PURPOSE: Studies show that LL-37 is a naturally occurring urinary defensin peptide that is up-regulated during urinary tract infections. Although normal urinary LL-37 levels are antimicrobial, we propose that increased LL-37 may trigger bladder inflammation. We further suggest that anti-inflammatory sulfated polysaccharides known as semi-synthetic glycosaminoglycan ether compounds can treat/prevent LL-37 mediated bladder inflammation. MATERIALS AND METHODS: C57BL/6 mice were catheterized/instilled with LL-37 (320 μM, 150 μl) for 45 minutes. Animals were sacrificed at 12 and 24 hours, and tissues were examined using hematoxylin and eosin. Separate experiments were performed for myeloperoxidase to quantify inflammation. GM-1111 semi-synthetic glycosaminoglycan ether treatments involved instillation of 10 mg/ml for 45 minutes directly before or after LL-37. Tissues were harvested at 24 hours. To compare semi-synthetic glycosaminoglycan ether efficacy, experiments were performed using 10 mg/ml heparin. Finally, tissue localization of semi-synthetic glycosaminoglycan ether was examined using a fluorescent GM-1111-Alexa Fluor® 633 conjugate. RESULTS: Profound bladder inflammation developed after LL-37. Greater tissue inflammation occurred after 24 hours compared to that at 12 hours. Myeloperoxidase assays revealed a 21 and 61-fold increase at 12 and 24 hours, respectively. Semi-synthetic glycosaminoglycan ether treatment after LL-37 showed mild attenuation of inflammation with myeloperoxidase 2.5-fold below that of untreated bladders. Semi-synthetic glycosaminoglycan ether treatment before LL-37 demonstrated almost complete attenuation of inflammation. Myeloperoxidase results mirrored those in controls. In heparin treated bladders minimal attenuation of inflammation occurred. Finally, instillation of GM-1111-Alexa Fluor 633 revealed urothelial coating, significant tissue penetration and binding to endovasculature. CONCLUSIONS: We developed what is to our knowledge a new model of inflammatory bladder disease by challenge with the naturally occurring urinary peptide LL-37. We also noted that a new class of anti-inflammatory sulfated polysaccharides prevents and mitigates bladder inflammation.
PURPOSE: Studies show that LL-37 is a naturally occurring urinary defensin peptide that is up-regulated during urinary tract infections. Although normal urinary LL-37 levels are antimicrobial, we propose that increased LL-37 may trigger bladder inflammation. We further suggest that anti-inflammatory sulfated polysaccharides known as semi-synthetic glycosaminoglycan ether compounds can treat/prevent LL-37 mediated bladder inflammation. MATERIALS AND METHODS: C57BL/6 mice were catheterized/instilled with LL-37 (320 μM, 150 μl) for 45 minutes. Animals were sacrificed at 12 and 24 hours, and tissues were examined using hematoxylin and eosin. Separate experiments were performed for myeloperoxidase to quantify inflammation. GM-1111 semi-synthetic glycosaminoglycan ether treatments involved instillation of 10 mg/ml for 45 minutes directly before or after LL-37. Tissues were harvested at 24 hours. To compare semi-synthetic glycosaminoglycan ether efficacy, experiments were performed using 10 mg/ml heparin. Finally, tissue localization of semi-synthetic glycosaminoglycan ether was examined using a fluorescent GM-1111-Alexa Fluor® 633 conjugate. RESULTS: Profound bladder inflammation developed after LL-37. Greater tissue inflammation occurred after 24 hours compared to that at 12 hours. Myeloperoxidase assays revealed a 21 and 61-fold increase at 12 and 24 hours, respectively. Semi-synthetic glycosaminoglycan ether treatment after LL-37 showed mild attenuation of inflammation with myeloperoxidase 2.5-fold below that of untreated bladders. Semi-synthetic glycosaminoglycan ether treatment before LL-37 demonstrated almost complete attenuation of inflammation. Myeloperoxidase results mirrored those in controls. In heparin treated bladders minimal attenuation of inflammation occurred. Finally, instillation of GM-1111-Alexa Fluor 633 revealed urothelial coating, significant tissue penetration and binding to endovasculature. CONCLUSIONS: We developed what is to our knowledge a new model of inflammatory bladder disease by challenge with the naturally occurring urinary peptide LL-37. We also noted that a new class of anti-inflammatory sulfated polysaccharides prevents and mitigates bladder inflammation.
Authors: Milan Chromek; Zuzana Slamová; Peter Bergman; László Kovács; L'udmila Podracká; Ingrid Ehrén; Tomas Hökfelt; Gudmundur H Gudmundsson; Richard L Gallo; Birgitta Agerberth; Annelie Brauner Journal: Nat Med Date: 2006-06-04 Impact factor: 53.440
Authors: Kenshi Yamasaki; Anna Di Nardo; Antonella Bardan; Masamoto Murakami; Takaaki Ohtake; Alvin Coda; Robert A Dorschner; Chrystelle Bonnart; Pascal Descargues; Alain Hovnanian; Vera B Morhenn; Richard L Gallo Journal: Nat Med Date: 2007-08-05 Impact factor: 53.440
Authors: Jonathan S Starkman; Magaly Martinez-Ferrer; Juan M Iturregui; Consolate Uwamariya; Roger R Dmochowski; Neil A Bhowmick Journal: J Urol Date: 2008-04-23 Impact factor: 7.450
Authors: Jennifer K Plichta; Casey J Holmes; Vanessa Nienhouse; Michelle Puszynski; Xiang Gao; Qunfeng Dong; Huaiying Lin; James Sinacore; Michael Zilliox; Evelyn Toh; David E Nelson; Richard L Gamelli; Katherine A Radek Journal: Crit Care Med Date: 2017-06 Impact factor: 7.598
Authors: Jeremiah A Alt; Xuan Qin; Abigail Pulsipher; Quinn Orb; Richard R Orlandi; Jianxing Zhang; Austin Schults; Wanjian Jia; Angela P Presson; Glenn D Prestwich; Siam Oottamasathien Journal: Int Forum Allergy Rhinol Date: 2015-09-08 Impact factor: 3.858
Authors: Abigail Pulsipher; Xuan Qin; Andrew J Thomas; Glenn D Prestwich; Siam Oottamasathien; Jeremiah A Alt Journal: Int Forum Allergy Rhinol Date: 2016-11-11 Impact factor: 3.858