Dataset Information

Adaptive design of mRNA-loaded extracellular vesicles for targeted immunotherapy of cancer

ABSTRACT: Extracellular vesicles (EVs) have emerged as promising delivery vehicles for RNA-based therapeutics because of their advantages over other mRNA delivery systems, including their excellent biosafety and biocompatibility, stability against degradation, and the ability to cross physiological barriers such as the blood-brain barrier. In this study, we report a nanosecond electroporation (nsEP) system with a microfluidic configuration. Applying millisecond and nanosecond pulses separately shifted the main impact of electroporation from the cell membrane to the membrane structure of cellular organelles. After being treated by a nsEP system, mouse embryonic fibroblasts (MEFs) demonstrated the capability to generate large quantities of small extracellular vesicles (sEVs). To gain a better understanding of the biological mechanisms underlying this nsEP-triggered sEV release, we conducted a proteomics analysis.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Fibroblast

SUBMITTER: Shiyan Dong

LAB HEAD: Wen Jiang

PROVIDER: PXD045239 | Pride | 2023-09-10

REPOSITORIES: Pride

ACCESS DATA

Similar Datasets

Project description:Electroporation is a versatile method for in vitro or in vivo delivery of different molecules into cells. However, no study so far has analysed the effects of electric pulses used in electrochemotherapy (ECT pulses) or electric pulses used in electrogene therapy (EGT pulses) on malignant cells. We studied the effect of ECT and EGT pulses in human malignant melanoma cells in vitro in order to understand and predict possible effect of electric pulses on gene expression and their possible effect on cell behaviour. We used microarrays with 2698 different oligonucleotides to obtain expression profile of genes involved in apoptosis and development of cancer in a malignant melanoma cell line (SK-MEL28) exposed to ECT pulses and EGT pulses. Cells exposed to ECT pulses showed 68.8% average survival rate and 31.4% average survival rate in cells exposed to EGT pulses. Only seven common genes were found differentially expressed in cells 16 h after exposure to ECT and EGT pulses. We found that ECT and EGT pulses induce HSP70 stress response mechanism, repress histone protein H4, a major protein involved in chromatin assembly, and down-regulate components involved in protein synthesis. Our results show that electroporation does not significantly change expression profile of major tumour suppressor genes or oncogenes of cell cycle. Moreover, electroporation also does not changes the expression of genes involved in stability of DNA, supporting the current evidence that electroporation is a safe method that does not promote tumorigenesis. However, in spite of being considered an isothermal method it does to some extent induce stress that resulted in expression of environmental stress response mechanism, HSP70. The difference in expression of genes involved in development of cancer was obtained by comparison of malignant melanoma cells exposed to EGT or ECT pulses against the same untreated malignant melanoma cells. In our experimental design, microarrays with 2698 different genes were used as a dual colour system in which exposed and non-exposed cellsâ mRNA were separately labelled, mixed and hybridised together on each array. Only microarrays expressing at least 50% of genes were used for further analysis. All oligonucleotides on the same array were spotted in quadruplicates and each microarray analysis was performed in duplicates, therefore obtaining eight measurements of the same oligonucleotide. The acquired data were analysed with Acuity 4.0 to select reliable signals. Only genes, present in both duplicated microarrays were considered for further processing.

Dataset Information

Adaptive design of mRNA-loaded extracellular vesicles for targeted immunotherapy of cancer

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure