Modified AAV-8 Capsid Insertion for Retinal Gene Delivery
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Solution Overview
Problem
Existing AAV-8 serotypes have low efficiency in translocating from the vitreous to the retina via intravitreal infusion, and there is a need for improved targeting and expression in ocular tissues to address ophthalmic diseases.
Innovation Solution
A modified AAV-8 capsid, AAV8-590RGD, is developed by inserting a 10-amino acid sequence comprising 7 random and 3 protective amino acids between positions 590 and 591 of the wild-type AAV-8 capsid, optimized for efficient infection of the retina and retinal pigment epithelial cells through intravitreal infusion.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If wild-type AAV-8 capsid is used for intravitreal infusion, then liver targeting is achieved, but retinal penetration efficiency is low
Solution Approach 1:
The patent applies local quality by introducing a specific 10-amino acid sequence (RGD motif) at a localized position (590-591) of the AAV-8 capsid protein. This localized modification enables the virus to specifically recognize and bind to integrin receptors on retinal cells, thereby improving retinal penetration efficiency without completely redesigning the entire capsid structure. The RGD sequence provides local functional enhancement for retinal targeting while maintaining the overall liver-tropic characteristics of AAV-8.
Solution Approach 2:
The patent employs parameter changes by modifying the amino acid sequence at specific positions (590-591) of the capsid protein. The introduction of the 10-amino acid sequence changes the biochemical parameters of the capsid, enabling new interaction capabilities with retinal cell receptors. This parameter modification allows the virus to maintain its basic AAV-8 properties while acquiring enhanced retinal penetration ability.
2Productivity
If AAV-8 is used for gene delivery, then liver cells are targeted, but immunogenicity increases
Solution Approach 1:
The patent applies local quality by introducing a specific 10-amino acid sequence (RGD motif) at a localized position (590-591) of the AAV-8 capsid protein. This localized modification enables the virus to specifically recognize and bind to integrin receivers on retinal cells, thereby improving retinal penetration efficiency without completely redesigning the entire capsid structure. The RGD sequence provides local functional enhancement for retinal targeting while maintaining the overall liver-tropic characteristics of AAV-8.
Solution Approach 2:
The patent employs parameter changes by modifying the amino acid sequence at specific positions (590-591) of the capsid protein. The introduction of the 10-amino acid sequence changes the biochemical parameters of the capsid, enabling new interaction capabilities with retinal cell receivers. This parameter modification allows the virus to maintain its basic AAV-8 properties while acquiring enhanced retinal penetration ability.
Data Source
AI summary
A modified vector of adenovirus-associated virus serotype 8 (AAV-8) for gene targeting and expression is provided, wherein the modified vector includes serotype coat amino acid sequence, insertion site and insertion amino acid sequence. A 10-amino acid sequence set forth in SEQ ID NO.3 is inserted between amino acids at positions 590 and 591 that are set forth in SEQ ID NO.2 of the AAV-8 serotype coat protein: LARGDSTKSA, wherein amino acids at positions 1, 2 and 10 are protective amino acids, and amino acids at positions 3 to 9 are screened amino acid sequences. In addition, the present invention also discloses construction method and application of the modified vector are also provided.