PMID- 37796762 OWN - NLM STAT- MEDLINE DCOM- 20231102 LR - 20231102 IS - 1545-7885 (Electronic) IS - 1544-9173 (Print) IS - 1544-9173 (Linking) VI - 21 IP - 10 DP - 2023 Oct TI - Significant decrease of maternal mitochondria carryover using optimized spindle-chromosomal complex transfer. PG - e3002313 LID - 10.1371/journal.pbio.3002313 [doi] LID - e3002313 AB - Mutations in mitochondrial DNA (mtDNA) contribute to a variety of serious multi-organ human diseases, which are strictly inherited from the maternal germline. However, there is currently no curative treatment. Attention has been focused on preventing the transmission of mitochondrial diseases through mitochondrial replacement (MR) therapy, but levels of mutant mtDNA can often unexpectedly undergo significant changes known as mitochondrial genetic drift. Here, we proposed a novel strategy to perform spindle-chromosomal complex transfer (SCCT) with maximal residue removal (MRR) in metaphase II (MII) oocytes, thus hopefully eliminated the transmission of mtDNA diseases. With the MRR procedure, we initially investigated the proportions of mtDNA copy numbers in isolated karyoplasts to those of individual oocytes. Spindle-chromosomal morphology and copy number variation (CNV) analysis also confirmed the safety of this method. Then, we reconstructed oocytes by MRR-SCCT, which well developed to blastocysts with minimal mtDNA residue and normal chromosomal copy numbers. Meanwhile, we optimized the manipulation order between intracytoplasmic sperm injection (ICSI) and SCC transfer and concluded that ICSI-then-transfer was conducive to avoid premature activation of reconstructed oocytes in favor of normal fertilization. Offspring of mice generated by embryos transplantation in vivo and embryonic stem cells derivation further presented evidences for competitive development competence and stable mtDNA carryover without genetic drift. Importantly, we also successfully accomplished SCCT in human MII oocytes resulting in tiny mtDNA residue and excellent embryo development through MRR manipulation. Taken together, our preclinical mouse and human models of the MRR-SCCT strategy not only demonstrated efficient residue removal but also high compatibility with normal embryo development, thus could potentially be served as a feasible clinical treatment to prevent the transmission of inherited mtDNA diseases. CI - Copyright: (c) 2023 Liao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. FAU - Liao, Xiaoyu AU - Liao X AUID- ORCID: 0000-0003-4755-6172 AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Li, Wenzhi AU - Li W AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Lin, Kaibo AU - Lin K AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Jin, Wei AU - Jin W AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Zhang, Shaozhen AU - Zhang S AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Wang, Yao AU - Wang Y AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Ma, Meng AU - Ma M AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Xie, Yating AU - Xie Y AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Yu, Weina AU - Yu W AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Yan, Zhiguang AU - Yan Z AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Gao, Hongyuan AU - Gao H AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Zhao, Leiwen AU - Zhao L AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Si, Jiqiang AU - Si J AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Wang, Yun AU - Wang Y AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Lin, Jiaying AU - Lin J AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Chen, Chen AU - Chen C AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Chen, Li AU - Chen L AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Kuang, Yanping AU - Kuang Y AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. FAU - Lyu, Qifeng AU - Lyu Q AUID- ORCID: 0000-0002-9885-6522 AD - Department of Assisted Reproduction, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China. LA - eng PT - Journal Article DEP - 20231005 PL - United States TA - PLoS Biol JT - PLoS biology JID - 101183755 RN - 0 (DNA, Mitochondrial) SB - IM MH - Male MH - Humans MH - Animals MH - Mice MH - *DNA Copy Number Variations/genetics MH - Semen MH - Mitochondria/genetics MH - DNA, Mitochondrial/genetics/analysis MH - *Mitochondrial Diseases/genetics/prevention & control MH - Oocytes PMC - PMC10553349 COIS- The authors have declared that no competing interests exist. EDAT- 2023/10/05 18:42 MHDA- 2023/11/02 12:42 PMCR- 2023/10/05 CRDT- 2023/10/05 13:23 PHST- 2023/04/19 00:00 [received] PHST- 2023/08/25 00:00 [accepted] PHST- 2023/11/02 12:42 [medline] PHST- 2023/10/05 18:42 [pubmed] PHST- 2023/10/05 13:23 [entrez] PHST- 2023/10/05 00:00 [pmc-release] AID - PBIOLOGY-D-23-01006 [pii] AID - 10.1371/journal.pbio.3002313 [doi] PST - epublish SO - PLoS Biol. 2023 Oct 5;21(10):e3002313. doi: 10.1371/journal.pbio.3002313. eCollection 2023 Oct.