Gene Editing for HIV Therapy Using HDR-Mediated Integration
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Solution Overview
Problem
Current anti-HIV therapies, such as CCR5 disruption, do not provide full resistance to HIV infection, especially for dual or X4-tropic viruses, and there is a need for alternative strategies to genetically modify cells for HIV resistance.
Innovation Solution
The development of methods and compositions for gene therapy that introduce mutations or donor sequences using nucleases like ZFNs, TALENs, and CRISPR/Cas systems to modify endogenous genes, including the CCR5 locus, to create cells resistant to HIV infection by enhancing HDR-mediated gene editing and introducing anti-HIV genes or proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If CCR5 disruption is used for HIV therapy, then resistance to R5-tropic HIV is improved, but resistance to dual or X4-tropic HIV is not achieved
Solution Approach 1:
The patent applies universality by designing gene editing approaches that can simultaneously target multiple HIV receptors (CCR5 and CXCR4) or introduce multi-functional anti-HIV genes. This allows a single therapy to provide broad-spectrum resistance against R5-tropic, X4-tropic, and dual-tropic HIV strains, making the treatment universally effective against diverse HIV variants rather than limited to a single tropism type.
2Duration of action of stationary object
If gene editing is applied to modify stem cells for HIV resistance, then long-term resistance is achieved, but the complexity of the treatment process increases
Solution Approach 1:
The patent applies preliminary action by performing gene editing on hematopoietic stem cells ex vivo before transplantation. The stem cells are pre-modified with anti-HIV genetic modifications in the laboratory, then transplanted into the patient. This preliminary genetic engineering ensures that all subsequent blood cells produced from these stem cells inherit the HIV resistance, providing long-term protection without requiring continuous treatment.
3Reliability
If donor sequences are introduced to enhance anti-HIV activity, then the level of HIV resistance is improved, but the precision of gene targeting is challenged
Solution Approach 1:
The patent applies the intermediary principle by using homology-directed repair (HDR) templates as mediators between the nuclease-induced double-strand break and the desired anti-HIV gene insertion. These donor sequences with homology arms serve as intermediaries that guide precise integration of anti-HIV genes at specific genomic loci, ensuring both high precision targeting and enhanced anti-HIV activity through the introduced donor sequences.
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
This approach allows for the modification of at least 50% of cells, including stem cells, to be resistant to HIV infection, providing a potential 'cure' by creating cells that can sustain an anti-HIV response and eliminating the need for HLA matching in transplants, with the modified cells retaining their resistance in secondary transplants.
Implementation Method 1
The nucleases act by creating double-stranded breaks (DSB) at a targeted DNA sequence
Implementation Method 2
the DNA break can be repaired instead using the cell's homology directed repair (HDR) pathways, where information is copied from a homologous 'donor sequence' that is also introduced into the cell
Implementation Method 3
if DSB repair occurs through the error-prone NHEJ pathway, the result can be small insertions and/or deletions of nucleotides at the break site
Data Source
AI summary
The present disclosure is in the field of genome engineering, particularly targeted integration of anti-HIV transgenes into the genome of a cell for the treatment and/or prevention of HIV.