The long-term objective of this proposal is to investigate the cellular and molecular mechanisms of growth factors mediated regeneration of acellular bladder matrix using both in vitro and in vivo models and its potential clinical use. The hypothesis is that growth factors (TGFb1, EGF, KGF and VEGF) will stimulate the process of regeneration of acellular matrix bladder when grafted to the host bladder (normal, diabetic and interstitial cystitis) after partial cystectomy. These growth factors will stimulate the process of ingrowth of smooth muscle, urothelial lining, vascularization and innervation in the regeneration of new functional bladder.
Specific Aim # 1. Effects of growth factors (TGFb1, EGF, KGF and VEGF) on regeneration of bladder morphology and growth using normal and diseased bladder. Under this specific aim, the investigators will test the hypothesis that growth factors will stimulate the process of regeneration of acellular matrix bladder when grafted to the host bladder (normal and diseased such as diabetic and interstitial cystitis) after partial cystectomy. To test this hypothesis following experiments will be conducted: (a). Effects of growth factors on epithelial migration and growth in normal and diseased bladder. (b). Effects of growth factors on smooth muscle cells migration and growth in normal and diseased bladder. (c). Effects of growth factors on invasion of blood vessels and nerves in normal and diseased bladder.
Specific Aim # 2. Functional studies of matrix-directed regenerated bladders in normal and diseased hosts treated with growth factors. Under this specific aim, they will test the hypothesis that the augmented bladder after acellular matrix grafting in normal and diseased hosts (diabetic and interstitial cystitis) will be re-innervated with cholinergic and adrenergic nerve fibers and perform normal voiding function through their receptors. To test this hypothesis, following experiments will be done: (a). In vivo bladder functional studies by micturition pattern. (b). In vitro bladder functional studies by tissue bath experiments. (c). Immunochemical studies of adrenergic and cholinergic nerves of regenerated bladder.
Specific Aim # 3. Cellular and molecular mechanisms of growth factor-induced and matrix directed bladder regeneration in normal and diseased hosts. Under this specific aim, they will test the hypothesis that gene and protein expression of growth factors (TGFb1, EGF, KGF and VEGF) will enhance reepithelization, vascularization and innervation of matrix-directed bladder regeneration in normal and diseased hosts (diabetic and interstitial cystitis). To test this hypothesis, following experiments will be done: (a). To analyze gene expression of growth factor TGFb1, EGF, KGF and VEGF in host and regenerated bladder.; (b). To analyze protein expression and localization of growth factors TGFb1, EGF, KGF and VEGF in host and re-generated bladder. (c). To analyze gene and protein expression of receptors for growth factors TGFb1, EGF, KGF and VEGF in host and re-generated bladder.
Specific Aim # 4. Clinical application of matrix-directed bladder regeneration. The investigators will initiate clinical trials for bladder augmentation and urethral stricture repair using organ specific acellular matrix.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK051101-08
Application #
6699655
Study Section
Special Emphasis Panel (ZRG1-UROL (02))
Program Officer
Mullins, Christopher V
Project Start
2000-12-01
Project End
2006-12-31
Budget Start
2004-01-01
Budget End
2006-12-31
Support Year
8
Fiscal Year
2004
Total Cost
$331,435
Indirect Cost
Name
University of California San Francisco
Department
Urology
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Youssif, Mohamed; Shiina, Hiroaki; Urakami, Shinji et al. (2005) Effect of vascular endothelial growth factor on regeneration of bladder acellular matrix graft: histologic and functional evaluation. Urology 66:201-7
Cayan, Selahitten; Chermansky, Christopher; Schlote, Norbert et al. (2002) The bladder acellular matrix graft in a rat chemical cystitis model: functional and histologic evaluation. J Urol 168:798-804
Baskin, L; DiSandro, M; Li, Y et al. (2001) Mesenchymal-epithelial interactions in bladder smooth muscle development: effects of the local tissue environment. J Urol 165:1283-8
Shiina, H; Igawa, M; Urakami, S et al. (2001) Alterations of beta- and gamma-catenin in N-butyl-N-(-4-hydroxybutyl)nitrosamine-induced murine bladder cancer. Cancer Res 61:7101-9
Wefer, J; Sievert, K D; Schlote, N et al. (2001) Time dependent smooth muscle regeneration and maturation in a bladder acellular matrix graft: histological studies and in vivo functional evaluation. J Urol 165:1755-9
Aboseif, S; El-Sakka, A; Young, P et al. (1999) Mesenchymal reprogramming of adult human epithelial differentiation. Differentiation 65:113-8
Wu, H Y; Baskin, L S; Liu, W et al. (1999) Understanding bladder regeneration: smooth muscle ontogeny. J Urol 162:1101-5
Baskin, L S; Hayward, S W; DiSandro, M S et al. (1999) Epithelial-mesenchymal interactions in the bladder. Implications for bladder augmentation. Adv Exp Med Biol 462:49-61
Cunha, G R; Forsberg, J G; Golden, R et al. (1999) New approaches for estimating risk from exposure to diethylstilbestrol. Environ Health Perspect 107 Suppl 4:625-30
Baskin, L S; Hayward, S W; Sutherland, R A et al. (1997) Cellular signaling in the bladder. Front Biosci 2:d592-5