Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

Di, Tiankai; Feng, Chao; Wang, Lulu; Xu, Jinlong; Du, Yang; Cheng, Baixiang; Chen, Yujiang; Wu, Lian

doi:10.1007/s12015-024-10695-6

Cited by 3 publications

References 52 publications

(44 reference statements)

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Single-cell RNA sequencing reveals vascularization-associated cell subpopulations in dental pulp: PDGFRβ+ DPSCs with activated PI3K/AKT pathway

Di,

Wang,

Cheng

et al. 2024

Stem Cells

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Background This study aims to address challenges in dental pulp regeneration therapy. The heterogeneity of DPSCs poses challenges, especially in stem cell transplantation for clinical use, particularly when sourced from donors of different ages and conditions. Methods Pseudotime analysis was employed to analyze single-cell sequencing data, and immunohistochemical studies were conducted to investigate the expression of fibronectin 1 (FN1). We performed in vitro sorting of PDGFRβ+ DPSCs using flow cytometry. A series of functional assays, including cell proliferation, scratch, and tube formation assays, were performed to experimentally validate the vasculogenic capabilities of the identified PDGFRβ+ DPSC subset. Furthermore, gene-edited mouse models were utilized to demonstrate the importance of PDGFRβ+ DPSCs. Transcriptomic sequencing was conducted to compare the differences between PDGFRβ+ DPSCs and P1-DPSCs. Results Single-cell sequencing analysis unveiled a distinct subset, PDGFRβ+ DPSCs, characterized by significantly elevated FN1 expression during dental pulp development. Subsequent cell experiments demonstrated that this subset possesses remarkable abilities to promote HUVEC proliferation, migration, and tube formation. Gene-edited mouse models confirmed the vital role of PDGFRβ+ DPSCs in dental pulp development. Transcriptomic sequencing and in vitro experiments demonstrated that the PDGFR/PI3K/AKT signaling pathway is a crucial factor mediating the proliferation rate and pro-angiogenic properties of PDGFRβ+ DPSCs. Conclusion We defined a new subset, PDGFRβ+ DPSCs, characterized by strong proliferative activity and pro-angiogenic capabilities, demonstrating significant clinical translational potential.

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Single-cell RNA sequencing reveals vascularization-associated cell subpopulations in dental pulp: PDGFRβ+ DPSCs with activated PI3K/AKT pathway

Di,

Wang,

Cheng

et al. 2024

Stem Cells

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Calcium‐sensing receptor regulates the angiogenic differentiation of LPS‐treated human dental pulp cells via the phosphoinositide 3‐kinase/Akt pathway in vitro

Yang,

Liu,

Qiu

et al. 2024

Int Endodontic J

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AimThe purpose of this study was to investigate the role of calcium‐sensing receptor (CaSR) in the angiogenic differentiation of lipopolysaccharide (LPS)‐treated human dental pulp cells (hDPCs).MethodologyThe LPS‐induced hDPCs were cultured in the medium with different combinations of CaSR agonist R568 and antagonist Calhex231. The cell proliferation, migration, and angiogenic capacity were measured by Cell Counting Kit‐8 (CCK‐8), scratch wound healing, and tube formation assays, respectively. Enzyme‐linked immunosorbent assay (ELISA), quantitative real‐time polymerase chain reaction (qRT‐PCR), and western blot were conducted to determine the gene/protein expression of CaSR, inflammatory mediators, and angiogenic‐associated markers. The activation of phosphoinositide 3‐kinase (PI3K) and protein kinase B (Akt) was assessed by western blot analysis.ResultsThe cell proliferation was elevated in response to R568 or Calhex231 exposure, but an enhanced cell migration was only found in cultures supplemented with Calhex231. Furthermore, R568 was found to potentiate the formation of vessel‐like structure, up‐regulated the protein expression of tumour necrosis factor (TNF)‐α, vascular endothelial growth factor (VEGF), and stromal cell‐derived factor (SDF)‐1; comparable influences were also observed in R568‐stimulated cells in the presence of PI3K inhibitor LY294002. In contrast, Calhex231 obviously inhibited the tube formation and VEGF protein level, whereas promoted the production of IL‐6, TNF‐α, and eNOS; however, in the presence of LY294002, Calhex231 showed a significant promotion on the protein expression of CaSR, VEGF, and SDF‐1. In addition, R568 exhibited a promotive action on the Akt phosphorylation, which can be reversed by LY294002.ConclusionsOur results demonstrated that CaSR can regulate the angiogenic differentiation of LPS‐treated hDPCs with an involvement of the PI3K/Akt signalling pathway.

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Mechanism of action of miR-330-5p targeting ITGA5 in the regulation of proliferation, migration, and invasion of gastric cancer

Wang,

Wei,

et al. 2024

Preprint

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BACKGROUND: ITGA5 is an oncogene that performs its biological function by integrating the intracellular structure and extracellular matrix. Our research group found that ITGA5 is a gastric cancer-related gene highly expressed in this tumor and is closely related to its proliferation and metastasis. The ITGA5 gene is regulated by multiple miRNAs during the occurrence and development of tumors. This study aimed to explore the role of targeting miRNAs upstream of ITGA5 in the regulation of the proliferation, invasion, and migration of gastric cancer cells. METHODS: The target miRNA molecules regulating the ITGA5 gene were predicted by four bioinformatics databases (TargetScan、miRDB、miRTarBase and mirDIP), and the unreported miRNAs with high correlation were selected and their expression in gastric cancer was assessed by qRT-PCR and western blot. The miRNAs with potential targeting abilities were further verified by dual luciferase reporter gene experiment. The effects of miR-330-5p and ITGA5 on the proliferation, invasion, and migration of gastric cancer cells were evaluated by CCK8, clonogenic assay, and Transwell chamber assay, respectively. RESULTS: Six miRNAs (miR-26a-5p、miR-92a-3p、miR-148a-3p、miR-148b-3p、miR-330-5p and miR-152-3p) with high stability and conservation were found, and miR-330-5p was the one targeting and regulating ITGA5. In vitro experiments demonstrated that miR-330-5p mimic significantly inhibited the proliferation, invasion, and migration of gastric cancer cells compared with the control group (p < 0.05). The transfection of miR-330-5p mimic into gastric cancer cells overexpressing ITGA5 (OE-ITGA5) resulted in a significant reversion of the promoting effect of OE-ITGA5 on the proliferation, invasion, and migration of gastric cancer cells (P < 0.05). In addition, miR-330-5p mimic reduced ITGA5 expression in gastric cancer cells and partially reversed the FAK/AKT signaling pathway activated by the ITGA5 gene. miR-330-5p inhibitor increased ITGA5 expression in gastric cancer cells, and they partially reversed the FAK/AKT signaling pathway blocked by sh-ITGA5. CONCLUSIONS: ITGA5 was promotive for GC tumor growth and cell biological behaviors, and miR-330-5p targeted 3'-UTR of ITGA5 and inhibited its expression. ITGA5 was expected to become a new molecular marker, with miR-330-5p representing a novel therapeutic target for GC. This discovery provides a theoretical basis to further understand the mechanism related to the occurrence and development of gastric cancer, improving the diagnosis and prognosis while discovering a new therapeutic target.

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Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

Cited by 3 publications

References 52 publications

Single-cell RNA sequencing reveals vascularization-associated cell subpopulations in dental pulp: PDGFRβ+ DPSCs with activated PI3K/AKT pathway

Single-cell RNA sequencing reveals vascularization-associated cell subpopulations in dental pulp: PDGFRβ+ DPSCs with activated PI3K/AKT pathway

Calcium‐sensing receptor regulates the angiogenic differentiation of LPS‐treated human dental pulp cells via the phosphoinositide 3‐kinase/Akt pathway in vitro

Mechanism of action of miR-330-5p targeting ITGA5 in the regulation of proliferation, migration, and invasion of gastric cancer

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