MP25-09 HIGH THROUGHPUT GENETIC SCREENING WITH DROSOPHILA MELANOGASTER IDENTIFIES NOVEL GENES ASSOCIATED WITH STONE FORMATION

Abstract

You have accessJournal of UrologyStone Disease: Basic Research II1 Apr 2014MP25-09 HIGH THROUGHPUT GENETIC SCREENING WITH DROSOPHILA MELANOGASTER IDENTIFIES NOVEL GENES ASSOCIATED WITH STONE FORMATION Thomas Chi, Man Su Kim, Sven Lang, Tiffany Zee, Gulinuer Muteliefu, Sarah Blaschko, David Killilea, Katja Bruckner, Arnold Kahn, Pankaj Kapahi, and Marshall Stoller Thomas ChiThomas Chi More articles by this author , Man Su KimMan Su Kim More articles by this author , Sven LangSven Lang More articles by this author , Tiffany ZeeTiffany Zee More articles by this author , Gulinuer MuteliefuGulinuer Muteliefu More articles by this author , Sarah BlaschkoSarah Blaschko More articles by this author , David KillileaDavid Killilea More articles by this author , Katja BrucknerKatja Bruckner More articles by this author , Arnold KahnArnold Kahn More articles by this author , Pankaj KapahiPankaj Kapahi More articles by this author , and Marshall StollerMarshall Stoller More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.311AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Vertebrate models for nephrolithiasis are limited in their ability to rapidly screen multiple and varied interventions that modulate urinary stone formation. A short lifespan and rapid stone formation makes Drosophila melanogaster an ideal system to screen large numbers of interventions to assess their effect on kidney stone formation. A method of screening single gene knockdown candidates for their ability to prevent the formation of urinary stones was successfully developed utilizing Drosophila. METHODS Microdissection facilitates identification and collection of fly stones within the lumen of the Drosophila Malpighian tubule (the functional equivalent of the human renal tubule). In a similar fashion as seen in humans, fly stones frequently obstruct the ureter and result in early death. Utilizing shortened lifespan as a surrogate for stone formation, an unbiased screen of orthologues for human genes known to be associated with nephrolithiasis was undertaken. A double knockdown unbiased screen to identify genes that lengthened lifespan and modulated the stone formation process was developed; microdissection confirmed these results. RESULTS Stone formation was associated with a significantly shortened fly lifespan (mean lifespan 60 days with no stones, 3 days with stones, p<0.05). An unbiased double knockdown screen of more than 80 genes was performed utilizing the GAL4-UAS RNAi system and identified 7 genes that rescued lifespan. Microdissection confirmed that three genes decreased stone formation in the fly, including genes encoding a salt transporter (SLC5A5), a zinc transporter (ZnT35C), and a regulator of oxidative stress (Cyp4d1). These lengthened lifespan to 9, 12, and 11 days (p<0.05) respectively and significantly modulated the stone formation process (see Table). CONCLUSIONS A Drosophila urinary stone model was leveraged to perform large scale genetic screens to identify novel genes that modulate calculi formation. Newly identified genes may represent novel targets that could translate to new therapies to prevent or reduce the formation of urinary stones. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e269-e270 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Thomas Chi More articles by this author Man Su Kim More articles by this author Sven Lang More articles by this author Tiffany Zee More articles by this author Gulinuer Muteliefu More articles by this author Sarah Blaschko More articles by this author David Killilea More articles by this author Katja Bruckner More articles by this author Arnold Kahn More articles by this author Pankaj Kapahi More articles by this author Marshall Stoller More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...