Merging organoid and organ-on-a-chip technology to generate complex multi-layer tissue models in a human Retina-on-a-Chip platform

Abstract

The devastating effects and incurable nature of hereditary and sporadic retinal diseases such as Stargardt disease, age-related macular degeneration or retinitis pigmentosa urgently require the development of new therapeutic strategies. Additionally, a high prevalence of retinal toxicities is becoming more and more an issue of novel targeted therapeutic agents. Ophthalmologic drug development, to date, largely relies on animal models, which often do not provide results that are translatable to human patients. Hence, the establishment of sophisticated human tissue-based in vitro models is of upmost importance. The discovery of self-forming retinal organoids (ROs) derived from human embryonic stem cells (hESCs) or human induced pluripotent stem cells (hiPSCs) is a promising approach to model the complex stratified retinal tissue. Yet, ROs lack vascularization and cannot recapitulate the important physiological interactions of matured photoreceptors and the retinal pigment epithelium (RPE). In this study, we present the retina-on-a-chip (RoC), a novel microphysiological model of the human retina integrating more than seven different essential retinal cell-types derived from hiPSCs. It provides vasculature-like perfusion and enables, for the first time, the recapitulation of the interaction of mature photoreceptor segments with RPE in vitro. We show that this interaction enhances the formation of outer segment-like structures and the establishment of in vivo-like physiological processes such as outer segment phagocytosis and calcium dynamics. In addition, we demonstrate the applicability of the RoC for drug testing, by reproducing the retinopathic side-effects of the anti-malaria drug chloroquine and the antibiotic gentamicin. The developed hiPSC-based RoC has the potential to promote drug development and provide new insights into the underlying pathology of retinal diseases.

Data availability

The authors declare that the main data supporting the findings of this study are available within the article and its Supplementary Information files.

Article and author information

Author details

  1. Kevin Achberger

    Institute of Neuroanatomy and Developmental Biology (INDB), Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Christopher Probst

    Attract Group Organ-on-a-Chip, Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Stuttgart, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Jasmin Haderspeck

    Institute of Neuroanatomy and Developmental Biology (INDB), Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Silvia Bolz

    Centre for Ophthalmology, Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Julia Rogal

    Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Stuttgart, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Johanna Chuchuy

    Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Stuttgart, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Marina Nikolova

    Institute of Neuroanatomy and Developmental Biology (INDB), Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Virginia Cora

    Institute of Neuroanatomy and Developmental Biology (INDB), Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Lena Antkowiak

    Institute of Neuroanatomy and Developmental Biology (INDB), Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  10. Wadood Haq

    Centre for Ophthalmology, Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  11. Nian Shen

    Department of Womens Health, Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  12. Katja Schenke-Layland

    Department of Womens Health, Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  13. Marius Ueffing

    Centre for Ophthalmology, Eberhard Karls University Tübingen, Tübingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  14. Stefan Liebau

    Institute of Neuroanatomy and Developmental Biology (INDB), Eberhard Karls University Tübingen, Tübingen, Germany
    For correspondence
    stefan.liebau@uni-tuebingen.de
    Competing interests
    The authors declare that no competing interests exist.
  15. Peter Loskill

    Attract Group Organ-on-a-Chip, Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Stuttgart, Germany
    For correspondence
    peter.loskill@igb.fraunhofer.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5000-0581

Funding

Fraunhofer-Gesellschaft

  • Peter Loskill

Deutsche Forschungsgemeinschaft

  • Katja Schenke-Layland
  • Stefan Liebau

Horizon 2020 Framework Programme

  • Peter Loskill

Ministerium für Wissenschaft, Forschung und Kunst Baden-Württemberg

  • Katja Schenke-Layland

National Centre for the Replacement, Refinement and Reduction of Animals in Research

  • Peter Loskill

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

Reviewing Editor

  1. Milica Radisic, University of Toronto, Canada

Version history

  1. Received: February 18, 2019
  2. Accepted: August 6, 2019
  3. Accepted Manuscript published: August 27, 2019 (version 1)
  4. Version of Record published: October 4, 2019 (version 2)

Copyright

© 2019, Achberger 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.

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  1. Kevin Achberger
  2. Christopher Probst
  3. Jasmin Haderspeck
  4. Silvia Bolz
  5. Julia Rogal
  6. Johanna Chuchuy
  7. Marina Nikolova
  8. Virginia Cora
  9. Lena Antkowiak
  10. Wadood Haq
  11. Nian Shen
  12. Katja Schenke-Layland
  13. Marius Ueffing
  14. Stefan Liebau
  15. Peter Loskill
(2019)
Merging organoid and organ-on-a-chip technology to generate complex multi-layer tissue models in a human Retina-on-a-Chip platform
eLife 8:e46188.
https://doi.org/10.7554/eLife.46188

Share this article

https://doi.org/10.7554/eLife.46188

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    Background:

    Viremia is a critical factor in understanding the pathogenesis of dengue infection, but limited data exist on viremia kinetics. This study aimed to investigate the kinetics of viremia and its effects on subsequent platelet count, severe dengue, and plasma leakage.

    Methods:

    We pooled data from three studies conducted in Vietnam between 2000 and 2016, involving 2340 dengue patients with daily viremia measurements and platelet counts after symptom onset. Viremia kinetics were assessed using a random effects model that accounted for left-censored data. The effects of viremia on subsequent platelet count and clinical outcomes were examined using a landmark approach with a random effects model and logistic regression model with generalized estimating equations, respectively. The rate of viremia decline was derived from the model of viremia kinetics. Its effect on the clinical outcomes was assessed by logistic regression models.

    Results:

    Viremia levels rapidly decreased following symptom onset, with variations observed depending on the infecting serotype. DENV-1 exhibited the highest mean viremia levels during the first 5–6 days, while DENV-4 demonstrated the shortest clearance time. Higher viremia levels were associated with decreased subsequent platelet counts from day 6 onwards. Elevated viremia levels on each illness day increased the risk of developing severe dengue and plasma leakage. However, the effect size decreased with later illness days. A more rapid decline in viremia is associated with a reduced risk of the clinical outcomes.

    Conclusions:

    This study provides comprehensive insights into viremia kinetics and its effect on subsequent platelet count and clinical outcomes in dengue patients. Our findings underscore the importance of measuring viremia levels during the early febrile phase for dengue studies and support the use of viremia kinetics as outcome for phase-2 dengue therapeutic trials.

    Funding:

    Wellcome Trust and European Union Seventh Framework Programme.