A single-domain i-body, AD-114, attenuates renal fibrosis through blockade of CXCR4

JCI Insight. 2022 Feb 22;7(4):e143018. doi: 10.1172/jci.insight.143018.


The G protein-coupled CXC chemokine receptor 4 (CXCR4) is a candidate therapeutic target for tissue fibrosis. A fully human single-domain antibody-like scaffold i-body AD-114-PA600 (AD-114) with specific high binding affinity to CXCR4 has been developed. To define its renoprotective role, AD-114 was administrated in a mouse model of renal fibrosis induced by folic acid (FA). Increased extracellular matrix (ECM) accumulation, macrophage infiltration, inflammatory response, TGF-β1 expression, and fibroblast activation were observed in kidneys of mice with FA-induced nephropathy. These markers were normalized or partially reversed by AD-114 treatment. In vitro studies demonstrated AD-114 blocked TGF-β1-induced upregulated expression of ECM, matrix metalloproteinase-2, and downstream p38 mitogen-activated protein kinase (p38 MAPK) and PI3K/AKT/mTOR signaling pathways in a renal proximal tubular cell line. Additionally, these renoprotective effects were validated in a second model of unilateral ureteral obstruction using a second generation of AD-114 (Fc-fused AD-114, also named AD-214). Collectively, these results suggest a renoprotective role of AD-114 as it inhibited the chemotactic function of CXCR4 as well as blocked CXCR4 downstream p38 MAPK and PI3K/AKT/mTOR signaling, which establish a therapeutic strategy for AD-114 targeting CXCR4 to limit renal fibrosis.

Keywords: Chemokines; Chronic kidney disease; Fibrosis; Nephrology; Therapeutics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Disease Models, Animal
  • Extracellular Matrix / metabolism
  • Fibrosis / genetics
  • Fibrosis / metabolism
  • Fibrosis / pathology
  • Gene Expression Regulation*
  • Humans
  • Kidney / metabolism
  • Kidney / pathology*
  • Kidney Diseases / genetics*
  • Kidney Diseases / metabolism
  • Kidney Diseases / pathology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Receptors, CXCR4 / biosynthesis
  • Receptors, CXCR4 / genetics*
  • Signal Transduction
  • Up-Regulation*


  • CXCR4 protein, mouse
  • Receptors, CXCR4