E2F1- deficient adipose-derived stem cells improve skin wound closure in mice by upregulating paracrine expression of VEGF and TGF-β1

Abstract

Background: Wound healing is a widespread health problem that imposes a financial burden on health systems. Cell therapy with genetically modified adipose-derived stem cells (ADSCs) is a promising strategy for dysregulated wound repair. E2F transcription factor 1 (E2F1) is a bidirectional regulator of cytokines. Here, the authors aimed to investigate the impact and potential mechanism of E2F1–/– ADSCs in promoting the wound healing process. Methods: Forty-five C57BL/6 mice (specific pathogen-free, male) with 10-mm full-thickness wounds were randomly treated with subcutaneous injection of 2 × 106 wild-type ADSCs, 2 × 106 E2F1–/– ADSCs, or phosphate-buffered saline. The wound closure rate was monitored at days 0, 3, 7, 10, and 14 after treatment. The collagen synthesis, angiogenesis, and wound contraction were calculated by Masson, immunohistochemistry, and immunofluorescent staining (CD31 and KI67), Western blotting (α-smooth muscle actin, collagen I, vascular endothelial growth factor, and transforming growth factor–β1) separately at day 14. In vitro, the conditioned media (CM) of wild-type ADSCs and E2F1–/– ADSCs were collected to evaluate the impact on proliferation, migration, and angiogenesis. Results: In vivo, the E2F1–/– ADSC group exhibited increased healing rate, proliferating vessels, and collagen synthesis compared with control at day 14 (P < 0.05). Moreover, E2F1–/– ADSCs showed enhanced vascular endothelial growth factor and transforming growth factor–β1 expression in the wound site and CM, and the CM from E2F1–/– ADSCs promoted the proliferation, migration, and tube formation of co-cultured cells in vitro (P < 0.05). Conclusion: The E2F1–/– ADSCs exhibited a strong paracrine ability to improve the vascularization process and collagen deposition, thereby accelerating wound healing in the rodent model. Clinical Relevance Statement: These findings show that targeting transcription factor E2F1 could regulate the paracrine function of ADSCs, developing E2F1-modified ADSCs as an effective therapeutic option for wound healing and regeneration.

Exosomes Derived from E2F1–/– Adipose-Derived Stem Cells Promote Skin Wound Healing via miR-130b-5p/TGFBR3 Axis

“… 38 Our previous research found that ADSCs from E2F1 –/– mice promoted wound healing via enhancing VEGF and TGF-β1 paracrine. 21 Besides, E2F1 knockout was protective in myocardial ischemia and skin injury. 39 , 40 MiRNAs participate in various biological processes through post-transcriptional regulation, and miRNA expression can be regulated by transcription factors.…”

“… 19 , 20 Besides, we also emphasized E2F1 deficiency could improve the ability of ADSC paracrine to promote wound healing. 21 Exosomal microRNAs (miRNAs) are considered to be important paracrine substances for ADSCs in promoting wound healing. 22 E2F1 can regulate the transcription of multiple miRNAs.…”

Adipose-Derived Stem Cells: Angiogenetic Potential and Utility in Tissue Engineering

“…Tissue healing: ADSCs can accelerate wound healing and/or improve healing by promoting angiogenesis, particularly in chronic wounds [154,161]. Their regenerative and immunomodulatory properties contribute to cell proliferation and accelerate wound healing and cutaneous regeneration [162][163][164][165].…”