Home Life Sciences In vitro modelling of familial amyloidotic polyneuropathy allows quantitative detection of transthyretin amyloid fibril-like structures in hepatic derivatives of patient-specific induced pluripotent stem cells
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In vitro modelling of familial amyloidotic polyneuropathy allows quantitative detection of transthyretin amyloid fibril-like structures in hepatic derivatives of patient-specific induced pluripotent stem cells

  • Jeannine Hoepfner , Mandy Kleinsorge , Oliver Papp , Susanne Alfken , Robin Heiringhoff , Andreas Pich , Vanessa Sauer , Andree Zibert , Gudrun Göhring , Hartmut Schmidt , Malte Sgodda and Tobias Cantz ORCID logo EMAIL logo
Published/Copyright: January 4, 2017
Biological Chemistry
From the journal Biological Chemistry Volume 398 Issue 8

Abstract

The transthyretin protein is thermodynamically destabilised by mutations in the transthyretin gene, promoting the formation of amyloid fibrils in various tissues. Consequently, impaired autonomic organ function is observed in patients suffering from transthyretin-related familial amyloidotic polyneuropathy (FAP). The influence of individual genetic backgrounds on fibril formation as a potential cause of genotype-phenotype variations needs to be investigated in order to ensure efficient patient-specific therapies. We reprogrammed FAP patient fibroblasts to induced pluripotent stem (iPS) cells and differentiated these cells into transthyretin-expressing hepatocyte-like cells (HLCs). HLCs differentiated from FAP iPS cells and healthy control iPS cells secreted the transthyretin protein in similar concentrations. Mass spectrometry revealed the presence of mutant transthyretin protein in FAP HLC supernatants. In comparison to healthy control iPS cells, we demonstrated the formation of transthyretin amyloid fibril-like structures in FAP HLC supernatants using the amyloid-specific dyes Congo red and thioflavin T. These dyes were also applicable for the quantitative determination of in vitro formed transthyretin fibril-like structures. Moreover, we confirmed the inhibition of fibril formation by the TTR kinetic stabiliser diclofenac. Thioflavin T fluorescence intensity measurements even allowed the quantification of amyloid fibril-like structures in 96-well plate formats as a prerequisite for patient-specific drug screening approaches.

Keywords: disease modelling; familial amyloidotic polyneuropathy; hepatic differentiation; induced pluripotent stem cells; transthyretin amyloid fibrils

Acknowledgements

This work was supported by funding from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) for the Cluster of Excellence REBIRTH (From Regenerative Biology to Reconstructive Therapy; EXC 62/3) and by the state of North Rhine-Westphalia through the initiative Translational Stem Cell Research (PtJ-Az: w1403ts011b).

  1. Conflict of interest statement:The authors declare that there is no conflict of interest regarding the publication of this article.

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Supplemental Material:

The online version of this article (DOI: https://doi.org/10.1515/hsz-2016-0258) offers supplementary material, available to authorized users.

Received: 2016-7-26
Accepted: 2016-12-22
Published Online: 2017-1-4
Published in Print: 2017-7-26

©2017 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/hsz-2016-0258
Keywords for this article
disease modelling; familial amyloidotic polyneuropathy; hepatic differentiation; induced pluripotent stem cells; transthyretin amyloid fibrils

Articles in the same Issue

  1. Frontmatter
  2. Guest Editorial
  3. Viruses and cancer: molecular relations and perspectives
  4. HIGHLIGHT: VIRUSES AND CANCER
  5. Chronic viral hepatitis and its association with liver cancer
  6. The biology of JC polyomavirus
  7. Rhadinoviral interferon regulatory factor homologues
  8. Human papillomavirus first and second generation vaccines–current status and future directions
  9. Employing RNA viruses to fight cancer: novel insights into oncolytic virotherapy
  10. Modulation of oncogenic signaling networks by Kaposi’s sarcoma-associated herpesvirus
  11. Mechanisms and strategies of papillomavirus replication
  12. Review
  13. MicroRNAs are important regulators of drug resistance in colorectal cancer
  14. Research Articles/Short Communications
  15. Molecular Medicine
  16. In vitro modelling of familial amyloidotic polyneuropathy allows quantitative detection of transthyretin amyloid fibril-like structures in hepatic derivatives of patient-specific induced pluripotent stem cells
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