Humanized C5 and C3 Rodent Models for Complement System Evaluation
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Solution Overview
Problem
Current methods for evaluating therapeutic molecules targeting human C5 or C3 proteins in non-human animals are inadequate due to species-specificity issues, where these molecules do not interact effectively with endogenous C5 or C3 proteins, hindering the assessment of pharmacokinetics and pharmacodynamics and therapeutic efficacy.
Innovation Solution
Development of genetically modified rodents with humanized C5 and C3 genes, where the rodent genome replaces endogenous C5 or C3 genes with human C5 or C3 sequences, allowing for the expression of human C5 or C3 proteins under control of rodent regulatory elements, mimicking human protein concentrations and functions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If therapeutic molecules targeting human C5 or C3 proteins are evaluated in non-human animals with endogenous C5 or C3 proteins, then the evaluation can be performed in standard animal models, but the therapeutic molecules do not interact effectively with the endogenous proteins due to species-specificity
Solution Approach 1:
The patent applies parameter changes by replacing the C5 or C3 protein sequences in the rodent genome with human sequences. This changes the fundamental parameter of protein species-specificity, allowing human-specific therapeutic molecules to bind and interact with the humanized complement proteins in the rodent model, thereby enabling reliable evaluation of pharmacokinetics and pharmacodynamics.
Solution Approach 2:
The patent creates a copy of the human C5 or C3 gene and integrates it into the rodent genome. This copied human gene replaces the endogenous rodent gene, producing human complement proteins in the rodent system. This allows the rodent model to accurately reflect human biological responses to human-specific therapeutic molecules.
2Measurement precision
If human-specific C5 or C3 protein antagonists are used in non-human animal models, then the molecular mechanism can be studied, but the antagonists do not interact with the endogenous C5 or C3 proteins
Solution Approach 1:
The patent changes the species origin parameter of the C5 or C3 proteins from rodent to human by gene replacement. This enables human-specific antagonists to bind with high precision to their target human proteins in the rodent model, allowing accurate measurement of therapeutic efficacy while maintaining the versatility of rodent as the animal model organism.
Solution Approach 2:
The humanized C5 or C3 proteins serve as an intermediary that bridges the human-specific antagonists and the rodent physiological system. The antagonists interact with the humanized proteins, which are expressed in the rodent complement system, thereby mediating the transmission of therapeutic effects through the rodent model.
3Reliability
If endogenous C5 or C3 genes are replaced with human C5 or C3 genes in rodents, then accurate evaluation of human-specific therapeutics is enabled, but the genetic modification complexity increases
Solution Approach 1:
The patent extracts the endogenous rodent C5 or C3 genes from the genome and removes them, then replaces them with human C5 or C3 genes. This extraction approach allows for clean replacement while maintaining the native genomic locus and regulatory elements, simplifying the overall genetic modification process compared to adding transgenes to unrelated loci.
Solution Approach 2:
The patent utilizes the universal functionality of the rodent genomic locus structure, where the endogenous C5 or C3 gene loci can accommodate human gene sequences while maintaining native regulatory control. This multi-functional approach allows the same locus to serve both as the target for gene replacement and as the source of regulatory elements for the humanized gene.
Data Source
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AI summary
Non-human animals comprising a human or humanized C3 and/or C5 nucleic acid sequence are provided as well as methods for using the same to identify compounds capable of modulating the complement system. Non-human animals that comprise a replacement of the endogenous C5 gene and/or C3 gene with a human or humanized C5 gene and/or C3 gene, and methods for making and using the non-human animals, are described. Non-human animals comprising a human or humanized C5 gene under control of non-human C5 regulatory elements is also provided, including non-human animals that have a replacement of non-human C5-encoding sequence with human C5-encoding sequence at an endogenous non-human C5 locus. Non-human animals comprising a human or humanized C3 gene under control of non-human C3 regulatory elements is also provided, including non-human animals that have a replacement of non-human C3 protein-encoding sequence with human or humanized C3 protein-encoding sequence at an endogenous non-human C3 locus. Non-human animals comprising human or humanized C3 and/or C5 sequences, wherein the non-human animals are rodents, e.g., mice or rats, are provided.