Reserpine maintains photoreceptor survival in retinal ciliopathy by resolving proteostasis imbalance and ciliogenesis defects

  1. Holly Y Chen
  2. Manju Swaroop
  3. Samantha Papal
  4. Anupam Mondal
  5. Hyun Beom Song
  6. Laura Campello
  7. Gregory Tawa
  8. Florian Regent
  9. Hiroko Shimada
  10. Kunio Nagashima
  11. Natalia de Val
  12. Samuel G Jacobson
  13. Wei Zheng
  14. Anand Swaroop  Is a corresponding author
  1. National Eye Institute, United States
  2. National Center for Advancing Translational Sciences, United States
  3. Frederick National Laboratory for Cancer Research, United States
  4. University of Pennsylvania, United States

Abstract

Ciliopathies manifest from sensory abnormalities to syndromic disorders with multi-organ pathologies, with retinal degeneration a highly penetrant phenotype. Photoreceptor cell death is a major cause of incurable blindness in retinal ciliopathies. To identify drug candidates to maintain photoreceptor survival, we performed an unbiased, high-throughput screening of over 6,000 bioactive small molecules using retinal organoids differentiated from induced pluripotent stem cells (iPSC) of rd16 mouse, which is a model of Leber congenital amaurosis (LCA) type 10 caused by mutations in the cilia-centrosomal gene CEP290. We identified five non-toxic positive hits, including the lead molecule reserpine, which maintained photoreceptor development and survival in rd16 organoids. Reserpine also improved photoreceptors in retinal organoids derived from induced pluripotent stem cells of LCA10 patients and in rd16 mouse retina in vivo. Reserpine-treated patient organoids revealed modulation of signaling pathways related to cell survival/death, metabolism, and proteostasis. Further investigation uncovered dysregulation of autophagy associated with compromised primary cilium biogenesis in patient organoids and rd16 mouse retina. Reserpine partially restored the balance between autophagy and the ubiquitin-proteasome system at least in part by increasing the cargo adaptor p62, resulting in improved primary cilium assembly. Our study identifies effective drug candidates in preclinical studies of CEP290 retinal ciliopathies through cross-species drug discovery using iPSC-derived organoids, highlights the impact of proteostasis in the pathogenesis of ciliopathies, and provides new insights for treatments of retinal neurodegeneration.

Data availability

All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. RNA-seq data are available through GEO accession #206959.

The following data sets were generated

Article and author information

Author details

  1. Holly Y Chen

    Neurobiology, Neurodegeneration and Repair Laboratory, National Eye Institute, Bethesda, United States
    Competing interests
    Holly Y Chen, Listed as inventor on a patent application related to the small molecules in this study by National Institutes of Health (PCT/US2021/040157).
  2. Manju Swaroop

    National Therapeutics for Rare and Neglected Diseases, National Center for Advancing Translational Sciences, Rockville, United States
    Competing interests
    Manju Swaroop, Listed as inventor on a patent application related to the small molecules in this study by National Institutes of Health (PCT/US2021/040157).
  3. Samantha Papal

    Neurobiology, Neurodegeneration and Repair Laboratory, National Eye Institute, Bethesda, United States
    Competing interests
    Samantha Papal, Listed as inventor on a patent application related to the small molecules in this study by National Institutes of Health (PCT/US2021/040157).
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9417-6215
  4. Anupam Mondal

    Neurobiology, Neurodegeneration and Repair Laboratory, National Eye Institute, Bethesda, United States
    Competing interests
    Anupam Mondal, Listed as inventor on a patent application related to the small molecules in this study by National Institutes of Health (PCT/US2021/040157).
  5. Hyun Beom Song

    Neurobiology, Neurodegeneration and Repair Laboratory, National Eye Institute, Bethesda, United States
    Competing interests
    No competing interests declared.
  6. Laura Campello

    Neurobiology, Neurodegeneration and Repair Laboratory, National Eye Institute, Bethesda, United States
    Competing interests
    No competing interests declared.
  7. Gregory Tawa

    National Therapeutics for Rare and Neglected Diseases, National Center for Advancing Translational Sciences, Rockville, United States
    Competing interests
    Gregory Tawa, Listed as inventor on a patent application related to the small molecules in this study by National Institutes of Health (PCT/US2021/040157).
  8. Florian Regent

    Neurobiology, Neurodegeneration and Repair Laboratory, National Eye Institute, Bethesda, United States
    Competing interests
    No competing interests declared.
  9. Hiroko Shimada

    Neurobiology, Neurodegeneration and Repair Laboratory, National Eye Institute, Bethesda, United States
    Competing interests
    No competing interests declared.
  10. Kunio Nagashima

    Electron Microscopy Laboratory, Frederick National Laboratory for Cancer Research, Frederick, United States
    Competing interests
    No competing interests declared.
  11. Natalia de Val

    Electron Microscopy Laboratory, Frederick National Laboratory for Cancer Research, Frederick, United States
    Competing interests
    No competing interests declared.
  12. Samuel G Jacobson

    Department of Ophthalmology, University of Pennsylvania, Philadelphia, United States
    Competing interests
    No competing interests declared.
  13. Wei Zheng

    National Therapeutics for Rare and Neglected Diseases, National Center for Advancing Translational Sciences, Rockville, United States
    Competing interests
    Wei Zheng, Listed as inventor on a patent application related to the small molecules in this study by National Institutes of Health (PCT/US2021/040157).
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1034-0757
  14. Anand Swaroop

    Neurobiology, Neurodegeneration and Repair Laboratory, National Eye Institute, Bethesda, United States
    For correspondence
    swaroopa@nei.nih.gov
    Competing interests
    Anand Swaroop, Listed as inventor on a patent application related to the small molecules in this study by National Institutes of Health (PCT/US2021/040157).
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1975-1141

Funding

National Eye Institute (Z01EY000546)

  • Anand Swaroop

National Eye Institute (Z01EY000450)

  • Anand Swaroop

National Center for Advancing Translational Sciences (ZIATR000018-06)

  • Wei Zheng

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Zhongjie Fu, Boston Children's Hospital, United States

Ethics

Animal experimentation: All animal procedures were approved by the Animal Care and Use committee of the National Eye Institutes (Animal study protocol NEI-650) and adhered to ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.

Version history

  1. Received: September 2, 2022
  2. Preprint posted: September 18, 2022 (view preprint)
  3. Accepted: March 23, 2023
  4. Accepted Manuscript published: March 28, 2023 (version 1)
  5. Version of Record published: April 21, 2023 (version 2)

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Metrics

  • 1,590
    Page views
  • 356
    Downloads
  • 10
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

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. Holly Y Chen
  2. Manju Swaroop
  3. Samantha Papal
  4. Anupam Mondal
  5. Hyun Beom Song
  6. Laura Campello
  7. Gregory Tawa
  8. Florian Regent
  9. Hiroko Shimada
  10. Kunio Nagashima
  11. Natalia de Val
  12. Samuel G Jacobson
  13. Wei Zheng
  14. Anand Swaroop
(2023)
Reserpine maintains photoreceptor survival in retinal ciliopathy by resolving proteostasis imbalance and ciliogenesis defects
eLife 12:e83205.
https://doi.org/10.7554/eLife.83205

Share this article

https://doi.org/10.7554/eLife.83205

Further reading

    1. Biochemistry and Chemical Biology
    Jake W Anderson, David Vaisar ... Natalie G Ahn
    Research Article

    Activation of the extracellular signal-regulated kinase-2 (ERK2) by phosphorylation has been shown to involve changes in protein dynamics, as determined by hydrogen-deuterium exchange mass spectrometry (HDX-MS) and NMR relaxation dispersion measurements. These can be described by a global exchange between two conformational states of the active kinase, named ‘L’ and ‘R,’ where R is associated with a catalytically productive ATP-binding mode. An ATP-competitive ERK1/2 inhibitor, Vertex-11e, has properties of conformation selection for the R-state, revealing movements of the activation loop that are allosterically coupled to the kinase active site. However, the features of inhibitors important for R-state selection are unknown. Here, we survey a panel of ATP-competitive ERK inhibitors using HDX-MS and NMR and identify 14 new molecules with properties of R-state selection. They reveal effects propagated to distal regions in the P+1 and helix αF segments surrounding the activation loop, as well as helix αL16. Crystal structures of inhibitor complexes with ERK2 reveal systematic shifts in the Gly loop and helix αC, mediated by a Tyr-Tyr ring stacking interaction and the conserved Lys-Glu salt bridge. The findings suggest a model for the R-state involving small movements in the N-lobe that promote compactness within the kinase active site and alter mobility surrounding the activation loop. Such properties of conformation selection might be exploited to modulate the protein docking interface used by ERK substrates and effectors.

    1. Biochemistry and Chemical Biology
    Anne E Hultgren, Nicole MF Patras, Jenna Hicks
    Feature Article

    Organizations that fund research are keen to ensure that their grant selection processes are fair and equitable for all applicants. In 2020, the Arnold and Mabel Beckman Foundation introduced blinding to the first stage of the process used to review applications for Beckman Young Investigator (BYI) awards: applicants were instructed to blind the technical proposal in their initial Letter of Intent by omitting their name, gender, gender-identifying pronouns, and institutional information. Here we examine the impact of this change by comparing the data on gender and institutional prestige of the applicants in the first four years of the new policy (BYI award years 2021–2024) with data on the last four years of the old policy (2017–2020). We find that under the new policy, the distribution of applicants invited to submit a full application shifted from those affiliated with institutions regarded as more prestigious to those outside of this group, and that this trend continued through to the final program awards. We did not find evidence of a shift in the distribution of applicants with respect to gender.