Exome sequencing in NICD driven mouse sarcomas

We have developed mice that overexpress Notch intracellular domain (NICD) specifically in their adipocytes (RosaNICD/NICD::AdipoqCre; PMID: 26137442 ). These mice develop lipodystrophy and concomitant diabetes, dyslipidemia and fatty liver disease. During aging and at a median age of 225 days the mice develop poorly differentiated high-grade sarcomas that are designated to be liposarcomas. This study aimed to investigate the mutational landscape of these tumors. To this end,genomic DNA was prepared from 5 tumors of AdNICD mice and 5 tails from the same animals using the QIAamp DNA mini kit (Qiagen). DNA was quantified by spectrophotometry using Nanodrop One and purity was assessed by the absorbance ratios at 260/280 and 260/230 nm. The integrity of the DNA was evaluated by the genomic DNA ScreenTape assay (Agilent) in the Agilent TapeStation system and the DNA integrity number (DIN) was calculated for each sample. Next generation sequencing services were provided by Admera Health LLC (South Plainfield, NJ). The SureSelect Mouse All Exon Kit (Agilent) was used for library preparation and the HiSeq sequencing system (Illumina, San Diego, CA) was used. Read length was 2x150bp (paired end) with mean on target coverage per sample 100x. The information that can be derived from this study is physiologically relevant given that some human sarcomas overexpress Notch and its target gene Hes1. It can also provide us what mutations are "favored" in a background of NICD overexpressing adipocytes.