Dados do Resumo

Título

Enhancing Antitumor Immune Response in Gene Therapy through Cell Membrane-Coated Adenoviral Vectors Expressing Targeted Peptides

Introdução

Cancer remains a significant global health challenge, driving the need for innovative therapeutic strategies. Gene therapy utilizing oncolytic viruses is a promising avenue, with non-replicative adenoviral vectors encoding p19Arf and interferon-β (IFNβ) showing potential in inducing immunogenic cell death and anti-tumor immunity. However, these vectors require improved specificity towards tumor antigens to enhance their therapeutic efficacy.

Objetivo

This study develops a synthetic biology approach to coat adenoviral vectors with cell membranes displaying tumor antigens via the tANCHOR system. By presenting antigens like the OVA peptide, this method aims to enhance immune responses against tumor cells, improving cancer gene therapy.

Métodos

B16 cells were cultured and harvested for membrane extraction, which was analyzed using Nanosight. The LacZ-expressing adenovirus was then coated onto these cell membranes through extrusion. Efficiency was assessed via ONPG absorbance, XGAL staining, electron microscopy (EM), and dot blot analysis to confirm successful membrane coating. Oligonucleotides encoding the OVA peptide were synthesized and inserted into the linearized, purified pEXPO63.1 vector of the ANCHOR system using standard recombinant DNA techniques. The construct was validated by sequencing and restriction mapping. Confocal microscopy was employed to visualize the membrane anchoring of the peptide in transfected B16 cells.

Resultados

ONPG assays indicated that enveloped viral particles exhibit a considerable degree of transduction compared to uncoated viruses. Electron microscopy and dot blot assays confirmed successful viral envelope formation. Furthermore, the construct of plasmid tAnchor vector was verified by sequencing and restriction mapping, along with transfection in B16 cells, as confirmed by microscopy and flow cytometry.

Conclusões

We are actively optimizing the adenovirus coating process and anticipate soon demonstrating that the tANCHOR system can effectively present antigens and direct the immune response. While results are still pending, our efforts are concentrated on harnessing this approach's potential to enhance the specificity and efficacy of cancer immunotherapy.

Financiador do resumo

FAPESP (São Paulo State Research Support Foundation) for the scholarship that has facilitated this research. Grant Process: 2022/04622-5.

Palavras Chave

Cancer Adenovirus Gene Therapy