Safety, efficacy, and molecular mechanism of claudin-1-specific peptides to enhance blood-nerve-barrier permeability.
The blood-nerve barrier consists of the perineurium and endoneurial vessels. The perineurial barrier is composed of a basal membrane and a layer of perineurial cells sealed by tight junction proteins preventing e.g. application of analgesics for selective regional pain control. One of the barrier-sealing proteins in the blood-nerve barrier is claudin-1. Therefore, the claudin-1-peptidomimetics (C1C2), derived from the first extracellular loop (ECL1) on claudin-1 was developed. In this study, we further evaluated the expression of tight junction proteins in the perineurium in Wistar rats and characterized the specificity, in vivo applicability, mechanism of action as well as the biocompatibility of C1C2. In the perineurium, claudin-19, tricellulin and ZO-1, but no claudin-2, 3, 8 and -11 were expressed. C1C2 specifically bound to the ECL1 of claudin-1 and fluorescent 5,6-carboxytetramethylrhodamine-C1C2 was rapidly internalized. Opening the perineurium with C1C2 reduced the mRNA and protein expression of claudin-1 and increased small and macromolecule permeability into the peripheral nerve. Application of C1C2 facilitated regional analgesia using μ-opioid receptor agonists like DAMGO or morphine without motor impairment in naïve rats as well as rats with hind paw inflammation. In contrast the control peptide C2C2 derived from ECL1 on claudin-2 did neither open the barrier nor facilitated opioid-mediated regional analgesia. C1C2 delivery was well tolerated and caused no morphological and functional nerve damage. C1C2 effects could be reversed by interference with the wnt-signal-transduction pathway, specifically the homeobox transcription factor cdx2, using a glycogen-synthase-kinase-3 inhibitor. In summary, we describe the composition of and a pathway to open the perineurial barrier employing a peptide to deliver hydrophilic substances to the peripheral nerve.
Dept. Anesthesiology, University Hospital of Wuerzburg, Wuerzburg, Germany.
Reine-Solange
RS
Sauer
Institute for Clinical Physiology, Charité Campus Benjamin Franklin, Berlin, Germany.
Susanne M
SM
Krug
Dept. Anesthesiology, University Hospital of Wuerzburg, Wuerzburg, Germany; Dept. Anesthesiology and Critical Care Medicine, Charité Campus Benjamin Franklin, Berlin, Germany.
Dagmar
D
Hackel
Leibniz Institute for Molecular Pharmacology, Berlin-Buch, Germany.
Christian
C
Staat
Dept. Anesthesiology, University Hospital of Wuerzburg, Wuerzburg, Germany.
Natalia
N
Konasin
Dept. Anesthesiology, University Hospital of Wuerzburg, Wuerzburg, Germany.
Shaobing
S
Yang
Dept. Anesthesiology, University Hospital of Wuerzburg, Wuerzburg, Germany.
Benedikt
B
Niedermirtl
Dept. Anesthesiology, University Hospital of Wuerzburg, Wuerzburg, Germany.
Judith
J
Bosten
Leibniz Institute for Molecular Pharmacology, Berlin-Buch, Germany.
Ramona
R
Günther
Leibniz Institute for Molecular Pharmacology, Berlin-Buch, Germany.
Sebastian
S
Dabrowski
Dept. Neurology, University Hospital of Wuerzburg, Wuerzburg, Germany.
Kathrin
K
Doppler
Dept. Neurology, University Hospital of Wuerzburg, Wuerzburg, Germany.
Claudia
C
Sommer
Leibniz Institute for Molecular Pharmacology, Berlin-Buch, Germany.
Ingolf E
IE
Blasig
Dept. Anesthesiology, University Hospital of Wuerzburg, Wuerzburg, Germany.
Alexander
A
Brack
Dept. Anesthesiology, University Hospital of Wuerzburg, Wuerzburg, Germany. Electronic address: rittner_h@ukw.de.
Heike L
HL
Rittner
D000595:Amino Acid Sequence; D000698:Analgesia; D000701:Analgesics, Opioid / Q000008:administration & dosage / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D062445:Claudin-1 / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology*; D010525:Peripheral Nerves / Q000187:drug effects* / Q000378:metabolism; D010539:Permeability / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D012584:Sciatic Nerve / Q000187:drug effects / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism
D000701:Analgesics, Opioid; D062445:Claudin-1; D010455:Peptides
Biocompatibility; Drug delivery; In vitro test; In vivo test; Peptide; Peripheral nerve