Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-Porter
Here, we report on validating a mitochondrial gene therapy by delivering nucleic acids to mitochondria of diseased cells by a MITO-Porter, a liposome-based carrier for mitochondrial delivery. We used cells derived from a patient with a mitochondrial disease with a G625A heteroplasmic mutation in the...
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doaj-cc64909ad8ef4925ab1babd22e0632a62020-11-25T03:15:00ZengElsevierMolecular Therapy: Nucleic Acids2162-25312020-06-0120687698Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-PorterEriko Kawamura0Minako Maruyama1Jiro Abe2Akira Sudo3Atsuhito Takeda4Shingo Takada5Takashi Yokota6Shintaro Kinugawa7Hideyoshi Harashima8Yuma Yamada9Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, JapanFaculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, JapanDepartment of Pediatrics, Hokkaido University Hospital, Kita-15, Nishi-7, Kita-ku, Sapporo 060-8638, JapanNire-no-kai Children’s Clinic, Atsubetsu-cho Shimonopporo-49, Atsubetsu-ku, Sapporo 004-0007, Japan; Department of Pediatrics, Sapporo City General Hospital, Kita-11, Nishi-13, Chuo-ku, Sapporo 060-8604, JapanDepartment of Pediatrics, Hokkaido University Hospital, Kita-15, Nishi-7, Kita-ku, Sapporo 060-8638, JapanDepartment of Cardiovascular Medicine, Graduate School of Medicine, Hokkaido University, Kita-15, Nishi-7, Kita-ku, Sapporo 060-8638, JapanDepartment of Cardiovascular Medicine, Graduate School of Medicine, Hokkaido University, Kita-15, Nishi-7, Kita-ku, Sapporo 060-8638, JapanDepartment of Cardiovascular Medicine, Graduate School of Medicine, Hokkaido University, Kita-15, Nishi-7, Kita-ku, Sapporo 060-8638, JapanFaculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, JapanFaculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan; Corresponding author: Yuma Yamada, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.Here, we report on validating a mitochondrial gene therapy by delivering nucleic acids to mitochondria of diseased cells by a MITO-Porter, a liposome-based carrier for mitochondrial delivery. We used cells derived from a patient with a mitochondrial disease with a G625A heteroplasmic mutation in the tRNAPhe of the mitochondrial DNA (mtDNA). It has been reported that some mitochondrial gene diseases are caused by heteroplasmic mutations, in which both mutated and wild-type (WT) genes are present, and the accumulation of pathological mutations leads to serious, intractable, multi-organ diseases. Therefore, the decrease of the mutated gene rate is considered to be a useful gene therapy strategy. To accomplish this, wild-type mitochondrial pre-tRNAPhe (pre-WT-tRNAPhe), prepared by in vitro transcription, was encapsulated in the MITO-Porter. The pre-WT-tRNAPhe encapsulated in the MITO-Porter was transfected into diseased mitochondrial cells, and the resulting mutant levels were examined by an amplification refractory mutation system (ARMS)-quantitative PCR. The mutation rate of tRNAPhe was decreased, and this therapeutic effect was sustained even on the 8th day after transfection. Furthermore, mitochondrial respiratory activity of the disease cells was increased after the transfection of therapeutic pre-WT-tRNAPhe. These results support the conclusion that the mitochondrial delivery of therapeutic nucleic acids represents a viable strategy for mitochondrial gene therapy.http://www.sciencedirect.com/science/article/pii/S2162253120301128mitochondrial deliveryMITO-Porternucleic acids medicinemitochondrial gene therapyheteroplasmic mutation |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Eriko Kawamura Minako Maruyama Jiro Abe Akira Sudo Atsuhito Takeda Shingo Takada Takashi Yokota Shintaro Kinugawa Hideyoshi Harashima Yuma Yamada |
spellingShingle |
Eriko Kawamura Minako Maruyama Jiro Abe Akira Sudo Atsuhito Takeda Shingo Takada Takashi Yokota Shintaro Kinugawa Hideyoshi Harashima Yuma Yamada Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-Porter Molecular Therapy: Nucleic Acids mitochondrial delivery MITO-Porter nucleic acids medicine mitochondrial gene therapy heteroplasmic mutation |
author_facet |
Eriko Kawamura Minako Maruyama Jiro Abe Akira Sudo Atsuhito Takeda Shingo Takada Takashi Yokota Shintaro Kinugawa Hideyoshi Harashima Yuma Yamada |
author_sort |
Eriko Kawamura |
title |
Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-Porter |
title_short |
Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-Porter |
title_full |
Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-Porter |
title_fullStr |
Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-Porter |
title_full_unstemmed |
Validation of Gene Therapy for Mutant Mitochondria by Delivering Mitochondrial RNA Using a MITO-Porter |
title_sort |
validation of gene therapy for mutant mitochondria by delivering mitochondrial rna using a mito-porter |
publisher |
Elsevier |
series |
Molecular Therapy: Nucleic Acids |
issn |
2162-2531 |
publishDate |
2020-06-01 |
description |
Here, we report on validating a mitochondrial gene therapy by delivering nucleic acids to mitochondria of diseased cells by a MITO-Porter, a liposome-based carrier for mitochondrial delivery. We used cells derived from a patient with a mitochondrial disease with a G625A heteroplasmic mutation in the tRNAPhe of the mitochondrial DNA (mtDNA). It has been reported that some mitochondrial gene diseases are caused by heteroplasmic mutations, in which both mutated and wild-type (WT) genes are present, and the accumulation of pathological mutations leads to serious, intractable, multi-organ diseases. Therefore, the decrease of the mutated gene rate is considered to be a useful gene therapy strategy. To accomplish this, wild-type mitochondrial pre-tRNAPhe (pre-WT-tRNAPhe), prepared by in vitro transcription, was encapsulated in the MITO-Porter. The pre-WT-tRNAPhe encapsulated in the MITO-Porter was transfected into diseased mitochondrial cells, and the resulting mutant levels were examined by an amplification refractory mutation system (ARMS)-quantitative PCR. The mutation rate of tRNAPhe was decreased, and this therapeutic effect was sustained even on the 8th day after transfection. Furthermore, mitochondrial respiratory activity of the disease cells was increased after the transfection of therapeutic pre-WT-tRNAPhe. These results support the conclusion that the mitochondrial delivery of therapeutic nucleic acids represents a viable strategy for mitochondrial gene therapy. |
topic |
mitochondrial delivery MITO-Porter nucleic acids medicine mitochondrial gene therapy heteroplasmic mutation |
url |
http://www.sciencedirect.com/science/article/pii/S2162253120301128 |
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