GDF-5 promotes epidermal stem cells proliferation via Foxg1-cyclin D1 signaling

Abstract Objective Epidermal stem cells (EpSCs) can self-renew, which are responsible for the long-term maintenance of the skin, and it also plays a critical role in wound re-epithelization, but the mechanism underlying EpSCs proliferation is unclear. GDF-5, also known as BMP-14, is a member of the BMP family and can be used as a self-renewal supporter. Here, we studied the effects of GDF-5 on mouse EpSCs proliferation mechanism in wound healing. Methods Firstly, the effects of GDF-5 on EpSCs proliferation was tested by using CCK8 reagent and PCNA expression was analyzed by Western blotting. Secondly, we screened genes that promote EpSCs proliferation in the FOX and cyclin family by qPCR, and then the protein expression level of the selected genes was further analyzed by Western blotting. Thirdly, siRNA plasmids and pAdEasy adenovirus were transfected or infected, respectively, into mouse EpSCs to detect the effect of target genes on GDF-5-induced cell proliferation. Furthermore, we injected GDF-5 to a deep partial thickness burn mouse model for finding out whether EpSCs proliferation can be detected by immunohistochemical. Finally, the relevant target genes were analyzed by qPCR, immunoblotting, and dual-luciferase reporter gene detection. Results We discovered that 100 ng/ml recombinant mouse GDF-5 was the optimal concentration for promoting mouse EpSCs proliferation. Through preliminary screened by qPCR, we found that Foxg1 and cyclin D1 could be the downstream molecules of GDF-5, and the results were confirmed by Western blotting. And the effect of GDF-5 on mouse EpSCs proliferation was adjusted by Foxg1/cyclin D1 in vitro and in vivo. Besides, GDF-5-induced transcription of cyclin D1 was regulated by Foxg1-mediated cyclin D1 promoter activity. Conclusion This paper showed that GDF-5 promotes mouse EpSCs proliferation via Foxg1-cyclin D1 signal pathway. It is suggested that GDF-5 may be a new approach to make EpSCs proliferation which can be used in wound healing.

Download Full-text

GDF-5 promotes Epidermal Stem Cells proliferation via Foxg1-cyclin D1 signaling

10.21203/rs.3.rs-102954/v1 ◽

2020 ◽

Author(s):

Xiaohong Zhao ◽

Ruyu Bian ◽

Fan Wang ◽

Ying Wang ◽

Xue Li ◽

...

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Cyclin D1 ◽

Western Blotting ◽

Target Genes ◽

Critical Role ◽

Luciferase Reporter ◽

Epidermal Stem Cells ◽

Luciferase Reporter Gene ◽

Partial Thickness Burn

Abstract Objective: Epidermal stem cells (EpSCs) can self-renew and are responsible for the long-term maintenance of the skin, it also plays a critical role in wound re-epithelization, but the mechanism underlying EpSCs proliferation is unclear. Here, we studied the effects of GDF-5 on mouse EpSCs proliferation mechanism in wound healing. Methods: Firstly, the effects of GDF-5 on EpSCs proliferation was tested by using CCK8 reagent and PCNA expression was analysed by western blotting. Secondly, we screened genes that promote EpSCs proliferation in the FOX family and cyclin by qPCR, and further analysed the protein expression level of the selected genes by Western blotting. Thirdly, siRNA plasmids and pAdEasy adenovirus were transfected or infected, respectively, into mouse EpSCs to detect the effect of target genes on GDF-5 induced cell proliferation. Furthermore, a deep partial thickness burn mouse model was used in which GDF-5 induced EpSCs proliferation was detected by immunohistochemical. Finally, the relationships target genes were analysed by qPCR, immunoblotting and dual luciferase reporter gene detection. Results: We discovered that 100 ng/ml recombinant mouse GDF-5 was the optimal concentration for promoting mouse EpSCs proliferation. Through preliminary screening by qPCR, we found that Foxg1 and cyclin D1 could be the downstream molecules of GDF-5, and the results were confirmed by Western blotting. And the effect of GDF-5 on mouse EpSCs proliferation was regulated by Foxg1/cyclin D1 in vitro and in vivo. Besides, GDF-5 induced transcription of cyclin D1 was regulated by Foxg1-mediated cyclin D1 promoter activity.Conclusion: This paper showed that GDF-5 promotes mouse EpSCs proliferation via Foxg1-cyclin D1 signal pathway. It is suggested that GDF-5 may be a new approach to yield EpSCs for wound healing.

Download Full-text

Epidermal Stem Cells in Skin Wound Healing

Advances in Wound Care ◽

10.1089/wound.2017.0728 ◽

2017 ◽

Vol 6 (9) ◽

pp. 297-307 ◽

Cited By ~ 30

Author(s):

Yuanyuan Li ◽

Jamie Zhang ◽

Jiping Yue ◽

Xuewen Gou ◽

Xiaoyang Wu

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Skin Wound ◽

Epidermal Stem Cells ◽

Skin Wound Healing

Download Full-text

Plasticity of Epidermal Stem Cells: The Future of Stem Cell-Based Therapeutics to Improve Cutaneous Wound Healing

Severe Trauma and Sepsis ◽

10.1007/978-981-13-3353-8_20 ◽

2019 ◽

pp. 357-389

Author(s):

Xiaoyan Sun ◽

Hua Qin ◽

Siming Yang ◽

Xiaobing Fu

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Stem Cell ◽

Cutaneous Wound Healing ◽

Epidermal Stem Cells ◽

Cutaneous Wound ◽

The Future

Download Full-text

Critical role of the endogenous renin-angiotensin system in maintaining self-renewal and regeneration potential of epidermal stem cells

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease ◽

10.1016/j.bbadis.2019.07.006 ◽

2019 ◽

Vol 1865 (10) ◽

pp. 2647-2656 ◽

Cited By ~ 2

Author(s):

Xuan Liao ◽

Jing Xiao ◽

Sheng-Hong Li ◽

Li-Ling Xiao ◽

Biao Cheng ◽

...

Keyword(s):

Stem Cells ◽

Critical Role ◽

Renin Angiotensin System ◽

Epidermal Stem Cells ◽

Regeneration Potential ◽

Self Renewal ◽

Angiotensin System ◽

Renin Angiotensin

Download Full-text

Curcumin promotes burn wound healing in mice by upregulating caveolin-1 in epidermal stem cells

Phytotherapy Research ◽

10.1002/ptr.6238 ◽

2018 ◽

Vol 33 (2) ◽

pp. 422-430 ◽

Cited By ~ 9

Author(s):

Ronghua Yang ◽

Jingru Wang ◽

Ziheng Zhou ◽

Shaohai Qi ◽

Shubin Ruan ◽

...

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Epidermal Stem Cells ◽

Burn Wound ◽

Caveolin 1 ◽

Burn Wound Healing

Download Full-text

Exosomes derived from mmu_circ_0000250-modified adipose-derived mesenchymal stem cells promote wound healing in diabetic mice by inducing miR-128-3p/SIRT1-mediated autophagy

AJP Cell Physiology ◽

10.1152/ajpcell.00041.2020 ◽

2020 ◽

Vol 318 (5) ◽

pp. C848-C856 ◽

Cited By ~ 5

Author(s):

Rongfeng Shi ◽

Yinpeng Jin ◽

Weiwei Hu ◽

Weishuai Lian ◽

Chuanwu Cao ◽

...

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Mesenchymal Stem Cells ◽

Therapeutic Effect ◽

Diabetic Rats ◽

Luciferase Reporter Assay ◽

Luciferase Reporter ◽

Reporter Assay ◽

High Concentration ◽

Promote Wound Healing

More and more evidence advises that circular RNAs (circRNAs) function critically in regulating different disease microenvironments. Our previous study found that autotransplantation of adipose-derived mesenchymal stem cells (ADSCs) promotes diabetes wound healing. Exosomes derived in ADSCs play an important regulatory role. This study aimed to characterize if mmu_circ_0000250 played a role in ADSC-exosome-mediated full-thickness skin wound repair in diabetic rats. Endothelial progenitor cells (EPCs) were selected to study the therapeutic mechanism of exosomes in high-glucose (HG)-induced cell damage and dysfunction. Analysis and luciferase reporter assay were utilized to explore the interaction among mmu_circ_0000250, miRNA (miR)-128-3p, and sirtuin (SIRT)1. The diabetic rats were used to confirm the therapeutic effect of mmu_circ_0000250 against exosome-mediated wound healing. Exosomes containing a high concentration of mmu_circ_0000250 had a greater therapeutic effect on restoration of the function of EPCs by promotion autophagy activation under HG conditions. Expression of mmu_circ_0000250 promoted SIRT1 expression by miR-128-3p adsorption, which was confirmed via luciferase reporter assay and bioinformatics analysis. In vivo, exosomes containing a high concentration of mmu_circ_0000250 had a more therapeutic effect on wound healing when compared with wild-type exosomes from ADSCs. Immunohistochemistry and immunofluorescence detection showed that mmu_circ_0000250 increased angiopoiesis with exosome treatment in wound skin and suppressed apoptosis by autophagy activation. In conclusion, we verified that mmu_circ_0000250 enhanced the therapeutic effect of ADSC-exosomes to promote wound healing in diabetes by absorption of miR-128-3p and upregulation of SIRT1. Therefore, these findings advocate targeting the mmu_circ_0000250/miR-128-3p/SIRT1 axis as a candidate therapeutic option for diabetic ulcers.

Download Full-text

A distal activation domain is critical in the regulation of the rat androgen receptor gene promoter

Biochemical Journal ◽

10.1042/bj2940779 ◽

1993 ◽

Vol 294 (3) ◽

pp. 779-784 ◽

Cited By ~ 19

Author(s):

C S Song ◽

S Her ◽

M Slomczynska ◽

S J Choi ◽

M H Jung ◽

...

Keyword(s):

Androgen Receptor ◽

Nucleotide Sequence ◽

Receptor Gene ◽

Critical Role ◽

Androgen Receptor Gene ◽

Luciferase Reporter ◽

Upstream Region ◽

Luciferase Reporter Gene ◽

Mobility Shift ◽

Cis Element

The far upstream region of the rat androgen receptor (AR) gene has been cloned, and the nucleotide sequence up to -2656 bp established. Nested deletion mutants of rat AR 5′ flanking sequences were ligated to the luciferase reporter gene, and their promoter activities were examined in transfected COS1 cells. Results show a critical cis-acting domain located between positions -960 and -940. Deletion of this cis element resulted in a greater than 90% decrease in the promoter activity. A nuclear protein that specifically binds to this 21-nucleotide sequence was identified by gel mobility shift analysis. The -960/-940 cis element has no identify to the binding sequence of any known transcription factor. Furthermore, the cognate binding protein is present in both rat and human (HeLa) cell nuclear extracts. We conclude that a novel trans-activator interacting at the -960/-940 region plays a critical role in the regulation of AR gene expression.

Download Full-text

Ezh2 Plays a Critical Role in the Progression of MLL-AF9-Induced Acute Myeloid Leukemia

Blood ◽

10.1182/blood.v118.21.57.57 ◽

2011 ◽

Vol 118 (21) ◽

pp. 57-57

Author(s):

Satomi Tanaka ◽

Goro Sashida ◽

Satoru Miyagi ◽

Koutaro Yokote ◽

Chiaki Nakaseko ◽

...

Keyword(s):

Stem Cells ◽

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Target Genes ◽

Nuclear Translocation ◽

Conflicts Of Interest ◽

Critical Role ◽

Histone H3 ◽

Chip Sequencing ◽

Acute Myeloid

Abstract Abstract 57 The polycomb group proteins function in gene silencing through histone modifications. They have been characterized as a general regulator of stem cells, but also play a critical role in cancer. EZH2 is a catalytic component of the polycomb repressive complex 2 (PRC2) and tri-methylates histone H3 at lysine 27 to transcriptionally repress the target genes. Although EZH2 is over-expressed in various cancers including hematological malignancies, it remains unknown how EZH2 contributes to the initiation and/or progression of acute myeloid leukemia (AML). To understand the role of EZH2 in AML, we transformed granulocyte macrophage progenitors (GMPs) from Cre-ERT;Ezh2+/+ and Cre-ERT;Ezh2flox/flox mice with the MLL-AF9 fusion gene. Then, Ezh2 was deleted by inducing nuclear translocation of Cre by adding tamoxifen to culture. We found that proliferation of Ezh2δ/δ transformed cells was severely compromised upon deletion of Ezh2 (Ezh2δ/δ) in liquid culture. They gave rise to a significantly reduced number of colonies in replating assays. Of note, while Ezh2+/+ cells formed compact colonies composed of immature myeloblasts, Ezh2δ/δ cells formed dispersed colonies composed of differentiated myeloid cells. We next transplanted Cre-ERT;Ezh2+/+ and Cre-ERT;Ezh2flox/flox GMPs transformed by MLL-AF9 into recipient mice. All the recipient mice developed AML by 3 weeks after transplantation. At 3 weeks after transplantation, we depleted Ezh2 by intraperitoneal injection of tamoxifen. Deletion of Ezh2 significantly prolonged the survival of the recipient mice (60 days vs. 76 days, p<0.0001), although all the mice eventually died of leukemia. Nonetheless, as was detected in vitro, Ezh2δ/δ AML cells in BM were apparently differentiated in morphology compared with the control. Ezh2δ/δ AML cells in BM gave rise to 10-fold fewer colonies in methylcellulose medium compared with Ezh2+/+ AML cells, and again showed an obvious tendency of differentiation. These observations imply that Ezh2 is critical for the progression of MLL-AF9 AML and maintains the immature state of AML cells. To elucidate the mechanism how Ezh2 promotes the progression of MLL-AF9-induced AML, we examined the genome-wide distribution of tri-methylation of histone H3 at lysine 27 (H3K27me3) by ChIP-sequencing and microarray-based expression analysis. ChIP-sequencing using Ezh2+/+ and Ezh2δ/δ BM AML cells identified 3525 and 89 genes exhibiting a ≧ 10-fold enrichment in H3K27me3 levels in Ezh2+/+ and Ezh2δ/δ AML cells, respectively, confirming a drastic reduction in the levels of global H3K27me3 in the absence of Ezh2. Microarray analysis using lineage marker (except for Mac1)−Sca-1−c-Kit+FcγRII/IIIhi BM AML cells revealed 252 upregulated and 154 downregulated genes (≧ 2-fold) in Ezh2δ/δ AML cells compared with Ezh2+/+ AML cells. Of interest, the absence of Ezh2 did not affect the transcriptional activation of the major target genes of MLL-AF9, including HoxA9 and Meis1. Because Ezh2 functions as transcriptional repressor, de-repressed genes could be direct targets of Ezh2. Based on these data, we are now engaged in further comprehensive analysis to narrow down the direct target genes of Ezh2 responsible for the progression of AML. Collectively, our findings suggest that Ezh2 is the major enzyme for H3K27me3 in AML and contributes to the progression of AML by regulating transcription a cohort of genes that are supposedly relevant to the self-renewal capacity and perturbed differentiation of AML stem cells. Disclosures: No relevant conflicts of interest to declare.

Download Full-text