Sulforaphane promotes brain organoids growth through promoting prefrontal development and neuronal differentiation
10.3760/cma.j.cn113661-20211103-00318
- VernacularTitle:萝卜硫素通过促进前额发育和神经元分化促进大脑类器官生长
- Author:
Ziwei TENG
1
;
Yuhan SU
1
;
Yue QIN
1
;
Jindong CHEN
1
;
Hui TANG
1
;
Renrong WU
1
;
Haishan WU
1
;
Jing HUANG
1
Author Information
1. 中南大学湘雅二医院精神科 国家精神疾病临床研究中心，长沙 410011
- Publication Type:Journal Article
- Keywords:
Multipotent stem cells;
Neurons;
Sulforaphane;
Brain organoids;
Prefrontal
- From:
Chinese Journal of Psychiatry
2022;55(3):204-213
- CountryChina
- Language:Chinese
-
Abstract:
Objective:By comparing the changes of brain differentiation and neuronal markers of brain organoids in different periods, this study aims to clarify the effect and possible mechanisms of sulforaphane on the development of brain organoids.Methods:Brain organoids differentiated from human induced pluripotent stem cells treated with sulforaphane were used in this study. The experimental group was treated with sulforaphane at different concentrations (1 μmol/L, 2 μmol/L, 5 μmol/L), and the control group was a blank group, with 20 organoids in each group. Immunofluorescence and hematoxylin-eosin staining (HE) were used to observe and compare the expression characteristics of brain markers and neuronal markers in different periods of the two groups after sulforaphane treatment. Ribonucleic Acid (RNA) sequencing analysis of differentially expressed genes, functional enrichment analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were conducted. Real-time Quantitative Reverse Transcriptase-mediated Polymerase Chain Reaction (qRT-PCR) was performed to further screen and verify the potential genes or possible targets that may be involved in sulforaphane in promoting the growth of brain organoids.Results:The sex determining region Y-box 2(SOX2), Forebrain protein forkhead box G1 (FOXG1), prefrontal cortex autism susceptibility candidate 2 (AUST2), and paired box 6 (PAX6) increased significantly in the brain organoids after 40 days of treatment with sulforaphane; the expression of neuron marker proteins like β-tubulin (TUJ1), neuronal nuclei (NeuN), recombinant microtubule associated protein 2 (MAP2) and T-brain-1 (TBR1) in the brain organoids also showed significant increase after 70 days of treatment with sulforaphane. RNA sequencing analysis revealed that compared with the control group, 105 up-regulated genes and 512 down-regulated genes were identified in the sulforaphane treatment group, which, in total, are 617 genes. qRT-PCR verified that the transcription levels of differentially expressed genes (SFTPC,AKR1C3,CXCR6,PRAP1,TMC8,GPR182,F2RL2,KCNJ10) are basically consistent with the sequencing results.Conclusion:AKR1C3,KCNJ10 and other genes may be involved in sulforaphane to promote the brain organoid prefrontal development and neuronal differentiation. This research helps to provide new experimental evidence that sulforaphane improves the cognition and symptoms of patients with mental illness and neurodegenerative diseases.
