Histone Trimethylations and HDAC5 Regulate Spheroid Subpopulation and Differentiation Signaling of Human Adipose-Derived Stem Cells-Reference-Cited by-同舟云学术

Histone Trimethylations and HDAC5 Regulate Spheroid Subpopulation and Differentiation Signaling of Human Adipose-Derived Stem Cells

Published:2023-12-30 Issue:3 Volume:13 Page:293-308
ISSN:2157-6564
Container-title:Stem Cells Translational Medicine
language:en
Short-container-title:

Author:

Chang Ming-Min¹²^ORCID,Hong Yi-Kai³⁴,Hsu Chao-Kai³⁴^ORCID,Harn Hans I-Chen⁵,Huang Bu-Miin¹,Liu Ya-Hsin⁶,Lu Fu-I⁷,Hsueh Yuan-Yu⁴⁸⁹^ORCID,Lin Shau-Ping¹⁰,Wu Chia-Ching¹⁴¹¹¹²^ORCID

Affiliation:

1. Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University , Tainan , Taiwan

2. Medical Device Innovation Center, National Cheng Kung University , Tainan , Taiwan

3. Department of Dermatology, College of Medicine, National Cheng Kung University , Tainan , Taiwan

4. International Center for Wound Repair and Regeneration, National Cheng Kung University , Tainan , Taiwan

5. Department of Pathology, Keck School of Medicine, University of Southern California , Los Angeles, CA , USA

6. Department of Life Sciences, College of Bioscience and Biotechnology, National Cheng Kung University , Tainan , Taiwan

7. Department of Biotechnology and Bioindustry Sciences, College of Bioscience and Biotechnology, National Cheng Kung University , Tainan , Taiwan

8. Division of Plastic and Reconstructive Surgery, Department of Surgery, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University , Tainan , Taiwan

9. Institute of Clinical Medicine, College of Medicine, National Cheng Kung University , Tainan , Taiwan

10. Institute of Biotechnology, College of Bio-Resources and Agriculture, National Taiwan University , Taipei , Taiwan

11. Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University , Tainan , Taiwan

12. Department of Biomedical Engineering, National Cheng Kung University , Tainan , Taiwan

Abstract

Abstract Human adipose-derived stem cells (ASCs) have shown immense potential for regenerative medicine. Our previous work demonstrated that chitosan nano-deposited surfaces induce spheroid formation and differentiation of ASCs for treating sciatic nerve injuries. However, the underlying cell fate and differentiation mechanisms of ASC-derived spheroids remain unknown. Here, we investigate the epigenetic regulation and signaling coordination of these therapeutic spheroids. During spheroid formation, we observed significant increases in histone 3 trimethylation at lysine 4 (H3K4me3), lysine 9 (H3K9me3), and lysine 27 (H3K27me3), accompanied by increased histone deacetylase (HDAC) activities and decreased histone acetyltransferase activities. Additionally, HDAC5 translocated from the cytoplasm to the nucleus, along with increased nuclear HDAC5 activities. Utilizing single-cell RNA sequencing (scRNA-seq), we analyzed the chitosan-induced ASC spheroids and discovered distinct cluster subpopulations, cell fate trajectories, differentiation traits, and signaling networks using the 10x Genomics platform, R studio/language, and the Ingenuity Pathway Analysis (IPA) tool. Specific subpopulations were identified within the spheroids that corresponded to a transient reprogramming state (Cluster 6) and the endpoint cell state (Cluster 3). H3K4me3 and H3K9me3 were discovered as key epigenetic regulators by IPA to initiate stem cell differentiation in Cluster 6 cells, and confirmed by qPCR and their respective histone methyltransferase inhibitors: SNDX-5613 (a KMT2A inhibitor for H3K4me3) and SUVi (an SUV39H1 inhibitor for H3K9me3). Moreover, H3K9me3 and HDAC5 were involved in regulating downstream signaling and neuronal markers during differentiation in Cluster 3 cells. These findings emphasize the critical role of epigenetic regulation, particularly H3K4me3, H3K9me3, and HDAC5, in shaping stem cell fate and directing lineage-specific differentiation.

Funder

National Science and Technology Council, Taiwan

National Health Research Institutes of Taiwan

Publisher

Oxford University Press (OUP)

Link

https://academic.oup.com/stcltm/article-pdf/13/3/293/56913956/szad090.pdf

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