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Area of Science:

  • Immunology
  • Oncology
  • Biotechnology

Background:

  • Hepatocellular carcinoma (HCC) presents a poor prognosis with limited treatment avenues.
  • Alpha-fetoprotein (AFP) is a key biomarker for HCC, often overexpressed in malignant tissues compared to nonmalignant liver.
  • Cancer-specific overexpression of AFP suggests its potential as a target for novel immunotherapies.

Purpose of the Study:

  • To verify AFP expression across various human liver tissues.
  • To engineer an affinity-optimized T-cell receptor (TCR) specifically targeting AFP in tumors.
  • To develop a TCR capable of differentiating between varying AFP expression levels in malignant and nonmalignant cells.

Main Methods:

  • AFP expression analysis using database searches, quantitative PCR (qPCR), and immunohistochemistry (IHC).
  • In vitro mutagenesis and screening to generate and optimize a TCR against the AFP/HLA-A*02-restricted peptide (AFP158-166).
  • Functional validation of TCR-transduced T cells against diverse normal and tumor cell types using alanine scan (X-scan) and human leukocyte antigen (HLA) allele analysis.

Main Results:

  • Confirmed differential expression of AFP in normal, diseased, and malignant liver tissues.
  • Generated a TCR with high affinity and specificity for the target AFP peptide presented by HLA-A*02.
  • Demonstrated that TCR-transduced T cells can distinguish between varying levels of AFP antigen on different cell types.

Conclusions:

  • A combination of computational, in vitro, and cell biology approaches successfully optimized a TCR for enhanced affinity and function.
  • The developed TCR exhibits properties suitable for discriminating between cancer cells and nonmalignant cells based on AFP expression levels.
  • T cells engineered with this TCR represent a promising foundation for adoptive T-cell immunotherapy trials in HCC patients.