TY - JOUR
T1 - Development and characterization of agonistic antibodies targeting the Ig-like 1 domain of MuSK
AU - Lim, Jamie L
AU - Augustinus, Roy
AU - Plomp, Jaap J
AU - Roya-Kouchaki, Kasra
AU - Vergoossen, Dana L E
AU - Fillié-Grijpma, Yvonne
AU - Struijk, Josephine
AU - Thomas, Rachel
AU - Salvatori, Daniela
AU - Steyaert, Christophe
AU - Blanchetot, Christophe
AU - Vanhauwaert, Roeland
AU - Silence, Karen
AU - van der Maarel, Silvère M
AU - Verschuuren, Jan J
AU - Huijbers, Maartje G
N1 - Funding Information:
We thank Dr. Christophe Borg and staff from the Borg laboratory in Besançon, France, for advice and assistance with the mice study. We thank Prof. Dr. Jelle Goeman for statistical advice, Prof. Dr. Boudewijn Lelieveldt for advice and help with data visualization, and Bernhardt Vankerckhoven for performing antibody affinity assays and stability tests.
Funding Information:
This study was funded by argenx BV and Prinses Beatrix Spierfonds. The authors are members of the European Reference Network for Rare Neuromuscular Diseases [ERN EURO-NMD] and the Netherlands Neuromuscular Center (NL-NMD). M.G.H. receives financial support from the LUMC (OIO, 2017 and a Gisela Their Fellowship 2021), Top Sector Life Sciences & Health to Samenwerkende Gezondheidsfondsen (LSHM18055-SGF and LSHM19130), Prinses Beatrix Spierfonds (W.OR-19.13) and the Dutch Science Organization NWO (VENI 0915016181 0040).
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/5/8
Y1 - 2023/5/8
N2 - Muscle-specific kinase (MuSK) is crucial for acetylcholine receptor (AChR) clustering and thereby neuromuscular junction (NMJ) function. NMJ dysfunction is a hallmark of several neuromuscular diseases, including MuSK myasthenia gravis. Aiming to restore NMJ function, we generated several agonist monoclonal antibodies targeting the MuSK Ig-like 1 domain. These activated MuSK and induced AChR clustering in cultured myotubes. The most potent agonists partially rescued myasthenic effects of MuSK myasthenia gravis patient IgG autoantibodies in vitro. In an IgG4 passive transfer MuSK myasthenia model in NOD/SCID mice, MuSK agonists caused accelerated weight loss and no rescue of myasthenic features. The MuSK Ig-like 1 domain agonists unexpectedly caused sudden death in a large proportion of male C57BL/6 mice (but not female or NOD/SCID mice), likely caused by a urologic syndrome. In conclusion, these agonists rescued pathogenic effects in myasthenia models in vitro, but not in vivo. The sudden death in male mice of one of the tested mouse strains revealed an unexpected and unexplained role for MuSK outside skeletal muscle, thereby hampering further (pre-) clinical development of these clones. Future research should investigate whether other Ig-like 1 domain MuSK antibodies, binding different epitopes, do hold a safe therapeutic promise.
AB - Muscle-specific kinase (MuSK) is crucial for acetylcholine receptor (AChR) clustering and thereby neuromuscular junction (NMJ) function. NMJ dysfunction is a hallmark of several neuromuscular diseases, including MuSK myasthenia gravis. Aiming to restore NMJ function, we generated several agonist monoclonal antibodies targeting the MuSK Ig-like 1 domain. These activated MuSK and induced AChR clustering in cultured myotubes. The most potent agonists partially rescued myasthenic effects of MuSK myasthenia gravis patient IgG autoantibodies in vitro. In an IgG4 passive transfer MuSK myasthenia model in NOD/SCID mice, MuSK agonists caused accelerated weight loss and no rescue of myasthenic features. The MuSK Ig-like 1 domain agonists unexpectedly caused sudden death in a large proportion of male C57BL/6 mice (but not female or NOD/SCID mice), likely caused by a urologic syndrome. In conclusion, these agonists rescued pathogenic effects in myasthenia models in vitro, but not in vivo. The sudden death in male mice of one of the tested mouse strains revealed an unexpected and unexplained role for MuSK outside skeletal muscle, thereby hampering further (pre-) clinical development of these clones. Future research should investigate whether other Ig-like 1 domain MuSK antibodies, binding different epitopes, do hold a safe therapeutic promise.
KW - Male
KW - Animals
KW - Mice
KW - Mice, SCID
KW - Receptor Protein-Tyrosine Kinases/metabolism
KW - Mice, Inbred C57BL
KW - Mice, Inbred NOD
KW - Myasthenia Gravis/metabolism
KW - Receptors, Cholinergic/metabolism
KW - Autoantibodies
KW - Muscle Weakness
KW - Acetylcholine
UR - http://www.scopus.com/inward/record.url?scp=85158156399&partnerID=8YFLogxK
U2 - 10.1038/s41598-023-32641-1
DO - 10.1038/s41598-023-32641-1
M3 - Article
C2 - 37156800
SN - 2045-2322
VL - 13
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 7478
ER -