Defective autophagy and increased apoptosis contribute toward the pathogenesis of FKRP-associated muscular dystrophies

Carolina Ortiz Cordero; Claudia Bincoletto; Neha Dhoke; Sridhar Selvaraj; Alessandro Magli; Haowen Zhou; Do Hyung Kim; Anne G. Bang; Rita C.R. Perlingeiro

doi:10.1016/j.stemcr.2021.09.009

Defective autophagy and increased apoptosis contribute toward the pathogenesis of FKRP-associated muscular dystrophies

Carolina Ortiz Cordero, Claudia Bincoletto, Neha Dhoke, Sridhar Selvaraj, Alessandro Magli, Haowen Zhou, Do Hyung Kim, Anne G. Bang, Rita C.R. Perlingeiro

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

Fukutin-related protein (FKRP) is a glycosyltransferase involved in glycosylation of alpha-dystroglycan (α-DG). Mutations in FKRP are associated with muscular dystrophies (MD) ranging from limb-girdle LGMDR9 to Walker-Warburg Syndrome (WWS), a severe type of congenital MD. Although hypoglycosylation of α-DG is the main hallmark of this group of diseases, a full understanding of the underlying pathophysiology is still missing. Here, we investigated molecular mechanisms impaired by FKRP mutations in pluripotent stem (PS) cell–derived myotubes. FKRP-deficient myotubes show transcriptome alterations in genes involved in extracellular matrix receptor interactions, calcium signaling, PI3K-Akt pathway, and lysosomal function. Accordingly, using a panel of patient-specific LGMDR9 and WWS induced PS cell–derived myotubes, we found a significant reduction in the autophagy-lysosome pathway for both disease phenotypes. In addition, we show that WWS myotubes display decreased ERK1/2 activity and increased apoptosis, which were restored in gene edited myotubes. Our results suggest the autophagy-lysosome pathway and apoptosis may contribute to the FKRP-associated MD pathogenesis.

Original language	English (US)
Pages (from-to)	2752-2767
Number of pages	16
Journal	Stem Cell Reports
Volume	16
Issue number	11
DOIs	https://doi.org/10.1016/j.stemcr.2021.09.009
State	Published - Nov 9 2021

Bibliographical note

Funding Information:
This project was supported by NIH grants R01 AR071439, AR055299 (R.C.R.P.), R35 GM130353 (DHK) and LGMD2I Research Funds (R.C.R.P. and A.G.B). PINN MICITT Costa Rica supported C.O.C. and C.B. was supported by the Funda??o de Amparo ? Pesquisa do Estado de S?o Paulo (FAPESP, #18/07633-2). We thank Peter Karachunski from the Paul and Shelia Wellstone Muscular Dystrophy Center at the University of Minnesota for patient samples and Carsten Bonnemann at the NINDS for sharing the B12-57 iPS cell line. We thank the Wellstone Muscular Dystrophy Specialized Research Center at University of Iowa for providing patient muscle cryosections. Cytogenetic analyses performed in the Cytogenomics Shared Resource at the University of Minnesota are supported by the National Center for Advancing Translational Sciences of NIH Award Number UL1-TR002494. Confocal imaging analysis was supported by the resources and staff at the University of Minnesota University Imaging Centers (UIC) SCR_020997. The antibodies to MHC and the IIH6 antibody were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa.

Funding Information:
This project was supported by NIH grants R01 AR071439 , AR055299 (R.C.R.P.), R35 GM130353 (DHK) and LGMD2I Research Funds (R.C.R.P. and A.G.B). PINN MICITT Costa Rica supported C.O.C., and C.B. was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, # 18/07633-2 ). We thank Peter Karachunski from the Paul and Shelia Wellstone Muscular Dystrophy Center at the University of Minnesota for patient samples and Carsten Bonnemann at the NINDS for sharing the B12-57 iPS cell line. We thank the Wellstone Muscular Dystrophy Specialized Research Center at University of Iowa for providing patient muscle cryosections. Cytogenetic analyses performed in the Cytogenomics Shared Resource at the University of Minnesota are supported by the National Center for Advancing Translational Sciences of NIH Award Number UL1-TR002494 . Confocal imaging analysis was supported by the resources and staff at the University of Minnesota University Imaging Centers (UIC) SCR_020997 . The antibodies to MHC and the IIH6 antibody were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa.

Publisher Copyright:
© 2021 The Authors

Keywords

dystroglycanopathies
iPS cells
in vitro modeling
skeletal muscle

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10.1016/j.stemcr.2021.09.009

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@article{7b6f0302c12641ad9c7931e796b5b16f,

title = "Defective autophagy and increased apoptosis contribute toward the pathogenesis of FKRP-associated muscular dystrophies",

abstract = "Fukutin-related protein (FKRP) is a glycosyltransferase involved in glycosylation of alpha-dystroglycan (α-DG). Mutations in FKRP are associated with muscular dystrophies (MD) ranging from limb-girdle LGMDR9 to Walker-Warburg Syndrome (WWS), a severe type of congenital MD. Although hypoglycosylation of α-DG is the main hallmark of this group of diseases, a full understanding of the underlying pathophysiology is still missing. Here, we investigated molecular mechanisms impaired by FKRP mutations in pluripotent stem (PS) cell–derived myotubes. FKRP-deficient myotubes show transcriptome alterations in genes involved in extracellular matrix receptor interactions, calcium signaling, PI3K-Akt pathway, and lysosomal function. Accordingly, using a panel of patient-specific LGMDR9 and WWS induced PS cell–derived myotubes, we found a significant reduction in the autophagy-lysosome pathway for both disease phenotypes. In addition, we show that WWS myotubes display decreased ERK1/2 activity and increased apoptosis, which were restored in gene edited myotubes. Our results suggest the autophagy-lysosome pathway and apoptosis may contribute to the FKRP-associated MD pathogenesis.",

keywords = "dystroglycanopathies, iPS cells, in vitro modeling, skeletal muscle",

author = "{Ortiz Cordero}, Carolina and Claudia Bincoletto and Neha Dhoke and Sridhar Selvaraj and Alessandro Magli and Haowen Zhou and Kim, {Do Hyung} and Bang, {Anne G.} and Perlingeiro, {Rita C.R.}",

note = "Funding Information: This project was supported by NIH grants R01 AR071439, AR055299 (R.C.R.P.), R35 GM130353 (DHK) and LGMD2I Research Funds (R.C.R.P. and A.G.B). PINN MICITT Costa Rica supported C.O.C. and C.B. was supported by the Funda??o de Amparo ? Pesquisa do Estado de S?o Paulo (FAPESP, #18/07633-2). We thank Peter Karachunski from the Paul and Shelia Wellstone Muscular Dystrophy Center at the University of Minnesota for patient samples and Carsten Bonnemann at the NINDS for sharing the B12-57 iPS cell line. We thank the Wellstone Muscular Dystrophy Specialized Research Center at University of Iowa for providing patient muscle cryosections. Cytogenetic analyses performed in the Cytogenomics Shared Resource at the University of Minnesota are supported by the National Center for Advancing Translational Sciences of NIH Award Number UL1-TR002494. Confocal imaging analysis was supported by the resources and staff at the University of Minnesota University Imaging Centers (UIC) SCR_020997. The antibodies to MHC and the IIH6 antibody were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa. Funding Information: This project was supported by NIH grants R01 AR071439 , AR055299 (R.C.R.P.), R35 GM130353 (DHK) and LGMD2I Research Funds (R.C.R.P. and A.G.B). PINN MICITT Costa Rica supported C.O.C., and C.B. was supported by the Funda{\c c}{\~a}o de Amparo {\`a} Pesquisa do Estado de S{\~a}o Paulo (FAPESP, # 18/07633-2 ). We thank Peter Karachunski from the Paul and Shelia Wellstone Muscular Dystrophy Center at the University of Minnesota for patient samples and Carsten Bonnemann at the NINDS for sharing the B12-57 iPS cell line. We thank the Wellstone Muscular Dystrophy Specialized Research Center at University of Iowa for providing patient muscle cryosections. Cytogenetic analyses performed in the Cytogenomics Shared Resource at the University of Minnesota are supported by the National Center for Advancing Translational Sciences of NIH Award Number UL1-TR002494 . Confocal imaging analysis was supported by the resources and staff at the University of Minnesota University Imaging Centers (UIC) SCR_020997 . The antibodies to MHC and the IIH6 antibody were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa. Publisher Copyright: {\textcopyright} 2021 The Authors",

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doi = "10.1016/j.stemcr.2021.09.009",

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T1 - Defective autophagy and increased apoptosis contribute toward the pathogenesis of FKRP-associated muscular dystrophies

AU - Ortiz Cordero, Carolina

AU - Bincoletto, Claudia

AU - Dhoke, Neha

AU - Selvaraj, Sridhar

AU - Magli, Alessandro

AU - Zhou, Haowen

AU - Kim, Do Hyung

AU - Bang, Anne G.

AU - Perlingeiro, Rita C.R.

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Y1 - 2021/11/9

N2 - Fukutin-related protein (FKRP) is a glycosyltransferase involved in glycosylation of alpha-dystroglycan (α-DG). Mutations in FKRP are associated with muscular dystrophies (MD) ranging from limb-girdle LGMDR9 to Walker-Warburg Syndrome (WWS), a severe type of congenital MD. Although hypoglycosylation of α-DG is the main hallmark of this group of diseases, a full understanding of the underlying pathophysiology is still missing. Here, we investigated molecular mechanisms impaired by FKRP mutations in pluripotent stem (PS) cell–derived myotubes. FKRP-deficient myotubes show transcriptome alterations in genes involved in extracellular matrix receptor interactions, calcium signaling, PI3K-Akt pathway, and lysosomal function. Accordingly, using a panel of patient-specific LGMDR9 and WWS induced PS cell–derived myotubes, we found a significant reduction in the autophagy-lysosome pathway for both disease phenotypes. In addition, we show that WWS myotubes display decreased ERK1/2 activity and increased apoptosis, which were restored in gene edited myotubes. Our results suggest the autophagy-lysosome pathway and apoptosis may contribute to the FKRP-associated MD pathogenesis.

AB - Fukutin-related protein (FKRP) is a glycosyltransferase involved in glycosylation of alpha-dystroglycan (α-DG). Mutations in FKRP are associated with muscular dystrophies (MD) ranging from limb-girdle LGMDR9 to Walker-Warburg Syndrome (WWS), a severe type of congenital MD. Although hypoglycosylation of α-DG is the main hallmark of this group of diseases, a full understanding of the underlying pathophysiology is still missing. Here, we investigated molecular mechanisms impaired by FKRP mutations in pluripotent stem (PS) cell–derived myotubes. FKRP-deficient myotubes show transcriptome alterations in genes involved in extracellular matrix receptor interactions, calcium signaling, PI3K-Akt pathway, and lysosomal function. Accordingly, using a panel of patient-specific LGMDR9 and WWS induced PS cell–derived myotubes, we found a significant reduction in the autophagy-lysosome pathway for both disease phenotypes. In addition, we show that WWS myotubes display decreased ERK1/2 activity and increased apoptosis, which were restored in gene edited myotubes. Our results suggest the autophagy-lysosome pathway and apoptosis may contribute to the FKRP-associated MD pathogenesis.

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KW - iPS cells

KW - in vitro modeling

KW - skeletal muscle

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Defective autophagy and increased apoptosis contribute toward the pathogenesis of FKRP-associated muscular dystrophies

Abstract

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Defective autophagy and increased apoptosis contribute toward the pathogenesis of FKRP-associated muscular dystrophies

Abstract

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Keywords

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