# Extracellular vesicles delivering TIMP-2 modulate MMP-1, MMP-2, and MMP-9 expression in human lung adenocarcinoma A549 cells

## TL;DR

This study explores using extracellular vesicles to deliver the TIMP-2 gene to lung cancer cells, which may help control cancer growth by modulating specific enzymes.

## Contribution

The novel contribution is demonstrating that EVs loaded with TIMP-2 can modulate specific matrix metalloproteinases in lung cancer cells.

## Key findings

- Electroporation effectively delivers plasmid DNA into extracellular vesicles.
- TIMP-2-loaded EVs significantly modulate MMP-2 and MMP-9 expression in A549 cells.
- The TIMP-2 cargo does not broadly alter MMP-1 levels in these cells.

## Abstract

Extracellular vesicles (EVs) carrying therapeutic cargos represent a promising strategy for cancer treatment by enabling the targeted delivery of genetic material directly to cancer cells. This study aimed to evaluate the effect of EVs loaded with the TIMP-2 gene on the expression of matrix metalloproteinases (MMPs 1, 2, and 9) in lung cancer cells (A549).

EVs derived from A549 cells were isolated by gradient centrifugation and ultracentrifugation. The coding sequence for TIMP-2 (tissue inhibitor of metalloproteinases 2) was amplified by PCR using cDNA derived from HUVEC cells. As-constructed plasmid (pTIMP-2) was introduced into the EVs by electroporation, and then the pTIMP-2-implanted EVs were subjected to PCR and NTA analysis. Additionally, the activity of MMP-1, MMP-2, and MMP-9 was determined by voltammetry in intact A549 cells and in A549 culture media.

Electroporation was found to demonstrate a good potential as an exogenous technique for uploading plasmid DNA into EVs. The results demonstrated that the as-uploaded EVs carrying the pTIMP-2 gene cargo do not broadly alter the overall balance of MMP-1 in pristine A549 cells. However, pTIMP-2-loaded EVs significantly modulate MMP-2 and MMP-9 expression in these cells, highlighting their potential as biological therapeutic moieties.

Our findings suggest a rational approach for exploring EV-based gene transfer targeting MMPs in lung cancer.