Elucidating the role of Agl in bladder carcinogenesis by generation and characterization of genetically engineered mice

Joseph L. Sottnik, Vandana Mallaredy, Ana Chauca-Diaz, Carolyn Ritterson Lew, Charles Owens, Garrett M. Dancik, Serena Pagliarani, Sabrina Lucchiari, Maurizio Moggio, Michela Ripolone, Giacomo P. Comi, Henry F. Frierson, David Clouthier, Dan Theodorescu

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Abstract

Amylo-α-1,6-glucosidase,4-α-glucanotransferase (AGL) is an enzyme primarily responsible for glycogen debranching. Germline mutations lead to glycogen storage disease type III (GSDIII). We recently found AGL to be a tumor suppressor in xenograft models of human bladder cancer (BC) and low levels of AGL expression in BC are associated with poor patient prognosis. However, the impact of low AGL expression on the susceptibility of normal bladder to carcinogenesis is unknown. We address this gap by developing a germline Agl knockout (Agl-/-) mouse that recapitulates biochemical and histological features of GSDIII. Agl-/-mice exposed to N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) had a higher BC incidence compared with wild-type mice (Agl +/+). To determine if the increased BC incidence observed was due to decreased Agl expression in the urothelium specifically, we developed a urothelium-specific conditional Agl knockout (Agl cko) mouse using a Uroplakin II-Cre allele. BBN-induced carcinogenesis experiments repeated in Agl cko mice revealed that Agl cko mice had a higher BC incidence than control (Agl fl/fl) mice. RNA sequencing revealed that tumors from Agl-/-mice had 19 differentially expressed genes compared with control mice. An â Agl Loss' gene signature was developed and found to successfully stratify normal and tumor samples in two BC patient datasets. These results support the role of AGL loss in promoting carcinogenesis and provide a rationale for evaluating Agl expression levels, or Agl Loss gene signature scores, in normal urothelium of populations at risk of BC development such as older male smokers.

Original languageEnglish
Pages (from-to)194-201
Number of pages8
JournalCarcinogenesis
Volume40
Issue number1
DOIs
Publication statusPublished - Mar 12 2019

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ASJC Scopus subject areas

  • Cancer Research

Cite this

Sottnik, J. L., Mallaredy, V., Chauca-Diaz, A., Ritterson Lew, C., Owens, C., Dancik, G. M., Pagliarani, S., Lucchiari, S., Moggio, M., Ripolone, M., Comi, G. P., Frierson, H. F., Clouthier, D., & Theodorescu, D. (2019). Elucidating the role of Agl in bladder carcinogenesis by generation and characterization of genetically engineered mice. Carcinogenesis, 40(1), 194-201. https://doi.org/10.1093/carcin/bgy139