Identification of synovium-specific homing peptides by in vivo phage display selection.
OBJECTIVE
To identify homing peptides specific for human synovium that could be used as targeting devices for delivering therapeutic/diagnostic agents to human joints.


METHODS
Human synovium and skin were transplanted into SCID mice. A disulfide-constrained 7-amino acid peptide phage display library was injected intravenously into the animals and synovial homing phage recovered from synovial grafts. Following 3-4 cycles of enrichment, DNA sequencing of homing phage clones allowed the identification of specific peptides that were synthesized by a-fluorenylmethyloxycarbonyl chemistry and used in competitive in vivo assays and immunohistochemistry analyses.


RESULTS
We isolated synovial homing phages displaying specific peptides that distinctively bound to synovial but not skin or mouse microvascular endothelium (MVE). They retained their tissue homing specificity in vivo, independently from the phage component, the original pathology of the transplanted tissue, and the degree of human/murine graft vascularization. One such peptide (CKSTHDRLC) maintained synovial homing specificity both when presented by the phage and as a free synthetic peptide. The synthetic peptide also competed with and inhibited in vivo the binding of the parent phage to the cognate synovial MVE ligand.


CONCLUSIONS
This is the first report describing peptides with homing properties specific for human synovial MVE. This was demonstrated using a novel approach targeting human tissues, transplanted into SCID mice, directly by in vivo phage display selection. The identification of such peptides opens the possibility of using these sequences to construct joint-specific drug delivery systems that may have considerable impact in the treatment of arthritic conditions.
Lee
Lewis
L
Guy's, King's, and St Thomas' School of Medicine, London, UK.
Buckley
Christopher
C
Blades
Mark C
MC
Panayi
Gabriel
G
George
Andrew J T
AJ
Pitzalis
Costantino
C
D000595:Amino Acid Sequence; D000818:Animals; D001172:Arthritis, Rheumatoid / Q000378:metabolism; D017104:Bacteriophage M13 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007275:Injections, Intravenous; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D009389:Neovascularization, Pathologic / Q000378:metabolism / Q000473:pathology; D010003:Osteoarthritis / Q000378:metabolism; D010446:Peptide Fragments / Q000008:administration & dosage / Q000302:isolation & purification / Q000378:metabolism*; D019151:Peptide Library; D020539:Sequence Analysis, Protein; D012867:Skin / Q000098:blood supply / Q000737:chemistry; D016038:Skin Transplantation; D013583:Synovial Membrane / Q000098:blood supply / Q000378:metabolism* / Q000637:transplantation
D010446:Peptide Fragments; D019151:Peptide Library