DAPK1 Ablation Promotes Wound Healing by Keratinocyte Modulation

Abstract

Death-associated protein kinase 1 (DAPK1), a Ca²+/calmodulin-regulated serine/threonine kinase, plays a pivotal role in epidermal homeostasis, tissue repair, and cutaneous wound healing. However, its role in oral mucosal repair remains unclear. In this study, we established a global Dapk1 knockout mouse model to create 1.5-mm circular palatal wounds in mice aged 8 to 12 wk. Our results demonstrated that DAPK1 deficiency significantly accelerated oral wound closure. In vitro experiments further confirmed that DAPK1 modulates the proliferation and migration of human oral keratinocytes. Mechanistic investigations through transcriptome sequencing revealed activation of the Wnt signaling pathway in Dapk1 knockout mice following injury, characterized by pronounced upregulation of Wnt3, Fosl1, and Ctnnd2, alongside downregulation of innate immune mediators, including Il7, Ccl28, Ccr2, Ccl5, and Cxcr4. These findings suggest that loss of DAPK1 enhances epithelial proliferation and migration while attenuating local inflammation. Furthermore, we developed a novel microneedle patch for drug delivery, consisting of GelMA tips encapsulating the DAPK1 inhibitor (HS-38) and a hyaluronic acid substrate. This system facilitated efficient mucosal penetration and localized delivery. Application of the microneedle patch significantly improved wound healing in both normal and diabetic mouse models. Collectively, these findings uncover a previously unrecognized role of DAPK1 in oral mucosal repair and highlight its potential as a molecular target to accelerate wound healing.

Keywords

death-associated protein kinase 1 oral wound healing HS-38 microneedle cell proliferation and migration diabetes

Get full access to this article

View all access options for this article.

References

American Diabetes Association Professional Practice Committee. 2024. 2. Diagnosis and classification of diabetes: standards of care in diabetes-2024. Diabetes Care. 47(Suppl 1):S20–S42. https://doi.org/10.2337/dc24-S002

Anjum

Roux

Ballif

Gygi

Blenis

2005. The tumor suppressor DAP kinase is a target of RSK-mediated survival signaling. Curr Biol. 15(19):1762–1767. https://doi.org/10.1016/j.cub.2005.08.050

Carlson

, et al. 2013. Fluorescence linked enzyme chemoproteomic strategy for discovery of a potent and selective DAPK1 and ZIPK inhibitor. ACS Chem Biol. 8(12):2715–2723. https://doi.org/10.1021/cb400407c

Chuang

Hsiao

Tong

Chisholm

. 2016. DAPK Interacts with Patronin and the microtubule cytoskeleton in epidermal development and wound repair. eLife. 5:e15833. https://doi.org/10.7554/elife.15833

Chuang

Fang

Lin-Feng

Chen

Lai

. 2008. The tumor suppressor death-associated protein kinase targets to TCR-stimulated NF-kappa B activation. J Immunol. 180(5):3238–3249.

Elbadawy

Usui

Yamawaki

Sasaki

2018. Novel functions of death-associated protein kinases through mitogen-activated protein kinase-related signals. Int J Mol Sci. 19(10):3031. https://doi.org/10.3390/ijms19103031

Eming

Krieg

Davidson

. 2007. Inflammation in wound repair: molecular and cellular mechanisms. J Invest Dermatol. 127(3):514–525. https://doi.org/10.1038/sj.jid.5700701

Gade

Roy

Nallar

Kalvakolanu

. 2008. Critical role for transcription factor C/EBP-β in regulating the expression of death-associated protein kinase 1. Mol Cell Biol. 28(8):2528–2548. https://doi.org/10.1128/mcb.00784-07

Graça

MFP

Miguel

Cabral

CSD

Correia

. 2020. Hyaluronic acid-based wound dressings: a review. Carbohydr Polym. 241:116364. https://doi.org/10.1016/j.carbpol.2020.116364

10.

Jervies

King

Suphioglu

2021. Apolipoprotein E4 as a novel treatment target for Alzheimer’s disease. Cell Physiol Biochem. 55(6):773–783. https://doi.org/10.33594/000000475

11.

Kawai

, et al. 1999. Death-associated protein kinase 2 is a new calcium/calmodulin-dependent protein kinase that signals apoptosis through its catalytic activity. Oncogene. 18(23):3471–3480. https://doi.org/10.1038/sj.onc.1202701

12.

, et al. 2023. Distinct fibroblast progenitor subpopulation expedites regenerative mucosal healing by immunomodulation. J Exp Med. 220(3):e20221350. https://doi.org/10.1084/jem.20221350

13.

, et al. 2025. Umbilical cord mesenchymal stem cell-derived extracellular vesicles attenuate oxidative damage via the miR-191-5p/DAPK1/AKT axis in type 2 diabetes. Biomater Res. 29:0224. https://doi.org/10.34133/bmr.0224

14.

Kolltveit

Tronstad

Olsen

2000. Systemic diseases caused by oral infection. Clin Microbiol Rev. 13(4):547–558. https://doi.org/10.1128/cmr.13.4.547

15.

, et al. 2006. DAPK1 variants are associated with Alzheimer’s disease and allele-specific expression. Hum Mol Genet. 15(17):2560–2568. https://doi.org/10.1093/hmg/ddl178

16.

MacDonald

Keiko

. 2009. Wnt/Beta-Catenin Signaling: Components, Mechanisms, and Diseases. Developmental Cell. 17(1): 9–26. https://doi.org/10.1016/j.devcel.2009.06.016

17.

Marino

, et al. 2022. Diabetes-induced cellular senescence and senescence-associated secretory phenotype impair cardiac regeneration and function independently of age. Diabetes. 71(5):1081–1098. https://doi.org/10.2337/db21-0536

18.

Martineau

Kirienko

Pujol

2021. Innate immunity in C. E=elegans. Curr Top Dev Biol. 144:309–351. https://doi.org/10.1016/bs.ctdb.2020.12.007

19.

Okuda

Ebihara

1998. Relationships between chronic oral infectious diseases and systemic diseases. Bull Tokyo Dent Coll. 39(3):165–174.

20.

Politis

Schoenaers

Jacobs

Agbaje

. 2016. Wound healing problems in the mouth. Front Physiol. 7:507. https://doi.org/10.3389/fphys.2016.00507

21.

Prausnitz

. 2017. Engineering microneedle patches for vaccination and drug delivery to skin. Annu Rev Chem Biomol Eng. 8:177–200. https://doi.org/10.1146/annurev-chembioeng-060816-101514

22.

Raja

Garcia

Isseroff

. 2007. Wound re-epithelialization: modulating kerationcyte migration in wound healing. Front Biosci. 12(8):2849–2868. https://doi.org/10.2741/2277

23.

Rennier

. 2015. Shear stress attenuates apoptosis due to TNFα, oxidative stress, and serum depletion via death-associated protein kinase (DAPK) expression. BMC Res Notes. 8:85. https://doi.org/10.1186/s13104-015-1037-8

24.

Rousselle

Braye

Dayan

2019. Re-epithelialization of adult skin wounds: cellular mechanisms and therapeutic strategies. Adv Drug Deliv Rev. 146:344–365. https://doi.org/10.1016/j.addr.2018.06.019

25.

Sulaiman Alsaadi

. 2019. Role of DAPK1 in neuronal cell death, survival and diseases in the nervous system. Int J Dev Neurosci. 74(1):11–17. https://doi.org/10.1016/j.ijdevneu.2019.02.003

26.

Toma

Fuller

Willett

Goudy

SL.

2021. Oral wound healing models and emerging regenerative therapies. Transl Res. 236:17–34. https://doi.org/10.1016/j.trsl.2021.06.003

27.

Tong

, et al. 2009. Negative regulation of Caenorhabditis elegans epidermal damage responses by death-associated protein kinase. Proc Natl Acad Sci. 106(5):1457–1461. https://doi.org/10.1073/pnas.0809339106

28.

Waasdorp

, et al. 2021. The bigger picture: why oral mucosa heals better than skin. Biomolecules. 11(8):1165. https://doi.org/10.3390/biom11081165

29.

Wynn

Vannella

KM.

2016. Macrophages in tissue repair, regeneration, and fibrosis. Immunity. 44(3):450–462. https://doi.org/10.1016/j.immuni.2016.02.015

30.

Xavier

, et al. 2015. Bioactive nanoengineered hydrogels for bone tissue engineering: a growth-factor-free approach. ACS Nano. 9(3):3109–3118. https://doi.org/10.1021/nn507488s

31.

Chisholm

AD.

2011. A Gαq-Ca²⁺ signaling pathway promotes actin-mediated epidermal wound closure in C. elegans. Curr Biol. 21(23):1960–1967. https://doi.org/10.1016/j.cub.2011.10.050

32.

Yetisen

, et al. 2016. Photonic hydrogel sensors. Biotechnol Adv. 34(3):250–271. https://doi.org/10.1016/j.biotechadv.2015.10.005

33.

Zafari

Shirian

Sadeghi

Teimourian

Bakhtiyari

2020. CD93 Hematopoietic stem cells improve diabetic wound healing by VEGF activation and downregulation of DAPK-1. J Cell Physiol. 235(3):2366–2376. https://doi.org/10.1002/jcp.29142

34.

Zhang

, et al. 2022. Death-associated protein kinase 1 (DAPK1) protects against myocardial injury induced by myocardial infarction in rats via inhibition of inflammation and oxidative stress. Dis Markers. 2022:9651092. https://doi.org/10.1155/2022/9651092

35.

Zhang

et al. 2025. DAPK1 Acts as a Positive Regulator of Hypertension via Induction of Vasoconstriction. Clinical Science (London, England: 1979). 139(12):667–81. https://doi.org/10.1042/CS20255840

36.

Zhao

, et al. 2016. Photocrosslinkable gelatin hydrogel for epidermal tissue engineering. Adv Healthc Mater. 5(1):108–118. https://doi.org/10.1002/adhm.201500005

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

2.34 MB