Stem cells and fluid flow drive cyst formation in an invertebrate excretory organ

  1. Hanh Thi-Kim Vu
  2. Jochen C Rink
  3. Sean A McKinney
  4. Melainia McClain
  5. Naharajan Lakshmanaperumal
  6. Richard Alexander
  7. Alejandro Sánchez Alvarado  Is a corresponding author
  1. Stowers Institute for Medical Research, United States
  2. Max Planck Institute of Molecular Cell Biology and Genetics, Germany

Abstract

Cystic kidney diseases (CKDs) affect millions of people worldwide. The defining pathological features are fluid-filled cysts developing from nephric tubules due to defective flow sensing, cell proliferation and differentiation. The underlying molecular mechanisms, however, remain poorly understood, and the derived excretory systems of established invertebrate models (C. elegans and D. melanogaster) are unsuitable to model CKDs. Systematic structure/function comparisons revealed that the combination of ultrafiltration and flow-associated filtrate modification that is central to CKD etiology is remarkably conserved between the planarian excretory system and the vertebrate nephron. Consistently, both RNA-mediated genetic interference (RNAi) of planarian orthologues of human CKD genes and inhibition of tubule flow led to tubular cystogenesis that share many features with vertebrate CKDs, suggesting deep mechanistic conservation. Our results demonstrate a common evolutionary origin of animal excretory systems and establish planarians as a novel and experimentally accessible invertebrate model for the study of human kidney pathologies.

Article and author information

Author details

  1. Hanh Thi-Kim Vu

    Stowers Institute for Medical Research, Kansas City, United States
    Competing interests
    No competing interests declared.
  2. Jochen C Rink

    Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
    Competing interests
    No competing interests declared.
  3. Sean A McKinney

    Stowers Institute for Medical Research, Kansas CIty, United States
    Competing interests
    No competing interests declared.
  4. Melainia McClain

    Stowers Institute for Medical Research, Kansas City, United States
    Competing interests
    No competing interests declared.
  5. Naharajan Lakshmanaperumal

    Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
    Competing interests
    No competing interests declared.
  6. Richard Alexander

    Stowers Institute for Medical Research, Kansas City, United States
    Competing interests
    No competing interests declared.
  7. Alejandro Sánchez Alvarado

    Stowers Institute for Medical Research, Kansas City, United States
    For correspondence
    asa@stowers.org
    Competing interests
    Alejandro Sánchez Alvarado, Reviewing editor, eLife.

Reviewing Editor

  1. Yukiko M Yamashita, University of Michigan, United States

Publication history

  1. Received: March 10, 2015
  2. Accepted: June 9, 2015
  3. Accepted Manuscript published: June 9, 2015 (version 1)
  4. Version of Record published: July 14, 2015 (version 2)

Copyright

© 2015, Thi-Kim Vu 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

  • 4,199
    Page views
  • 898
    Downloads
  • 48
    Citations

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

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. Hanh Thi-Kim Vu
  2. Jochen C Rink
  3. Sean A McKinney
  4. Melainia McClain
  5. Naharajan Lakshmanaperumal
  6. Richard Alexander
  7. Alejandro Sánchez Alvarado
(2015)
Stem cells and fluid flow drive cyst formation in an invertebrate excretory organ
eLife 4:e07405.
https://doi.org/10.7554/eLife.07405
  1. Further reading

Further reading

    1. Developmental Biology
    2. Stem Cells and Regenerative Medicine
    Marta Perera et al.
    Research Article

    During embryonic development cells acquire identity at the same time as they are proliferating, implying that an intrinsic facet of cell fate choice requires coupling lineage decisions to rates of cell division. How is the cell cycle regulated to promote or suppress heterogeneity and differentiation? We explore this question combining time lapse imaging with single cell RNA-seq in the contexts of self-renewal, priming and differentiation of mouse embryonic stem cells (ESCs) towards the Primitive Endoderm lineage (PrE). Since ESCs are derived from the Inner Cell Mass of the mammalian blastocyst, ESCs in standard culture conditions are transcriptionally heterogeneous containing subfractions that are primed for either of the two ICM lineages, Epiblast and PrE. These subfractions represent dynamic states that can readily interconvert in culture, and the PrE subfraction is functionally primed for endoderm differentiation. Here we find that differential regulation of cell cycle can tip the balance between these primed populations, such that naïve ESC culture conditions promote Epiblast-like expansion and PrE differentiation stimulates the selective survival and proliferation of PrE-primed cells. In endoderm differentiation, we find that this change is accompanied by a counter-intuitive increase in G1 length that also appears replicated in vivo. While FGF/ERK signalling is a known key regulator of ESCs and PrE differentiation, we find it is not just responsible for ESCs heterogeneity, but also cell cycle synchronisation, required for the inheritance of similar cell cycles between sisters and cousins. Taken together, our results point to a tight relationship between transcriptional heterogeneity and cell cycle regulation in the context of lineage priming, with primed cell populations providing a pool of flexible cell types that can be expanded in a lineage-specific fashion while allowing plasticity during early determination.

    1. Stem Cells and Regenerative Medicine
    Rute A Tomaz et al.
    Tools and Resources

    Production of large quantities of hepatocytes remains a major challenge for a number of clinical applications in the biomedical field. Directed differentiation of human pluripotent stem cells (hPSCs) into hepatocyte-like cells (HLCs) provides an advantageous solution and a number of protocols have been developed for this purpose. However, these methods usually follow different steps of liver development in vitro, which is time consuming and requires complex culture conditions. In addition, HLCs lack the full repertoire of functionalities characterising primary hepatocytes. Here, we explore the interest of forward programming to generate hepatocytes from hPSCs and to bypass these limitations. This approach relies on the overexpression of three hepatocyte nuclear factors (HNF1A, HNF6, and FOXA3) in combination with different nuclear receptors expressed in the adult liver using the OPTi-OX platform. Forward programming allows for the rapid production of hepatocytes (FoP-Heps) with functional characteristics using a simplified process. We also uncovered that the overexpression of nuclear receptors such as RORc can enhance specific functionalities of FoP-Heps thereby validating its role in lipid/glucose metabolism. Together, our results show that forward programming could offer a versatile alternative to direct differentiation for generating hepatocytes in vitro.