Delay of EGF-Stimulated EGFR Degradation in Myotonic Dystrophy Type 1 (DM1)

Eva Alegre-Cortés; Alberto Giménez-Bejarano; Elisabet Uribe-Carretero; Marta Paredes-Barquero; André R A Marques; Mafalda Lopes-da-Silva; Otília V Vieira; Saray Canales-Cortés; Pedro J Camello; Guadalupe Martínez-Chacón; Ana Aiastui; Roberto Fernández-Torrón; Adolfo López de Munain; Patricia Gomez-Suaga; Mireia Niso-Santano; Rosa A González-Polo; José M Fuentes; Sokhna M S Yakhine-Diop

doi:10.3390/cells11193018

Delay of EGF-Stimulated EGFR Degradation in Myotonic Dystrophy Type 1 (DM1)

Cells. 2022 Sep 27;11(19):3018. doi: 10.3390/cells11193018.

Authors

Eva Alegre-Cortés^{1

2}, Alberto Giménez-Bejarano^{1

2}, Elisabet Uribe-Carretero^{1

2

3}, Marta Paredes-Barquero^{1

2}, André R A Marques⁴, Mafalda Lopes-da-Silva⁴, Otília V Vieira⁴, Saray Canales-Cortés^{1

2}, Pedro J Camello^{5

6}, Guadalupe Martínez-Chacón^{1

2

3}, Ana Aiastui^{3

7

8}, Roberto Fernández-Torrón^{3

8

9

10

11}, Adolfo López de Munain^{3

8

9

10

11}, Patricia Gomez-Suaga^{1

2}, Mireia Niso-Santano^{1

2

3}, Rosa A González-Polo^{1

2

3}, José M Fuentes^{1

2

3}, Sokhna M S Yakhine-Diop^{2

3}

Affiliations

¹ Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Enfermería y Terapia Ocupacional, Universidad de Extremadura, Avda de la universidad s/n, 10003 Cáceres, Spain.
² Instituto de Investigación Biosanitaria de Extremadura (INUBE), 06071 Cáceres, Spain.
³ Centro de Investigación Biomédica en Red de Enfermedades (CIBERNED), 28031 Madrid, Spain.
⁴ iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade Nova de Lisboa, 1169-056 Lisboa, Portugal.
⁵ Departamento de Fisiología, Facultad de Veterinaria, Universidad de Extremadura, 10003 Cáceres, Spain.
⁶ Instituto Universitario de Biomarcadores de Patologías Metabólicas, 10003 Cáceres, Spain.
⁷ Cell Culture Platform, Biodonostia Health Research Institute, 20014 San Sebastián, Spain.
⁸ Neuroscience Area of Biodonostia Health Research Institute, Donostia University Hospital, 20014 San Sebastián, Spain.
⁹ Department of Neurology, Donostia University Hospital, 20014 Osakidetza, Spain.
¹⁰ Ilundain Foundation, 20014 San Sebastian, Spain.
¹¹ Department of Neurosciences, University of the Basque Country UPV-EHU, 48940 San Sebastián, Spain.

Abstract

Myotonic dystrophy type 1 (DM1) is an autosomal dominant disease caused by a CTG repeat expansion in the 3' untranslated region of the dystrophia myotonica protein kinase gene. AKT dephosphorylation and autophagy are associated with DM1. Autophagy has been widely studied in DM1, although the endocytic pathway has not. AKT has a critical role in endocytosis, and its phosphorylation is mediated by the activation of tyrosine kinase receptors, such as epidermal growth factor receptor (EGFR). EGF-activated EGFR triggers the internalization and degradation of ligand-receptor complexes that serve as a PI3K/AKT signaling platform. Here, we used primary fibroblasts from healthy subjects and DM1 patients. DM1-derived fibroblasts showed increased autophagy flux, with enlarged endosomes and lysosomes. Thereafter, cells were stimulated with a high concentration of EGF to promote EGFR internalization and degradation. Interestingly, EGF binding to EGFR was reduced in DM1 cells and EGFR internalization was also slowed during the early steps of endocytosis. However, EGF-activated EGFR enhanced AKT and ERK1/2 phosphorylation levels in the DM1-derived fibroblasts. Therefore, there was a delay in EGF-stimulated EGFR endocytosis in DM1 cells; this alteration might be due to the decrease in the binding of EGF to EGFR, and not to a decrease in AKT phosphorylation.

Keywords: AKT; DMPK; LBPA; autophagy; endosomes; lysosomes; muscle atrophy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

3' Untranslated Regions
Epidermal Growth Factor* / genetics
Epidermal Growth Factor* / pharmacology
ErbB Receptors / metabolism
Humans
Ligands
Myotonic Dystrophy* / genetics
Phosphatidylinositol 3-Kinases / genetics
Proto-Oncogene Proteins c-akt / metabolism

Substances

3' Untranslated Regions
Ligands
Epidermal Growth Factor
EGFR protein, human
ErbB Receptors
Proto-Oncogene Proteins c-akt

Grants and funding

This research was supported by the Isabel Gemio Foundation (P18–13) and was also partially supported by the “Fondo Europeo de Desarrollo Regional” (FEDER) from the European Union. E.A.-C. was supported by a pre-doctoral fellowship of Valhondo Calaff Foundation. S.C.-C. and E.U.-C. were supported by FPU fellowships (FPU19/04435 and FPU16/00684, respectively) from the Ministerio de Ciencia, Innovación y Universidades, Spain. M.P.-B. and A.G.-B. received fellowships from the “Plan Propio de Iniciación a la Investigación, Desarrollo Tecnológico e Innovación (Universidad de Extremadura). M.N.-S. was supported by the “Ramon y Cajal” Program (RYC-2016–20883), and P.G.-S., was funded by “Juan de la Cierva Incorporación” Program (IJC2019–039229-I), Spain. S.M.S.Y.-D. was supported by the Isabel Gemio Foundation and CIBERNED (CB06/05/0041). J.M.F received research support from the Isabel Gemio Foundation and the “Instituto de Salud Carlos” III, CIBERNED (CB06/05/0041).