Inhibition of AXL/GAS6 signaling in the treatment of liver fibrosis
Compositions and methods are provided for alleviating endometriosis, kidney disease, inflammatory disease and/or transplant rejection in a mammal by administering a therapeutic dose of a pharmaceutical composition that inhibits AXL, MER or Tyro3 protein activity, for example by competitive or non-competitive inhibition of the binding interaction between AXL, MER or Tyro3 and its ligand GAS6.
1. A method of reducing liver fibrosis in a mammalian patient, the method comprising:
administering an effective dose of a soluble AXL variant polypeptide wherein said soluble AXL variant polypeptide:
lacks the AXL transmembrane domain,
lacks a functional fibronectin (FN) domain,
has an Ig1 domain, and an Ig2 domain,
has a set of amino acid substitutions at positions relative to SEQ ID NO:1: G32, A72, D87, V92 and G127; or at positions G32, D87, V92 and G127,
comprises an Fc domain linked to the AXL variant polypeptide by a linker comprising one or more (GLY) 4 SER (SEQ ID NO:10) units; and
wherein said AXL-variant polypeptide exhibits increased affinity of binding to GAS6 compared to wild-type AXL (SEQ ID NO:1).
2. The method of claim 1 , wherein said soluble AXL variant polypeptide comprises a set of amino acid substitutions where glycine 32 is replaced with a serine residue, aspartic acid 87 is replaced with a glycine residue, valine 92 is replaced with an alanine residue, and glycine 127 is replaced with an arginine residue, and optionally alanine 72 is replaced with a valine residue.
3. The method of claim 1 , wherein said soluble AXL variant polypeptide has an affinity of at least about 1×10 −8 M, 1×10 −9 M, 1×10 −10 M, 1×10 −11 M or 1×10 −12 M for GAS6.
4. The method of claim 1 , wherein said soluble AXL variant polypeptide exhibits an affinity to GAS6 that is at least about 5-fold stronger, at least about 10-fold stronger or at least about 20-fold stronger than the affinity of the wild-type AXL polypeptide.