Premium
A new patient‐derived iPSC model for dystroglycanopathies validates a compound that increases glycosylation of α‐dystroglycan
Author(s) -
Kim Jihee,
Lana Beatrice,
Torelli Silvia,
Ryan David,
Catapano Francesco,
Ala Pierpaolo,
Luft Christin,
Stevens Elizabeth,
Konstantinidis Evangelos,
Louzada Sandra,
Fu Beiyuan,
ParedesRedondo Amaia,
Chan AW Edith,
Yang Fengtang,
Stemple Derek L,
Liu Pentao,
Ketteler Robin,
Selwood David L,
Muntoni Francesco,
Lin YungYao
Publication year - 2019
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.15252/embr.201947967
Subject(s) - glycosylation , dystroglycan , biology , hek 293 cells , laminin , induced pluripotent stem cell , microbiology and biotechnology , gene , extracellular matrix , genetics , embryonic stem cell
Dystroglycan, an extracellular matrix receptor, has essential functions in various tissues. Loss of α‐dystroglycan‐laminin interaction due to defective glycosylation of α‐dystroglycan underlies a group of congenital muscular dystrophies often associated with brain malformations, referred to as dystroglycanopathies. The lack of isogenic human dystroglycanopathy cell models has limited our ability to test potential drugs in a human‐ and neural‐specific context. Here, we generated induced pluripotent stem cells ( iPSC s) from a severe dystroglycanopathy patient with homozygous FKRP (fukutin‐related protein gene) mutation. We showed that CRISPR /Cas9‐mediated gene correction of FKRP restored glycosylation of α‐dystroglycan in iPSC ‐derived cortical neurons, whereas targeted gene mutation of FKRP in wild‐type cells disrupted this glycosylation. In parallel, we screened 31,954 small molecule compounds using a mouse myoblast line for increased glycosylation of α‐dystroglycan. Using human FKRP ‐ iPSC ‐derived neural cells for hit validation, we demonstrated that compound 4‐(4‐bromophenyl)‐6‐ethylsulfanyl‐2‐oxo‐3,4‐dihydro‐1H‐pyridine‐5‐carbonitrile (4 BPPN it) significantly augmented glycosylation of α‐dystroglycan, in part through upregulation of LARGE 1 glycosyltransferase gene expression. Together, isogenic human iPSC ‐derived cells represent a valuable platform for facilitating dystroglycanopathy drug discovery and therapeutic development.