1. Cell Biology
  2. Stem Cells and Regenerative Medicine

NAD+ enhances ribitol and ribose rescue of α-dystroglycan functional glycosylation in human FKRP-mutant myotubes

  1. Carolina Ortiz Cordero
  2. Alessandro Magli
  3. Neha Dhoke
  4. Taylor Kuebler
  5. Nelio AJ Oliveira
  6. Haowen Zhou
  7. Yuk Y Sham
  8. Anne G Bang
  9. Rita CR Perlingeiro  Is a corresponding author
  1. University of Minnesota, United States
  2. Sanford Burnham Prebys Medical Discovery Institute, United States
https://doi.org/10.7554/eLife.65443
Share this article
Cite this article
  1. Carolina Ortiz Cordero
  2. Alessandro Magli
  3. Neha Dhoke
  4. Taylor Kuebler
  5. Nelio AJ Oliveira
  6. Haowen Zhou
  7. Yuk Y Sham
  8. Anne G Bang
  9. Rita CR Perlingeiro
(2021)
NAD+ enhances ribitol and ribose rescue of α-dystroglycan functional glycosylation in human FKRP-mutant myotubes
eLife 10:e65443.
https://doi.org/10.7554/eLife.65443
  2. Download BibTeX
  3. Download .RIS

Abstract

Mutations in the fukutin-related protein (FKRP) cause Walker-Warburg Syndrome (WWS), a severe form of congenital muscular dystrophy. Here we established a WWS human induced pluripotent stem cell-derived myogenic model that recapitulates hallmarks of WWS pathology. We used this model to investigate the therapeutic effect of metabolites of the pentose phosphate pathway in human WWS. We show that functional recovery of WWS myotubes is promoted not only by ribitol but also its precursor ribose. Moreover, we found that the combination of each of these metabolites with NAD+ results in a synergistic effect, as demonstrated by rescue of α-dystroglycan glycosylation and laminin binding capacity. Mechanistically, we find that FKRP residual enzymatic capacity, characteristic of many recessive FKRP mutations, is required for rescue as supported by functional and structural mutational analysis. These findings provide the rationale for testing ribose/ribitol in combination with NAD+ to treat WWS and other diseases associated with FKRP mutations.

Data availability

Complete Images for blots and analyzed data is available at Dryad, Dataset, (https://doi.org/10.5061/dryad.x3ffbg7hx).

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Carolina Ortiz Cordero

    Department of Integrative Biology and Physiology and Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6953-0366

  2. Alessandro Magli

    Lillehei Heart Institute, University of Minnesota, Minneapolis, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3874-2838

  3. Neha Dhoke

    Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Taylor Kuebler

    Bioinformatics and Computational Biology Program, University of Minnesota, Minneapolis, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Nelio AJ Oliveira

    Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. Haowen Zhou

    Conrad Prebys Center for Chemical Genomics, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.

  7. Yuk Y Sham

    Department of Integrative Biology and Bioinformatics and Computational Biology Program, University of Minnesota, Minneapolis, United States
    Competing interests
    The authors declare that no competing interests exist.

  8. Anne G Bang

    Conrad Prebys Center for Chemical Genomics, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.

  9. Rita CR Perlingeiro

    Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, United States
    For correspondence
    perli032@umn.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9412-1118

Funding

National Institute of Arthritis and Musculoskeletal and Skin Diseases (AR055299)

  • Rita CR Perlingeiro

National Institute of Arthritis and Musculoskeletal and Skin Diseases (AR071439)

  • Rita CR Perlingeiro

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

Ethics

Animal experimentation: Animal experiments were carried out according to protocols (protocol ID: 2002-37833A) approved by the University of Minnesota Institutional Animal Care and Use Committee.

Copyright

© 2021, Ortiz Cordero 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

  • 1,863
    views
  • 267
    downloads
  • 10
    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. Carolina Ortiz Cordero
  2. Alessandro Magli
  3. Neha Dhoke
  4. Taylor Kuebler
  5. Nelio AJ Oliveira
  6. Haowen Zhou
  7. Yuk Y Sham
  8. Anne G Bang
  9. Rita CR Perlingeiro
(2021)
NAD+ enhances ribitol and ribose rescue of α-dystroglycan functional glycosylation in human FKRP-mutant myotubes
eLife 10:e65443.
https://doi.org/10.7554/eLife.65443

Share this article

https://doi.org/10.7554/eLife.65443

Categories and tags

Research organism

Further reading

    1. Cell Biology
    2. Computational and Systems Biology

    Unbiased identification of cell identity in dense mixed neural cultures

    Sarah De Beuckeleer, Tim Van De Looverbosch ... Winnok H De Vos
    Research Article

    Induced pluripotent stem cell (iPSC) technology is revolutionizing cell biology. However, the variability between individual iPSC lines and the lack of efficient technology to comprehensively characterize iPSC-derived cell types hinder its adoption in routine preclinical screening settings. To facilitate the validation of iPSC-derived cell culture composition, we have implemented an imaging assay based on cell painting and convolutional neural networks to recognize cell types in dense and mixed cultures with high fidelity. We have benchmarked our approach using pure and mixed cultures of neuroblastoma and astrocytoma cell lines and attained a classification accuracy above 96%. Through iterative data erosion, we found that inputs containing the nuclear region of interest and its close environment, allow achieving equally high classification accuracy as inputs containing the whole cell for semi-confluent cultures and preserved prediction accuracy even in very dense cultures. We then applied this regionally restricted cell profiling approach to evaluate the differentiation status of iPSC-derived neural cultures, by determining the ratio of postmitotic neurons and neural progenitors. We found that the cell-based prediction significantly outperformed an approach in which the population-level time in culture was used as a classification criterion (96% vs 86%, respectively). In mixed iPSC-derived neuronal cultures, microglia could be unequivocally discriminated from neurons, regardless of their reactivity state, and a tiered strategy allowed for further distinguishing activated from non-activated cell states, albeit with lower accuracy. Thus, morphological single-cell profiling provides a means to quantify cell composition in complex mixed neural cultures and holds promise for use in the quality control of iPSC-derived cell culture models.

    1. Cell Biology

    Endocytic recycling is central to circadian collagen fibrillogenesis and disrupted in fibrosis

    Joan Chang, Adam Pickard ... Karl E Kadler
    Research Article

    Collagen-I fibrillogenesis is crucial to health and development, where dysregulation is a hallmark of fibroproliferative diseases. Here, we show that collagen-I fibril assembly required a functional endocytic system that recycles collagen-I to assemble new fibrils. Endogenous collagen production was not required for fibrillogenesis if exogenous collagen was available, but the circadian-regulated vacuolar protein sorting (VPS) 33b and collagen-binding integrin α11 subunit were crucial to fibrillogenesis. Cells lacking VPS33B secrete soluble collagen-I protomers but were deficient in fibril formation, thus secretion and assembly are separately controlled. Overexpression of VPS33B led to loss of fibril rhythmicity and overabundance of fibrils, which was mediated through integrin α11β1. Endocytic recycling of collagen-I was enhanced in human fibroblasts isolated from idiopathic pulmonary fibrosis, where VPS33B and integrin α11 subunit were overexpressed at the fibrogenic front; this correlation between VPS33B, integrin α11 subunit, and abnormal collagen deposition was also observed in samples from patients with chronic skin wounds. In conclusion, our study showed that circadian-regulated endocytic recycling is central to homeostatic assembly of collagen fibrils and is disrupted in diseases.

    1. Cell Biology

    Hypersensitive intercellular responses of endometrial stromal cells drive invasion in endometriosis

    Chun-Wei Chen, Jeffery B Chavez ... Bruce J Nicholson
    Research Article Updated

    Endometriosis is a debilitating disease affecting 190 million women worldwide and the greatest single contributor to infertility. The most broadly accepted etiology is that uterine endometrial cells retrogradely enter the peritoneum during menses, and implant and form invasive lesions in a process analogous to cancer metastasis. However, over 90% of women suffer retrograde menstruation, but only 10% develop endometriosis, and debate continues as to whether the underlying defect is endometrial or peritoneal. Processes implicated in invasion include: enhanced motility; adhesion to, and formation of gap junctions with, the target tissue. Endometrial stromal (ESCs) from 22 endometriosis patients at different disease stages show much greater invasiveness across mesothelial (or endothelial) monolayers than ESCs from 22 control subjects, which is further enhanced by the presence of EECs. This is due to the enhanced responsiveness of endometriosis ESCs to the mesothelium, which induces migration and gap junction coupling. ESC-PMC gap junction coupling is shown to be required for invasion, while coupling between PMCs enhances mesothelial barrier breakdown.