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

  1. Guangliang Wang
  2. Surendra K Rajpurohit
  3. Fabien Delaspre
  4. Steven L Walker
  5. David T White
  6. Alexis Ceasrine
  7. Rejji Kuruvilla
  8. Ruo-jing Li
  9. Joong S Shim
  10. Jun O Liu
  11. Michael J Parsons
  12. Jeff S Mumm  Is a corresponding author
  1. Johns Hopkins University, United States
  2. Georgia Regents University, United States
  3. University of Macau, China

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). Here, 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 FDA-approved drugs that increased the number of insulin-producing β cells in the pancreas. Twenty-four 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.

Article and author information

Author details

  1. Guangliang Wang

    McKusick-Nathans Institute for Genetic Medicine, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.
  2. Surendra K Rajpurohit

    Department of Cellular Biology and Anatomy, Georgia Regents University, Augusta, United States
    Competing interests
    No competing interests declared.
  3. Fabien Delaspre

    McKusick-Nathans Institute for Genetic Medicine, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.
  4. Steven L Walker

    Department of Cellular Biology and Anatomy, Georgia Regents University, Augusta, United States
    Competing interests
    No competing interests declared.
  5. David T White

    Wilmer Eye Institute, Johns Hopkins University, Augusta, United States
    Competing interests
    No competing interests declared.
  6. Alexis Ceasrine

    Department of Biology, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.
  7. Rejji Kuruvilla

    Department of Biology, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.
  8. Ruo-jing Li

    Pharmacology and Molecular Sciences, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.
  9. Joong S Shim

    Faculty of Health Sciences, University of Macau, Macau, China
    Competing interests
    No competing interests declared.
  10. Jun O Liu

    Pharmacology and Molecular Sciences, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.
  11. Michael J Parsons

    McKusick-Nathans Institute for Genetic Medicine, Johns Hopkins University, Baltimore, United States
    Competing interests
    No competing interests declared.
  12. Jeff S Mumm

    Wilmer Eye Institute, Johns Hopkins University, Baltimore, United States
    For correspondence
    jmumm3@jhmi.edu
    Competing interests
    Jeff S Mumm, acts as a consultant for, Luminomics Inc., a company which uses drug discovery techniques applied in thetext.

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved animal care and use committee (ACUC) protocols of Johns Hopkins University and Georgia Regents University

Copyright

© 2015, Wang et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 6,528
    views
  • 1,271
    downloads
  • 80
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Guangliang Wang
  2. Surendra K Rajpurohit
  3. Fabien Delaspre
  4. Steven L Walker
  5. David T White
  6. Alexis Ceasrine
  7. Rejji Kuruvilla
  8. Ruo-jing Li
  9. Joong S Shim
  10. Jun O Liu
  11. Michael J Parsons
  12. Jeff S Mumm
(2015)
First quantitative high-throughput screen in zebrafish identifies novel pathways for increasing pancreatic β-cell mass
eLife 4:e08261.
https://doi.org/10.7554/eLife.08261

Share this article

https://doi.org/10.7554/eLife.08261

Further reading

    1. Cell Biology
    Rachel Pudlowski, Lingyi Xu ... Jennifer T Wang
    Research Advance

    Centrioles have a unique, conserved architecture formed by three linked, ‘triplet’, microtubules arranged in ninefold symmetry. The mechanisms by which these triplet microtubules are formed remain unclear but likely involve the noncanonical tubulins delta-tubulin and epsilon-tubulin. Previously, we found that human cells lacking delta-tubulin or epsilon-tubulin form abnormal centrioles, characterized by an absence of triplet microtubules, lack of central core protein POC5, and a futile cycle of centriole formation and disintegration (Wang et al., 2017). Here, we show that human cells lacking either TEDC1 or TEDC2 have similar abnormalities. Using ultrastructure expansion microscopy, we observed that mutant centrioles elongate to the same length as control centrioles in G2 phase and fail to recruit central core scaffold proteins. Remarkably, mutant centrioles also have an expanded proximal region. During mitosis, these mutant centrioles further elongate before fragmenting and disintegrating. All four proteins physically interact and TEDC1 and TEDC2 can form a subcomplex in the absence of the tubulins, supporting an AlphaFold Multimer model of the tetramer. TEDC1 and TEDC2 localize to centrosomes and are mutually dependent on each other and on delta-tubulin and epsilon-tubulin for localization. Our results demonstrate that delta-tubulin, epsilon-tubulin, TEDC1, and TEDC2 function together to promote robust centriole architecture, laying the foundation for future studies on the mechanisms underlying the assembly of triplet microtubules and their interactions with centriole structure.

    1. Cancer Biology
    2. Cell Biology
    Zuzana Outla, Gizem Oyman-Eyrilmez ... Martin Gregor
    Research Article

    The most common primary malignancy of the liver, hepatocellular carcinoma (HCC), is a heterogeneous tumor entity with high metastatic potential and complex pathophysiology. Increasing evidence suggests that tissue mechanics plays a critical role in tumor onset and progression. Here, we show that plectin, a major cytoskeletal crosslinker protein, plays a crucial role in mechanical homeostasis and mechanosensitive oncogenic signaling that drives hepatocarcinogenesis. Our expression analyses revealed elevated plectin levels in liver tumors, which correlated with poor prognosis for HCC patients. Using autochthonous and orthotopic mouse models we demonstrated that genetic and pharmacological inactivation of plectin potently suppressed the initiation and growth of HCC. Moreover, plectin targeting potently inhibited the invasion potential of human HCC cells and reduced their metastatic outgrowth in the lung. Proteomic and phosphoproteomic profiling linked plectin-dependent disruption of cytoskeletal networks to attenuation of oncogenic FAK, MAPK/Erk, and PI3K/Akt signatures. Importantly, by combining cell line-based and murine HCC models, we show that plectin inhibitor plecstatin-1 (PST) is well-tolerated and potently inhibits HCC progression. In conclusion, our study demonstrates that plectin-controlled cytoarchitecture is a key determinant of HCC development and suggests that pharmacologically induced disruption of mechanical homeostasis may represent a new therapeutic strategy for HCC treatment.