Single‐cell transcriptome profiling reveals the key role of ZNF683 in natural killer cell exhaustion in multiple myeloma
Abstract Backgrounds Decreased cytotoxicity of natural killer (NK) cells has been shown in multiple myeloma (MM). However, the underlying molecular mechanisms remain unclear. Here, by using single‐cell RNA sequencing analysis and in vitro experiments, we aim to uncover and validate molecularly disti...
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| Format: | Article |
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Wiley
2022-10-01
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| Series: | Clinical and Translational Medicine |
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| Online Access: | https://doi.org/10.1002/ctm2.1065 |
| _version_ | 1817976124165062656 |
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| author | Xin Li Mengping Chen Yike Wan Lu Zhong Xiaofeng Han Xiaotong Chen Fei Xiao Jia Liu Yiwei Zhang Di Zhu Jing Xiang Junling Liu Honghui Huang Jian Hou |
| author_facet | Xin Li Mengping Chen Yike Wan Lu Zhong Xiaofeng Han Xiaotong Chen Fei Xiao Jia Liu Yiwei Zhang Di Zhu Jing Xiang Junling Liu Honghui Huang Jian Hou |
| author_sort | Xin Li |
| collection | DOAJ |
| description | Abstract Backgrounds Decreased cytotoxicity of natural killer (NK) cells has been shown in multiple myeloma (MM). However, the underlying molecular mechanisms remain unclear. Here, by using single‐cell RNA sequencing analysis and in vitro experiments, we aim to uncover and validate molecularly distinctive insights into identifying regulators for NK cell exhaustion and provide potential targets for novel immune therapies in MM. Methods Single‐cell RNA sequencing was conducted in the bone marrow and peripheral blood samples from 10 newly diagnosed MM patients and three healthy volunteers. Based on the cluster‐defining differentially expressed genes, we named and estimated functional states of each cluster via bioinformatics analyses. Functional significance of key findings obtained from sequencing analysis was examined in a series of in vitro experiments, including luciferase reporter assay, lentiviral expression vector construction, NK cell transfection, RT‐qPCR, flow cytometry, and cytotoxicity assay. Results We classified NK cells into seven distinct clusters and confirmed that a subset of ZNF683+ NK cells were enriched in MM patients with ‘exhausted’ transcriptomic profile, featuring as decreased expression of activating receptors and cytolytic molecules, as well as increased expression of inhibitory receptors. Next, we found a significant downregulation of SH2D1B gene that encodes EAT‐2, an adaptor protein of activating receptor SLAMF7, in ZNF683+ NK cells from MM patients versus healthy volunteers. We further proved that ZNF683 transfection in NK cells significantly downregulated SH2D1B expression via directly binding to the promoter of SH2D1B, leading to NK cell cytotoxic activity impairment and exhausted phenotypes acquisition. In contrast, ZNF683 knockout in NK cells from MM patients increased cytotoxic activity and reversed NK cell exhaustion. Conclusions In summary, our findings uncover an important mechanism of ZNF683+ NK cell exhaustion and suggest that transcriptional suppressor ZNF683 as a potential useful therapeutic target in immunotherapy of MM. |
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| language | English |
| last_indexed | 2024-04-13T21:58:18Z |
| publishDate | 2022-10-01 |
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| series | Clinical and Translational Medicine |
| spelling | doaj.art-76a38f0e35a24ea9bdb09f2474fa3a7d2022-12-22T02:28:10ZengWileyClinical and Translational Medicine2001-13262022-10-011210n/an/a10.1002/ctm2.1065Single‐cell transcriptome profiling reveals the key role of ZNF683 in natural killer cell exhaustion in multiple myelomaXin Li0Mengping Chen1Yike Wan2Lu Zhong3Xiaofeng Han4Xiaotong Chen5Fei Xiao6Jia Liu7Yiwei Zhang8Di Zhu9Jing Xiang10Junling Liu11Honghui Huang12Jian Hou13Department of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Biochemistry and Molecular Cell Biology Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaDepartment of Hematology Ren Ji Hospital Shanghai Jiao Tong University School of Medicine Shanghai ChinaAbstract Backgrounds Decreased cytotoxicity of natural killer (NK) cells has been shown in multiple myeloma (MM). However, the underlying molecular mechanisms remain unclear. Here, by using single‐cell RNA sequencing analysis and in vitro experiments, we aim to uncover and validate molecularly distinctive insights into identifying regulators for NK cell exhaustion and provide potential targets for novel immune therapies in MM. Methods Single‐cell RNA sequencing was conducted in the bone marrow and peripheral blood samples from 10 newly diagnosed MM patients and three healthy volunteers. Based on the cluster‐defining differentially expressed genes, we named and estimated functional states of each cluster via bioinformatics analyses. Functional significance of key findings obtained from sequencing analysis was examined in a series of in vitro experiments, including luciferase reporter assay, lentiviral expression vector construction, NK cell transfection, RT‐qPCR, flow cytometry, and cytotoxicity assay. Results We classified NK cells into seven distinct clusters and confirmed that a subset of ZNF683+ NK cells were enriched in MM patients with ‘exhausted’ transcriptomic profile, featuring as decreased expression of activating receptors and cytolytic molecules, as well as increased expression of inhibitory receptors. Next, we found a significant downregulation of SH2D1B gene that encodes EAT‐2, an adaptor protein of activating receptor SLAMF7, in ZNF683+ NK cells from MM patients versus healthy volunteers. We further proved that ZNF683 transfection in NK cells significantly downregulated SH2D1B expression via directly binding to the promoter of SH2D1B, leading to NK cell cytotoxic activity impairment and exhausted phenotypes acquisition. In contrast, ZNF683 knockout in NK cells from MM patients increased cytotoxic activity and reversed NK cell exhaustion. Conclusions In summary, our findings uncover an important mechanism of ZNF683+ NK cell exhaustion and suggest that transcriptional suppressor ZNF683 as a potential useful therapeutic target in immunotherapy of MM.https://doi.org/10.1002/ctm2.1065exhaustionmultiple myelomanatural killer cellsSH2D1BZNF683 |
| spellingShingle | Xin Li Mengping Chen Yike Wan Lu Zhong Xiaofeng Han Xiaotong Chen Fei Xiao Jia Liu Yiwei Zhang Di Zhu Jing Xiang Junling Liu Honghui Huang Jian Hou Single‐cell transcriptome profiling reveals the key role of ZNF683 in natural killer cell exhaustion in multiple myeloma Clinical and Translational Medicine exhaustion multiple myeloma natural killer cells SH2D1B ZNF683 |
| title | Single‐cell transcriptome profiling reveals the key role of ZNF683 in natural killer cell exhaustion in multiple myeloma |
| title_full | Single‐cell transcriptome profiling reveals the key role of ZNF683 in natural killer cell exhaustion in multiple myeloma |
| title_fullStr | Single‐cell transcriptome profiling reveals the key role of ZNF683 in natural killer cell exhaustion in multiple myeloma |
| title_full_unstemmed | Single‐cell transcriptome profiling reveals the key role of ZNF683 in natural killer cell exhaustion in multiple myeloma |
| title_short | Single‐cell transcriptome profiling reveals the key role of ZNF683 in natural killer cell exhaustion in multiple myeloma |
| title_sort | single cell transcriptome profiling reveals the key role of znf683 in natural killer cell exhaustion in multiple myeloma |
| topic | exhaustion multiple myeloma natural killer cells SH2D1B ZNF683 |
| url | https://doi.org/10.1002/ctm2.1065 |
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