Recombinant MPT Protein Targeting TBK1 and HK2 for Tuberculosis
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Solution Overview
Problem
Current understanding of Mycobacterium tuberculosis interactions with host proteins is limited, particularly regarding MPT63 and MPT64, which are immunodominant secreted antigens, hindering effective prevention and treatment strategies for tuberculosis.
Innovation Solution
Development of a recombinant MPT protein incorporating specific regions of MPT63 and MPT64 that bind to TBK1, p47phox, or HK2, designed to target Mycobacterium tuberculosis-infected macrophages, increasing ROS production and inflammatory cytokine expression to induce bacterial death.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If MPT63 and MPT64 are used as tuberculosis vaccine candidates, then immune response activation is improved, but the mechanism of interaction with host proteins remains unclear limiting effectiveness
Solution Approach 1:
The patent segments MPT63 and MPT64 into distinct functional domains: MPT63 contains an N-terminal region (amino acids 1-50) that binds to TBK1 and a C-terminal region (amino acids 100-200) that binds to p47phox. Similarly, MPT64 contains an N-terminal region binding to TBK1 and a C-terminal region binding to HK2. This segmentation allows for targeted manipulation of specific interaction mechanisms while maintaining overall vaccine function.
Solution Approach 2:
The patent introduces a recombinant MPT protein as an intermediary that combines specific binding regions of both MPT63 and MPT64. This recombinant protein serves as a mediator that simultaneously engages TBK1, p47phox, and HK2 host proteins, thereby elucidating and exploiting the interaction mechanisms to enhance vaccine reliability.
2Reliability
If MPT63 induces apoptosis and MPT64 reduces inflammation, then immune regulation is improved, but the combined effect mechanism is not understood
Solution Approach 1:
The patent merges the N-terminal region of MPT63 (TBK1-binding), the C-terminal region of MPT63 (p47phox-binding), the N-terminal region of MPT64 (TBK1-binding), and the C-terminal region of MPT64 (HK2-binding) into a single recombinant MPT protein. This merging creates a unified molecule that can simultaneously mediate apoptosis induction through TBK1 and inflammation regulation through p47phox and HK2 interactions.
Solution Approach 2:
The recombinant MPT protein achieves multi-functionality by incorporating domains from both MPT63 and MPT64. It can simultaneously bind to TBK1 (inducing apoptosis), p47phox (regulating inflammation), and HK2 (metabolic regulation), thereby consolidating multiple immune regulation functions into a single vaccine candidate with enhanced reliability.
3Reliability
If host proteins TBK1, p47phox, and HK2 are targeted, then bacterial death is induced, but the complexity of identifying and targeting multiple proteins increases
Solution Approach 1:
The patent combines multiple targeting functions into a single recombinant MPT protein that simultaneously targets TBK1, p47phox, and HK2. Rather than requiring separate therapeutic agents for each host protein, the merged recombinant protein provides multi-targeted bacterial killing through its integrated domain structure, reducing the complexity of the targeting system while maintaining high reliability.
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 recombinant MPT protein effectively reduces Mycobacterium tuberculosis viability by modulating host immune responses, enhancing vaccine efficacy when combined with BCG, and demonstrating low cytotoxicity and no side effects.
Implementation Method 1
MPT63 and MPT64 are immunodominant secretory antigens of Mycobacterium tuberculosis... MPT63 induces apoptosis of macrophages... MPT64 encoded in the RD2 region is known to induce IFN-γ production and increase TGF-β expression in rat macrophages
Implementation Method 2
Increased expression of inflammatory cytokines and chemokines and production of reactive oxygen species through activation of the NF-κB signaling pathway are essential for removing active MTB from the host
Implementation Method 3
Increased expression of inflammatory cytokines and chemokines and production of reactive oxygen species through activation of the NF-κB signaling pathway
Implementation Method 4
MPT63 induces apoptosis of macrophages through host pH-dependent compatibility switches
Data Source
AI summary
The present disclosure is the first to identify a host cell protein and its function with which MPT63 and MPT64, secreted antigens of Mycobacterium tuberculosis, interact, and to construct a recombinant MPT protein including each domain of MPT63 and MPT64 interacting with the host cell protein, and the recombinant MPT protein may be applied to a use for the prevention and treatment of tuberculosis by confirming that the recombinant MPT protein targets the Mycobacterium tuberculosis-infected macrophages and increases the ROS level and inflammatory cytokine expression in macrophages, thereby inducing the death of Mycobacterium tuberculosis. And MPT protein of the present disclosure can improve the vaccine effect by the BCG vaccine so that it can be used as a tuberculosis vaccine and/or vaccine adjuvant either alone or together with known tuberculosis vaccines.