Engineered Meganucleases Targeting HAO1 Gene for Primary Hyperoxaluria Treatment
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Solution Overview
Problem
Primary hyperoxaluria type 1 (PH1) lacks effective therapeutic options, as current treatments primarily focus on managing symptoms and kidney transplantation, with no approved medications available to address the underlying genetic mutation causing excessive oxalate production.
Innovation Solution
Engineered meganucleases are designed to specifically bind and cleave the recognition sequence within the HAO1 gene, reducing HAO1 protein expression and increasing serum glycolate levels by introducing modifications such as insertions or deletions at the HAO 25-26 recognition site, thereby mitigating oxalate accumulation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional treatments (symptom management and transplantation) are used, then patient survival is maintained, but the underlying genetic cause of excessive oxalate production is not addressed
Solution Approach 1:
The patent extracts and eliminates the harmful function of the HAO1 gene by using engineered meganucleases to specifically target and disrupt the gene. The meganucleases recognize and cleave the HAO1 gene sequence, preventing the production of hydroxyacid oxidase 1 enzyme, thereby stopping the conversion of glycolate to glyoxylate and subsequently preventing excessive oxalate formation.
Solution Approach 2:
The patent changes the molecular parameters of the HAO1 gene by introducing specific mutations through meganuclease-mediated cleavage. This results in frameshift mutations or premature stop codons that alter the gene's reading frame and prevent production of functional protein, thereby changing the metabolic parameter of oxalate production from excessive to normal levels.
2Productivity
If no therapeutic intervention is applied, then the natural metabolic pathway operates, but oxalate accumulation causes kidney stones and nephrocalcinosis
Solution Approach 1:
The patent converts the harmful metabolic pathway into a beneficial one by blocking the production of glyoxylate (the harmful intermediate) through HAO1 gene disruption. This prevents the downstream conversion to oxalate, thereby eliminating the root cause of kidney stones and nephrocalcinosis while maintaining overall metabolic function through alternative pathways.
3Reliability
If liver-kidney transplantation is performed, then renal function is restored, but the genetic mutation causing PH1 remains and requires lifelong management
Solution Approach 1:
The patent applies preliminary action by using gene therapy to correct the underlying HAO1 mutation before it causes irreversible organ damage. By delivering meganucleases or nucleic acid sequences that disrupt the mutant HAO1 gene early in the disease course, the treatment prevents the progression to end-stage renal disease, thereby avoiding the need for transplantation and its associated lifelong management complexity.
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 engineered meganucleases effectively reduce oxalate levels and increase glycolate levels, providing a potential gene therapy approach for treating PH1 by disrupting the HAO1 gene function, thus alleviating the symptoms associated with excessive oxalate production.
Implementation Method 1
Homing endonucleases are a group of naturally-occurring nucleases which recognize 15-40 base-pair cleavage sites commonly found in the genomes of plants and fungi. They naturally promote homologous recombination or gene insertion at specific locations in the host genome by producing a double-stranded break in the chromosome
Implementation Method 2
They naturally promote homologous recombination or gene insertion at specific locations in the host genome by producing a double-stranded break in the chromosome, which recruits the cellular DNA-repair machinery
Implementation Method 3
The cleavage site is repaired by non-homologous end joining resulting in the modified target sequence
Data Source
AI summary
Disclosed are engineered meganucleases that bind and cleave a recognition sequence within a hydroxyacid oxidase 1 (HAO1) gene. The present invention also encompasses methods of using such engineered meganucleases to make genetically-modified cells. Further, the invention encompasses pharmaceutical compositions comprising engineered meganuclease proteins, or nucleic acids encoding engineered meganucleases of the invention, and the use of such compositions for treatment of primary hyperoxaluria type I (PH1).