Bicistronic retroviral vectors for combining myeloprotection with cell-surface marking

Area of Science:

• Gene Therapy
• Cancer Research
• Molecular Biology

Background:

• Developing effective cancer therapies requires strategies to overcome multidrug resistance.
• Cell-surface markers are crucial for tracking genetically modified cells in vivo and in vitro.

Purpose of the Study:

• To create a retroviral vector for coexpressing multidrug resistance 1 (MDR1) and truncated low-affinity nerve growth factor receptor (DeltaLNGFR).
• To optimize coexpression of MDR1 and DeltaLNGFR in hematopoietic cells for cancer drug resistance and cell tracking.

Main Methods:

• Constructed retroviral vectors based on the FMEV backbone with different coexpression strategies (splice signals, IRES, internal promoter).
• Evaluated vector performance using two-color flow cytometry and confocal laser microscopy.
• Transduced primary human CD34+ cells and assessed multidrug resistance and enrichment capabilities.

Main Results:

• The SF1mSdelta vector, utilizing retroviral splice signals, demonstrated the best coexpression correlation between MDR1 and DeltaLNGFR.
• Simultaneous expression of both genes was confirmed at the single-cell level.
• Transduced hematopoietic progenitor cells, including CD34+ cells, acquired multidrug resistance and could be enriched via DeltaLNGFR expression.

Conclusions:

• The SF1mSdelta vector enables efficient coexpression of cancer drug resistance and cell-surface markers in hematopoietic cells.
• This vector facilitates in vitro enrichment of chemotherapy-resistant cells without exposing them to cytotoxic agents.
• The developed vector system simplifies monitoring and analysis of chemotherapy-resistant cells in cancer research and therapy.