Stabilizing Poxvirus Transgenes via Nucleotide Shuffling

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Solution Overview

Problem

Recombinant poxviruses used in cancer immunotherapy face instability issues during successive passaging, leading to decreased expression of MUC1 and CEA transgenes, which hinders production and purity of cancer vaccines.

Innovation Solution

The development of recombinant poxviruses with modified MUC1, CEA, and TRICOM transgenes, where the MUC1 peptide has shuffled and codon-optimized Variable N-Terminal Repeat (VNTR) domains, and CEA and TRICOM nucleic acids have specific nucleotide substitutions in repetitive regions, enhancing genetic stability through successive passaging.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If recombinant poxviruses are used for expressing MUC1 and CEA transgenes in cancer immunotherapy, then therapeutic efficacy is improved, but genetic stability through successive passaging deteriorates

Engineering Contradiction:

Improvetherapeutic efficacyVSAvoidgenetic stability

Core Design Contradiction:

ReliabilityVSStability of the object's composition

Solution Approach 1:

The patent applies parameter changes by modifying nucleotide sequences in repetitive regions of transgenes (MUC1, CEA, and TRICOM) to reduce homology and prevent recombination events. This includes changing codon usage and modifying repetitive nucleotide patterns while maintaining the encoded protein sequences, thereby improving genetic stability without compromising therapeutic efficacy

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent creates optimized copies of the transgenes with modified nucleotide sequences. These are 'copy' versions of MUC1, CEA, and TRICOM that have been redesigned at the nucleotide level to eliminate instability-causing repetitive patterns while preserving the functional protein-coding sequences, allowing stable propagation through multiple passages

Inventive Principle:

Principle #26Copying

2Reliability

If transgenes are expressed in recombinant poxviruses, then vaccine immunogenicity is improved, but manufacturing precision deteriorates due to decreased expression consistency

Engineering Contradiction:

Improvevaccine immunogenicityVSAvoidexpression consistency

Core Design Contradiction:

ReliabilityVSManufacturing precision

Solution Approach 1:

The patent modifies nucleotide parameters in the transgene sequences, specifically changing repetitive nucleotide patterns and codon usage to eliminate recombination-prone regions. This ensures consistent gene expression across multiple passages, improving manufacturing precision while maintaining immunogenicity

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent performs preliminary optimization of the nucleotide sequences before virus production and passaging. By pre-modifying the transgenes to eliminate instability-causing patterns, the patent ensures consistent expression from the outset, preventing degradation of expression consistency during subsequent manufacturing and passaging steps

Inventive Principle:

Principle #10Preliminary action

Data Source

PatentUS11654189B2Compositions and methods for enhancing the stability of transgenes in poxviruses

Publication Date: 2023.05.23 BAVARIAN NORDIC AS

AI summary

Provided herein are recombinant poxviruses that are stable through successive passaging of the recombinant poxviruses. More particularly, the recombinant poxviruses comprise one or more modified nucleic acids encoding MUC1, CEA, and/or TRICOM antigens, wherein the recombinant poxviruses are stable through successive passaging. Also, provided herein are compositions and method related thereto.