title: Normalization of proliferation and tight junction formation in bladder epithelial cells from patients with interstitial cystitis/painful bladder syndrome by d-proline and d-pipecolic acid derivatives of antiproliferative factor.
abstract: Interstitial cystitis/painful bladder syndrome is a chronic bladder disorder with epithelial thinning or ulceration, pain, urinary frequency and urgency, for which there is no reliably effective therapy. We previously reported that interstitial cystitis/painful bladder syndrome bladder epithelial cells make a glycopeptide antiproliferative factor or 'APF' (Neu5Acα2-3Galβ1-3GalNAcα-O-TVPAAVVVA) that induces abnormalities in normal cells similar to those in interstitial cystitis/painful bladder syndrome cells in vitro, including decreased proliferation, decreased tight junction formation, and increased paracellular permeability. We screened inactive APF derivatives for their ability to block antiproliferative activity of asialylated-APF ('as-APF') in normal bladder cells and determined the ability of as-APF-blocking derivatives to normalize tight junction protein expression, paracellular permeability, and/or proliferation of interstitial cystitis/painful bladder syndrome cells. Only two of these derivatives [Galβ1-3GalNAcα-O-TV-(d-pipecolic acid)-AAVVVA and Galβ1-3GalNAcα-O-TV-(d-proline)-AAVVVA] blocked as-APF antiproliferative activity in normal cells (p < 0.001 for both). Both of these antagonists also 1) significantly increased mRNA expression of ZO-1, occludin, and claudins 1, 4, 8, and 12 in interstitial cystitis/painful bladder syndrome cells by qRT-PCR; 2) normalized interstitial cystitis/painful bladder syndrome epithelial cell tight junction protein expression and tight junction formation by confocal immunofluorescence microscopy; and 3) decreased paracellular permeability of (14) C-mannitol and (3) H-inulin between confluent interstitial cystitis/painful bladder syndrome epithelial cells on Transwell plates, suggesting that these potent APF antagonists may be useful for the development as interstitial cystitis/painful bladder syndrome therapies.
authors.affiliation: Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, USA. skeay@medicine.umaryland.edu
authors.forename: Susan
authors.initials: S
authors.lastname: Keay
authors.forename: Piotr
authors.initials: P
authors.lastname: Kaczmarek
authors.forename: Chen-Ou
authors.initials: CO
authors.lastname: Zhang
authors.forename: Kristopher
authors.initials: K
authors.lastname: Koch
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Szekely
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Barchi
authors.forename: Christopher
authors.initials: C
authors.lastname: Michejda
mesh_terms: D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D002463:Cell Membrane Permeability / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018856:Cystitis, Interstitial / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005786:Gene Expression Regulation / Q000187:drug effects; D006023:Glycoproteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D010875:Pipecolic Acids / Q000737:chemistry; D011392:Proline / Q000737:chemistry; D019108:Tight Junctions / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D001743:Urinary Bladder / Q000166:cytology*
chemical_list: D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D010875:Pipecolic Acids; C490708:antiproliferative factor APF, human; D011392:Proline; C031345:pipecolic acid