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CRISPR-enhanced human adipocyte “browning” as cell therapy for metabolic disease

View ORCID ProfileEmmanouela Tsagkaraki, Sarah Nicoloro, Tiffany De Souza, View ORCID ProfileJavier Solivan-Rivera, Anand Desai, Yuefei Shen, Mark Kelly, Adilson Guilherme, View ORCID ProfileFelipe Henriques, View ORCID ProfileRaed Ibraheim, View ORCID ProfileNadia Amrani, View ORCID ProfileKevin Luk, Stacy Maitland, Randall H. Friedline, Lauren Tauer, Xiaodi Hu, View ORCID ProfileJason K. Kim, View ORCID ProfileScot A. Wolfe, View ORCID ProfileErik J. Sontheimer, View ORCID ProfileSilvia Corvera, View ORCID ProfileMichael P. Czech
doi: https://doi.org/10.1101/2020.10.13.337923
Emmanouela Tsagkaraki
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
6University of Crete School of Medicine, Crete 71003
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  • ORCID record for Emmanouela Tsagkaraki
Sarah Nicoloro
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Tiffany De Souza
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Javier Solivan-Rivera
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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  • ORCID record for Javier Solivan-Rivera
Anand Desai
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Yuefei Shen
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Mark Kelly
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Adilson Guilherme
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Felipe Henriques
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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  • ORCID record for Felipe Henriques
Raed Ibraheim
2RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester MA 01605
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Nadia Amrani
2RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester MA 01605
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Kevin Luk
4Department of Molecular,Cell and Cancer Biology, University of Massachusetts Medical School, Worcester MA 01605
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Stacy Maitland
4Department of Molecular,Cell and Cancer Biology, University of Massachusetts Medical School, Worcester MA 01605
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Randall H. Friedline
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Lauren Tauer
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Xiaodi Hu
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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Jason K. Kim
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
3Division of Endocrinology, Metabolism and Diabetes, Department of Medicine, University of Massachusetts Medical School Worcester, MA 01605
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Scot A. Wolfe
4Department of Molecular,Cell and Cancer Biology, University of Massachusetts Medical School, Worcester MA 01605
5Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, MA 01605
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Erik J. Sontheimer
2RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester MA 01605
5Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, MA 01605
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Silvia Corvera
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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  • For correspondence: Michael.Czech@umassmed.edu Silvia.Corvera@umassmed.edu
Michael P. Czech
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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  • ORCID record for Michael P. Czech
  • For correspondence: Michael.Czech@umassmed.edu Silvia.Corvera@umassmed.edu
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Abstract

Obesity and type 2 diabetes (T2D) are associated with poor tissue responses to insulin1,2, disturbances in glucose and lipid fluxes3–5 and comorbidities including steatohepatitis6 and cardiovascular disease7,8. Despite extensive efforts at prevention and treatment9,10, diabetes afflicts over 400 million people worldwide11. Whole body metabolism is regulated by adipose tissue depots12–14, which include both lipid-storing white adipocytes and less abundant “brown” and “brite/beige” adipocytes that express thermogenic uncoupling protein UCP1 and secrete factors favorable to metabolic health15–18. Application of clustered regularly interspaced short palindromic repeats (CRISPR) gene editing19,20 to enhance “browning” of white adipose tissue is an attractive therapeutic approach to T2D. However, the problems of cell-selective delivery, immunogenicity of CRISPR reagents and long term stability of the modified adipocytes are formidable. To overcome these issues, we developed methods that deliver complexes of SpyCas9 protein and sgRNA ex vivo to disrupt the thermogenesis suppressor gene NRIP121,22 with near 100% efficiency in human or mouse adipocytes. NRIP1 gene disruption at discrete loci strongly ablated NRIP1 protein and upregulated expression of UCP1 and beneficial secreted factors, while residual Cas9 protein and sgRNA were rapidly degraded. Implantation of the CRISPR-enhanced human or mouse brown-like adipocytes into high fat diet fed mice decreased adiposity and liver triglycerides while enhancing glucose tolerance compared to mice implanted with unmodified adipocytes. These findings advance a therapeutic strategy to improve metabolic homeostasis through CRISPR-based genetic modification of human adipocytes without exposure of the recipient to immunogenic Cas9 or delivery vectors.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted October 13, 2020.
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CRISPR-enhanced human adipocyte “browning” as cell therapy for metabolic disease
Emmanouela Tsagkaraki, Sarah Nicoloro, Tiffany De Souza, Javier Solivan-Rivera, Anand Desai, Yuefei Shen, Mark Kelly, Adilson Guilherme, Felipe Henriques, Raed Ibraheim, Nadia Amrani, Kevin Luk, Stacy Maitland, Randall H. Friedline, Lauren Tauer, Xiaodi Hu, Jason K. Kim, Scot A. Wolfe, Erik J. Sontheimer, Silvia Corvera, Michael P. Czech
bioRxiv 2020.10.13.337923; doi: https://doi.org/10.1101/2020.10.13.337923
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CRISPR-enhanced human adipocyte “browning” as cell therapy for metabolic disease
Emmanouela Tsagkaraki, Sarah Nicoloro, Tiffany De Souza, Javier Solivan-Rivera, Anand Desai, Yuefei Shen, Mark Kelly, Adilson Guilherme, Felipe Henriques, Raed Ibraheim, Nadia Amrani, Kevin Luk, Stacy Maitland, Randall H. Friedline, Lauren Tauer, Xiaodi Hu, Jason K. Kim, Scot A. Wolfe, Erik J. Sontheimer, Silvia Corvera, Michael P. Czech
bioRxiv 2020.10.13.337923; doi: https://doi.org/10.1101/2020.10.13.337923

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