Role in tumor growth of a glycogen debranching enzyme lost in glycogen storage disease

Sunny Guin, Courtney Pollard, Yuanbin Ru, Carolyn Ritterson Lew, Jason E. Duex, Garrett Dancik, Charles Owens, Andrea Spencer, Scott Knight, Heather Holemon, Sounak Gupta, Donna Hansel, Marc Hellerstein, Pawel Lorkiewicz, Andrew N. Lane, Teresa W.M. Fan, Dan Theodorescu

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

Background Bladder cancer is the most common malignancy of the urinary system, yet our molecular understanding of this disease is incomplete, hampering therapeutic advances. Methods Here we used a genome-wide functional short-hairpin RNA (shRNA) screen to identify suppressors of in vivo bladder tumor xenograft growth (n = 50) using bladder cancer UMUC3 cells. Next-generation sequencing was used to identify the most frequently occurring shRNAs in tumors. Genes so identified were studied in 561 patients with bladder cancer for their association with stratification of clinical outcome by Kaplan-Meier analysis. The best prognostic marker was studied to determine its mechanism in tumor suppression using anchorage-dependent and-independent growth, xenograft (n = 20), and metabolomic assays. Statistical significance was determined using two-sided Student t test and repeated-measures statistical analysis. Results We identified the glycogen debranching enzyme AGL as a prognostic indicator of patient survival (P =. 04) and as a novel regulator of bladder cancer anchorage-dependent (P <. 001), anchorage-independent (mean ± standard deviation, 180 ± 23.1 colonies vs 20±9.5 in control, P <. 001), and xenograft growth (P <. 001). Rescue experiments using catalytically dead AGL variants revealed that this effect is independent of AGL enzymatic functions. We demonstrated that reduced AGL enhances tumor growth by increasing glycine synthesis through increased expression of serine hydroxymethyltransferase 2. Conclusions Using an in vivo RNA interference screen, we discovered that AGL, a glycogen debranching enzyme, has a biologically and statistically significant role in suppressing human cancer growth.

Original languageEnglish (US)
Article numberdju062
JournalJournal of the National Cancer Institute
Volume106
Issue number5
DOIs
StatePublished - May 14 2014
Externally publishedYes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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