RESUMO
Regenerative capabilities of the endothelium rely on vessel-resident progenitors termed endothelial colony forming cells (ECFCs). This study aimed to investigate if these progenitors are impacted by conditions (i.e., obesity or atherosclerosis) characterized by increased serum levels of oxidized low-density lipoprotein (oxLDL), a known inducer of Endothelial-to-Mesenchymal Transition (EndMT). Our investigation focused on understanding the effects of EndMT on the self-renewal capabilities of progenitors and the associated molecular alterations. In the presence of oxLDL, ECFCs displayed classical features of EndMT, through reduced endothelial gene and protein expression, function as well as increased mesenchymal genes, contractility, and motility. Additionally, ECFCs displayed a dramatic loss in self-renewal capacity in the presence of oxLDL. RNA-sequencing analysis of ECFCs exposed to oxLDL validated gene expression changes suggesting EndMT and identified SOX9 as one of the highly differentially expressed genes. ATAC sequencing analysis identified SOX9 binding sites associated with regions of dynamic chromosome accessibility resulting from oxLDL exposure, further pointing to its importance. EndMT phenotype and gene expression changes induced by oxLDL in vitro or high fat diet (HFD) in vivo were reversed by the silencing of SOX9 in ECFCs or the endothelial-specific conditional knockout of Sox9 in murine models. Overall, our findings support that EndMT affects vessel-resident endothelial progenitor's self-renewal. SOX9 activation is an early transcriptional event that drives the mesenchymal transition of endothelial progenitor cells. The identification of the molecular network driving EndMT in vessel-resident endothelial progenitors presents a new avenue in understanding and preventing a range of condition where this process is involved.
Assuntos
Lipoproteínas LDL , Fatores de Transcrição SOX9 , Lipoproteínas LDL/metabolismo , Lipoproteínas LDL/farmacologia , Animais , Fatores de Transcrição SOX9/metabolismo , Fatores de Transcrição SOX9/genética , Camundongos , Humanos , Aterosclerose/metabolismo , Aterosclerose/patologia , Aterosclerose/genética , Transição Epitelial-Mesenquimal , Camundongos Endogâmicos C57BL , Masculino , Células Progenitoras Endoteliais/metabolismo , Células Progenitoras Endoteliais/citologia , Autorrenovação Celular , Células Endoteliais/metabolismoRESUMO
The aggressiveness of solid cancers, such as melanoma, relies on their metastatic potential. It has become evident that this key cause of mortality is largely conferred by the tumour-associated stromal cells, especially endothelial cells. In addition to their essential role in the formation of the tumour vasculature, endothelial cells significantly contribute to the establishment of the tumour microenvironment, thus enabling the dissemination of cancer cells. Melanoma tumour vascularization occurs through diverse biological processes. Vasculogenesis is the formation of de novo blood vessels from endothelial progenitor cells (EPCs), and recent research has shown the role of EPCs in melanoma tumour vascularization. A more detailed understanding of the complex role of EPCs and how they contribute to the abnormal vessel structures in tumours is of importance. Moreover, anti-angiogenic drugs have a limited effect on melanoma tumour vascularization, and the role of these drugs on EPCs remains to be clarified. Overall, targeting cancer vasculature remains a challenge, and the role of anti-angiogenic drugs and combination therapies in melanoma, a focus of this review, is an area of extensive exploration.
RESUMO
Endothelial to mesenchymal transition (EndMT) is a leading cause of fibrosis and disease, however its mechanism has yet to be elucidated. The endothelium possesses a profound regenerative capacity to adapt and reorganize that is attributed to a population of vessel-resident endovascular progenitors (EVP) governing an endothelial hierarchy. Here, using fate analysis, we show that two transcription factors SOX9 and RBPJ specifically affect the murine EVP numbers and regulate lineage specification. Conditional knock-out of Sox9 from the vasculature (Sox9fl/fl/Cdh5-CreER RosaYFP) depletes EVP while enhancing Rbpj expression and canonical Notch signalling. Additionally, skin wound analysis from Sox9 conditional knock-out mice demonstrates a significant reduction in pathological EndMT resulting in reduced scar area. The converse is observed with Rbpj conditionally knocked-out from the murine vasculature (Rbpjfl/fl/Cdh5-CreER RosaYFP) or inhibition of Notch signaling in human endothelial colony forming cells, resulting in enhanced Sox9 and EndMT related gene (Snail, Slug, Twist1, Twist2, TGF-ß) expression. Similarly, increased endothelial hedgehog signaling (Ptch1fl/fl/Cdh5-CreER RosaYFP), that upregulates the expression of Sox9 in cells undergoing pathological EndMT, also results in excess fibrosis. Endothelial cells transitioning to a mesenchymal fate express increased Sox9, reduced Rbpj and enhanced EndMT. Importantly, using topical administration of siRNA against Sox9 on skin wounds can substantially reduce scar area by blocking pathological EndMT. Overall, here we report distinct fates of EVPs according to the relative expression of Rbpj or Notch signalling and Sox9, highlighting their potential plasticity and opening exciting avenues for more effective therapies in fibrotic diseases.
Assuntos
Células Endoteliais/metabolismo , Endotélio/metabolismo , Proteína de Ligação a Sequências Sinal de Recombinação J de Imunoglobina/metabolismo , Fatores de Transcrição SOX9/metabolismo , Transdução de Sinais/genética , Animais , Diferenciação Celular/genética , Linhagem da Célula , Endotélio/citologia , Feminino , Técnicas de Inativação de Genes , Proteínas Hedgehog/metabolismo , Humanos , Proteína de Ligação a Sequências Sinal de Recombinação J de Imunoglobina/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , RNA Interferente Pequeno , Receptores Notch/metabolismo , Fatores de Transcrição SOX9/genética , Fator de Crescimento Transformador beta/metabolismo , Cicatrização/genéticaRESUMO
PURPOSE: Dentin hypersensitivity is characterized by short, sharp pain arising from exposed dentin in response to external stimuli. Several modalities have been suggested for treatment of this condition such as low-level laser therapy (LLLT) and application of dentin bonding agents. The aim of this study was to compare the clinical efficacy of diode laser, Nd:YAG laser and dentin bonding agent for treatment of dentin hypersensitivity. MATERIALS AND METHODS: In this study, 135 teeth of 22 patients diagnosed with dentin hypersensitivity were divided into three groups: In group 1, the teeth were irradiated by diode laser with 810 nm wavelength for 30 seconds and in group 2, the teeth were irradiated by Nd:YAG laser with 1064 nm wavelength for 40 seconds. CLEARFIL SE BOND was applied on teeth in group 3. LLLT was carried out in 3 sessions with 7-day intervals between sessions, during a period of 3 consecutive weeks. Hypersensitivity was assessed by cold test according to the criteria proposed by Uchida at baseline, immediately after treatment and at 1, 3 and 6 months, postoperatively. RESULTS: Reduction of dentin hypersensitivity was observed at 3 and 6 months following the use of Nd:YAG laser (p < 0.001). Reduction in dentin hypersensitivity was observed immediately after treatment in all groups. Statistically significant differences in level of dentin hypersensitivity were found between groups at 3 and at 6 months (p ≤ 0.001). The reduction in dentin hypersensitivity by Nd:YAG laser was significantly superior to that in other groups at this time. CONCLUSION: The efficacy of Nd:YAG laser in reduction of dentin hypersensitivity was significantly superior to that of other modalities at 3 and 6 months.