Masked TGF-Beta Polypeptide Constructs for Local Signaling Control
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Solution Overview
Problem
Existing approaches to regulate TGF-β signaling are limited in effectively modulating its activity to treat diseases such as autoimmune disorders and inflammation, with potential adverse effects on cells and tissues.
Innovation Solution
Development of masked TGF-β constructs and complexes using a scaffold polypeptide and masking polypeptide sequences to control TGF-β activity, allowing reversible masking and binding to heteromeric cell surface receptors, with sequence variations to prevent off-target interactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If TGF-β activity is directly activated to treat autoimmune disorders and inflammation, then therapeutic effectiveness is improved, but adverse effects on cells and tissues increase
Solution Approach 1:
The patent uses a masking polypeptide as an intermediary between the TGF-β signal and the target cells. This masking polypeptide temporarily binds to the TGF-β, preventing direct interaction with cells until the complex reaches the target tissue, where proteolytic cleavage releases the active TGF-β locally. This mediator approach ensures therapeutic effectiveness while minimizing systemic adverse effects on non-target cells and tissues.
Solution Approach 2:
The TGF-β is pre-complexed with the masking polypeptide in an inactive state before administration. This preliminary masking action protects the TGF-β from premature activation and allows controlled delivery to target sites. The active TGF-β is only released when needed at the target tissue through proteolytic cleavage by matrix metalloproteinases or other proteases present in the inflammatory microenvironment.
2Reliability
If TGF-β signaling is strongly activated to achieve therapeutic outcomes, then disease treatment effectiveness is improved, but off-target interactions and unwanted effects increase
Solution Approach 1:
The patent employs protease-specific cleavage sites in the masking polypeptide that are selectively activated by proteases present in the inflammatory microenvironment (such as matrix metalloproteinases MMP-2, MMP-9, or cathepsins). This creates local quality control where TGF-β is only activated in tissues with high protease activity associated with inflammation, ensuring therapeutic effectiveness at target sites while preventing off-target interactions in healthy tissues with lower protease activity.
Solution Approach 2:
The patent utilizes changes in the local biochemical environment (protease activity levels, pH, redox state) at the disease site to trigger selective activation of TGF-β. The masking polypeptide is designed with cleavage sites that respond to these parameter changes, allowing the TGF-β to remain inactive during circulation and only become active when environmental parameters indicate the presence of disease-associated proteases, thereby avoiding off-target effects.
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 masked TGF-β constructs and complexes provide controlled TGF-β signaling, reducing adverse effects and enhancing therapeutic outcomes in treating autoimmune and inflammatory diseases.
Implementation Method 1
wherein the masking polypeptide sequence and the TGF-β polypeptide sequence bind to each other
Implementation Method 2
wherein the intersspecific dimerization sequence and the counterpart intersspecific dimerization sequence interact with each other
Implementation Method 3
The TGF-β may become activated by the action of such proteases that release the latent complex from the matrix, which is followed by proteolysis of the LAP to release TGF-β to its receptors
Implementation Method 4
Activation appears to occur by inducing conformational changes to the latent TGF-β1 complex and hence releasing the active TGF-β1
Implementation Method 5
ROS are thought to alter the interaction between LAP and TGF-β, leading to its activation
Implementation Method 6
TSP-1 is believed to activate latent TGF-β by forming direct interactions with the latent TGF-β complex and preventing it from binding to the matured TGF-β
Data Source
Figure 1
Figure 2A~2D
Figure 2D~2F
AI summary
The present disclosure provides Transforming Growth Factor Beta (TGF-β) polypeptide constructs and complexes that find use in, for example, therapeutic treatment of diseases including autoimmune diseases. Also described are nucleic acids that encode the constructs and complexes and methods of preparing the constructs and complexes in cell-based expression systems.