Induction of stress granules alleviates programmed cell death induced by lysosomal damage during NK cell cryopreservation PMID: 42098076

ADC Targets in Oncology

2026-04-10

Induction of stress granules alleviates programmed cell death induced by lysosomal damage during NK cell cryopreservation

Cell Death Discov. 2026 May 7;12(1):286. doi: 10.1038/s41420-026-03149-0.

Yang Liu ^# ¹ ² , Xiaolong Liu ^# ¹ ² , Guangyuan Wu ^# ¹ ² , Xin Wang ¹ ² , Kuo Yu ³ , Hailong Wang ¹ ² , Xin Sun ¹ ⁴ , Zhongyao An ¹ ² , Hongrui Xu ¹ ⁵ , Linlin Zhao ⁶ , Ce Shi ¹ ² , Song Guo Zheng ⁷ , Mingli Huang ⁸ ⁹ , Zhiren Zhang ¹⁰ ¹¹ ¹² , Zhenkun Wang ¹³ ¹⁴

Affiliations

1. NHC Key Laboratory of Cell Transplantation, Harbin Medical University, Harbin, China.
2. Central Laboratory, First Affiliated Hospital, Harbin Medical University, Harbin, China.
3. Key Laboratory of Animal Cellular and Genetics Engineering of Heilongjiang Province, College of Life Science, Northeast Agricultural University, Harbin, China.
4. Obstetrical Department, First Affiliated Hospital, Harbin Medical University, Harbin, China.
5. Department of Child Dental, First Affiliated Hospital, Harbin Medical University, Harbin, China.
6. Department of Blood Transfusion, First Affiliated Hospital, Harbin Medical University, Harbin, China.
7. Department of Immunology, School of Cell and Gene Therapy, Songjiang Research Institute, Shanghai Songjiang District Central Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
8. NHC Key Laboratory of Cell Transplantation, Harbin Medical University, Harbin, China. [email protected].
9. Obstetrical Department, First Affiliated Hospital, Harbin Medical University, Harbin, China. [email protected].
10. NHC Key Laboratory of Cell Transplantation, Harbin Medical University, Harbin, China. [email protected].
11. Central Laboratory, First Affiliated Hospital, Harbin Medical University, Harbin, China. [email protected].
12. Department of Cardiology and Pharmacy and Breast Cancer Surgery, Harbin Medical University Cancer Hospital, Institute of Metabolic Disease, Heilongjiang Academy of Medical Science, Heilongjiang Key Laboratory for Metabolic Disorder and Cancer Related Cardiovascular Diseases, Harbin, China. [email protected].
13. NHC Key Laboratory of Cell Transplantation, Harbin Medical University, Harbin, China. [email protected].
14. Central Laboratory, First Affiliated Hospital, Harbin Medical University, Harbin, China. [email protected].
# Contributed equally.

Abstract

Natural killer (NK) cell-based therapies are under assessment for the treatment of various cancers due to their intrinsic ability to distinguish between malignant and healthy cells in an allogeneic context, enabling off-the-shelf manufacturing possibilities. However, cryopreservation reduces both the recovery and function of NK cells, thereby limiting their therapeutic feasibility. In this study, we evaluated three cryoprotectants (CryoStor 10; ZKCELL FM-01; FBS + DMSO) for the cryopreservation of NK cells. Post-thaw viability, ATP levels, and cytotoxicity were assessed and found to have persistent differences between cryopreserved and fresh cells. Transmission electron microscopy, flow cytometry, and Western blot analysis revealed a complex mode of cell death in cryopreserved cells, which could be partially mitigated by adding some death inhibitors. We further investigated the effects of centrifugation on thawed cells, identifying lysosomal stability as a key determinant of cell death. Pretreatment with low-dose LLOMe prior to cryopreservation induced stress granule formation, stabilizing lysosomes and improving cell recovery rates without compromising effector functional capacity. These findings offer new insights for optimizing NK cell cryopreservation and facilitating their clinical application.