AAV Vector Hybrid HGF Gene Intron Removal

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Solution Overview

Problem

Current gene delivery systems for hepatocyte growth factor (HGF) isoforms, such as flHGF and dHGF, face inefficiencies in delivery and expression, particularly when using adeno-associated virus (AAV) vectors, limiting their therapeutic potential for conditions like diabetic peripheral neuropathy and amyotrophic lateral sclerosis.

Innovation Solution

A downsized mutant hybrid HGF gene sequence is introduced into an AAV vector, optimizing nucleotide sequence delivery and expression efficiency by removing parts of the intron 4 fragment, enhancing gene delivery and protein expression compared to previous sequences like HGF-X8.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If a full-length hybrid HGF gene is used for gene therapy, then the complete HGF isoforms (flHGF and dHGF) can be expressed, but the gene delivery efficiency using AAV vectors is limited due to the large size of the gene sequence

Engineering Contradiction:

Improvegene delivery efficiencyVSAvoidgene sequence size

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent extracts and removes the intron 4 fragment from the hybrid HGF gene sequence, creating a downsized version that can be efficiently delivered by AAV vectors while maintaining the ability to express functional HGF isoforms through selective splicing

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent modifies the nucleotide sequence parameters of the hybrid HGF gene by removing specific intronic regions, changing the physical size of the gene while preserving its functional capacity to produce flHGF and dHGF isoforms through alternative splicing mechanisms

Inventive Principle:

Principle #35Parameter changes

2Productivity

If the hybrid HGF gene is downsized by removing intron 4 fragment, then gene delivery efficiency is improved, but there may be concerns about maintaining proper splicing for flHGF and dHGF isoform production

Engineering Contradiction:

Improvegene delivery and expression efficiencyVSAvoidisoform expression accuracy

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent segments the hybrid HGF gene into distinct functional regions, removing only the intron 4 fragment while preserving the essential splicing sites and regulatory elements needed for producing both flHGF and dHGF isoforms through alternative splicing

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent designes the downsized hybrid HGF gene to include necessary splicing regulatory elements that act as intermediaries, enabling the splicing machinery to correctly produce both full-length and deleted HGF isoforms despite the removal of intron 4

Inventive Principle:

Principle #24Intermediary (Mediator)

Data Source

PatentUS11634728B2Adeno-associated virus (AAV) vector having hybrid HGF gene introduced thereto

Publication Date: 2023.04.25 HELIXMITH CO LTD

AI summary

The present invention relates to an AAV vector carrying a predetermined hybrid HGF gene sequence. Use of the AAV vector of the present invention allows a hybrid HGF gene to be delivered to a subject at a high delivery yield.