Dataset Information

Protein palmitoylation and sphingolipid metabolism cooperatively 1 control regulated exocytosis in cytotoxic lymphocytes

ABSTRACT: The cytotoxic activity of lymphocytes is particularly dependent on the regulated and polarised delivery of lytic granules to infected or malignant cells. Although genetic and mechanistic studies have identified a number of factors that regulate exocytosis in cytotoxic lymphocytes, a systematic mapping of the relevant factors and their relationships is lacking. Here, using a genome-scale CRISPR knockout screen in human natural killer cells, we characterise a complex genetic network regulating cytotoxic granule exocytosis, with lipid metabolism and protein lipidation among the most prominent pathways. By combining global protein lipidation and membrane lipid composition studies, we further uncover the critical role of ZDHHC17 in SNAP23 palmitoylation and targeting of palmitoylated SNAP23 to the membrane for cytotoxic granule fusion and release. Using ODYA-17 labelling of Ctrl and sgZDHHC17 NK-92 cells followed by pulldown and proteomic analysis, we investigated the palmitoylome of the NK-92 model cell line. By comparing the abundance of ODYA-17 labelled proteins in the lysates of Ctrl and sgZDHHC17 NK-92 cells, we defined the molecules that depend on the activity of the palmitoyl transferase ZDHHC17.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Cell, Blood

SUBMITTER: Andrea Rukavina

LAB HEAD: Kaan Boztug

PROVIDER: PXD045419 | Pride | 2025-09-18

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	CC_KBo_135_Sep2021.pdStudy	Other
	CC_Kbo_135-M986-B04-B08-O536-P11898-1-CarboxyNorm-SunInd-20211104.pdResult	Other
	CC_Kbo_135-M986-B04-B08-O536-P11898-1-CarboxyNorm-SunInd-20211104_filtered.xlsx	Xlsx
	CC_Kbo_135-M986-B04-B08-O536-P11898-1.msf	Msf
	CC_Kbo_135-M986-B04-O536-P11898-1.raw	Raw

Items per page:

1 - 5 of 12

Similar Datasets

Project description:NK-92 cells and their CAR-modified derivatives exhibit strong cytotoxic activity against tumors and are promising "off-the-shelf" therapeutics compared to CAR-T cells. In order to ensure safety and prevent the occurrence of secondary tumors, (CAR-)NK-92 cells need to be inactivated before transfusion. With increasing clinical use, there is a growing demand for enhanced safety and an efficient method that stops cell proliferation, but better maintains the cytotoxic effector functions of the cells compared to commonly used gamma irradiation. Recently, low-energy electron irradiation (LEEI) was successfully applied for inactivation of bacteria and viruses for vaccine production, and was described as a method for NK-92 inactivation for the first time. In the present publication, data on extensive characterization of LEEI and its comparison with gamma irradiation for the inactivation of parental NK-92 cells as well as CD123-directed CAR-NK-92 cells are provided. Our results show that both irradiation methods cause a progressive decrease in cell viability and are therefore suitable for inhibition of proliferation. Notably, cytotoxicity of the NK cells was significantly reduced by gamma irradiation, but not by LEEI three days after irradiation, compared to non-irradiated cells. Both gamma irradiation as well as LEEI led to substantial DNA-damage and an accumulation of irradiated cells in the G2/M phase. In addition, transcriptomic analysis of cells irradiated with both methods revealed approximately 12-fold more differentially expressed genes after gamma irradiation compared to LEEI. Analysis of surface molecules revealed an irradiation-induced decrease in CD56 expression but no changes in the levels of the activating receptors NKp46, NKG2D, and NKp30. Conclusions: The present data show that LEEI efficiently inactivates (CAR )NK-92 cells and sustains their cytotoxic capacity better than gamma irradiation. Taking into account additional logistic advantages of LEEI proves a potent alternative in the manufacturing of (CAR-)NK-92 cells for clinical application.

Dataset Information

Protein palmitoylation and sphingolipid metabolism cooperatively 1 control regulated exocytosis in cytotoxic lymphocytes

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets