Urothelial expression of neuropilins and VEGF receptors in control and interstitial cystitis patients

Departments of 1 Physiology and 5 Urology, University of Oklahoma Health Sciences Center, and 2 Oklahoma Medical Research Foundation (OMRF), Imaging Core Facility, Oklahoma City, Oklahoma; 3 VirtualScopics, Incorporated, Rochester; 4 Departments of Urology and Pathology, New York University Medical School, New York, New York; and 6 SibTech, Incoporated, Brookfield, Connecticut Submitted 3 June 2008 ; accepted in final form 19 September 2008 Interstitial cystitis (IC) is a chronic and painful bladder syndrome of unknown cause with no reliable biological marker or effective therapy. Vascular endothelial growth factor (VEGF), which plays a key role in bladder inflammation, is closely associated with the vascular alterations observed in patients with IC. However, our recent findings of VEGF receptors (VEGF-Rs) and VEGF coreceptors on nonendothelial cells in human and mouse urothelium suggest that additional VEGF targets and functions are possible in IC bladders. We report here that VEGF-Rs and coreceptors (neuropilins; NRP) are strongly expressed in both the human bladder urothelium and in the human bladder cancer cell line (J82) and that the expression of NRP2 and VEGF-R1 is significantly downregulated in IC compared with control subjects. In addition, treatment of J82 cells with bacillus Calmette-Guérin (BCG), a novel treatment strategy for IC, upregulates the messages for NRPs and VEGF-Rs. Furthermore, intravesical instillation of an internalizable VEGF fluorescent tracer (scVEGF/Cy5.5) into mouse urinary bladders results in a marked ligand accumulation in the urothelium and bladder parenchyma, indicating that urothelial VEGF-Rs are functionally active and capable of ligand interaction and internalization. Our results suggest that the VEGF pathway is altered in IC, that urinary VEGF may gain access to the bladder wall via these receptors, and that BCG treatment may replenish the missing VEGF-Rs/NRP receptors. Together, these results suggest that levels of NRPs, VEGF-Rs, and VEGF are new putative markers for the diagnosis of IC and that modulating these receptors can be exploited as therapeutic strategies. BCG (bacillus Calmette-Guérin); bladder cancer cell line (J82); molecular imaging; NIRF (near-infrared fluorescence); translational research; ChIP Address for reprint requests and other correspondence: R. Saban, College of Medicine, Univ. of Oklahoma Health Sciences Center (OUHSC), Urinary Tract Physiological Genomics Laboratory, 800 Research Pkwy., Rm. 41