Characterization of HIF-1α Knockout Primary Human Natural Killer Cells Including Populations in Allogeneic Glioblastoma
Abstract
:1. Introduction
2. Results
2.1. Induction of HIF1α Knockout Human PB-Derived NK Cells
2.2. Anti-Tumor Effects of HIF-1α Knockout Human Primary NK Cells Including Populations on Allogeneic GBM Cells in Normoxic and Hypoxic Conditions
2.3. HIF-1α Knockout Human NK Cells Including Populations Enhances Apoptosis Induction of Spheroids Derived from Allogeneic GBM Cells in Hypoxic Conditions
2.4. RNA-Sequencing (RNAseq) Analysis of HIF-1α Knockout NK Cells Including Populations in Hypoxic Conditions
3. Discussion
4. Materials and Methods
4.1. Ethics
4.2. Cell Lines
4.3. Single Guide RNAs
4.4. Induction of HIF-1α Knockout NK Cells Including Populations from Human PB
4.5. Efficacy of CRISPR/Cas9 Gene Disruption
4.6. RNAseq
4.7. Western Blotting
4.8. Growth Inhibition Assays
4.9. GBM Spheroid Invasion Assay
4.10. Cytotoxicity Assays for Spheroids
4.11. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Nakazawa, T.; Morimoto, T.; Maeoka, R.; Yamada, K.; Matsuda, R.; Nakamura, M.; Nishimura, F.; Yamada, S.; Park, Y.-S.; Tsujimura, T.; et al. Characterization of HIF-1α Knockout Primary Human Natural Killer Cells Including Populations in Allogeneic Glioblastoma. Int. J. Mol. Sci. 2024, 25, 5896. https://doi.org/10.3390/ijms25115896
Nakazawa T, Morimoto T, Maeoka R, Yamada K, Matsuda R, Nakamura M, Nishimura F, Yamada S, Park Y-S, Tsujimura T, et al. Characterization of HIF-1α Knockout Primary Human Natural Killer Cells Including Populations in Allogeneic Glioblastoma. International Journal of Molecular Sciences. 2024; 25(11):5896. https://doi.org/10.3390/ijms25115896
Chicago/Turabian StyleNakazawa, Tsutomu, Takayuki Morimoto, Ryosuke Maeoka, Kengo Yamada, Ryosuke Matsuda, Mitsutoshi Nakamura, Fumihiko Nishimura, Shuichi Yamada, Young-Soo Park, Takahiro Tsujimura, and et al. 2024. "Characterization of HIF-1α Knockout Primary Human Natural Killer Cells Including Populations in Allogeneic Glioblastoma" International Journal of Molecular Sciences 25, no. 11: 5896. https://doi.org/10.3390/ijms25115896