The CD33 short isoform is a gain-of-function variant that enhances Aβ1–42 phagocytosis in microglia
Abstract Background CD33 is genetically linked to Alzheimer’s disease (AD) susceptibility through differential expression of isoforms in microglia. The role of the human CD33 short isoform (hCD33m), preferentially encoded by an AD-protective CD33 allele (rs12459419T), is unknown. Here, we test wheth...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-03-01
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| Series: | Molecular Neurodegeneration |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13024-021-00443-6 |
| _version_ | 1818574327843389440 |
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| author | Abhishek Bhattacherjee Jaesoo Jung Sameera Zia Madelene Ho Ghazaleh Eskandari-Sedighi Chris D. St. Laurent Kelli A. McCord Arjun Bains Gaurav Sidhu Susmita Sarkar Jason R. Plemel Matthew S. Macauley |
| author_facet | Abhishek Bhattacherjee Jaesoo Jung Sameera Zia Madelene Ho Ghazaleh Eskandari-Sedighi Chris D. St. Laurent Kelli A. McCord Arjun Bains Gaurav Sidhu Susmita Sarkar Jason R. Plemel Matthew S. Macauley |
| author_sort | Abhishek Bhattacherjee |
| collection | DOAJ |
| description | Abstract Background CD33 is genetically linked to Alzheimer’s disease (AD) susceptibility through differential expression of isoforms in microglia. The role of the human CD33 short isoform (hCD33m), preferentially encoded by an AD-protective CD33 allele (rs12459419T), is unknown. Here, we test whether hCD33m represents a loss-of-function or gain-of-function variant. Methods We have developed two models to test the role of hCD33m. The first is a new strain of transgenic mice expressing hCD33m in the microglial cell lineage. The second is U937 cells where the CD33 gene was disrupted by CRISPR/Cas9 and complemented with different variants of hCD33. Primary microglia and U937 cells were tested in phagocytosis assays and single cell RNA sequencing (scRNAseq) was carried out on the primary microglia. Furthermore, a new monoclonal antibody was developed to detect hCD33m more efficiently. Results In both primary microglia and U937 cells, we find that hCD33m enhances phagocytosis. This contrasts with the human CD33 long isoform (hCD33M) that represses phagocytosis, as previously demonstrated. As revealed by scRNAseq, hCD33m+ microglia are enriched in a cluster of cells defined by an upregulated expression and gene regulatory network of immediate early genes, which was further validated within microglia in situ. Using a new hCD33m-specific antibody enabled hCD33m expression to be examined, demonstrating a preference for an intracellular location. Moreover, this newly discovered gain-of-function role for hCD33m is dependent on its cytoplasmic signaling motifs, dominant over hCD33M, and not due to loss of glycan ligand binding. Conclusions These results provide strong support that hCD33m represents a gain-of-function isoform and offers insight into what it may take to therapeutically capture the AD-protective CD33 allele. |
| first_indexed | 2024-12-15T00:25:08Z |
| format | Article |
| id | doaj.art-9d903e7fcaaf46188f4b75d343f185d2 |
| institution | Directory Open Access Journal |
| issn | 1750-1326 |
| language | English |
| last_indexed | 2024-12-15T00:25:08Z |
| publishDate | 2021-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Neurodegeneration |
| spelling | doaj.art-9d903e7fcaaf46188f4b75d343f185d22022-12-21T22:42:12ZengBMCMolecular Neurodegeneration1750-13262021-03-0116112210.1186/s13024-021-00443-6The CD33 short isoform is a gain-of-function variant that enhances Aβ1–42 phagocytosis in microgliaAbhishek Bhattacherjee0Jaesoo Jung1Sameera Zia2Madelene Ho3Ghazaleh Eskandari-Sedighi4Chris D. St. Laurent5Kelli A. McCord6Arjun Bains7Gaurav Sidhu8Susmita Sarkar9Jason R. Plemel10Matthew S. Macauley11Department of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaNeuroscience and Mental Health Institute, University of AlbertaNeuroscience and Mental Health Institute, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaDepartment of Chemistry, University of AlbertaNeuroscience and Mental Health Institute, University of AlbertaDepartment of Chemistry, University of AlbertaAbstract Background CD33 is genetically linked to Alzheimer’s disease (AD) susceptibility through differential expression of isoforms in microglia. The role of the human CD33 short isoform (hCD33m), preferentially encoded by an AD-protective CD33 allele (rs12459419T), is unknown. Here, we test whether hCD33m represents a loss-of-function or gain-of-function variant. Methods We have developed two models to test the role of hCD33m. The first is a new strain of transgenic mice expressing hCD33m in the microglial cell lineage. The second is U937 cells where the CD33 gene was disrupted by CRISPR/Cas9 and complemented with different variants of hCD33. Primary microglia and U937 cells were tested in phagocytosis assays and single cell RNA sequencing (scRNAseq) was carried out on the primary microglia. Furthermore, a new monoclonal antibody was developed to detect hCD33m more efficiently. Results In both primary microglia and U937 cells, we find that hCD33m enhances phagocytosis. This contrasts with the human CD33 long isoform (hCD33M) that represses phagocytosis, as previously demonstrated. As revealed by scRNAseq, hCD33m+ microglia are enriched in a cluster of cells defined by an upregulated expression and gene regulatory network of immediate early genes, which was further validated within microglia in situ. Using a new hCD33m-specific antibody enabled hCD33m expression to be examined, demonstrating a preference for an intracellular location. Moreover, this newly discovered gain-of-function role for hCD33m is dependent on its cytoplasmic signaling motifs, dominant over hCD33M, and not due to loss of glycan ligand binding. Conclusions These results provide strong support that hCD33m represents a gain-of-function isoform and offers insight into what it may take to therapeutically capture the AD-protective CD33 allele.https://doi.org/10.1186/s13024-021-00443-6Alzheimer’s diseaseCD33ImmunomodulatoryIsoformMicrogliaPhagocytosis |
| spellingShingle | Abhishek Bhattacherjee Jaesoo Jung Sameera Zia Madelene Ho Ghazaleh Eskandari-Sedighi Chris D. St. Laurent Kelli A. McCord Arjun Bains Gaurav Sidhu Susmita Sarkar Jason R. Plemel Matthew S. Macauley The CD33 short isoform is a gain-of-function variant that enhances Aβ1–42 phagocytosis in microglia Molecular Neurodegeneration Alzheimer’s disease CD33 Immunomodulatory Isoform Microglia Phagocytosis |
| title | The CD33 short isoform is a gain-of-function variant that enhances Aβ1–42 phagocytosis in microglia |
| title_full | The CD33 short isoform is a gain-of-function variant that enhances Aβ1–42 phagocytosis in microglia |
| title_fullStr | The CD33 short isoform is a gain-of-function variant that enhances Aβ1–42 phagocytosis in microglia |
| title_full_unstemmed | The CD33 short isoform is a gain-of-function variant that enhances Aβ1–42 phagocytosis in microglia |
| title_short | The CD33 short isoform is a gain-of-function variant that enhances Aβ1–42 phagocytosis in microglia |
| title_sort | cd33 short isoform is a gain of function variant that enhances aβ1 42 phagocytosis in microglia |
| topic | Alzheimer’s disease CD33 Immunomodulatory Isoform Microglia Phagocytosis |
| url | https://doi.org/10.1186/s13024-021-00443-6 |
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