title: Mitochondrial replacement in human oocytes carrying pathogenic mitochondrial DNA mutations.
abstract: Maternally inherited mitochondrial (mt)DNA mutations can cause fatal or severely debilitating syndromes in children, with disease severity dependent on the specific gene mutation and the ratio of mutant to wild-type mtDNA (heteroplasmy) in each cell and tissue. Pathogenic mtDNA mutations are relatively common, with an estimated 778 affected children born each year in the United States. Mitochondrial replacement therapies or techniques (MRT) circumventing mother-to-child mtDNA disease transmission involve replacement of oocyte maternal mtDNA. Here we report MRT outcomes in several families with common mtDNA syndromes. The mother's oocytes were of normal quality and mutation levels correlated with those in existing children. Efficient replacement of oocyte mutant mtDNA was performed by spindle transfer, resulting in embryos containing >99% donor mtDNA. Donor mtDNA was stably maintained in embryonic stem cells (ES cells) derived from most embryos. However, some ES cell lines demonstrated gradual loss of donor mtDNA and reversal to the maternal haplotype. In evaluating donor-to-maternal mtDNA interactions, it seems that compatibility relates to mtDNA replication efficiency rather than to mismatch or oxidative phosphorylation dysfunction. We identify a polymorphism within the conserved sequence box II region of the D-loop as a plausible cause of preferential replication of specific mtDNA haplotypes. In addition, some haplotypes confer proliferative and growth advantages to cells. Hence, we propose a matching paradigm for selecting compatible donor mtDNA for MRT.
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Eunju
authors.initials: E
authors.lastname: Kang
authors.affiliation: Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Wu
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Nuria Marti
authors.initials: NM
authors.lastname: Gutierrez
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Amy
authors.initials: A
authors.lastname: Koski
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Tippner-Hedges
authors.affiliation: Department of Cell Biology School of Osteopathic Medicine, Rowan University, 2 Medical Center Drive, Stratford, New Jersey 08084, USA.
authors.forename: Karen
authors.initials: K
authors.lastname: Agaronyan
authors.affiliation: Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.
authors.forename: Aida
authors.initials: A
authors.lastname: Platero-Luengo
authors.affiliation: Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.
authors.forename: Paloma
authors.initials: P
authors.lastname: Martinez-Redondo
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Ma
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Yeonmi
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Tomonari
authors.initials: T
authors.lastname: Hayama
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Crystal
authors.initials: C
authors.lastname: Van Dyken
authors.affiliation: Division of Human Genetics, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, Ohio 45229, USA.
authors.forename: Xinjian
authors.initials: X
authors.lastname: Wang
authors.affiliation: Division of Human Genetics, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, Ohio 45229, USA.
authors.forename: Shiyu
authors.initials: S
authors.lastname: Luo
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Riffat
authors.initials: R
authors.lastname: Ahmed
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Reproductive Medical Centre, Anhui Medical University, No 218, Jixi Rd, Shushan District, Heifei, Anhui 230022, China.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Ji
authors.affiliation: IviGen Los Angeles, 406 Amapola Avenue, Suite 215, Torrance, California 90501, USA.
authors.forename: Refik
authors.initials: R
authors.lastname: Kayali
authors.affiliation: IviGen Los Angeles, 406 Amapola Avenue, Suite 215, Torrance, California 90501, USA.
authors.forename: Cengiz
authors.initials: C
authors.lastname: Cinnioglu
authors.affiliation: Research Cytogenetics Laboratory, Oregon Health &Science University, 3181 SW Sam Jackson Park Road, Portland, Oregon 97239, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Olson
authors.affiliation: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Jensen
authors.affiliation: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.
authors.forename: David
authors.initials: D
authors.lastname: Battaglia
authors.affiliation: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.
authors.forename: David
authors.initials: D
authors.lastname: Lee
authors.affiliation: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.
authors.forename: Diana
authors.initials: D
authors.lastname: Wu
authors.affiliation: Division of Human Genetics, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, Ohio 45229, USA.
authors.forename: Taosheng
authors.initials: T
authors.lastname: Huang
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.
authors.forename: Don P
authors.initials: DP
authors.lastname: Wolf
authors.affiliation: Department of Cell Biology School of Osteopathic Medicine, Rowan University, 2 Medical Center Drive, Stratford, New Jersey 08084, USA.
authors.forename: Dmitry
authors.initials: D
authors.lastname: Temiakov
authors.affiliation: Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.
authors.forename: Juan Carlos Izpisua
authors.initials: JC
authors.lastname: Belmonte
authors.affiliation: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.
authors.forename: Paula
authors.initials: P
authors.lastname: Amato
authors.affiliation: Center for Embryonic Cell and Gene Therapy, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Division of Reproductive &Developmental Sciences, Oregon National Primate Research Center, Oregon Health &Science University, 505 NW 185th Avenue, Beaverton, Oregon 97006, USA.|Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.|Knight Cardiovascular Institute, Oregon Health &Science University, 3181 SW Sam Jackson Park Road, Portland, Oregon 97239, USA.|Department of Biomedical Engineering, Oregon Health &Science University, 3303 SW Bond Avenue, Portland, Oregon 97239, USA.
authors.forename: Shoukhrat
authors.initials: S
authors.lastname: Mitalipov
mesh_terms: D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D017124:Conserved Sequence / Q000235:genetics; D004272:DNA, Mitochondrial / Q000096:biosynthesis / Q000235:genetics* / Q000627:therapeutic use*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D006239:Haplotypes / Q000235:genetics; D006801:Humans; D008297:Male; D000072741:Maternal Inheritance / Q000235:genetics*; D008540:Meiosis; D028361:Mitochondrial Diseases / Q000235:genetics* / Q000378:metabolism / Q000473:pathology* / Q000517:prevention & control; D000069321:Mitochondrial Replacement Therapy / Q000379:methods*; D009154:Mutation; D018587:Oocyte Donation; D009865:Oocytes / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D010085:Oxidative Phosphorylation; D010375:Pedigree; D011110:Polymorphism, Genetic
chemical_list: D004272:DNA, Mitochondrial