Small tumor necrosis factor-α receptor (TNFR) biologics selectively inhibit the TNF-p38 signalling axis and inflammation — ASN Events
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Small tumor necrosis factor-α receptor (TNFR) biologics selectively inhibit the TNF-p38 signalling axis and inflammation (#248)

Violet R Mukaro 1 2 , Michelle E Gahan 3 , Bernadette Boog 1 , Alex Quach 1 2 , Zia H Huang 1 2 , Xiuhui Gao 1 2 , Carol Haddad 1 , Suresh Mahalingam 4 , Charles S Hii 1 2 , Antonio Ferrante 1 2 5 6
  1. SA Pathology at Women's and Children's Hospital, North Adelaide, SA, Australia
  2. School of Paediatrics and Reproductive Medicine, Robinson Research Institute, University of Adelaide, Adelaide, South Australia, Australia
  3. Virus and Inflammation Research Group, Faculty of Applied Science, University of Canberra, Canberra, Australian Capital Territory, Australia
  4. Emerging Viruses and Inflammation Research Group, Institute of Glycomics, Griffith University, Gold Coast, Queensland, Austalia
  5. Microbiology and Immunology, University of Adelaide, Adelaide, South Australia, Australia
  6. School of Pharmacy and Medical Science, Univeristy of South Australia, Adelaide, South Australia, Australia

INTRODUCTION: Tumor necrosis factor (TNF) neutralising biologics, antibodies and soluble TNF receptors (TNFR), have revolutionised treatment of diseases such as rheumatoid arthritis. However, targeting a cytokine with a key role in immunity to infection and malignancy continues to raise serious concerns. Attempts to target the pro-inflammatory p38 MAP kinase, downstream of TNFR, have ceased owing to poor efficacy and unacceptable toxicity. The aim of this research was to develop biologics that could selectively inhibit the TNFR-p38 signalling axis and inflammation.

METHODS: Two TNF-derived peptides, TNF70-80 and TNF132-150 were investigated for their effects on neutrophil activation (lucigenin-dependent chemiluminescence assay), tumour cell cytotoxicity (MTT assay) and activation of MAP kinases (western blot). A pre-B cell line transfected with truncated TNFRI mutants was used to identify the region on TNFRI which binds TNF70-80. Peptides to this region of TNFRI were made and investigated in mice for their ability to inhibit LPS-induced peritoneal inflammation, carrageenan- and antigen-induce paw swelling/thickness and respiratory syncytial virus -induced lung inflammation, by assessing cell types and number and cytokines present at the inflammatory sites.

RESULTS: We have previously demonstrated that, TNF70-80 and, TNF132-150 had distinct biological properties but which were collectively representative of the parent TNF. Our data show that these differences reflect the types of signalling pathways they stimulate via TNFRI. Of interest was that TNF70-80 selectively stimulated p38 activation. Using this peptide and truncated TNFR mutants we identified the sequence in TNFRI which is coupled selectively to p38 activation. Peptides of this TNFRI sequence bind TNF/TNF70-80 and selectively inhibited their ability to induced p38 activation, TNF-induced neutrophil stimulation and inflammation in all four experimental models.

CONCLUSIONS: A TNFRI peptide which binds to TNF and selectively inhibits the TNFRI-p38 signalling axis and inflammation has been developed as potentially a new approach to targeting TNF in inflammatory diseases.