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Generation of a sensitive TNFR2-specific murine assays system
Abstract:
Tumor necrosis factor (TNF)/TNF receptors (TNFR1/TNFR2) are considered to be potential drug targets to treat refractory diseases, including autoimmune diseases and malignant tumors. However, their specific functions, especially in the case of TNFR2, are poorly understood. In this study, we constructed a mouse TNFR2 (mTNFR2)-mediated biological assay system that shows no effects of mouse TNFR1 (mTNFR1) in order to screen mTNFR2-selective stimulating agents. Mouse TNFR1(-/-)R2(-/-) preadipocytes were transfected with the gene encoding the mTNFR2/mouse Fas (mFas) chimeric receptor in which the extracellular and transmembrane domains of mTNFR2 were fused to the intracellular domain of mFas. Our results demonstrated that this cell line exhibits highly sensitive mTNFR2-mediated cytotoxic effects. We propose that this mTNFR2-mediated biological assay system would be a useful tool to screen for mTNFR2-selective stimulating agents.
Insights
Researchers developed a new assay to specifically target mouse Tumor Necrosis Factor Receptor 2 (mTNFR2). This system enables the screening of agents that stimulate mTNFR2, crucial for understanding its role in diseases.
Area of Science:
- Immunology and Molecular Biology
- Drug Discovery and Development
Background:
- Tumor Necrosis Factor (TNF) and its receptors (TNFR1/TNFR2) are implicated as therapeutic targets for autoimmune diseases and cancers.
- The precise functions of TNF receptors, particularly TNFR2, remain incompletely elucidated, hindering targeted therapeutic development.
Purpose of the Study:
- To establish a novel biological assay system for screening mouse TNFR2 (mTNFR2)-selective stimulating agents.
- To develop a system that isolates mTNFR2 activity, excluding the influence of mouse TNFR1 (mTNFR1).
Main Methods:
- Construction of a mouse TNFR1 knockout and TNFR2 knockout (mTNFR1(-/-)R2(-/-)) preadipocyte cell line.
- Transfection of the cell line with a chimeric receptor gene encoding mTNFR2 fused with the intracellular domain of mouse Fas (mFas).
- Evaluation of the cell line's sensitivity to mTNFR2-mediated cytotoxic effects.
Main Results:
- The developed cell line demonstrated highly sensitive cytotoxic effects mediated specifically by mTNFR2.
- The assay system effectively screened for mTNFR2-selective stimulating agents, independent of mTNFR1 activity.
Conclusions:
- The novel mTNFR2-mediated biological assay system is a valuable tool for identifying mTNFR2-selective agonists.
- This system facilitates further research into TNFR2 functions and the development of targeted therapies for refractory diseases.
