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Establishment and comprehensive analysis of a new human cell line (NK-NJ) with NK-cell characteristics established from extranodal natural killer cell lymphoma/leukemia

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Abstract

Extranodal NK/T cell lymphoma, nasal type (ENKTL) is an aggressive and heterogeneous disease. With standard treatment containing pegaspargase-based regimen, patients who were resistant to pegaspargase have rapidly disease progression and worse prognosis. Thus, there is an urgent requirement for constructing ENKTL cell line model to explore the mechanism of pegaspargase resistance and new molecular targeted drugs to improve prognosis. We report here on the establishment of a novel ENKTL cell line, NK-NJ. The cells were isolated from a 52-year-old Chinese man who was diagnosed with relapse/refractory (R/R) ENKTL and grow steadily in vitro. The NK-NJ cells express CD56, CD2, CD45RA with no expression of CD3, CD16, CD57, CD4, CD8, CD26, CD28, CD5, TCR, CD45RO and CD161 and showed a TCR gene unrearrangement, which suggested an origin in the NK-lineage but not T-lineage. The immunophenotypes of NK-NJ cells were consistent with the patient. Moreover, short tandem repeat (STR) profiling results also demonstrated that NK-NJ originated from the patient. NK-NJ showed complex karyotype. Target sequencing method indicated that the main mutation genes of the first-time disease progression of lymph nodal were the same as main mutation genes of the primary nasal lesions. Moreover, NK-NJ was recognized as latency I with EBER positivity and carried high EBV-DNA viral load. The chemosensitivity results suggested synthetic lethality of epigenetic drugs and PD-1 inhibitor for ENKTL patients by reasons of epigenetic drugs promoting PD-L1 expression. In conclusion, we established a new ENKTL cell line in the era of new targeted drugs. We hope that this cell line can help to further understand underlying pathogenesis of ENKTL especially for advanced ENKTL and the functional role of EBV in ENKTL pathogenetic process.

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Data availability

Details data in this study are available from the corresponding author upon request.

Abbreviations

CNV:

Copy number variations

CR:

Complete remission

ENKTL:

Extranodal NK/T cell lymphoma, nasal type

FCM:

Flow cytometry

IFN:

Interferon

OS:

Overall survival

QC:

Quality control

TNF:

Tumor necrosis factor

WES:

Whole exon sequencing

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Acknowledgements

The authors would like to thank the patient for the willingness to contribute to medical research.

Funding

This study was supported by National Natural Science Foundation of China (81770166, 81720108002), Jiangsu Province’s Medical Elite Programme (ZDRCA2016022), Project of National Key Clinical Specialty, Jiangsu Provincial Special Program of Medical Science (BE2017751) and National Science and Technology Major Project (2018ZX09734007).

Author information

Author notes

  1. Jin-Hua Liang and Wei-Ting Wang contributed equally to this work.

Authors and Affiliations

  1. Department of Hematology, The First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing, 210029, China

    Jin-Hua Liang, Wei-Ting Wang, Kai-Xin Du, Tong-Yao Xing, Yan Wang, Hui Wang, Lu Liu, Rui Guo, Yue Li, Hao-Rui Shen, Li Wang, Jian-Yong Li & Wei Xu

  2. Key Laboratory of Hematology, Nanjing Medical University, Nanjing, 210029, China

    Jin-Hua Liang, Wei-Ting Wang, Kai-Xin Du, Tong-Yao Xing, Yan Wang, Hui Wang, Lu Liu, Rui Guo, Yue Li, Hao-Rui Shen, Li Wang, Jian-Yong Li & Wei Xu

  3. Collaborative Innovation Center for Cancer Personalized Medicine, Nanjing, 210029, China

    Jin-Hua Liang, Wei-Ting Wang, Kai-Xin Du, Tong-Yao Xing, Yan Wang, Hui Wang, Lu Liu, Rui Guo, Yue Li, Hao-Rui Shen, Li Wang, Jian-Yong Li & Wei Xu

  4. Nanjing Geneseeq Technology Inc, Nanjing, Jiangsu, China

    Yang Shao & Junheng Liang

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  1. Jin-Hua Liang
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Contributions

JL and TW: performed the experiments, analyzed data, wrote the article and made the figures. KD: and TX: analyzed data. YW, HW, LL, RG, YS and JL: performed the experiments. YL, HS and LW: analyzed data. JL and WX: supervised the project, drafted and revised the paper.

Corresponding author

Correspondence to Wei Xu.

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Conflict of interest

The authors declare no competing interests.

Ethical approval

The patient gave consent which was approved by Institutional Review Boards of the First Affiliated Hospital of Nanjing Medical University.

Informed consent

The patient gave consent which was approved by Institutional Review Boards of the First Affiliated Hospital of Nanjing Medical University.

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Supplementary Information

Below is the link to the electronic supplementary material.

13577_2022_841_MOESM1_ESM.tif

Supplementary file1 Supplement Figure 1. Karyotype analysis and immunophenotype for this ENKTL patient (A) Karyotype analysis of this R/R ENKTL patient stimulated by CpG-oligodeoxynucleotide. (B) The immunophenotype of this R/R ENKTL patient by flow cytometry (FCM). (TIF 3847 KB)

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Liang, JH., Wang, WT., Du, KX. et al. Establishment and comprehensive analysis of a new human cell line (NK-NJ) with NK-cell characteristics established from extranodal natural killer cell lymphoma/leukemia. Human Cell 36, 835–846 (2023). https://doi.org/10.1007/s13577-022-00841-y

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