Cold Atmospheric Plasma Jet Promotes Wound Healing Through CK2-Coordinated PI3K/AKT and MAPK Signaling Pathways

• Published in: Molecular & cellular proteomics Vol. 24; no. 5; p. 100962
• Main Authors: Wu, Pei-Shan, Wong, Tzu-Hsuan, Hou, Chun-Wei, Chu, Teng-Ping, Lee, Jyh-Wei, Lou, Bih-Show, Lin, Miao-Hsia
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.05.2025; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Casein Kinase II - metabolism; Cell Movement - drug effects; Cell Proliferation - drug effects; CK2 kinase; cold atmospheric plasma jet (CAPJ); Humans; Keratinocytes - drug effects; Keratinocytes - metabolism; MAP Kinase Signaling System - drug effects; Phosphatidylinositol 3-Kinases - metabolism; Phosphorylation - drug effects; PI3K/AKT and MAPK signaling; plasma activated medium (PAM); Plasma Gases - pharmacology; Proteome - metabolism; Proteomics; Proto-Oncogene Proteins c-akt - metabolism; Signal Transduction; wound healing; Wound Healing - drug effects; CAPJ; PPI; RTK; CK2; DMEM; LC-MS/MS; SDC; PI3K; FDR; mTOR; plasma activated medium (PAM); SLS; PI3K/AKT and MAPK signaling; AGC; PHOXTRACK; AA; CK2 kinase; cold atmospheric plasma jet (CAPJ); BCA; wound healing; RONS; MS; AKT; ACN; MAPK; DEP; ECM; PCA; TEABC; FA; FC; PAM
• ISSN: 1535-9476; 1535-9484; 1535-9484
• DOI: 10.1016/j.mcpro.2025.100962

The promising role of cold atmospheric plasma jet (CAPJ) treatment in promoting wound healing has been widely documented in therapeutic implications. However, the fact that not all subjects respond equally to CAPJ necessitates the investigation of the underlying cellular mechanisms, which have been rarely understood so far. Given that wound healing is a complex and prolonged process, post plasma-activated medium (PAM) treated keratinocytes were collected at two time points, 2 h (receiving) and 24 h (recovery), for (phospho)proteomic analysis to systematically dissect the molecular basis of CAPJ-promoted wound healing. The receiving (phospho)proteomics datasets, referred to the time point of 2 h, revealed an apparent increase in the phosphorylation of CK2 and its-mediated PI3K/AKT and MAPK signaling pathways, accompanied by a prompted downstream physiological response of cell migration. Additionally, incorporating the network analysis of predicted kinases and their direct interactors, we reiterated that CAPJ influenced cell growth and migration, thereby paving the way for its role in subsequent wound healing processes. Further determining the proteome profiles at recovery phase, which is the time point of 24 h, displayed a totally different view from the receiving proteome which had almost no change. The upregulation of ROBOs/SLITs expression and vesicle trafficking and fusion-related proteins, along with the abundant presence of 14-3-3 family proteins, indicated that the persistent effect of PAM on the wound healing process could potentially promote keratinocyte-fibroblast cross talk and stimulate extracellular matrix synthesis upon epithelialization. Consistent with proteome patterns, CAPJ-treated wound tissues indeed showed a denser and well-organized extracellular matrix architecture, implying hastened epithelialization during wound healing. Collectively, we delineated the molecular basis of CAPJ-accelerated wound healing at early and late responses, providing valuable insights for treatment selection and the development of therapeutic strategies to achieve better outcomes. [Display omitted] •Protein phosphorylation acts as the immediate responders to CAPJ treatment.•CAPJ-activated PI3K/AKT and MAPK signaling directs keratinocyte migration.•CK2-coordinated PI3K/AKT and MAPK pathways are key to CAPJ-enhanced wound closure.•Persistent CAPJ effects enhanced ECM remodeling during wound epithelialization. (Phospho)proteomics analyses have revealed that CAPJ enhances wound healing via CK2 coordinated PI3K/AKT and MAPK phosphorylation events, which boosts keratinocyte migration. Furthermore, CAPJ consistently influences the late phase of wound closure by upregulating 14-3-3 proteins expression, which facilitates keratinocyte-fibroblast cross talk and promotes ECM remodeling at wound area during epithelialization. This study unveils the molecular mechanisms behind CAPJ's therapeutic effects, offering valuable insights for better predictions and treatment outcomes.