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Understanding insulin-like peptide 5 (INSL5) and relaxin family peptide receptor 4 (RXFP4): structure, signalling, and function.

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Engineering a long acting, non-biased relaxin agonist using Protein-in-Protein technology.

Irina U Agoulnik1, Elena M Kaftanovskaya1, Courtney Myhr1

  • 1Department of Human and Molecular Genetics, Herbert Wertheim College of Medicine, Florida International University, Miami, FL 33199, USA.

Biochemical Pharmacology
|June 30, 2024
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Researchers developed a long-acting relaxin biologic using Protein-in-Protein technology to target fibrosis. This novel approach enhances relaxin

Keywords:
FibrosisProtein-in-ProteinRXFP1Relaxin

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Area of Science:

  • Endocrinology and Pharmacology
  • Biotechnology and Protein Engineering
  • Fibrosis Research

Background:

  • The peptide hormone relaxin is crucial for tissue remodeling via RXFP1 receptor activation.
  • Relaxin's matrix-modifying properties suggest potential for treating fibrotic diseases.
  • Short half-life of recombinant relaxin peptides limits clinical efficacy in conditions like heart failure.

Purpose of the Study:

  • To engineer a long-acting relaxin biologic using Protein-in-Protein (PiP) technology.
  • To evaluate the pharmacokinetic and pharmacodynamic properties of the novel relaxin-PiP construct.
  • To assess the therapeutic potential of relaxin-PiP in a liver fibrosis mouse model.

Main Methods:

  • A single-chain human relaxin was inserted into an IgG backbone using PiP technology.
  • In vitro assays measured receptor binding, activation (cAMP, cGMP, pERK), and signal transduction.
  • A carbon tetrachloride-induced mouse model of liver fibrosis was used to evaluate R2-PiP efficacy.

Main Results:

  • Relaxin-PiP constructs exhibited a half-life of approximately 4-5 days in mice.
  • Relaxin-PiPs demonstrated full agonist activity on human and mouse RXFP1 receptors without signal bias.
  • R2-PiP treatment reduced liver lesions, collagen accumulation, and Collagen1a1 gene expression, while increasing hepatic cell proliferation.

Conclusions:

  • Protein-in-Protein technology successfully created a long-acting relaxin biologic (relaxin-PiP).
  • Relaxin-PiP acts as a potent RXFP1 agonist with therapeutic potential for fibrotic diseases.
  • These findings support further investigation into relaxin/RXFP1 signaling for treating human fibrotic conditions.