rAAV Antisense CYP2J2 Vector for Tumor Inhibition

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current treatments for tumors lack effective methods to inhibit the proliferation and metastasis of tumor cells, as conventional drugs have limited success and gene therapy faces challenges with vector selection and immunogenicity.

Innovation Solution

A recombinant adeno-associated virus (rAAV) expressing antisense human CYP2J2 gene is developed, which is packaged and prepared using specific plasmids and the calcium phosphate precipitation technique, and transfected into tumor cells to inhibit CYP2J2 gene expression, thereby blocking the synthesis of epoxyeicosatrienoic acids and reducing tumor growth and metastasis.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional drugs are used for tumor treatment, then treatment simplicity is maintained, but treatment effectiveness is insufficient

Engineering Contradiction:

Improvetreatment effectivenessVSAvoidtreatment complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent replaces conventional chemical drug treatment with gene therapy using recombinant adeno-associated virus (rAAV) vectors. The virus delivers antisense CYP2J2 gene sequences into tumor cells, utilizing molecular biology mechanisms rather than traditional pharmacology to inhibit tumor proliferation and metastasis.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the fundamental mechanism of action from chemical inhibition to genetic modulation. By introducing antisense RNA that binds to and inhibits CYP2J2 gene expression, the treatment fundamentally alters the biological parameters controlling tumor growth, achieving superior effectiveness.

Inventive Principle:

Principle #35Parameter changes

2Reliability

If gene therapy vectors are used for tumor treatment, then treatment effectiveness is improved, but vector selection and immunogenicity issues arise

Engineering Contradiction:

Improvetreatment effectivenessVSAvoidimmunogenicity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent employs adeno-associated virus (AAV) vectors, which are non-pathogenic and do not replicate in human cells. The viral genome remains as a stable plasmid without producing harmful proteins, eliminating immunogenicity concerns while maintaining effective gene delivery and long-term expression.

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent converts the potential harm of viral vectors into a benefit by selecting AAV, which has lost its pathogenicity. The virus serves as an effective delivery vehicle without causing disease or immune reactions, turning a potentially harmful mechanism into a safe and effective treatment approach.

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

3Reliability

If CYP2J2 gene expression is inhibited in tumor cells, then tumor proliferation is reduced, but gene delivery efficiency must be maintained

Engineering Contradiction:

Improvetumor growth inhibitionVSAvoidgene delivery efficiency

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The patent uses rAAV vectors that can efficiently deliver genetic material to diverse cell types including tumor cells, endothelial cells, and other target cells. The vector system provides universal applicability across different tissue types while maintaining high delivery efficiency and stable long-term gene expression.

Inventive Principle:

Principle #6Universality (Multi-functionality)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The rAAV-anti2J2 significantly inhibits tumor cell proliferation, migration, and apoptosis, providing a promising approach for tumor treatment with minimal toxicity and long-term gene expression, overcoming limitations of existing gene therapy vectors.

Implementation Method 1

using calcium phosphate precipitation technique, pXX2, pXX6, and pXXUF1-anti2J2 are cotransfected into 293 cell to pack and prepare the recombinant adeno-associated virus expressing antisense human CYP2J2 (rAAV-anti2J2)

Methodology Applied

Scientific EffectCalcium phosphate precipitation: Precipitation

Data Source

PatentUS8273344B2Recombinant adeno-associated virus expressing human antisense gene CyP2J2 and its preparation methods

Publication Date: 2012.09.25 XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV

AI summary

A recombinant adeno-associated virus (rAAV) expressing antisense human CYP2J2 gene produced by a method of co-transfection into 293 cell by calcium phosphate precipitation, the rAAV having the following three plasmids: a) pXX2: a packing plasmid having a nucleotide sequence encoding AAV Rep and Cap protein, wherein the nucleotide sequence is operably linked upstream and downstream to a p5 promoter respectively to increase expression 15-fold; b) pXXUF1-anti2J2: an eukaryotic expression vector having a CMV promoter, a NotI site for insertion of target genes, the CYP2J2 gene oriented in the 3′ to 5′ direction, and inverted terminal repeats (ITRs); and c) pXX6: a helper plasmid deleted of the pathogenic gene sequences of adenovirus and having an E1A, an E2A, and a VA1 RNA gene of adenovirus. Also provided is a method of preparing the rAAV.