Single-cell analysis on specification of mammalian germline and its role in health and diseases
Author: Luo, Qing
Date: 2024-04-19
Location: Nils Ringertz, Biomedicum, Solnavägen 9, Karolinska Institutet, Solna
Time: 09.00
Department: Inst för fysiologi och farmakologi / Dept of Physiology and Pharmacology
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Thesis (2.500Mb)
Abstract
Germ cell specification is the first step for developing reproductive cells. The specification requires formative pluripotent stem cells as precursors. Previous attempts to capture formative pluripotency used opposite manipulation of Wnt signaling, activating or inhibiting, and achieved two distinctive states that indicated two ends of the formative pluripotency spectrum. Study I explored the role of Wnt signaling in the formative pluripotency spectrum. We produced a new form of formative pluripotency in epiblast-like stem cells (EpiLSCs) with activation of Wnt. We developed a computational single-cell method for transcriptionally aligning various cell lines within the formative pluripotency spectrum. Our analysis highlighted that EpiLSCs filled the gap in the pluripotency spectrum between previously published cell lines. Additionally, we revealed context-dependent roles of Wnt signaling in sustaining pluripotency and facilitating differentiation at the two ends of the formative pluripotency spectrum.
Female germ cell specification in humans generally exhibits lower efficiencies than males in vitro. Human X chromosome inactivation has a large extent of incompleteness, resulting in escapees. Whether the female lower efficiencies are related to X-linked escapees is unknown. Study II investigated the influence of Xlinked escapees on germ cell specification in females and individuals with Klinefelter syndrome (KS), a condition typically characterized by an extra copy of the X chromosome and infertility. Through RNA sequencing and functional assays, we identified critical X-linked escapees, CHRDL1, IGSF1, and USP9X, inhibiting germ cell specification in females and KS. We found that USP9X elevated SOX2 to repress oxidative phosphorylation, promote mitochondria fusion and clustering, and perturb SOX17's regulation, exerting a profound reduction in germ cell specification.
Germ cells carry both genetic and epigenetic information. Sperm's small RNA composition is shaped by the soma-to-germline communication pathway and, therefore, is responsive to environmental exposure. Polycystic ovary syndrome (PCOS) is an epigenetically heritable disorder affecting female offspring. The possibility of PCOS equivalent in males has prompted questions about the potential impact on male offspring and whether sperm small RNA plays a role in it. In study III, using a Swedish registered cohort, a Chile longitudinal cohort, and a mouse model, we found that women with PCOS transgenerationally transmitted reproductive and metabolic dysfunction into their male offspring. Using small RNA sequencing, we identified transgenerationally altered sperm small RNA in the mice, which overlapped with changes in the sons of women with PCOS, suggesting potential mechanistic parallels of inheritance between mice and humans.
In this thesis, we uncover critical insights into the Wnt's context-dependent roles in the formative pluripotency spectrum, the repression of XCI escapees on germ cell development, and sperm small RNAs' transgenerational transmission. Additionally, this research paves the way for further exploration into germ cell development for individuals with KS and the inheritance of PCOS in male offspring, offering potential avenues for therapeutic interventions.
Female germ cell specification in humans generally exhibits lower efficiencies than males in vitro. Human X chromosome inactivation has a large extent of incompleteness, resulting in escapees. Whether the female lower efficiencies are related to X-linked escapees is unknown. Study II investigated the influence of Xlinked escapees on germ cell specification in females and individuals with Klinefelter syndrome (KS), a condition typically characterized by an extra copy of the X chromosome and infertility. Through RNA sequencing and functional assays, we identified critical X-linked escapees, CHRDL1, IGSF1, and USP9X, inhibiting germ cell specification in females and KS. We found that USP9X elevated SOX2 to repress oxidative phosphorylation, promote mitochondria fusion and clustering, and perturb SOX17's regulation, exerting a profound reduction in germ cell specification.
Germ cells carry both genetic and epigenetic information. Sperm's small RNA composition is shaped by the soma-to-germline communication pathway and, therefore, is responsive to environmental exposure. Polycystic ovary syndrome (PCOS) is an epigenetically heritable disorder affecting female offspring. The possibility of PCOS equivalent in males has prompted questions about the potential impact on male offspring and whether sperm small RNA plays a role in it. In study III, using a Swedish registered cohort, a Chile longitudinal cohort, and a mouse model, we found that women with PCOS transgenerationally transmitted reproductive and metabolic dysfunction into their male offspring. Using small RNA sequencing, we identified transgenerationally altered sperm small RNA in the mice, which overlapped with changes in the sons of women with PCOS, suggesting potential mechanistic parallels of inheritance between mice and humans.
In this thesis, we uncover critical insights into the Wnt's context-dependent roles in the formative pluripotency spectrum, the repression of XCI escapees on germ cell development, and sperm small RNAs' transgenerational transmission. Additionally, this research paves the way for further exploration into germ cell development for individuals with KS and the inheritance of PCOS in male offspring, offering potential avenues for therapeutic interventions.
List of papers:
I. QING LUO, Han-Pin Pui, Jiayu Chen, Leqian Yu, Paulo R Jannig, Yu Pei, Linxuan Zhao, Xingqi Chen, Sophie Petropoulos, Jorge L Ruas, Jun Wu, Qiaolin Deng. Epiblast-like stem cells established by Wnt/β-catenin signaling manifest distinct features of formative pluripotency and germline competence. Cell Reports. 2023 Jan 31;42(1):112021. (QL and HP contributed equally to this work.)
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II. Wenteng He, QING LUO, Jian Zhao, Allan Zhao, Luohua Feng, Ahmed Reda, Eva Lindgren, Jan-Bernd Strukenborg, Qiaolin Deng. SOX2 upregulation as downstream of X-linked gene dosage affects the specification of human primordial germ cell-like cells. (WH and QL contributed equally to this work.) [Manuscript]
III. Sanjiv Risal , Congru Li, QING LUO, Romina Fornes, Haojiang Lu, Gustaw Eriksson, Maria Manti, Claes Ohlsson, Eva Lindgren, Nicolas Crisosto, Manuel Maliqueo, Barbara Echiburú, Sergio Recabarren, Teresa Sir Petermann, Anna Benrick, Nele Brusselaers, Jie Qiao, Qiaolin Deng, Elisabet Stener-Victorin. Transgenerational transmission of reproductive and metabolic dysfunction in the male progeny of polycystic ovary syndrome. Cell Reports Medicine. 2023 May 16;4(5):101035. (SR, CL, QL and RF contributed equally to this work.)
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I. QING LUO, Han-Pin Pui, Jiayu Chen, Leqian Yu, Paulo R Jannig, Yu Pei, Linxuan Zhao, Xingqi Chen, Sophie Petropoulos, Jorge L Ruas, Jun Wu, Qiaolin Deng. Epiblast-like stem cells established by Wnt/β-catenin signaling manifest distinct features of formative pluripotency and germline competence. Cell Reports. 2023 Jan 31;42(1):112021. (QL and HP contributed equally to this work.)
Fulltext (DOI)
Pubmed
View record in Web of Science®
II. Wenteng He, QING LUO, Jian Zhao, Allan Zhao, Luohua Feng, Ahmed Reda, Eva Lindgren, Jan-Bernd Strukenborg, Qiaolin Deng. SOX2 upregulation as downstream of X-linked gene dosage affects the specification of human primordial germ cell-like cells. (WH and QL contributed equally to this work.) [Manuscript]
III. Sanjiv Risal , Congru Li, QING LUO, Romina Fornes, Haojiang Lu, Gustaw Eriksson, Maria Manti, Claes Ohlsson, Eva Lindgren, Nicolas Crisosto, Manuel Maliqueo, Barbara Echiburú, Sergio Recabarren, Teresa Sir Petermann, Anna Benrick, Nele Brusselaers, Jie Qiao, Qiaolin Deng, Elisabet Stener-Victorin. Transgenerational transmission of reproductive and metabolic dysfunction in the male progeny of polycystic ovary syndrome. Cell Reports Medicine. 2023 May 16;4(5):101035. (SR, CL, QL and RF contributed equally to this work.)
Fulltext (DOI)
Pubmed
View record in Web of Science®
Institution: Karolinska Institutet
Supervisor: Deng, Qiaolin
Co-supervisor: Stener-Victorin, Elisabet; Chen, Xingqi
Issue date: 2024-03-25
Rights:
Publication year: 2024
ISBN: 978-91-8017-330-8
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