Harnessing autophagy for cell fate control gene therapy

Area of Science:

• Immunology
• Cell Biology
• Gene Therapy

Background:

• T-Rapa cells are investigated in allogeneic hematopoietic cell transplantation.
• Lentiviral (LV) vectors are used for transgene delivery in gene therapy.
• Autophagy modulation is a potential strategy to enhance gene transfer efficiency.

Purpose of the Study:

• To investigate if rapamycin-induced autophagy can improve lentiviral (LV)-based transgene delivery to human T-Rapa cells.
• To assess the impact of rapamycin on T-Rapa cell phenotype and transgene expression.
• To evaluate the safety of this gene therapy approach using a cell-fate control system.

Main Methods:

• T-Rapa cells were treated with rapamycin and exposed to an LV vector encoding CD19-DTYMKΔ.
• Autophagy was assessed via mitochondrial autophagy, MAP1LC3B, and SQSTM1 expression.
• The effect of the autophagy inhibitor 3-MA was evaluated.
• Transgene expression and cell viability were assessed in vitro and in vivo (xenogeneic transplantation model).

Main Results:

• Rapamycin treatment induced mitochondrial autophagy and an antiapoptotic phenotype in T-Rapa cells, reversible by 3-MA.
• Changes in MAP1LC3B and SQSTM1 expression were observed, also reversed by 3-MA.
• Increased expression of the CD19-DTYMKΔ fusion protein was achieved despite reduced T-Rapa cell proliferation.
• Transgene-expressing cells showed susceptibility to AZT-induced cell death in vitro and in vivo.

Conclusions:

• Rapamycin induction of T cell autophagy facilitates LV-based gene therapy.
• The CD19-DTYMKΔ cell-fate control axis enhances the safety of T cell immunotherapy.
• This approach offers a promising strategy for improving the safety and efficacy of T cell gene therapy.