Engineering a potent receptor superagonist or antagonist from a novel IL-6 family cytokine ligand

Abstract

Interleukin-6 (IL-6) family cytokines signal through multimeric receptor complexes, providing unique opportunities to create novel ligand-based therapeutics. The cardiotrophin-like cytokine factor 1 (CLCF1) ligand has been shown to play a role in cancer, osteoporosis, and atherosclerosis. Once bound to ciliary neurotrophic factor receptor (CNTFR), CLCF1 mediates interactions to coreceptors glycoprotein 130 (gp130) and leukemia inhibitory factor receptor (LIFR). By increasing CNTFR-mediated binding to these coreceptors we generated a receptor superagonist which surpassed the potency of natural CNTFR ligands in neuronal signaling. Through additional mutations, we generated a receptor antagonist with increased binding to CNTFR but lack of binding to the coreceptors that inhibited tumor progression in murine xenograft models of nonsmall cell lung cancer. These studies further validate the CLCF1–CNTFR signaling axis as a therapeutic target and highlight an approach of engineering cytokine activity through a small number of mutations.

Structural insights into the assembly of gp130 family cytokine signaling complexes

“…Like F152 and K155 of CNTF, the two conserved CLCF1 residues F178 and K181 also serve as key anchor points engaging LIFR residues (S310, N313, V315, I322, G324, and V326) (Fig. 2H), consistent with previous mutagenesis studies ( 18 , 20 ). CLCF1 helix B does not make significant contacts with the N-terminal loop of LIFR D4 as CNTF helix B does, because CLCF1 helix B is slightly shorter than CNTF helix B at the C-terminus.…”

The IL6-like Cytokine Family: Role and Biomarker Potential in Breast Cancer

“…This might greatly impact the selectiveness of a given inhibitor's effect on downstream signalling, so that finer blockade of the effects of a specific cytokine might be achieved [305]. Also in line with this, a recent study showed that targeted mutagenesis of different residues in CLC, which mediate interactions in the CNTF-CNTFR-LIFR-gp130/IL6ST signalling complex, can yield novel recombinant variants with distinct functions [306].…”

The Role of Interleukin-6 Family Members in Cardiovascular Diseases

“…Kim, Jun W et al have engineered CLCF1 variants that can inhibit or activate CNTFR. The application of it in atherosclerosis might be meaningful research ( 80 ).…”