A targeting delivery system for effective genome editing in leukemia cells to reverse malignancy

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Hematology

Background:

• Therapy resistance and relapse are primary causes of mortality in acute myeloid leukemia (AML).
• Targeting specific cellular pathways, like CXCR4, presents a potential strategy to overcome resistance in AML.
• CRISPR-Cas9 genome editing offers precise tools for genetic modification in cancer therapy.

Purpose of the Study:

• To develop and evaluate a dual-targeting CRISPR-Cas9 genome editing nanosystem for CXCR4 knockout in AML.
• To investigate the efficacy of this nanosystem in reversing leukemia cell malignancy.
• To establish an ex vivo model using patient peripheral blood for treatment evaluation.

Main Methods:

• Construction of a dual-targeting nanosystem with MUC1 aptamer-alginate and T22-NLS peptide for CXCR4 targeting.
• Utilizing protamine complexed with CRISPR-Cas9 plasmid as the core for genome editing.
• In vitro studies on THP-1 cells and ex vivo studies on AML patient peripheral blood.

Main Results:

• The nanosystem successfully mediated CXCR4 knockout in vitro, inducing apoptosis, cell cycle arrest, and inhibiting migration/adhesion in THP-1 cells.
• Ex vivo studies confirmed effective CXCR4 knockout in leukemia cells from AML patients, with reduced protein and mRNA levels.
• The system demonstrated potential as a delivery vector for AML gene therapy and validated peripheral blood ex vivo studies.

Conclusions:

• The developed dual-targeting CRISPR-Cas9 nanosystem is a promising delivery vector for gene therapy in AML.
• CRISPR-Cas9 mediated CXCR4 knockout can reverse leukemia cell malignancy and overcome therapy resistance.
• Peripheral blood-based ex vivo studies provide an effective strategy for evaluating AML treatment efficiency.