Advanced paternal age exacerbates neuroinflammation in offspring via m6A modification-mediated intergenerational inheritance

Yiting Mao; Yicong Meng; Kexin Zou; Ningxin Qin; Yinyu Wang; Jing Yan; PinJia Chen; Yi Cheng; Weihui Shi; Chengliang Zhou; Huixi Chen; Jianzhong Sheng; Xinmei Liu; Jiexue Pan; Hefeng Huang

doi:10.1186/s12974-024-03248-8

Advanced paternal age exacerbates neuroinflammation in offspring via m6A modification-mediated intergenerational inheritance

J Neuroinflammation. 2024 Oct 4;21(1):249. doi: 10.1186/s12974-024-03248-8.

Authors

Yiting Mao^#^{1

2

3}, Yicong Meng^#^{2

4}, Kexin Zou^{2

5}, Ningxin Qin^{2

5}, Yinyu Wang^{2

4}, Jing Yan^{1

2}, PinJia Chen^{1

2}, Yi Cheng^{1

2}, Weihui Shi^{1

2}, Chengliang Zhou^{2

4}, Huixi Chen^{2

4}, Jianzhong Sheng^{6

2}, Xinmei Liu^{7

8

9}, Jiexue Pan^{10

11

12}, Hefeng Huang^{13

14

15

16}

Affiliations

¹ Obstetrics and Gynecology Hospital, Institute of Reproduction and Development, Fudan University, Shanghai, 200011, China.
² Shanghai Key Laboratory of Reproduction and Development, Shanghai, China.
³ Research Units of Embryo Original Diseases, Chinese Academy of Medical Sciences, (No. 2019RU056), Shanghai, China.
⁴ The International Peace Maternal and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
⁵ Shanghai First Maternity and Infant Hospital, School of Medicine, Tongji University, Shanghai, China.
⁶ Key Laboratory of Reproductive Genetics (Ministry of Education), Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, China.
⁷ Obstetrics and Gynecology Hospital, Institute of Reproduction and Development, Fudan University, Shanghai, 200011, China. liuxinmei@fudan.edu.cn.
⁸ Shanghai Key Laboratory of Reproduction and Development, Shanghai, China. liuxinmei@fudan.edu.cn.
⁹ Research Units of Embryo Original Diseases, Chinese Academy of Medical Sciences, (No. 2019RU056), Shanghai, China. liuxinmei@fudan.edu.cn.
¹⁰ Obstetrics and Gynecology Hospital, Institute of Reproduction and Development, Fudan University, Shanghai, 200011, China. panjiexue@fudan.edu.cn.
¹¹ Shanghai Key Laboratory of Reproduction and Development, Shanghai, China. panjiexue@fudan.edu.cn.
¹² Research Units of Embryo Original Diseases, Chinese Academy of Medical Sciences, (No. 2019RU056), Shanghai, China. panjiexue@fudan.edu.cn.
¹³ Obstetrics and Gynecology Hospital, Institute of Reproduction and Development, Fudan University, Shanghai, 200011, China. huanghefg@fudan.edu.cn.
¹⁴ Key Laboratory of Reproductive Genetics (Ministry of Education), Department of Reproductive Endocrinology, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, China. huanghefg@fudan.edu.cn.
¹⁵ Shanghai Key Laboratory of Reproduction and Development, Shanghai, China. huanghefg@fudan.edu.cn.
¹⁶ Research Units of Embryo Original Diseases, Chinese Academy of Medical Sciences, (No. 2019RU056), Shanghai, China. huanghefg@fudan.edu.cn.

^# Contributed equally.

Abstract

Background: The trend of postponing childbearing age is prevalent worldwide. Advanced paternal age (APA) is associated with adverse pregnancy outcomes and offspring health. However, the underlying mechanism by which paternal aging affects the risk of offspring neuropsychiatric disorders is unclear. Our study aims to explore the behavioral phenotypes and the pathologic epigenetic alterations of APA offspring inherited from aging sperm.

Methods: Behavioral tests, ELISA assay, immunofluorescence and western blotting were performed on offspring mice. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA immunoprecipitation sequencing (RIP-seq) were used to investigate the modified N6-methyladenosine (m6A) profiles of paternal sperm and offspring hippocampus. Intervention of gene expression by lentivirus and adeno-associated virus in both vivo and vitro examined the potential therapeutic targets of intergenerational inherited neuroinflammation.

Results: In our study, APA offspring exhibit cognitive impairment and autism-like behavior. An increase in neuroinflammation in APA offspring is associated with microglial overactivation, which manifests as abnormal morphology and augmented engulfment. MeRIP-seq of F0 sperm and F1 hippocampus reveal that Nr4a2 is hypermethylated with decreased expression in APA offspring involving in synaptic plasticity and microglial function. In addition, Ythdc1, an m6A reader protein, is markedly elevated in aging sperm and remains elevated in adult hippocampus of APA group. Enhanced Ythdc1 recognizes and suppresses the hypermethylated Nr4a2, thereby contributing to the abnormal phenotype in offspring. The overexpression of Ythdc1 triggers microglial activation in vitro and its suppression in the hippocampus of APA progeny alleviates behavioral aberrations and attenuates neuroinflammation.

Conclusion: Our study provides additional evidence of the abnormal behavioral phenotypes of APA offspring and reveals potential epigenetic inheritance signatures and targeted genes for future research.

Keywords: Advanced paternal age; Autism; Cognitive deficits; Inheritance; Microglia; m6A.

MeSH terms

Adenosine / analogs & derivatives
Adenosine / metabolism
Aging / genetics
Animals
Epigenesis, Genetic
Female
Hippocampus / metabolism
Hippocampus / pathology
Male
Mice
Mice, Inbred C57BL
Neuroinflammatory Diseases* / genetics
Neuroinflammatory Diseases* / metabolism
Pregnancy

Substances

N-methyladenosine
Adenosine

Abstract

MeSH terms

Substances

Grants and funding