KAT8 Knockdown Reverses hucMSC Senescence and Enhances Diabetic Wound
                    Healing Efficacy

Ruijing Chen; Xiaoyue Zhang; Jun Chen

doi:10.1055/a-2684-5975

Experimental and Clinical Endocrinology & Diabetes, Table of Contents

Exp Clin Endocrinol Diabetes 2025; 133(09): 441-452
DOI: 10.1055/a-2684-5975

Article

KAT8 Knockdown Reverses hucMSC Senescence and Enhances Diabetic Wound Healing Efficacy

Authors

Ruijing Chen

¹Department of Endocrinology and Metabolism, Qilu Hospital of Shandong University, Jinan, China (Ringgold ID: RIN91623)
Xiaoyue Zhang

¹Department of Endocrinology and Metabolism, Qilu Hospital of Shandong University, Jinan, China (Ringgold ID: RIN91623)
Jun Chen

¹Department of Endocrinology and Metabolism, Qilu Hospital of Shandong University, Jinan, China (Ringgold ID: RIN91623)

Abstract

Background

The histone acetyltransferase KAT8 has been implicated in stem cell biology, but its specific role in human umbilical cord mesenchymal stem cells (hucMSCs) senescence and therapeutic efficacy for diabetic wounds is unclear.

Aims

This study aimed to investigate the role of KAT8 in hucMSC senescence and to determine if KAT8 knockdown could reverse senescence and enhance the efficacy of hucMSCs in promoting diabetic wound healing.

Materials and Methods

hucMSCs were extracted from the umbilical cord, and senescence was induced. KAT8 expression was assessed in senescent and non-senescent hucMSCs. Senescence markers (senescence-associated β-galactosidase [SA-β-gal] staining, P16, P21 expression), proliferation (Cell Counting Kit-8 [CCK-8], colony formation), cell migration (cell scratch), and differentiation potential (alizarin red and oil red O staining) were evaluated in vitro. Western blotting and quantitative polymerase chain reaction were performed to detect protein expression and mRNA expression levels, respectively. For in vivo studies, a type 1 diabetes mellitus mouse wound healing model was established. Mice received local injections of phosphate-buffered saline (PBS), hucMSCs transduced with a negative control vector (NC hucMSCs), or KAT8-knockdown hucMSCs. Wound closure rates were monitored, and histological analyses (Hematoxylin and eosin [H&E], Masson staining).

Results

KAT8 expression decreased during hucMSCs senescence. Knockdown of KAT8 downregulated senescence-associated genes (e.g., P21 and P16) while enhancing hucMSCs proliferation, migration, and survival without altering surface stem cell marker expression. In vivo experiments further confirm that KAT8-knockdown hucMSCs significantly promoted wound healing in a type 1 diabetic mouse model, exhibiting superior therapeutic efficacy.

Conclusion

Knockdown of KAT8 effectively reverses senescence in hucMSCs and enhances their therapeutic potential for diabetic wound healing.

Keywords

KAT8 - mesenchymal stem cells - senescence - epigenetic modification - wound healing - diabetic

Full Text

References

References
1 Pittenger MF, Mackay AM, Beck SC. et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143-147
2 Dash R, Ali MC, Jahan I. et al. Emerging potential of cannabidiol in reversing proteinopathies. Ageing Res Rev 2021; 65: 101209
3 Ren R, Ocampo A, Liu GH. et al. Regulation of stem cell aging by metabolism and epigenetics. Cell Metab 2017; 26: 460-474
4 Wang Y, Yuan Q, Xie L. Histone modifications in aging: the underlying mechanisms and implications. Curr Stem Cell Res Ther 2018; 13: 125-135
5 Zhu Y, Song X, Han F. et al. Alteration of histone acetylation pattern during long-term serum-free culture conditions of human fetal placental mesenchymal stem cells. PLoS One 2015; 10: e0117068
6 Ye L, Fan Z, Yu B. et al. Histone demethylases KDM4B and KDM6B promotes osteogenic differentiation of human MSCs. Cell Stem Cell 2012; 11: 50-61
7 Jing H, Liao L, An Y. et al. Suppression of EZH2 prevents the shift of osteoporotic MSC fate to adipocyte and enhances bone formation during osteoporosis. Mol Ther 2016; 24: 217-229
8 Cakouros D, Isenmann S, Cooper L. et al. Twist-1 induces Ezh2 recruitment regulating histone methylation along the Ink4A/Arf locus in mesenchymal stem cells. Mol Cell Biol 2012; 32: 1433-1441
9 Deng P, Yuan Q, Cheng Y. et al. Loss of KDM4B exacerbates bone-fat imbalance and mesenchymal stromal cell exhaustion in skeletal aging. Cell Stem Cell 2021; 28: 1057-1073. e7
10 Wilson C, Krieg AJ. KDM4B: A nail for every hammer?. Genes 2019; 10: 134
11 Wang C, Wang J, Li J. et al. KDM5A controls bone morphogenic protein 2-induced osteogenic differentiation of bone mesenchymal stem cells during osteoporosis. Cell Death Dis 2016; 7: e2335-e2335
12 Kirtana R, Manna S, Patra SK. Molecular mechanisms of KDM5A in cellular functions: Facets during development and disease. Exp Cell Res 2020; 396: 112314
13 Yin B, Yu F, Wang C. et al. Epigenetic control of mesenchymal stem cell fate decision via histone methyltransferase Ash1l. Stem Cells 2019; 37: 115-127
14 Gupta A, Hunt CR, Pandita RK. et al. T-cell-specific deletion of Mof blocks their differentiation and results in genomic instability in mice. Mutagenesis 2013; 28: 263-270
15 Li X, Li L, Pandey R. et al. The histone acetyltransferase MOF is a key regulator of the embryonic stem cell core transcriptional network. Cell Stem Cell 2012; 11: 163-178
16 Khoa LTP, Tsan YC, Mao F. et al. Histone acetyltransferase MOF blocks acquisition of quiescence in ground-state ESCs through activating fatty acid oxidation. Cell Stem Cell 2020; 27: 441-458. e10
17 Chen J, Liu D, Chen B. et al. The histone acetyltransferase Mof regulates Runx2 and Osterix for osteoblast differentiation. Cell Tissue Res 2023; 393: 265-279
18 Pun FW, Leung GHD, Leung HW. et al. Hallmarks of aging-based dual-purpose disease and age-associated targets predicted using PandaOmics AI-powered discovery engine. Aging (Albany NY) 2022; 14: 2475
19 Detamore MS. Human umbilical cord mesenchymal stromal cells in regenerative medicine. Stem Cell Res Ther 2013; 4: 1-3
20 Can A, Balci D. Isolation, culture, and characterization of human umbilical cord stroma-derived mesenchymal stem cells. Methods Mol Biol 2011; 698: 51-62
21 Ding DC, Chang YH, Shyu WC. et al. Human umbilical cord mesenchymal stem cells: a new era for stem cell therapy. Cell Transplant 2015; 24: 339-347
22 Xu J, Liu G, Wang X. et al. hUC-MSCs: Evaluation of acute and long-term routine toxicity testing in mice and rats. Cytotechnology 2022; 74: 17-29
23 Chen Y, Yu Q, Hu Y. et al. Current research and use of mesenchymal stem cells in the therapy of autoimmune diseases. Curr Stem Cell Res Ther 2019; 14: 579-582
24 Yang Y, Cao TT, Tian ZM. et al. Subarachnoid transplantation of human umbilical cord mesenchymal stem cell in rodent model with subacute incomplete spinal cord injury: Preclinical safety and efficacy study. Exp Cell Res 2020; 395: 112184
25 Liu W, Yuan F, Bai H. et al. hUC-MSCs attenuate acute graft-versus-host disease through Chi3l1 repression of Th17 differentiation. Stem Cells Int 2022; 2022: 1052166
26 Sun W, Yan S, Yang C. et al. Mesenchymal stem cells-derived exosomes ameliorate lupus by inducing M2 macrophage polarization and regulatory T cell expansion in MRL/lpr mice. Immunol Invest 2022; 51: 1785-1803
27 Jia Y, Cao N, Zhai J. et al. HGF mediates clinical-grade human umbilical cord-derived mesenchymal stem cells improved functional recovery in a senescence-accelerated mouse model of Alzheimer's disease. Adv Sci 2020; 7: 1903809
28 Wang S, Cheng H, Dai G. et al. Umbilical cord mesenchymal stem cell transplantation significantly improves neurological function in patients with sequelae of traumatic brain injury. Brain Res 2013; 1532: 76-84
29 Chai NL, Zhang XB, Chen SW. et al. Umbilical cord-derived mesenchymal stem cells alleviate liver fibrosis in rats. World J Gastroenterol 2016; 22: 6036
30 He Y, Guo X, Lan T. et al. Human umbilical cord-derived mesenchymal stem cells improve the function of liver in rats with acute-on-chronic liver failure via downregulating Notch and Stat1/Stat3 signaling. Stem Cell Res Ther 2021; 12: 1-19
31 Aierken A, Li B, Liu P. et al. Melatonin treatment improves human umbilical cord mesenchymal stem cell therapy in a mouse model of type II diabetes mellitus via the PI3K/AKT signaling pathway. Stem Cell Res Ther 2022; 13: 164
32 Bharti D, Shivakumar SB, Park JK. et al. Comparative analysis of human Wharton’s jelly mesenchymal stem cells derived from different parts of the same umbilical cord. Cell Tissue Res 2018; 372: 51-65
33 Dabrowski FA, Burdzinska A, Kulesza A. et al. Comparison of the paracrine activity of mesenchymal stem cells derived from human umbilical cord, amniotic membrane and adipose tissue. J Obstet Gynaecol Res 2017; 43: 1758-1768
34 Bi ZM, Zhou QF, Geng Y. et al. Human umbilical cord mesenchymal stem cells ameliorate experimental cirrhosis through activation of keratinocyte growth factor by suppressing microRNA-199. Eur Rev Med Pharmacol Sci 2016; 20: 4905-4912
35 Sun X, Hao H, Han Q. et al. Human umbilical cord-derived mesenchymal stem cells ameliorate insulin resistance by suppressing NLRP3 inflammasome-mediated inflammation in type 2 diabetes rats. Stem Cell Res Ther 2017; 8: 1-14
36 Yao J, Zheng J, Cai J. et al. Extracellular vesicles derived from human umbilical cord mesenchymal stem cells alleviate rat hepatic ischemia-reperfusion injury by suppressing oxidative stress and neutrophil inflammatory response. FASEB J 2019; 33: 1695-1710
37 Zhou C, Zhang B, Yang Y. et al. Stem cell-derived exosomes: emerging therapeutic opportunities for wound healing. Stem Cell Res Ther 2023; 14: 107
38 Ma H, Siu WS, Leung PC. The potential of MSC-based cell-free therapy in wound healing—a thorough literature review. Int J Mol Sci 2023; 24: 9356
39 Ikebe C, Suzuki K. Mesenchymal stem cells for regenerative therapy: optimization of cell preparation protocols. BioMed Res Int 2014; 2014: 951512
40 Zhang Y, Li Y, Li W. et al. Therapeutic effect of human umbilical cord mesenchymal stem cells at various passages on acute liver failure in rats. Stem Cells Int 2018; 2018
41 Jung JW, Lee S, Seo MS. et al. Histone deacetylase controls adult stem cell aging by balancing the expression of polycomb genes and jumonji domain containing 3. Cell Mol Life Sci 2010; 67: 1165-1176
42 Lee S, Park JR, Seo MS. et al. Histone deacetylase inhibitors decrease proliferation potential and multilineage differentiation capability of human mesenchymal stem cells. Cell Prolif 2009; 42: 711-720
43 Zhang L, Qi M, Chen J. et al. Impaired autophagy triggered by HDAC9 in mesenchymal stem cells accelerates bone mass loss. Stem Cell Res Ther 2020; 11: 1-16
44 Suka N, Suka Y, Carmen AA. et al. Highly specific antibodies determine histone acetylation site usage in yeast heterochromatin and euchromatin. Mol Cell 2001; 8: 473-479
45 Zhang P, Liu Y, Jin C. et al. Histone acetyltransferase GCN5 regulates osteogenic differentiation of mesenchymal stem cells by inhibiting NF-κB. J Bone Miner Res 2016; 31: 391-402
46 Jing H, Su X, Gao B. et al. Epigenetic inhibition of Wnt pathway suppresses osteogenic differentiation of BMSCs during osteoporosis. Cell Death Dis 2018; 9: 176
47 Herrera JE, Bergel M, Yang XJ. et al. The histone acetyltransferase activity of human GCN5 and PCAF is stabilized by coenzymes. J Biol Chem 1997; 272: 27253-27258
48 Zhang P, Liu Y, Jin C. et al. Histone H3K9 acetyltransferase PCAF is essential for osteogenic differentiation through bone morphogenetic protein signaling and may be involved in osteoporosis. Stem Cells 2016; 34: 2332-2341
49 Li Z, Liu C, Xie Z. et al. Epigenetic dysregulation in mesenchymal stem cell aging and spontaneous differentiation. PloS One 2011; 6: e20526
50 Liu G, Chen H, Liu H. et al. Emerging roles of SIRT6 in human diseases and its modulators. Med Res Rev 2021; 41: 1089-1137
51 Xie B, Zhang M, Li J. et al. KAT8-catalyzed lactylation promotes eEF1A2-mediated protein synthesis and colorectal carcinogenesis. Proc Natl Acad Sci 2024; 121: e2314128121
52 Li P, Yang L, Park SY. et al. Stabilization of MOF (KAT8) by USP10 promotes esophageal squamous cell carcinoma proliferation and metastasis through epigenetic activation of ANXA2/Wnt signaling. Oncogene 2024; 43: 899-917
53 Wu Y, Zhou L, Zou Y. et al. Disrupting the phase separation of KAT8–IRF1 diminishes PD-L1 expression and promotes antitumor immunity. Nat Cancer 2023; 4: 382-400
54 Qiu B, Li S, Li M. et al. KAT8 acetylation-controlled lipolysis affects the invasive and migratory potential of colorectal cancer cells. Cell Death Dis 2023; 14: 164
55 Krishnan V, Chow MZ, Wang Z. et al. Histone H4 lysine 16 hypoacetylation is associated with defective DNA repair and premature senescence in Zmpste24-deficient mice. Proc Natl Acad Sci 2011; 108: 12325-12330
56 Sheikh B, Bechtel-Walz W, Lucci J. et al. MOF maintains transcriptional programs regulating cellular stress response. Oncogene 2016; 35: 2698-2710

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