Modified SEVs with Surface OSM for Fibrosis Treatment
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Solution Overview
Problem
Current therapeutic strategies are inadequate for effectively addressing excessive fibrosis in conditions like cardiac fibrosis, where there is a need for modulation of extracellular matrix deposition and fibroblast activation to prevent tissue remodeling and heart failure.
Innovation Solution
Development of Small Extracellular Vesicles (SEVs) derived from genetically modified mesenchymal stromal cells that overexpress Oncostatin M (OSM) on their surface, enhancing their anti-fibrotic properties by modifying the OSM sequence to prevent intra and extracellular lysis and maintain stability, allowing targeted delivery and increased therapeutic potency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If SEVs are derived from genetically modified mesenchymal stromal cells to overexpress OSM on their surface, then anti-fibrotic therapeutic potency is enhanced, but device complexity and manufacturing difficulty increase
Solution Approach 1:
The patent modifies the OSM protein sequence by changing a single amino acid (arginine to glycine at position 116) to prevent proteolytic cleavage. This parameter change in the protein structure maintains the anti-fibrotic potency while enabling stable expression on SEV surface, thereby resolving the contradiction between therapeutic effectiveness and manufacturing complexity
Solution Approach 2:
The patent uses genetically modified mesenchymal stromal cells to produce SEVs that carry the modified OSM protein. Instead of directly administering the protein, the cells are engineered to continuously produce and secrete the therapeutic agent on their extracellular vesicles, simplifying the overall therapeutic delivery system while maintaining high potency
2Stability of the object's composition
If OSM sequence is modified to prevent intra and extracellular lysis, then protein stability is improved, but manufacturing precision requirements increase
Solution Approach 1:
The patent introduces a single point mutation (R116G) in the OSM sequence to eliminate the proteolytic cleavage site. This minimal parameter change achieves the goal of preventing protein degradation without requiring complex manufacturing modifications, thus improving stability while keeping manufacturing precision requirements manageable
Solution Approach 2:
The patent specifically targets and removes the vulnerable arginine residue at position 116 that is susceptible to proteolytic cleavage. By extracting this single critical amino acid and replacing it with glycine, the patent eliminates the weakness in the protein structure without altering the rest of the functional domains, thereby achieving high stability with minimal manufacturing complexity
3Adaptability or versatility
If SEVs are used to target fibrotic processes, then therapeutic specificity is improved, but loss of substance increases due to protein degradation
Solution Approach 1:
The patent modifies the OSM protein sequence by substituting arginine with glycine at position 116 to eliminate the proteolytic cleavage site. This parameter change prevents protein degradation in the extracellular environment, reducing substance loss while maintaining the targeted anti-fibrotic therapeutic specificity
Solution Approach 2:
The patent performs preliminary genetic modification of the OSM sequence before the protein is secreted on SEV surface. By pre-engineering the protein to be resistant to degradation, the patent ensures that the therapeutic substance remains stable and active throughout its therapeutic lifecycle, preventing loss of substance while maintaining therapeutic specificity
Data Source
Figure 1a~1c
Figure 1d
Figure 2
AI summary
The present invention refers to an isolated Small Extracellular Vesicle (SEV) with a mutated form of functional Oncostatin M expressed on its surface. Further, pharmaceutical compositions comprising one or more SEVs, as active ingredient, are also contemplated. The invention also refers to an in vitro method for producing the SEVs of the invention, comprising the steps of a) immortalizing a human MSC line by telomerase overexpression, b) introducing in the immortalized MSCs a vector expression constituted by a genetic construct, comprising the modified OSM sequence, selected from SEQ ID NO 1 or SEQ ID NO 2, fused to CD81 sequence, and an inducible promoter, which responds to doxycycline, c) culturing the genetically modified MSCs obtained in b) in a medium culture under conditions permitting their expansion, d) adding doxocycline to the culture medium, and e) Collecting Small Extracellular vesicles (SEVs) from the culture supernatant.