Exosome RNA Therapeutics Loading and Stabilization
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Solution Overview
Problem
Current methods for delivering nucleic acid-based therapeutics, such as mRNA, face challenges with rapid clearance, nuclease activity, limited organ-specific distribution, and low cellular uptake, particularly for non-hepatic targets, and existing systems like TAMEL struggle with efficient loading and bioactive delivery of nucleic acids into target cells due to high affinity binding of RNA-binding proteins, leading to instability and immunological responses.
Innovation Solution
The development of novel extracellular vesicle (EV) engineering technology that utilizes polyA binding proteins and fusion polypeptides to efficiently load and stabilize nucleic acid cargo, ensuring high loading efficiency, stability, and controlled release into target cells, while avoiding immunological responses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If high affinity RNA-binding proteins are used to load nucleic acids into EVs, then loading efficiency is improved, but nucleic acid release and translation are prevented
Solution Approach 1:
The patent changes the binding affinity parameter by using engineered RNA-binding proteins with tunable, moderate affinity instead of very high affinity proteins. This allows the nucleic acids to be loaded efficiently into EVs while maintaining the ability to release and translate upon delivery to target cells
Solution Approach 2:
The patent introduces engineered RNA-binding proteins as intermediaries that facilitate nucleic acid loading into EVs without permanently sequestering the nucleic acids. These proteins act as temporary carriers that can release their cargo under appropriate conditions, enabling both efficient loading and subsequent translation
2Speed
If nucleic acids are delivered without stabilization, then delivery speed is improved, but half-life and stability are reduced
Solution Approach 1:
The patent applies preliminary stabilization actions by engineering nucleic acids with enhanced stability features before delivery. This includes modifying nucleic acid structures and using stabilized formulations that protect against degradation while maintaining rapid delivery capabilities
Solution Approach 2:
The patent creates composite delivery systems combining nucleic acids with stabilizing components within EVs. This composite approach provides both rapid delivery and extended half-life by protecting the nucleic acid cargo through the EV structure and associated proteins
3Reliability
If specialized delivery vehicles are used for nucleic acid delivery, then cellular uptake and distribution are improved, but device complexity increases
Solution Approach 1:
The patent employs extracellular vesicles as natural, self-organizing delivery vehicles that leverage cellular machinery for their formation and delivery. This self-service approach reduces the need for complex external delivery systems while maintaining effective cellular uptake and distribution
Solution Approach 2:
The patent uses extracellular vesicles as universal delivery platforms that can transport multiple types of nucleic acid therapeutics (mRNA, siRNA, etc.) to various target cells and tissues. This multi-functional approach simplifies the overall delivery system architecture while maintaining versatility
4Strength
If MS2 protein is used for RNA binding, then binding affinity is improved, but RNA release is prevented
Solution Approach 1:
The patent changes the binding strength parameter by using engineered RNA-binding proteins with moderate, tunable affinity rather than the very high affinity of MS2 protein. This parameter adjustment enables both efficient loading and subsequent release of RNA cargo
Solution Approach 2:
The patent inverts the binding strategy by using proteins that bind RNA with appropriate strength for loading but allow release under physiological conditions. Instead of using extremely tight-binding proteins like MS2, the patent employs proteins designed for reversible, controlled binding
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 achieves significant improvements in the loading and bioactive delivery of nucleic acid therapeutics, enhancing their stability, efficacy, and translation efficiency within target cells, with up to 95% of EVs loaded with nucleic acid cargo and increased half-life, leading to more effective therapeutic outcomes.
Implementation Method 1
at least one polyA binding protein (PABP) or a fragment or domain thereof and at least one NA cargo molecule comprising a contiguous stretch of adenine nucleotides
Implementation Method 2
PCT/GB2017/051479 teaches improved methods for endogenous loading of various types of RNA therapeutics with the aid of RNA-binding proteins, which drag RNA of interest into EVs forming within parental cells
Data Source
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AI summary
The present invention pertains to extracellular vesicle (EV) therapeutics, wherein the EVs comprise nucleic acid (NA)-based therapeutics such as mRNAs, circular RNAs, miRNAs, shRNAs, and/or DNA molecules. The NA therapeutics are loaded into EVs using inventive protein and NA engineering strategies which stabilize the cargo NAs and enhance loading into EVs, thereby enhancing therapeutic activity of the cargo NA molecules half-life.