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A pan-cancer single-cell panorama of human natural killer cells
Open accessAbstract
dendritic cells, appear to mediate the regulation of NK cell anti-tumor immunity. Our study provides insights into NK-cell-based cancer immunity and highlights potential clinical utilities of NK cell subsets as therapeutic targets.
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Metascape provides a biologist-oriented resource for the analysis of systems-level datasets
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Scenic: single-cell regulatory network inference and clustering.
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“We then identified activated regulons for both tumor-infiltrating CD56brightCD16lo and CD56dim CD16hi NK cell subsets using SCENIC.49 Importantly, the tumor-enriched c6-DNAJB1 subset exhibitedmuch higher expression of transcription factors such as KLF6 and EGR3, which are associated with the inhibition of cytotoxicity…”
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The interaction of TIGIT with PVR and PVRL2 inhibits human NK cell cytotoxicity
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“This pattern mirrors the stress response observed in T cells in the context of immunotherapy resistance 34 and the poor cytotoxicity seen in tumor-associated NK cells in a recent pan-cancer single-cell atlas of human NK cells 28 .”
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“…cells characterized by reduced cytotoxicity and high expression of stress-associated proteins ( DNAJB1 and HSP gene family), inhibitory MHC-class-I-specific receptors (members of the KIR family) and several transcription factors that control the fitness of NK cells such as NR4A1, EGR3 and KLF6 29 .”
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“We then confirmed the robustness of the classification of human blood NK cells into the NK1, NK2 and NK3 clusters by applying the derived transcriptional signatures to blood NK cells from other available datasets 11 (Extended Data Figs.”
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“8e), consistent with what was reported in hepatocellular carcinoma 46 .”
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“Using a pan-cancer, single-cell atlas approach, Tang et al. 16 identified a tumor-enriched dysfunctional CD56 dim CD16 hi NK cell population interacting with LAMP3 + dendritic cells in the TME.”
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“Recent studies also report that CCR7 + DCs may engage other immune cells in tumours, including CXCL13 + CD4 + T cells, regulatory T cells and NK cells 38,50 – 52 , and may reside in TLS 12,15 .”
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“Although accumulating evidence suggests that NK cells, as non-speci fi c immune effectors, play animportantrole inregulating thetumor microenvironment and antitumor immunity, their molecular markers and prognostic signi fi cance remain insuf fi ciently characterized 30 – 32 .”
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CREM is a regulatory checkpoint of CAR and IL-15 signalling in NK cells
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“Recent studies have reported distinct subsets of NK cells with high CREM expression 8,48 , although the functional implications were not explored.”
Engineered natural killer cells for cancer therapy
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“The advent of single-cell analysis has provided insights into the landscape and dynamics of immune cell populations within cancer tissues [16, 17].”
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“Notably, TaNK cells express higher levels of inhibitory receptors (e.g., KIRs) and lower levels of cytotoxic markers, suggesting a potentially “ dysfunctional ” phenotype with impaired tumor-killing capacity (100).”
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“CD56 dim CD16 hi NK cells exert potent cytotoxicity via perforin and granzyme B secretion, whereas CD56 bright CD16 lo NK cells produce substantial amounts of IFN-γ and other cytokines that are crucial for immunomodulation [75].”
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“Single-cell RNA-seq (scRNA-seq) technologies empower the high-resolution characterization of cellular heterogeneity in the TME, bringing new opportunities to i l luminate the spectrum of TECs [18 ,22 ,23 ].”
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“To validate these NK signatures, we applied the gene-sets to a pan-cancer human NK cell atlas developed by Tang et al., 2023 35 (Supplementary Figure S6B).”
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“17,21 In addition, the development of advanced single-cell technologies, such as scRNA-seq and CITE (Cellular Indexing of Transcriptomes and Epitopes)-seq, has accelerated the dissection of NK cell subset heterogeneity, revealing an intricate and nuanced NK cell landscape as well as different…”
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“In keeping with this, three recent studies described an association between NK cells and lack of response to immunotherapy in different cancer types (6–8).”
InfluentialXiao, M., Deng, Y., Guo, H., Ren, Z., He, Y., Ren, X., Huang, L., Zhang, W., Chen, H., Yang, S., et al. (2025) · Translational Lung Cancer Research · doi:10.21037/tlcr-2025-244 · Cited by 31
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“Single-cell RNA sequencing coupled with spatial profiling has further shown that such immunoregulatory NK populations preferentially localize within T cell-rich tumor regions, positioning them as key modulators of T cell responses in immune-inflamed tumors [56,57].”
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“In tumors, NR4A2, along with NR4A1 and NR4A3, has been shown to drive exhaustion and limit effector functions in chimeric antigen receptor T cells 27,28 , whereas in tumor-associated NK cells, NR4A2 expression is linked to reduced cytotoxic activity and a dysfunctional state 43 .”
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“In a pan-cancer scRNA-seq study [122], CD56 dim CD16 hi c6-DNAJB1 NK cells were designated as tumor-associated NK (TaNK) cells, which were enriched in tumors and exhibited impaired cytotoxicity, indicative of a terminal state.”
BackgroundZheng, Y., He, D., Zuo, W., Wang, W., Wu, K., Wu, H., Yuan, Y., Huang, Y.-J., Li, H., Lu, Y., et al. (2025) · EBioMedicine · doi:10.1016/j.ebiom.2025.105738 · Cited by 31
“37 The de fi nition of “ viral load ” in a cell is the percentage of UMIs mapped to one of eight viral segments in the total mapped UMIs.”
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“Additionally, TaNKs and iCAF were significantly correlated with resistance to immunotherapy [46, 134].”
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“…cell (EC) (VWF) [35], lymphatic endothelial cell (LEC) (TFF3) [35]; immune cells: T cell (CD3D) [19, 35], B cell (CD79A) [19, 35], NK cell (KLRB1) [51], monocyte (CD14) [35, 52], mast cell (TPSB2) [35]; cells in cell cycle (cycling) (TOP2A) [53] and red blood cell (RBC) (HBA2) [54] (Fig.”
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Single-cell meta-analysis of T cells reveals clonal dynamics of response to checkpoint immunotherapy
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“…in ICI-treated patients with Richter syndrome 41 and to the revival of pre-existing ZNF683 + clones in patients with HNSCC responding to ICI treatment, 42 this finding emphasizes the importance of the contextual co-expression of genes in relation to clinical outcome, as well as differences…”
Zhang, J., Zhang, M., Lou, J., Wu, L., Zhang, S., Liu, X., Ke, Y., Zhao, S., Song, Z., Bai, X., et al. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms252312715 · Cited by 21
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“34,35 As depicted in Figure S6, these markers were highly expressed in our identi fi ed NK cell clusters.”
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“Thus in some solid organ cancers, CD56dim CD16high HSP40+ tumour-asssociated NK cells display a dysfunctional state with reduced perforin and granzyme B [29].”
The Problem with Syngeneic Mouse Tumor Models
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Pan-Cancer Analyses Refine the Single-Cell Portrait of Tumor-Infiltrating Dendritic Cells
Ma, T., Chu, X., Wang, J., Li, X., Zhang, Y., Tong, D., Xu, W., Dang, G., Qi, L., Miao, Y., et al. (2025) · Cancer Research · doi:10.1158/0008-5472.can-24-3595 · Cited by 20
He, R., Lu, J., Feng, J., Lu, Z., Shen, K., Xu, K., Luo, H., Yang, G., Chi, H., & Huang, S. (2024) · Frontiers in Immunology · doi:10.3389/fimmu.2024.1435187 · Cited by 20
“Additionally, single-cell analysis can elucidate changes in systemic immunity during tumor progression; researchers identi fi ed an adaptive NK cell subgroup characterized by upregulation of speci fi c pro-in fl ammatory cytokines and MHC class II genes in the peripheral blood of cancer patients (52).”
BackgroundUnveiling novel insights in acute myeloid leukemia through single-cell RNA sequencing
Zhou, J., & Chng, W. J. (2024) · Frontiers in Oncology · doi:10.3389/fonc.2024.1365330 · Cited by 22
Wang, J., Shi, Y., Gao, Y., Zhu, N., Liu, Y., Zhang, Y., Chen, X., & Yang, R. (2026) · Journal for ImmunoTherapy of Cancer · doi:10.1136/jitc-2025-013697 · Cited by 18
Jiao, J.-Z., Zhang, Y., Zhang, W., He, M., Meng, M., Liu, T., Ma, Q., Xu, Y., Gao, P., Chen, C., et al. (2024) · Acta Pharmacologica Sinica · doi:10.1038/s41401-024-01260-5 · Cited by 16
“However, NK cells exhibit tissue heterogeneity across cancer types [34].”
BackgroundThe stress response regulator HSF1 modulates natural killer cell anti-tumour immunity
Hockemeyer, K., Sakellaropoulos, T., Chen, X., Ivashkiv, O., Sirenko, M., Zhou, H., Gambi, G., Battistello, E., Avrampou, K., Sun, Z., et al. (2024) · Nature Cell Biology · doi:10.1038/s41556-024-01490-z · Cited by 17
“For analysis of the pan-cancer atlas of human NK cells 39 , analysis was performed using Seurat (v.4.3.0.1) in R v.4.2.2 and Scanpy v.1.6.0 in Python v.3.7.8.”
InfluentialZhou, T., Yan, H., Deng, Y., Zhu, Y., Xia, X., Wu, W., Zhang, W., Chen, H., Hu, J., Zhou, Z., et al. (2024) · Interdisciplinary medicine · doi:10.1002/inmd.20240018 · Cited by 17
Mao, C., Chen, Y., Xing, D., Zhang, T., Lin, Y., Long, C., Yu, J., Luo, Y., Ming, T., Mei, D., et al. (2024) · eLife · doi:10.7554/elife.97201 · Cited by 17
Zheng, X., Wu, W., Zhao, Z., Zhang, X., & Yu, S. (2024) · Journal of Cancer Research and Clinical Oncology · doi:10.1007/s00432-024-05787-2 · Cited by 16
“6D, E, F) (Tang et al. 2023).”
Cheng, H., Yan, Y., Zhang, B., Ma, Z., Fu, S., Ji, Z. L., Zou, Z., & Wang, Q. (2025) · PLoS ONE · doi:10.1371/journal.pone.0312764 · Cited by 19
“TaNK cells have been associated with poor patient outcomes and resistance to immunotherapy in different cancer types [39].”
BackgroundInfluentialJiang, Q., Yu, W., Ma, J., Zhao, M., Zou, J., Mir, S. A., Zhang, J., Germain, R. N., & Hassan, R. (2025) · Science Advances · doi:10.1126/sciadv.adt9932 · Cited by 15
“The NK5 cluster was characterized by high expression of cell division genes, including STMN1 , RRM2 , and ZWINT , along with the tumor-infiltrating NK marker RGS1 ( 27 ), suggesting that they may correspond to the Ki67 + NK population observed in immunofluorescence imaging.”
Ozmen, F., Ozmen, T. Y., Ors, A., Janghorban, M., Rames, M. J., Li, X., Doe, A. R., Behbod, F., Mills, G. B., & Mohammed, H. (2025) · npj Breast Cancer · doi:10.1038/s41523-025-00808-w · Cited by 21
“Afterintegration,NKcells,myeloidcells,Tcells,Bcells,and fi broblasts werefurtherclassi fi edusingthefollowingpubliclyavailablesingle-cellRNA seq datasets: NK cells (GSE212890) 115 , T, myeloid, and B cells (GSE169426) 116 , and fi broblasts(GSE103322, GSE132465, GSE154778, and GSE212966) 86 .”
Chen, Y., Meng, Z., & Xiang, Z. (2024) · Frontiers in Immunology · doi:10.3389/fimmu.2024.1415435 · Cited by 15
“The comprehensive mapping of TME components in HCC is continuously evolving, encompassing T cells (15), neutrophils (16), NK cells (17), as well as various potentially signi fi cant subsets and driver genes previously undiscovered, such as the function and clinical relevance of exhausted CD8+ T…”
BackgroundRedefining the tumor microenvironment with emerging therapeutic strategies
Xu, S., Li, X., & Ma, W. (2024) · Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics · doi:10.32604/or.2024.055161 · Cited by 15
“Recent studies using single-cell RNA sequencing have uncovered a range of cell types in the TME, highlighting the importance of treatment strategies [5 – 7].”
BackgroundZhang, H., Wang, J., & Li, F. (2023) · Frontiers in Immunology · doi:10.3389/fimmu.2023.1286986 · Cited by 15
“Further transcriptomic feature analysis in this study has shown that a subset of tumor-associated NK cells is enriched in tumors and displays exhaustion features and impaired antitumor functions in multiple cancers, including lung cancer (58).”
BackgroundRAC1high NK cell-based immunotherapy in hepatocellular carcinoma via STAT3-NKG2D axis
Shi, X., Chen, W., Yin, Y., Cao, H., Wang, X., Jiang, W., Li, Q., Li, X., Yu, Y., & Wang, X. (2024) · Cancer Letters · doi:10.1016/j.canlet.2024.216909 · Cited by 14
Choi, W. S., Kwon, H., Yi, E., Lee, H., Kim, J. M., Park, H. J., Choi, E. J., Choi, M. E., Sung, Y. H., Won, C. H., et al. (2024) · Advanced Science · doi:10.1002/advs.202400920 · Cited by 14
Xu, W. V., Birch, G., Meliki, A., Moritz, V., Bharadwaj, M., Schindler, N. R., Labaki, C., Saliby, R. M., Dinh, K., Horst, J. T., et al. (2025) · ESMO Open · doi:10.1016/j.esmoop.2024.104105 · Cited by 14
Kim, S., Son, E., Park, H.-R., Kim, M., & Yang, H. W. (2025) · Journal of Clinical Investigation · doi:10.1172/jci188839 · Cited by 14
Single‐Cell and Spatial Omics: Methods and Applications
Cen, X., Huang, X., Deng, E., Gong, X., Tan, N., Ye, J. T., Wang, Y., Eils, R., Luo, Q., Li, Y., et al. (2026) · MedComm · doi:10.1002/mco2.70713 · Cited by 14
“…single-cell atlases encompassing more than 700 cases have delineated conserved and divergent immune, stromal, and malignant programs across tumors, identifying recurrent T cell exhaustion states [ 324 ], natural killer cell phenotypes [ 325 ], and cancer-associated B cell populations [ 326 ].”
Yang, Y., Liu, Z., Wang, Z., Fu, X., Li, Z., Li, J., Xu, Z., & Cen, B. (2025) · Biology Direct · doi:10.1186/s13062-025-00640-z · Cited by 16
OR7A10 GPCR engineering boosts CAR-NK therapy against solid tumours
Yang, L., Renauer, P. A., Tang, K., Saskin, J., Zhou, L., Zou, C., Lee, S.-H., Fox, M. L., Johnson-Noya, S., Weiss, B., et al. (2026) · Nature · doi:10.1038/s41586-026-10149-8 · Cited by 15
“…subsets were each characterized by established NK cell markers from molecular and SCT studies of tumour-infiltrating NK cells, including those for proliferation ( MKI67 ), NK differentiation ( BATF3 ), IL32 , memory-like NK phenotype ( KLRC2 ), stress ( CREM ) and NK cell dysfunction ( EGR2 ) 74 .”
Li, J., Zhang, Y., Liang, C., Yan, X., Hui, X., & Liu, Q. (2024) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2024.1396836 · Cited by 13
Yu, Y., You, Y., Duan, Y., Kang, M., Zhou, B., Yang, J., Yin, K.-L., Ye, W., Xu, R., Wang, H., et al. (2025) · Hepatology · doi:10.1097/hep.0000000000001352 · Cited by 12
Qu, W., Zhu, G., Yang, R., Chu, T.-H., Guan, Z.-Q., Huang, R., Tian, M., Jiang, X., Tao, C.-Y., Fang, Y., et al. (2025) · Science Advances · doi:10.1126/sciadv.ads8597 · Cited by 14
Netskar, H., Pfefferle, A., Goodridge, J. P., Sohlberg, E., Dufva, O., Teichmann, S. A., Clancy, T., Horowitz, A., & Malmberg, K. (2023) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2023.10.26.564050 · Cited by 12
“Using a pan-cancer single-cell atlas approach, Tang et al. 16 identified a tumor-enriched dysfunctional CD56 dim CD16 hi NK cell population interacting with LAMP3 + dendritic cells in the tumor microenvironment (TME).”
MethodResultInfluentialGuedes, J., Woldmar, N., Szasz, A. M., Wieslander, E., Pawłowski, K., Horvatovich, P., Malm, J., Szadai, L., Németh, I. B., Marko-Varga, G., et al. (2025) · Journal of Proteomics · doi:10.1016/j.jprot.2025.105493 · Cited by 16
Chen, W., Liao, K., Chen, H., Xiao, Z.-X., Meng, Q., Liu, Z., Lu, Y., Sheng, H., Mo, H., Wu, Q.-N., et al. (2025) · Signal Transduction and Targeted Therapy · doi:10.1038/s41392-025-02221-y · Cited by 13
“Considering that NK cells in the TME have been reported to exhibit different phenotypes, including dysfunctional states, 18,47 we examined NK cells isolated from orthotopic tumors and performed Smart-seq2 to ensure that the elevated tumor-in fi ltrating NK cells induced by ACAT1 were activated.”
Engineering NK and T cells with metabolite-sensing receptors to target solid tumors
Kim, Y., Tsai, M. K., Sun, C., Laveroni, O., Akana, R. V., Frombach, K. R., & Jerby‐Arnon, L. (2026) · Nature Immunology · doi:10.1038/s41590-026-02473-y · Cited by 14
Yin, Y., Wang, Y., Yu, X., Li, Y., Zhao, Y., & Liu, Z. (2024) · Cancer Letters · doi:10.1016/j.canlet.2024.217045 · Cited by 11
Innovative treatment of age-related hearing loss using MSCs and EVs with Apelin
Xu, S., Liu, D., Zhang, F., & Tian, Y. (2025) · Cell Biology and Toxicology · doi:10.1007/s10565-025-09988-4 · Cited by 12
Luo, R., Liu, J., Wang, T., Zhao, W., Wang, Y., Wen, J., Wang, H., Ding, S., & Zhou, X. (2025) · Cancer Letters · doi:10.1016/j.canlet.2025.217591 · Cited by 13
NKG7 is a Stable Marker of Cytotoxicity Across Immune Contexts and Within the Tumor Microenvironment
Turiello, R., Ng, S. S., Tan, E., van der Voort, G., Salim, N., Yong, M. C. R., Khassenova, M., Oldenburg, J., Rühl, H., Hasenauer, J., et al. (2025) · European Journal of Immunology · doi:10.1002/eji.202551885 · Cited by 12
Masters of adaptation: How cancer and immune cell plasticity mediates tumor progression
Rosenbaum, S. R., Fields, K. M., & Ford, H. L. (2025) · PLoS Biology · doi:10.1371/journal.pbio.3003301 · Cited by 12
“…analysis of patient samples across 24 cancer types, which demonstrated that NK cell heterogeneity was specific to tumor type and that tumor-infiltrating NK cells had reduced cytotoxic features compared to blood-derived NK cells [148], suggesting that NK cells shift their phenotype upon tumor entry.”
Metadata
- Authors
- Tang, F., Li, J., Qi, L., Liu, D., Bo, Y., Qin, S., Miao, Y., Yu, K., Hou, W., Li, J., Peng, J., Tian, Z., Zhu, L., Peng, H., Wang, D., & Zhang, Z.
- Affiliations
- Peking University (CN)
- Capital Medical University (CN)
- Shenzhen Bay Laboratory (CN)
- University of Science and Technology of China (CN)
- Corresponding author
- Zhu, Linnan
- Peng, Hui
- Wang, Dongfang
- Zhang, Zemin
- ORCID
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 2023-09
- Volume
- 186
- Issue
- 19
- Pages
- 4235-4251.e20
- ISSN
- 0092-8674 (ISSN-L)
- OpenAlex
- W4386034505
- Cited by
- 450
- References
- 91
- Citations by year
- 2026: 136 · 2025: 181 · 2024: 120 · 2023: 11
- FWCI
- 50.09
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Cell (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Greater South Information System
- Funders
- National Natural Science Foundation of China (31991171, 592259205, 62203019, 81988101, 91959000, 92159305)
- Peking University
- Beijing Municipal Science and Technology Commission, Adminitrative Commission of Zhongguancun Science Park (Z221100007022002)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Immune Cell Function and Interaction
- Health Sciences › Medicine › Oncology › Cancer Immunotherapy and Biomarkers
- Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
- Keywords
- natural killer cells, pan-cancer, single-cell transcriptomics, NK cell subsets, cancer immunotherapy, dendritic cells, antitumor immunity
- MeSH terms
- Single-Cell Gene Expression Analysis, Dendritic Cells, Humans, Immunity, Innate, Immunotherapy, Killer Cells, Natural, Neoplasms, Myeloid Cells, Tumor Microenvironment
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