Structural basis and targeting of the interaction between fibroblast growth factor-inducible 14 and tumor necrosis factor-like weak inducer of apoptosis.
Deregulation of the TNF-like weak inducer of apoptosis (TWEAK)-fibroblast growth factor-inducible 14 (Fn14) signaling pathway is observed in many diseases, including inflammation, autoimmune diseases, and cancer. Activation of Fn14 signaling by TWEAK binding triggers cell invasion and survival and therefore represents an attractive pathway for therapeutic intervention. Based on structural studies of the TWEAK-binding cysteine-rich domain of Fn14, several homology models of TWEAK were built to investigate plausible modes of TWEAK-Fn14 interaction. Two promising models, centered on different anchoring residues of TWEAK (tyrosine 176 and tryptophan 231), were prioritized using a data-driven strategy. Site-directed mutagenesis of TWEAK at Tyr(176), but not Trp(231), resulted in the loss of TWEAK binding to Fn14 substantiating Tyr(176) as the anchoring residue. Importantly, mutation of TWEAK at Tyr(176) did not disrupt TWEAK trimerization but failed to induce Fn14-mediated nuclear factor κ-light chain enhancer of activated B cell (NF-κB) signaling. The validated structural models were utilized in a virtual screen to design a targeted library of small molecules predicted to disrupt the TWEAK-Fn14 interaction. 129 small molecules were screened iteratively, with identification of molecules producing up to 37% inhibition of TWEAK-Fn14 binding. In summary, we present a data-driven in silico study revealing key structural elements of the TWEAK-Fn14 interaction, followed by experimental validation, serving as a guide for the design of small molecule inhibitors of the TWEAK-Fn14 ligand-receptor interaction. Our results validate the TWEAK-Fn14 interaction as a chemically tractable target and provide the foundation for further exploration utilizing chemical biology approaches focusing on validating this system as a therapeutic target in invasive cancers.
From the Translational Genomics Research Institute, Phoenix, Arizona 85004.
Harshil
H
Dhruv
the Mayo Clinic, Scottsdale, Arizona 85259.
Joseph C
JC
Loftus
the Mayo Clinic, Scottsdale, Arizona 85259.
Pooja
P
Narang
the Mayo Clinic, Scottsdale, Arizona 85259.
Joachim L
JL
Petit
From the Translational Genomics Research Institute, Phoenix, Arizona 85004.
Maureen
M
Fameree
From the Translational Genomics Research Institute, Phoenix, Arizona 85004.
Julien
J
Burton
From the Translational Genomics Research Institute, Phoenix, Arizona 85004.
Giresse
G
Tchegho
From the Translational Genomics Research Institute, Phoenix, Arizona 85004.
Donald
D
Chow
From the Translational Genomics Research Institute, Phoenix, Arizona 85004.
Holly
H
Yin
the Center for Molecular Innovation, Feinstein Institute for Medical Research, Manhasset, New York 11030.
Yousef
Y
Al-Abed
From the Translational Genomics Research Institute, Phoenix, Arizona 85004.
Michael E
ME
Berens
From the Translational Genomics Research Institute, Phoenix, Arizona 85004,. Electronic address: ntran@tgen.org.
Nhan L
NL
Tran
the Mayo Clinic, Scottsdale, Arizona 85259. Electronic address: meurice.nathalie@mayo.edu.
Nathalie
N
Meurice
D019943:Amino Acid Substitution; D045744:Cell Line, Tumor; D000074049:Cytokine TWEAK; D057809:HEK293 Cells; D006801:Humans; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009369:Neoplasms / Q000737:chemistry / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D017434:Protein Structure, Tertiary; D018124:Receptors, Tumor Necrosis Factor / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000074050:TWEAK Receptor; D000079424:Tumor Necrosis Factor Inhibitors; D048069:Tumor Necrosis Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D000074049:Cytokine TWEAK; D009363:Neoplasm Proteins; D018124:Receptors, Tumor Necrosis Factor; C000613297:TNFRSF12A protein, human; C109743:TNFSF12 protein, human; D000074050:TWEAK Receptor; D000079424:Tumor Necrosis Factor Inhibitors; D048069:Tumor Necrosis Factors
Cancer; Cell Signaling; Cell Surface Receptor; Docking; Drug Development; Drug Discovery; Drug Screening; Protein-Protein Interactions