Zebrafish ApoB Reporter Screening for In Vivo Drug Discovery
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Solution Overview
Problem
Current methods for identifying modulators of Apolipoprotein B (ApoB) are limited by their inability to effectively screen across all cell and tissue types in vivo, which is crucial for understanding its role in metabolic diseases like diabetes and cardiovascular disease.
Innovation Solution
A high-throughput phenotypic screening method using larval zebrafish expressing an ApoB-reporter fusion protein, such as the ApoB-NanoLuc luciferase fusion, to monitor and modulate ApoB expression in a whole animal, allowing for the identification of compounds that enhance or inhibit ApoB levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If in vitro or cell-free systems are used to screen ApoB modulators, then the screening process is simpler and more controllable, but the ability to screen across all cell and tissue types in vivo is lost
Solution Approach 1:
The patent creates a transgenic zebrafish model with an ApoB-reporter fusion gene that copies the expression pattern of endogenous ApoB across all cell and tissue types. This reporter system allows in vivo screening to proceed with the versatility of whole-organism screening while maintaining the controllability and simplicity of in vitro assays, as the reporter provides a direct readout of ApoB expression levels throughout the organism.
2Adaptability or versatility
If in vivo screening in whole animals is performed, then comprehensive screening across all cell and tissue types is achieved, but the complexity of the screening system increases
Solution Approach 1:
The patent replaces complex anatomical and physiological systems with a simplified reporter-based readout. Instead of directly measuring ApoB expression through complex tissue analysis, the ApoB-reporter fusion provides a straightforward optical or biochemical signal that can be detected across the whole organism, dramatically reducing the complexity of the screening system while maintaining comprehensive screening capability.
3Productivity
If transgenic zebrafish with ApoB-reporter fusion are used, then high-throughput in vivo screening is enabled, but the complexity of generating and maintaining the transgenic model increases
Solution Approach 1:
The patent performs the complex work of generating the transgenic zebrafish model with the ApoB-reporter fusion gene in advance, before the actual high-throughput screening begins. This preliminary action creates a ready-to-use screening platform where the transgenic line can be propagated and used for multiple screening campaigns, amortizing the initial complexity investment across many high-throughput experiments.
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 enables the identification of viable pre-therapeutic leads for compounds that modulate ApoB levels, potentially treating metabolic diseases by providing a comprehensive in vivo screening method that maximizes the likelihood of finding effective drug targets.
Implementation Method 1
An in vivo high-throughput screen (HTS) for modulators of ApoB using transgenic zebrafish larvae carrying an optical reporter of ApoB has been carried out. The larval zebrafish is ideal for this study as it (i) recapitulates all major aspects of vertebrate metabolism in a small, rapidly developing organism, (ii) is the only vertebrate system conducive to HTS, and (iii) has a proven to be a powerful model for drug discovery owing to remarkably conserved physiology and pharmacology with humans.
Data Source
AI summary
We describe a high-throughput, phenotypic screening method for one or more modulator(s) of Apolipoprotein B (ApoB) in larval zebrafish. The modulator(s) may be enhancers or inhibitors of ApoB expression. This represents a remarkable opportunity to investigate drug targets in every cell and tissue type of a whole animal without bias, thus maximizing the likelihood of identifying viable pre-therapeutic leads for compounds or biologics in a subject (e.g., human).