Advances in Next-Generation Sequencing and Single-Cell Transcriptomics for Profiling NK Cell Heterogeneity and Functional Diversity in Cancer

Natural Killer (NK) cells, as the prototypic member of innate lymphocytes, play a key role in recognizing and killing malignant cells without the need for prior desensitization. NK cell-based therapies have been considered as a promising approach for the treatment of a wide range of hematological and solid malignancies. However, identification and characterization of highly functional populations of NK cells is essential for the advancement of this field. Recent advances in next-generation sequencing (NGS) and single-cell transcriptomics have enabled high-resolution profiling of NK cell diversity at the molecular level. This review explores methods for assessing NK cell function, including chromium-51 release assays, flow cytometry, impedance-based cell analysis systems, microfluidic platforms, bioluminescence imaging, PET, and multiplex imaging systems. Applications of NGS in NK cell research, including KIR receptor genotyping, exome and genome sequencing, and RNA-seq, are also discussed. Herein we examined the impact of NK cell heterogeneity on cancer treatment strategies and personalized therapeutic approaches, and analyzes the challenges and future directions in this field.