Title: The Function of Peroxisome Proliferator-Activated Receptor-Gamma in the Urothelium
Description: The urothelium is a stratified epithelium that serves as a barrier between the urinary tract and blood. It consists of terminally differentiated umbrella cells, which are specialized for synthesizing and assembling uroplakins into a tough apical plaque and responsible for the barrier function; intermediate cells which are few in number but serve as umbrella cell progenitors; and unipotent basal cells, which populate the majority of the urothelium. The urothelium is one of the most quiescent epithelia in the body but can rapidly regenerate in response to damage. The urothelium is also a source of cells that give rise to bladder cancer. Patients with chronic inflammation caused by indwelling catheters or repeated urinary tract infections have a higher risk of developing bladder cancer. Bladder cancers with squamous histological features are considered to be more aggressive with poor prognosis and the majority are categorized as basal subtype. The expression of Peroxisome Proliferator-Activated Receptor-gamma (PPARG) is strongly down regulated in the basal subtype of bladder cancer, suggesting that its removal might be essential in tumorigenesis.

PPARG is a nuclear hormone receptor that was originally described as a master regulator of adipogenesis but could also promote cellular differentiation in a number of epithelium. PPARG also serves as an important regulator in anti-inflammatory activity after a variety of injuries, acting in part by antagonizing the NF-B pathway. In urothelial cells, it has been shown that PPARG promotes urothelial differentiation in vitro, but its function in vivo remains unexplored. To determine the role of PPARG in vivo, we used Cre-Lox recombination to conditionally delete the Pparg gene in the mouse urothelium using the ShhCre driver, which drives recombination in basal and intermediate cells, and their respective daughters. Interestingly, ShhCre;Ppargfl/fl mutants lack umbrella and intermediate cells which normally populate the luminal and sub-luminal layers, and are instead populated with an abnormal cell population negative for classical urothelial markers. The basal compartment, which in wild type mice is largely populated by P63+ KRT5+ basal cells with a small sub-population expressing KRT14; has an increased number of KRT14-expressing cells in the mutants and exhibits squamous features that are not present in the normal urothelium.

In wild type animals, urinary tract infection (UTI) with uropathogenic E.coli results in a transient innate immune response, followed by proliferation and repair, which is largely complete within 2 weeks. When ShhCre;Ppargfl/fl mutants were challenged with urinary tract infection, the innate immune response was not resolved even after several weeks, as characterized by persistent NF-B activity, excessive influx of neutrophils and macrophages, and massive granulation tissue in the stroma. In addition, the Pparg-knockout urothelium exhibited squamous metaplasia. The Krt14+ basal cell population, which is considered to be the cells of origin of bladder cancer, greatly expanded in the Pparg-deleted urothelium after infection, and some lesions progressed to acquire invasive features. Together these findings suggest that PPARG is essential for the normal differentiation of the urothelium and is a potent regulator of the inflammatory response after UTI. Understanding the link between the loss of PPARG, chronic inflammation and tumorigenesis in the urothelium could shed light on the urothelial differentiation network and pave the way for the development of therapeutic approaches to various urinary diseases.