Sperm and offspring production in a nonobstructive azoospermia mouse model via testicular mRNA delivery using lipid nanoparticles

Mashiko, Daisuke; Emori, Chihiro; Hatanaka, Yuki; Motooka, Daisuke; Pan, Chen; Kaneda, Yuki; Matzuk, Martin M.; Ikawa, Masahito

doi:https://doi.org/10.1073/pnas.2516573122

Sperm and offspring production in a nonobstructive azoospermia mouse model via testicular mRNA delivery using lipid nanoparticles

Mashiko, Daisuke; Emori, Chihiro; Hatanaka, Yuki et al.

Proceedings of the National Academy of Sciences of the United States of America, 2025, 122(42), e2516573122

Number of Access:37（2026-02-05 22:24 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/103681

Link to primary information

File	Format	Terms of use	Size	Views	Date.Available	Description	information
ProcNatlAcadSciUSA_122_42_e2516573122	pdf	None	3.74 MB	8	2026.01.30

Creative Commons : Attribution

Item Information

Output File Export EndNote Basic Export Mendeley

Title	Sperm and offspring production in a nonobstructive azoospermia mouse model via testicular mRNA delivery using lipid nanoparticles
Creator	ORCID 0000-0003-3927-3076 0000-0003-3927-3076 researchmap permalink mashikodaisuke mashikodaisuke Scopus Author ID 55953432500 55953432500 CreatorName Mashiko, Daisuke
	NRID 1000010868136 1000010868136 ORCID 0000-0001-8040-2517 0000-0001-8040-2517 researchmap permalink cemori cemori Scopus Author ID 55937469900 55937469900 CreatorName Emori, Chihiro
	NRID 1000070719450 1000070719450 ORCID 0000-0003-1339-8612 0000-0003-1339-8612 researchmap permalink y.hatanaka y.hatanaka Scopus Author ID 36494602600 36494602600 CreatorName Hatanaka, Yuki
	NRID 1000010636830 1000010636830 researchmap permalink daisukem daisukem Scopus Author ID 35315737900 35315737900 CreatorName Motooka, Daisuke
	CreatorName Pan, Chen
	ORCID 0000-0001-6300-4234 0000-0001-6300-4234 Scopus Author ID 57218423045 57218423045 CreatorName Kaneda, Yuki
	CreatorName Matzuk, Martin M.
	NRID 1000020304066 1000020304066 ORCID 0000-0001-9859-6217 0000-0001-9859-6217 researchmap permalink read0054910 read0054910 Scopus Author ID 16689054500 16689054500 CreatorName Ikawa, Masahito
Subject	azoospermia
	germline therapy
	lipid nanoparticle
	LNP therapy
	sterility
Description	Mashiko D., Emori C., Hatanaka Y., et al. Sperm and offspring production in a nonobstructive azoospermia mouse model via testicular mRNA delivery using lipid nanoparticles. Proceedings of the National Academy of Sciences of the United States of America 122, e2516573122 (2025); https://doi.org/10.1073/pnas.2516573122.
Abstract	Microsurgical testicular sperm extraction (microTESE) with intracytoplasmic sperm injection (ICSI) represents the current standard treatment for nonobstructive azoospermia (NOA). However, cures remain unavailable for NOA patients lacking retrievable haploid cells. mRNA supplementation could be a potential treatment for genetic defects leading to impaired spermatogenesis. Lipid nanoparticles (LNPs) have emerged as mRNA delivery vehicles with minimal risk of genome integration; however, their ability to selectively deliver mRNA to specific cell types remains limited. To overcome this, microRNA (miRNA) target sequences were incorporated into mRNA constructs to restrict expression specifically to germ cells. Using pyruvate dehydrogenase E1 subunit alpha 2 (PDHA2) knockout mice as an NOA model with meiotic arrest, we demonstrate that LNP-mediated delivery of Pdha2 mRNA enables the resumption and completion of meiosis, restores sperm production, and facilitates the generation of healthy fertile offspring via ICSI. Whole-genome sequencing of the offspring confirmed the absence of large-scale genomic abnormalities. Our results provide proof of concept for a safe and effective chemically synthesized LNP-based mRNA therapy with miRNA-regulated germ cell specificity, offering a promising therapeutic approach to treating male infertility caused by spermatogenesis arrest.
Publisher	National Academy of Sciences
Source Title	Proceedings of the National Academy of Sciences of the United States of America
Volume (Issue)	122(42)
Page	e2516573122
Date of Issued	2025-10-13
Language	English
Handle URL	https://hdl.handle.net/11094/103681
PISSN	00278424
eISSN	10916490
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1073/pnas.2516573122
Access Rights	open access
CopyRight	This article is licensed under a Creative Commons Attribution 4.0 International License.
oaire:version	Version of Record
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCMI Type	Text
DCTERMS.bibliographicCitation	Proceedings of the National Academy of Sciences of the United States of America. 2025, 122(42), p. e2516573122
DC.title	Sperm and offspring production in a nonobstructive azoospermia mouse model via testicular mRNA delivery using lipid nanoparticles
DC.creator	Mashiko, Daisuke
	Emori, Chihiro
	Hatanaka, Yuki
	Motooka, Daisuke
	Pan, Chen
	Kaneda, Yuki
	Matzuk, Martin M.
	Ikawa, Masahito
DC.publisher	National Academy of Sciences
DC.language' scheme='DCTERMS.RFC1766	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2025-10-13
DC.identifier	https://doi.org/10.1073/pnas.2516573122
DC.subject	azoospermia
	germline therapy
	lipid nanoparticle
	LNP therapy
	sterility
DC.description	Mashiko D., Emori C., Hatanaka Y., et al. Sperm and offspring production in a nonobstructive azoospermia mouse model via testicular mRNA delivery using lipid nanoparticles. Proceedings of the National Academy of Sciences of the United States of America 122, e2516573122 (2025); https://doi.org/10.1073/pnas.2516573122.
DCTERMS.abstract	Microsurgical testicular sperm extraction (microTESE) with intracytoplasmic sperm injection (ICSI) represents the current standard treatment for nonobstructive azoospermia (NOA). However, cures remain unavailable for NOA patients lacking retrievable haploid cells. mRNA supplementation could be a potential treatment for genetic defects leading to impaired spermatogenesis. Lipid nanoparticles (LNPs) have emerged as mRNA delivery vehicles with minimal risk of genome integration; however, their ability to selectively deliver mRNA to specific cell types remains limited. To overcome this, microRNA (miRNA) target sequences were incorporated into mRNA constructs to restrict expression specifically to germ cells. Using pyruvate dehydrogenase E1 subunit alpha 2 (PDHA2) knockout mice as an NOA model with meiotic arrest, we demonstrate that LNP-mediated delivery of Pdha2 mRNA enables the resumption and completion of meiosis, restores sperm production, and facilitates the generation of healthy fertile offspring via ICSI. Whole-genome sequencing of the offspring confirmed the absence of large-scale genomic abnormalities. Our results provide proof of concept for a safe and effective chemically synthesized LNP-based mRNA therapy with miRNA-regulated germ cell specificity, offering a promising therapeutic approach to treating male infertility caused by spermatogenesis arrest.
DC.rights	This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	103681
citation_title	Sperm and offspring production in a nonobstructive azoospermia mouse model via testicular mRNA delivery using lipid nanoparticles
citation_author	Mashiko, Daisuke
	Emori, Chihiro
	Hatanaka, Yuki
	Motooka, Daisuke
	Pan, Chen
	Kaneda, Yuki
	Matzuk, Martin M.
	Ikawa, Masahito
citation_publisher	National Academy of Sciences
citation_language	English
citation_date	2025-10-13
citation_journal_title	Proceedings of the National Academy of Sciences of the United States of America
citation_volume	122
citation_issue	42
citation_firstpage	e2516573122
citation_doi	https://doi.org/10.1073/pnas.2516573122
citation_public_url	https://hdl.handle.net/11094/103681

Sperm and offspring production in a nonobstructive azoospermia mouse model via testicular mRNA delivery using lipid nanoparticles

Link to primary information

Item Information

Show details