In order to identify novel approaches for the treat ment of tumors associated with TSC, we used two mod els of TSC related tumors in a series of preclinical studies. Tsc2 mice were Dovitinib clinical trial used to compare Inhibitors,Modulators,Libraries disease severity of kidney disease Inhibitors,Modulators,Libraries in two different mouse strains, evaluate the age related progression of kidney disease, and compare three dif ferent dosing schedules of rapamycin. We used a subcutaneous Tsc2 tumor model to evaluate the efficacy of two VEGF inhi bitors, asparaginase, and a microtubule inhibitor. Methods Baseline tumor burden for untreated A J versus C57BL 6 Tsc2 mice and age related kidney disease in A J Tsc2 mice The Tsc2 mouse is heterozygous for a deletion of exons 1 2 as previously described. In order to determine the baseline tumor burden for untreated Tsc2 in the A J and C57BL 6 backgrounds, strain specific colonies of each background were created.

Strain speci fic colonies were created for both the A J and C57BL 6 background by backcrossing female Tsc2 heterozygous offspring with their pure strain Inhibitors,Modulators,Libraries Tsc2 wildtype fathers until the N5 generation was reached. Mice from the N5 generations were assigned to cohorts based on age, gen der, and genotype. The cohorts were Tsc2 9 months consisting of 8 males and 8 females, Tsc2 9 months consisting of 2 males and 2 females, Tsc2 12 months consisting of 4 males and 4 females, and Tsc2 12 months consisting of 2 males and 2 females. To deter mine the age related kidney disease in the A J back ground, A J Tsc2 mice were assigned to three additional cohorts. The cohorts were A J Tsc2 3 months, A J Tsc2 5 months, and A J Tsc2 7 months.

Each cohort contained Inhibitors,Modulators,Libraries 4 mice. Mice were sacrificed according to age and cohort assignment. Upon sacrifice, kidneys, livers, and lungs were examined. All animals in Tsc2 cohorts had gross kidney lesions. There were no obvious liver tumors. Three A J Tsc2 animals had gross lung abnormalities and one mouse, from the cohort treated with weekly rapamycin 12 weeks, had a superficial tail tumor. Since non kid ney tumors were rare events, these were not studied further. We also looked at Tsc2 cohorts at nine and twelve months of age and observed no gross or micro scopic kidney lesions. Quantification of kidney cystadenomas in Tsc2 mice For histological Inhibitors,Modulators,Libraries quantification of kidney cystadenomas, each kidney was prepared as previously described.

All cystadenomas were counted, measured, and scored according to the scale shown in Additional File Regorafenib CAS 1 by a blinded researcher. Since the kidney cysta denomas of these Tsc2 mice can be divided into the subgroups cystic, pre papillary, papillary and solid lesions, we use kidney cystadenomas to refer to the entire spectrum of kidney lesions observed. In addition to analyzing data according to all cystadenomas, a sub group analysis was also done by coding cystic, pre papil lary, papillary, and solid kidney lesions separately.