Functional analysis of iPSC-derived myocytes from a patient with carnitine palmitoyltransferase II deficiency.
BACKGROUND
Carnitine palmitoyltransferase II (CPT II) deficiency is an inherited disorder involving β-oxidation of long-chain fatty acids (FAO), which leads to rhabdomyolysis and subsequent acute renal failure. The detailed mechanisms of disease pathogenesis remain unknown; however, the availability of relevant human cell types for investigation, such as skeletal muscle cells, is limited, and the development of novel disease models is required.


METHODS
We generated human induced pluripotent stem cells (hiPSCs) from skin fibroblasts of a Japanese patient with CPT II deficiency. Mature myocytes were differentiated from the patient-derived hiPSCs by introducing myogenic differentiation 1 (MYOD1), the master transcriptional regulator of myocyte differentiation. Using an in vitro acylcarnitine profiling assay, we investigated the effects of a hypolipidemic drug, bezafibrate, and heat stress on mitochondrial FAO in CPT II-deficient myocytes and controls.


RESULTS
CPT II-deficient myocytes accumulated more palmitoylcarnitine (C16) than did control myocytes. Heat stress, induced by incubation at 38°C, leads to a robust increase of C16 in CPT II-deficient myocytes, but not in controls. Bezafibrate reduced the amount of C16 in control and CPT II-deficient myocytes.


CONCLUSIONS
In this study, we induced differentiation of CPT II-deficient hiPSCs into mature myocytes in a highly efficient and reproducible manner and recapitulated some aspects of the disease phenotypes of CPT II deficiency in the myocyte disease models. This approach addresses the challenges of modeling the abnormality of FAO in CPT II deficiency using iPSC technology and has the potential to revolutionize translational research in this field.
Division of Nephrology and Rheumatology, Department of Internal Medicine, Fukuoka University School of Medicine, Fukuoka, Japan. Electronic address: yasuno9584@fukuoka-u.ac.jp.
Tetsuhiko
T
Yasuno
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Kenji
K
Osafune
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Hidetoshi
H
Sakurai
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Isao
I
Asaka
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Akihito
A
Tanaka
Department of Pediatrics, Shimane University School of Medicine, Izumo, Shimane, Japan.
Seiji
S
Yamaguchi
Department of Pediatrics, Shimane University School of Medicine, Izumo, Shimane, Japan.
Kenji
K
Yamada
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Hirofumi
H
Hitomi
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Sayaka
S
Arai
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
Yuko
Y
Kurose
Faculty of Sports and Health Science, Fukuoka University, Japan.
Yasuki
Y
Higaki
Faculty of Sports and Health Science, Fukuoka University, Japan.
Mizuki
M
Sudo
Faculty of Sports and Health Science, Fukuoka University, Japan.
Soichi
S
Ando
Division of Nephrology and Rheumatology, Department of Internal Medicine, Fukuoka University School of Medicine, Fukuoka, Japan.
Hitoshi
H
Nakashima
Division of Nephrology and Rheumatology, Department of Internal Medicine, Fukuoka University School of Medicine, Fukuoka, Japan; General Medical Research Center, Fukuoka University School of Medicine, Japan.
Takao
T
Saito
Division of Nephrology and Rheumatology, Department of Internal Medicine, Fukuoka University School of Medicine, Fukuoka, Japan; Division of Medical Sciences, Fukuoka University School of Nursing, Japan.
Hidetoshi
H
Kaneoka
D001629:Bezafibrate / Q000494:pharmacology; D002331:Carnitine / Q000031:analogs & derivatives / Q000378:metabolism; D002334:Carnitine O-Palmitoyltransferase / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000473:pathology; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D008661:Metabolism, Inborn Errors / Q000378:metabolism* / Q000473:pathology*; D032342:Muscle Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D010172:Palmitoylcarnitine / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism / Q000473:pathology*; D055815:Young Adult
C116917:acylcarnitine; D010172:Palmitoylcarnitine; D002334:Carnitine O-Palmitoyltransferase; D002331:Carnitine; D001629:Bezafibrate
Bezafibrate; Carnitine palmitoyltransferase II deficiency; Disease modeling; Rhabdomyolysis; iPSC