First quantitative high-throughput screen in zebrafish identifies novel pathways for increasing pancreatic β-cell mass

Abstract

Whole-organism chemical screening can circumvent bottlenecks that impede drug discovery. However, in vivo screens have not attained throughput capacities possible with in vitro assays. We therefore developed a method enabling in vivo high-throughput screening (HTS) in zebrafish, termed automated reporter quantification in vivo (ARQiv). In this study, ARQiv was combined with robotics to fully actualize whole-organism HTS (ARQiv-HTS). In a primary screen, this platform quantified cell-specific fluorescent reporters in >500,000 transgenic zebrafish larvae to identify FDAapproved (Federal Drug Administration) drugs that increased the number of insulin-producing β cells in the pancreas. 24 drugs were confirmed as inducers of endocrine differentiation and/or stimulators of β-cell proliferation. Further, we discovered novel roles for NF-κB signaling in regulating endocrine differentiation and for serotonergic signaling in selectively stimulating β-cell proliferation. These studies demonstrate the power of ARQiv-HTS for drug discovery and provide unique insights into signaling pathways controlling β-cell mass, potential therapeutic targets for treating diabetes.

This work was supported by the NIDDK, NIH (1RC4DK090816). Other support was as follows: GW and MJP—Juvenile Diabetes Research Foundation (17-2012-408), and the NIH (R01DK080730); FD -MSCRF 2013 Postdoctoral Fellowship; SKR and JSM—Diabetic Complications Consortium, NIDDK, NIH (12GHSU209). JSS—Science and Technology Development Fund (FDCT) of Macau SAR (FDCT/119/2013/A3); RK—NINDS (R01 NS073751); JOL—FAMRI, ITCR and Prostate Cancer Foundation. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.