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Updated: Jun 9, 2025
Generation of Human Chimeric Antigen Receptor Regulatory T Cells
Published on: January 3, 2025
LIN28 upregulation in primary human T cells impaired CAR T antitumoral activity
Patricia Garcia-Rodriguez1,2, Laura Hidalgo1, Miguel Angel Rodriguez-Milla1
1Cellular Biotechnology Unit, Instituto de Salud Carlos III, Madrid, Spain.
Abstract:
LIN28, a highly conserved RNA-binding protein that acts as a posttranscriptional modulator, plays a vital role in the regulation of T-cell development, reprogramming, and immune activity in infectious diseases and T-cell-based immunotherapies. LIN28 inhibit the expression of let-7 miRNAs, the most prevalent family of miRNAs in lymphocytes. Recently it has been suggested that let-7 enhances murine anti-tumor immune responses. Here, we investigated the impact of LIN28 upregulation on human T cell functions, focusing on its influence on CAR T cell therapy. LIN28 lentiviral transduction of human T cells led to a stable expression of LIN28 that significantly downregulated the let-7 miRNA family without affecting cell viability or expansion potential. LIN28 overexpression maintained human T cell phenotype markers and functionality but impaired the antitumoral cytotoxicity of NKG2D-CAR T cells both in vitro and in vivo. These findings highlight the intricate relationship between LIN28/let-7 axis and human T cell functionality, including in CAR T cell therapy.
Insights
LIN28 upregulation in human T cells inhibits let-7 miRNAs, maintaining cell function but impairing CAR T cell therapy
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- LIN28 is a conserved RNA-binding protein regulating gene expression post-transcriptionally.
- LIN28 inhibits let-7 microRNAs (miRNAs), crucial for T cell development and immune responses.
- The let-7 miRNA family is implicated in enhancing anti-tumor immunity.
Purpose of the Study:
- To investigate the effect of LIN28 upregulation on human T cell functions.
- To assess the impact of LIN28 on CAR T cell therapy efficacy.
Main Methods:
- LIN28 lentiviral transduction of human T cells.
- Quantification of let-7 miRNA family expression.
- Assessment of T cell phenotype, viability, expansion, and anti-tumor cytotoxicity (in vitro and in vivo) of NKG2D-CAR T cells.
Main Results:
- Stable LIN28 expression was achieved without affecting T cell viability or expansion.
- LIN28 overexpression significantly downregulated the let-7 miRNA family.
- LIN28 maintained T cell phenotype and functionality but impaired NKG2D-CAR T cell anti-tumor cytotoxicity.
Conclusions:
- The LIN28/let-7 axis critically influences human T cell functionality.
- LIN28 upregulation negatively impacts the anti-tumor efficacy of CAR T cell therapy.
- Findings underscore the complexity of LIN28/let-7 interactions in T cell-based immunotherapies.
