microglia in Amyotrophic Lateral Sclerosis |
| title_sort | single nucleus rna sequencing reveals the spatiotemporal dynamics of disease associated microglia in amyotrophic lateral sclerosis |
| url | https://spj.science.org/doi/10.34133/research.0548 |
| work_keys_str_mv | AT luxichen singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT meidizhang singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT haifengxu singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT haiqinye singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT dianfuchen singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT peishanwang singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT zhiweibao singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT shengmeizou singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT yongtinglv singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT zhiyingwu singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis AT hongfuli singlenucleusrnasequencingrevealsthespatiotemporaldynamicsofdiseaseassociatedmicrogliainamyotrophiclateralsclerosis |