Phenotypic features of dystrophin gene knockout pigs harboring a human artificial chromosome containing the entire dystrophin gene
Mammalian artificial chromosomes have enabled the introduction of extremely large amounts of genetic information into animal cells in an autonomously replicating, nonintegrating format. However, the evaluation of human artificial chromosomes (HACs) as novel tools for curing intractable hereditary di...
| Main Authors: | , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-09-01
|
| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253123001944 |
| _version_ | 1797752482685779968 |
|---|---|
| author | Masahito Watanabe Hitomaru Miyamoto Kazutoshi Okamoto Kazuaki Nakano Hitomi Matsunari Kanako Kazuki Koki Hasegawa Ayuko Uchikura Shuko Takayanagi Kazuhiro Umeyama Yosuke Hiramuki Elisabeth Kemter Nikolai Klymuik Mayuko Kurome Barbara Kessler Eckhard Wolf Yasuhiro Kazuki Hiroshi Nagashima |
| author_facet | Masahito Watanabe Hitomaru Miyamoto Kazutoshi Okamoto Kazuaki Nakano Hitomi Matsunari Kanako Kazuki Koki Hasegawa Ayuko Uchikura Shuko Takayanagi Kazuhiro Umeyama Yosuke Hiramuki Elisabeth Kemter Nikolai Klymuik Mayuko Kurome Barbara Kessler Eckhard Wolf Yasuhiro Kazuki Hiroshi Nagashima |
| author_sort | Masahito Watanabe |
| collection | DOAJ |
| description | Mammalian artificial chromosomes have enabled the introduction of extremely large amounts of genetic information into animal cells in an autonomously replicating, nonintegrating format. However, the evaluation of human artificial chromosomes (HACs) as novel tools for curing intractable hereditary disorders has been hindered by the limited efficacy of the delivery system. We generated dystrophin gene knockout (DMD-KO) pigs harboring the HAC bearing the entire human DMD via a somatic cell cloning procedure (DYS-HAC-cloned pig). Restored human dystrophin expression was confirmed by immunofluorescence staining in the skeletal muscle of the DYS-HAC-cloned pigs. Viability at the first month postpartum of the DYS-HAC-cloned pigs, including motor function in the hind leg and serum creatinine kinase level, was improved significantly when compared with that in the original DMD-KO pigs. However, decrease in systemic retention of the DYS-HAC vector and limited production of the DMD protein might have caused severe respiratory impairment with general prostration by 3 months postpartum. The results demonstrate that the use of transchromosomic cloned pigs permitted a straightforward estimation of the efficacy of the DYS-HAC carried in affected tissues/organs in a large-animal disease model, providing novel insights into the therapeutic application of exogenous mammalian artificial chromosomes. |
| first_indexed | 2024-03-12T17:04:04Z |
| format | Article |
| id | doaj.art-7a8c635119da410397f678e585a6364e |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-03-12T17:04:04Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-7a8c635119da410397f678e585a6364e2023-08-07T04:04:44ZengElsevierMolecular Therapy: Nucleic Acids2162-25312023-09-0133444453Phenotypic features of dystrophin gene knockout pigs harboring a human artificial chromosome containing the entire dystrophin geneMasahito Watanabe0Hitomaru Miyamoto1Kazutoshi Okamoto2Kazuaki Nakano3Hitomi Matsunari4Kanako Kazuki5Koki Hasegawa6Ayuko Uchikura7Shuko Takayanagi8Kazuhiro Umeyama9Yosuke Hiramuki10Elisabeth Kemter11Nikolai Klymuik12Mayuko Kurome13Barbara Kessler14Eckhard Wolf15Yasuhiro Kazuki16Hiroshi Nagashima17Meiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, JapanDepartment of Chromosome Biomedical Engineering, School of Life Science, Faculty of Medicine, Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, JapanLaboratory of Medical Bioengineering, Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, JapanMeiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, JapanMeiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, JapanChromosome Engineering Research Center (CERC), Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, JapanLaboratory of Medical Bioengineering, Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, JapanMeiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, JapanMeiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, JapanMeiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, JapanChromosome Engineering Research Center (CERC), Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, JapanChair for Molecular Animal Breeding and Biotechnology, Gene Center and Department of Veterinary Sciences, LMU Munich, 81377 Munich, Germany; Center for Innovative Medical Models (CiMM), LMU Munich, 85764 Oberschleissheim, GermanyChair for Molecular Animal Breeding and Biotechnology, Gene Center and Department of Veterinary Sciences, LMU Munich, 81377 Munich, Germany; Center for Innovative Medical Models (CiMM), LMU Munich, 85764 Oberschleissheim, GermanyMeiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan; Chair for Molecular Animal Breeding and Biotechnology, Gene Center and Department of Veterinary Sciences, LMU Munich, 81377 Munich, Germany; Center for Innovative Medical Models (CiMM), LMU Munich, 85764 Oberschleissheim, GermanyChair for Molecular Animal Breeding and Biotechnology, Gene Center and Department of Veterinary Sciences, LMU Munich, 81377 Munich, Germany; Center for Innovative Medical Models (CiMM), LMU Munich, 85764 Oberschleissheim, GermanyMeiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan; Chair for Molecular Animal Breeding and Biotechnology, Gene Center and Department of Veterinary Sciences, LMU Munich, 81377 Munich, Germany; Center for Innovative Medical Models (CiMM), LMU Munich, 85764 Oberschleissheim, GermanyDepartment of Chromosome Biomedical Engineering, School of Life Science, Faculty of Medicine, Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, Japan; Chromosome Engineering Research Center (CERC), Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, Japan; Chromosome Engineering Research Group, The Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan; Corresponding author: Yasuhiro Kazuki, Department of Chromosome Biomedical Engineering, School of Life Science, Faculty of Medicine, Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, Japan.Meiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan; Laboratory of Medical Bioengineering, Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan; Corresponding author: Hiroshi Nagashima, Meiji University International Institute for Bio-Resource Research, 1-1-1 Higashimita, Tama-ku, Kawasaki, Kanagawa 214-8571, Japan.Mammalian artificial chromosomes have enabled the introduction of extremely large amounts of genetic information into animal cells in an autonomously replicating, nonintegrating format. However, the evaluation of human artificial chromosomes (HACs) as novel tools for curing intractable hereditary disorders has been hindered by the limited efficacy of the delivery system. We generated dystrophin gene knockout (DMD-KO) pigs harboring the HAC bearing the entire human DMD via a somatic cell cloning procedure (DYS-HAC-cloned pig). Restored human dystrophin expression was confirmed by immunofluorescence staining in the skeletal muscle of the DYS-HAC-cloned pigs. Viability at the first month postpartum of the DYS-HAC-cloned pigs, including motor function in the hind leg and serum creatinine kinase level, was improved significantly when compared with that in the original DMD-KO pigs. However, decrease in systemic retention of the DYS-HAC vector and limited production of the DMD protein might have caused severe respiratory impairment with general prostration by 3 months postpartum. The results demonstrate that the use of transchromosomic cloned pigs permitted a straightforward estimation of the efficacy of the DYS-HAC carried in affected tissues/organs in a large-animal disease model, providing novel insights into the therapeutic application of exogenous mammalian artificial chromosomes.http://www.sciencedirect.com/science/article/pii/S2162253123001944MT: Delivery Strategieshuman artificial chromosomecloned pigDuchenne muscular dystrophysomatic cell cloningdystrophin gene knockout pig |
| spellingShingle | Masahito Watanabe Hitomaru Miyamoto Kazutoshi Okamoto Kazuaki Nakano Hitomi Matsunari Kanako Kazuki Koki Hasegawa Ayuko Uchikura Shuko Takayanagi Kazuhiro Umeyama Yosuke Hiramuki Elisabeth Kemter Nikolai Klymuik Mayuko Kurome Barbara Kessler Eckhard Wolf Yasuhiro Kazuki Hiroshi Nagashima Phenotypic features of dystrophin gene knockout pigs harboring a human artificial chromosome containing the entire dystrophin gene Molecular Therapy: Nucleic Acids MT: Delivery Strategies human artificial chromosome cloned pig Duchenne muscular dystrophy somatic cell cloning dystrophin gene knockout pig |
| title | Phenotypic features of dystrophin gene knockout pigs harboring a human artificial chromosome containing the entire dystrophin gene |
| title_full | Phenotypic features of dystrophin gene knockout pigs harboring a human artificial chromosome containing the entire dystrophin gene |
| title_fullStr | Phenotypic features of dystrophin gene knockout pigs harboring a human artificial chromosome containing the entire dystrophin gene |
| title_full_unstemmed | Phenotypic features of dystrophin gene knockout pigs harboring a human artificial chromosome containing the entire dystrophin gene |
| title_short | Phenotypic features of dystrophin gene knockout pigs harboring a human artificial chromosome containing the entire dystrophin gene |
| title_sort | phenotypic features of dystrophin gene knockout pigs harboring a human artificial chromosome containing the entire dystrophin gene |
| topic | MT: Delivery Strategies human artificial chromosome cloned pig Duchenne muscular dystrophy somatic cell cloning dystrophin gene knockout pig |
| url | http://www.sciencedirect.com/science/article/pii/S2162253123001944 |
| work_keys_str_mv | AT masahitowatanabe phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT hitomarumiyamoto phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT kazutoshiokamoto phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT kazuakinakano phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT hitomimatsunari phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT kanakokazuki phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT kokihasegawa phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT ayukouchikura phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT shukotakayanagi phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT kazuhiroumeyama phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT yosukehiramuki phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT elisabethkemter phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT nikolaiklymuik phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT mayukokurome phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT barbarakessler phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT eckhardwolf phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT yasuhirokazuki phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene AT hiroshinagashima phenotypicfeaturesofdystrophingeneknockoutpigsharboringahumanartificialchromosomecontainingtheentiredystrophingene |