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Structural, functional, and immunogenicity implications of F9 gene recoding.

Upendra K Katneni1, Aikaterini Alexaki1, Ryan C Hunt1

  • 1Division of Plasma Protein Therapeutics, Hemostasis Branch, Office of Tissues and Advanced Therapies, Center for Biologics Evaluation & Research, US Food and Drug Administration (FDA), Silver Spring, MD.

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|April 12, 2022
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Summary
This summary is machine-generated.

Codon optimization for recombinant factor IX (FIX) in hemophilia B treatment may alter protein structure, function, and immunogenicity. This gene recoding technique can lead to unintended changes, impacting therapeutic protein safety and efficacy.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Immunology

Background:

  • Hemophilia B is a bleeding disorder due to deficient coagulation factor IX (FIX).
  • Recombinant FIX proteins and gene therapies are used for treatment.
  • Codon optimization aims to enhance protein expression but assumes no change in protein characteristics.

Purpose of the Study:

  • To investigate the impact of gene recoding via codon optimization on F9 (FIX) protein properties.
  • To explore potential alterations in protein structure, function, and immunogenicity.
  • To assess the implications for therapeutic recombinant protein development.

Main Methods:

  • Evaluation of multiple recoded F9 variants.
  • Ribosome profiling to assess translation kinetics.
  • MHC-associated peptide proteomics to analyze peptide presentation.
  • Posttranslational modification analysis.

Main Results:

  • Recoded F9 variants exhibited altered conformations and distinct local translation kinetics.
  • Differential presentation of FIX-derived peptides bound to MHC class II molecules was observed.
  • Overexpression from gene recoding led to suboptimal posttranslational processing.

Conclusions:

  • Gene recoding of F9 can lead to unintended changes in protein structure, function, and immunogenicity.
  • Codon optimization may pose risks for therapeutic recombinant proteins.
  • Findings have broad implications for other gene-recoded therapeutic proteins.