title: Genetic and pharmacological correction of aberrant dopamine synthesis using patient iPSCs with BH4 metabolism disorders.
abstract: Dopamine (DA) is a neurotransmitter in the brain, playing a central role in several disease conditions, including tetrahydrobiopterin (BH4) metabolism disorders and Parkinson's disease (PD). BH4 metabolism disorders present a variety of clinical manifestations including motor disturbance via altered DA metabolism, since BH4 is a cofactor for tyrosine hydroxylase (TH), a rate-limiting enzyme for DA synthesis. Genetically, BH4 metabolism disorders are, in an autosomal recessive pattern, caused by a variant in genes encoding enzymes for BH4 synthesis or recycling, including 6-pyruvoyltetrahydropterin synthase (PTPS) or dihydropteridine reductase (DHPR), respectively. Although BH4 metabolism disorders and its metabolisms have been studied, it is unclear how gene variants cause aberrant DA synthesis in patient neurons. Here, we generated induced pluripotent stem cells (iPSCs) from BH4 metabolism disorder patients with PTPS or DHPR variants, corrected the gene variant in the iPSCs using the CRISPR/Cas9 system, and differentiated the BH4 metabolism disorder patient- and isogenic control iPSCs into midbrain DA neurons. We found that by the gene correction, the BH4 amount, TH protein level and extracellular DA level were restored in DA neuronal culture using PTPS deficiency iPSCs. Furthermore, the pharmacological correction by BH4 precursor sepiapterin treatment also improved the phenotypes of PTPS deficiency. These results suggest that patient iPSCs with BH4 metabolism disorders provide an opportunity for screening substances for treating aberrant DA synthesis-related disorders.
authors: Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, Japan.|Sumitomo Dainippon Pharma, 3-1-98 Kasugadenaka, Konohana-ku, Osaka, Japan.
authors: Taizo
authors: T
authors: Ishikawa
authors: Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, Japan.
authors: Keiko
authors: K
authors: Imamura
authors: Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, Japan.
authors: Takayuki
authors: T
authors: Kondo
authors: Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, Japan.|Department of Neurology, Kyoto University Graduate School of Medicine, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, Japan.
authors: Yasushi
authors: Y
authors: Koshiba
authors: School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
authors: Satoshi
authors: S
authors: Hara
authors: School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
authors: Hiroshi
authors: H
authors: Ichinose
authors: Department of Pediatrics, Okayama Medical Center, National Hospital Organization, Okayama, Japan.
authors: Mahoko
authors: M
authors: Furujo
authors: Department of Neurology, Utano National Hospital, National Hospital Organization, Kyoto, Japan.
authors: Masako
authors: M
authors: Kinoshita
authors: Department of Neurology, Utano National Hospital, National Hospital Organization, Kyoto, Japan.
authors: Tomoko
authors: T
authors: Oeda
authors: Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, Japan.
authors: Jun
authors: J
authors: Takahashi
authors: Department of Neurology, Kyoto University Graduate School of Medicine, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, Japan.
authors: Ryosuke
authors: R
authors: Takahashi
authors: Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, Japan.
authors: Haruhisa
authors: H
authors: Inoue
mesh_terms: D001708:Biopterins / Q000031:analogs & derivatives* / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D004298:Dopamine / Q000096:biosynthesis / Q000235:genetics*; D059290:Dopaminergic Neurons / Q000378:metabolism / Q000473:pathology; D005838:Genotype; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D059785:Karyotype; D008659:Metabolic Diseases / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D010300:Parkinson Disease / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D011622:Pterins / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
chemical_list: D011622:Pterins; D001708:Biopterins; D014446:Tyrosine 3-Monooxygenase; C003402:sapropterin; D004298:Dopamine