Jong-Hyuk Sung’s research while affiliated with R&D Systems and other places

What is this page?


This page lists works of an author who doesn't have a ResearchGate profile or hasn't added the works to their profile yet. It is automatically generated from public (personal) data to further our legitimate goal of comprehensive and accurate scientific recordkeeping. If you are this author and want this page removed, please let us know.

Publications (112)


Figure 4. The expression and regulation of CXCL12 in human dermal papilla cells (DPCs). (A,B) Testosterone propionate (TP, A) and dihydrotestosterone (DHT, B) increased the CXCL12 level in human DPCs. (C) AR knockdown (AR-KD) in DPCs using AR-specific siRNA reduced the expression of CXCL12 induced by TP (100 nM) and DHT (100 nM). * p < 0.05; ** p < 0.01 vs. control.
Figure 5. The expression and regulation of CXCL12 in human dermal sheath cup cells (DSCs). (A,B) Testosterone propionate (TP, A) and dihydrotestosterone (DHT, B) increased the CXCL12 level in human DPCs. (C) AR knockdown (AR-KD) in DSCs using AR-specific siRNA reduced the expression of CXCL12 induced by TP and DHT. * p < 0.05; ** p < 0.01 vs. control.
Differential Expression of CXCL12 in Human and Mouse Hair: Androgens Induce CXCL12 in Human Dermal Papilla and Dermal Sheath Cup
  • Article
  • Full-text available

December 2024

·

10 Reads

International Journal of Molecular Sciences

·

·

In Guk Park

·

[...]

·

Jong-Hyuk Sung

We previously demonstrated that C-X-C Motif Chemokine Ligand 12 (CXCL12) is primarily secreted by dermal fibroblasts in response to androgens and induces hair miniaturization in the mouse androgenic alopecia (AGA) model. However, the direct effects of androgen-induced CXCL12 on dermal papilla cells (DPCs) and dermal sheath cup cells (DSCs) have not been demonstrated. First, we compared single-cell RNA sequencing data between mouse and human skin, and the results show that CXCL12 is highly co-expressed with the androgen receptor (AR) in the DPCs and DSCs of only human hair. Immunohistochemistry also showed that CXCL12 is co-expressed with the AR in the DPCs and DSCs of human hair follicles. In human hair organ culture, androgens also increased CXCL12 expression in DPCs and DSCs and reduced hair length, while the CXCL12 antibody increased hair length via AR inactivation. CXCL12 mRNA was upregulated by androgen treatment in primary human DPCs and DSCs. On the contrary, AR inhibitors or siRNA treatment reduced CXCL12 expression. Collectively, these results suggest that CXCL12 is co-expressed with the AR in the DPCs and DSCs of human hair follicles; therefore, inhibition of CXCL12 using antibodies is a promising strategy for AGA treatment.

Download


scRNA-seq of CXCL12 Ab-treated AA mouse model. (A) Workflow of scRNA-seq analysis. (B–D) t-SNE representation of scRNA-seq results from (B) Neg, (C) AA, and (D) AA + Ab groups, colored by annotated cell types. (E) Expression levels for selected marker genes of each cell type. (F) Proportion of each cell type among total cells for each group. (G–I) Proportion of (G) immune cells, (H) T cells, and (I) DC/Mac among total cells for each group. Statistical significance was assessed using the binomial test. *P < 0.05, ***P < 0.001.
Pseudobulk RNA-seq analysis of CXCL12 Ab-treated AA mouse model. (A, B) Identification of DEGs: (A) Neg vs. AA and (B) AA vs. AA + Ab. Genes with a more than twofold increase are shown in red, and those with a decrease are shown in blue, with the number of each DEG indicated. (C) Heatmap showing the normalized Z-scores of all DEGs. (D) Venn diagram depicting the identification of 153 DEGs (AA up & AA + Ab down). (E) STRING network based on protein-protein interactions of the 153 DEGs. (F) Three major clusters are identified from community detection based on weighted edge betweenness. (G) Gene ontology (GO) enrichment analysis results for each cluster. Significant results with P value < 0.001 are shown. (H) Pre-ranked GSEA results using log2 fold change from each comparison as input.
Expression profile of CXCL12 and its receptors in the AA mouse model. (A) Heatmap showing the relative importance of each cell type based on network centrality measures calculated from the CXCL signaling network using CellChat. (B) Expression levels of Cxcl12 and genes of its receptors Cxcr4 and Ackr3. (C) Normalized expression of Cxcl12 in fibroblasts and ECs of each group. (D) Interaction strength between various cell types within the CXCL signaling pathway. The thickness of the lines connecting cell types is proportional to the calculated interaction strength. (E) Significant ligand-receptor pairs contributing to Cxcl12 signaling from fibroblasts. Dot color and size represent calculated communication probability and P values from a one-sided permutation test. (F) Coexpression patterns of Cxcr4 with DEGs related to immune cell activation in each group. In the t-SNE representation, cells expressing Cxcr4 are shown in red, cells expressing Ifng, Cd8a, Ccl4, Ccl5, Il21r, or Ccr5 are shown in green, and cells expressing both are shown in yellow. The percentage at the bottom left represents the proportion of cells expressing both Cxcr4 and the selected genes among Cxcr4-expressing cells.
T cell subpopulations in the AA mouse model. (A) t-SNE representation of T cell subpopulations colored by detailed cell types. (B) Expression levels for selected marker genes of each cell type. (C) t-SNE representation of T cells colored by group (Neg, AA, or AA + Ab). (D) Proportion of each cell type among total T cells. Statistical significance was assessed using the binomial test. *** P < 0.001. (E) Pseudotime analysis of naïve-like T cells and CD8+ T cells. ‘s’ indicates the designated starting point (root) on the trajectory constructed based on UMAP representation. (F) Expression levels of selected marker genes over pseudotime. (G) Density of cells at each pseudotime by group. (H) Coexpression patterns of Cd8a with Jak/Stat-related genes in each group. In the t-SNE representation, cells expressing Cd8a are shown in red, cells expressing Jak2, Stat3, or Stat5a are shown in green, and cells expressing both are shown in yellow. The percentage at the bottom left represents the proportion of cells expressing both Cd8a and the selected genes among Cd8a-expressing cells.
Humanized CXCL12 antibody delays onset and modulates immune response in alopecia areata mice: insights from single-cell RNA sequencing

October 2024

·

16 Reads

·

1 Citation

It has been demonstrated that CXCL12 inhibits hair growth via CXCR4, and its neutralizing antibody (Ab) increases hair growth in alopecia areata (AA). However, the molecular mechanisms have not been fully elucidated. In the present study, we further prepared humanized CXCL12 Ab for AA treatment and investigated underlying molecular mechanisms using single-cell RNA sequencing. Subcutaneous injection of humanized CXCL12 Ab significantly delayed AA onset in mice, and dorsal skin was analyzed. T cells and dendritic cells/macrophages were increased in the AA model, but decreased after CXCL12 Ab treatment. Pseudobulk RNA sequencing identified 153 differentially expressed genes that were upregulated in AA model and downregulated after Ab treatment. Gene ontology analysis revealed that immune cell chemotaxis and cellular response to type II interferon were upregulated in AA model but downregulated after Ab treatment. We further identified key immune cell-related genes such as Ifng, Cd8a, Ccr5, Ccl4, Ccl5, and Il21r, which were colocalized with Cxcr4 in T cells and regulated by CXCL12 Ab treatment. Notably, CD8+ T cells were significantly increased and activated via Jak/Stat pathway in the AA model but inactivated after CXCL12 Ab treatment. Collectively, these results indicate that humanized CXCL12 Ab is promising for AA treatment via immune modulatory effects.


The structure of the skin and the movement of subcutaneously administered mAbs. The hypodermis consists of adipose and connective tissues interspersed with blood and lymphatic vessels. The connective tissues of the hypodermis are made up of highly polymerized macromolecular networks, and the vascular capillaries in the hypodermis are impermeable to large molecules (>50 kDa). Therefore, it is generally assumed that reaching from the hypodermis to systemic circulation results from lymphatic drainage for most mAbs. However, lymph flow is slower than 0.2% of blood flow, the systemic absorption of mAbs from injection site is limited.
Multi-functional role of CXCL12 for AGA therapy. Androgen hormones secrete CXCL12 from dermal fibroblasts (DF) via androgen receptor (AR). Secreted CXCL12 activates AR in DF and dermal papilla cells (DPCs), and miniaturized hair follicle. On the contrary, CXCL12 Ab inactivates AR in DF and DPCs, and inhibits inflammation and fibrosis surrounding hair follicle.
Recent approaches of antibody therapeutics in androgenetic alopecia

Therapeutic antibodies (Abs) have been anticipated as promising alternatives to conventional treatments such as topical minoxidil and oral finasteride for androgenetic alopecia (AGA). Due to the high molecular weight of typical Abs, the half-life of subcutaneous Abs exceeds 2 weeks, allowing an administration intervals of once a month or longer. Direct injection into the areas of hair loss is also feasible, potentially enhancing treatment efficacy while minimizing systemic side effects. However, therapeutic Abs are rarely developed for AGA therapy due to the requirement to be responsiveness to androgens and to exist in the extracellular fluid or cell surface surrounding the hair follicle. In this review, we introduce recent progress of antibody therapeutics in AGA targeting the prolactin receptor, Interleukin-6 receptor, C-X-C motif chemokine ligand 12, and dickkopf 1. As therapeutic Abs for AGA are still in the early stages, targets need further validation and optimization for clinical application.


Delivery Strategies of siRNA Therapeutics for Hair Loss Therapy

July 2024

·

39 Reads

·

1 Citation

International Journal of Molecular Sciences

Therapeutic needs for hair loss are intended to find small interfering ribonucleic acid (siRNA) therapeutics for breakthrough. Since naked siRNA is restricted to meet a druggable target in clinic,, delivery systems are indispensable to overcome intrinsic and pathophysiological barriers, enhancing targetability and persistency to ensure safety, efficacy, and effectiveness. Diverse carriers repurposed from small molecules to siRNA can be systematically or locally employed in hair loss therapy, followed by the adoption of new compositions associated with structural and environmental modification. The siRNA delivery systems have been extensively studied via conjugation or nanoparticle formulation to improve their fate in vitro and in vivo. In this review, we introduce clinically tunable siRNA delivery systems for hair loss based on design principles, after analyzing clinical trials in hair loss and currently approved siRNA therapeutics. We further discuss a strategic research framework for optimized siRNA delivery in hair loss from the scientific perspective of clinical translation.


Endogenous stem cell mobilization and localized immunosuppression synergistically ameliorate DSS-induced Colitis in mice

June 2024

·

127 Reads

Stem Cell Research & Therapy

Background Stem cell therapy is a promising alternative for inflammatory diseases and tissue injury treatment. Exogenous delivery of mesenchymal stem cells is associated with instant blood-mediated inflammatory reactions, mechanical stress during administration, and replicative senescence or change in phenotype during long-term culture in vitro. In this study, we aimed to mobilize endogenous hematopoietic stem cells (HSCs) using AMD-3100 and provide local immune suppression using FK506, an immunosuppressive drug, for the treatment of inflammatory bowel diseases. Methods Reactive oxygen species (ROS)-responsive FK506-loaded thioketal microspheres were prepared by emulsification solvent-evaporation method. Thioketal vehicle based FK506 microspheres and AMD3100 were co-administered into male C57BL6/J mice with dextran sulfate sodium (DSS) induced colitis. The effect of FK506-loaded thioketal microspheres in colitis mice were evaluated using disease severity index, myeloperoxidase activity, histology, flow cytometry, and gene expression by qRT-PCR. Results The delivery of AMD-3100 enhanced mobilization of HSCs from the bone marrow into the inflamed colon of mice. Furthermore, targeted oral delivery of FK506 in an inflamed colon inhibited the immune activation in the colon. In the DSS-induced colitis mouse model, the combination of AMD-3100 and FK506-loaded thioketal microspheres ameliorated the disease, decreased immune cell infiltration and activation, and improved body weight, colon length, and epithelial healing process. Conclusion This study shows that the significant increase in the percentage of mobilized hematopoietic stem cells in the combination therapy of AMD and oral FK506 microspheres may contribute to a synergistic therapeutic effect. Thus, low-dose local delivery of FK506 combined with AMD3100 could be a promising alternative treatment for inflammatory bowel diseases.


Stem cell factor and cKIT modulate endothelial glycolysis in hypoxia

March 2024

·

27 Reads

·

3 Citations

Cardiovascular Research

Aims In hypoxia, endothelial cells (ECs) proliferate, migrate, and form new vasculature in a process called angiogenesis. Recent studies have suggested that ECs rely on glycolysis to meet metabolic needs for angiogenesis in ischaemic tissues, and several studies have investigated the molecular mechanisms integrating angiogenesis and endothelial metabolism. Here, we investigated the role of stem cell factor (SCF) and its receptor, cKIT, in regulating endothelial glycolysis during hypoxia-driven angiogenesis. Methods and results SCF and cKIT signalling increased the glucose uptake, lactate production, and glycolysis in human ECs under hypoxia. Mechanistically, SCF and cKIT signalling enhanced the expression of genes encoding glucose transporter 1 (GLUT1) and glycolytic enzymes via Akt- and ERK1/2-dependent increased translation of hypoxia inducible factor 1A (HIF1A). In hypoxic conditions, reduction of glycolysis and HIF-1α expression using chemical inhibitors significantly reduced the SCF-induced in vitro angiogenesis in human ECs. Compared with normal mice, mice with oxygen-induced retinopathy (OIR), characterized by ischaemia-driven pathological retinal neovascularization, displayed increased levels of SCF, cKIT, HIF-1α, GLUT1, and glycolytic enzymes in the retina. Moreover, cKIT-positive neovessels in the retina of mice with OIR showed elevated expression of GLUT1 and glycolytic enzymes. Further, blocking SCF and cKIT signalling using anti-SCF neutralizing IgG and cKIT mutant mice significantly reduced the expression of HIF-1α, GLUT1, and glycolytic enzymes and decreased the pathological neovascularization in the retina of mice with OIR. Conclusion We demonstrated that SCF and cKIT signalling regulate angiogenesis by controlling endothelial glycolysis in hypoxia and elucidated the SCF/cKIT/HIF-1α axis as a novel metabolic regulation pathway during hypoxia-driven pathological angiogenesis.




Figure 2. Androgen treatment enhances CXCL12 expression in dermal fibroblasts. (A) Dermal fibroblasts (DFs) were treated with various concentrations of TP (1, 10, and 100 nM) and DHT (1, 10, and 100 nM) for 24 h, and the expression of CXCL12 was assessed using qRT-PCR. $ p < 0.05 vs. control. (B) After treating DFs with different concentrations of TP and DHT for 48 h, the culture medium was collected and the secreted CXCL12 levels were quantified using ELISA. $ p < 0.05, $$ p < 0.01 vs. control. The dollar sign ($) indicates differences in one-way ANOVA. (C) Immunostaining revealed that the translocation of the AR (green) in DFs increased after TP (100 nM) and DHT (100 nM) treatment for 1 h, as indicated by the white arrows. DAPI staining (blue) marks the cell nuclei. The scale bar is set at 50 µm. (D) After AR-CRISPR/Cas9 knockout (AR-KO) for 48 h, DFs were treated with TP and DHT for an additional 48 h to collect the culture medium for ELISA analysis. AR-KO significantly reduced the secretion of CXCL12 from DFs. Western blot analysis indicated differences in AR expression between the control and AR-KO groups. ### p < 0.001 vs. control, ** p < 0.01, *** p < 0.001 vs. TP or DHT treatment. The asterisk and sharp symbols indicate statistical differences using Student's t-test.
Figure 3. CXCL12 secreted from DFs induces AR and CXCR4 in DPCs. The effects of rCXCL12 on the expression of AR and CXCR4 in DPCs were observed using qRT-PCR (A,B) and Western blot analysis (C). rCXCL12 increased the mRNA and protein expression of the AR and CXCR4 in DPCs. $ p < 0.05, $$ p < 0.01 vs. Control. (D) DFs were treated with 100 nM TP or DHT for 48 h and the culture medium (CM) was collected. CM from DFs treated with TP and DHT (DFCM TP and DFCM DHT ) significantly reduced hair length in human hair organ culture. $ p < 0.05 vs. DFCM, n = 10. The dollar sign ($) indicates differences in a one-way ANOVA.
Figure 5. CXCL12 neutralization prevents alopecia areata onset. (A) In this experimental design, 10-week-old C3H female mice were treated with αCXCL12 (20 µg) through subcutaneous injections
Primary antibodies used for immunofluorescence analysis and western blot.
CXCL12 Neutralizing Antibody Promotes Hair Growth in Androgenic Alopecia and Alopecia Areata

January 2024

·

185 Reads

·

5 Citations

International Journal of Molecular Sciences

We had previously investigated the expression and functional role of C-X-C Motif Chemokine Ligand 12 (CXCL12) during the hair cycle progression. CXCL12 was highly expressed in stromal cells such as dermal fibroblasts (DFs) and inhibition of CXCL12 increased hair growth. Therefore, we further investigated whether a CXCL12 neutralizing antibody (αCXCL12) is effective for androgenic alopecia (AGA) and alopecia areata (AA) and studied the underlying molecular mechanism for treating these diseases. In the AGA model, CXCL12 is highly expressed in DFs. Subcutaneous (s.c.) injection of αCXCL12 significantly induced hair growth in AGA mice, and treatment with αCXCL12 attenuated the androgen-induced hair damage in hair organ culture. Androgens increased the secretion of CXCL12 from DFs through the androgen receptor (AR). Secreted CXCL12 from DFs increased the expression of the AR and C-X-C Motif Chemokine Receptor 4 (CXCR4) in dermal papilla cells (DPCs), which induced hair loss in AGA. Likewise, CXCL12 expression is increased in AA mice, while s.c. injection of αCXCL12 significantly inhibited hair loss in AA mice and reduced the number of CD8+, MHC-I+, and MHC-II+ cells in the skin. In addition, injection of αCXCL12 also prevented the onset of AA and reduced the number of CD8+ cells. Interferon-γ (IFNγ) treatment increased the secretion of CXCL12 from DFs through the signal transducer and activator of transcription 3 (STAT3) pathway, and αCXCL12 treatment protected the hair follicle from IFNγ in hair organ culture. Collectively, these results indicate that CXCL12 is involved in the progression of AGA and AA and antibody therapy for CXCL12 is promising for hair loss treatment.


Citations (79)


... Androgens increase CXCL12 secretion by DFs and upregulate the expression of the androgen receptor (AR) and CXCR4 in dermal papilla cells (DPCs) in a paracrine manner, which is responsible for hair miniaturization in the mouse AGA model. However, the direct effects of androgens on CXCL12 and its expression in DPCs and dermal sheath cup cells (DSCs) have not been reported in AGA because CXCL12 is rarely expressed in mouse DPCs and DSCs [8,9]. ...

Reference:

Differential Expression of CXCL12 in Human and Mouse Hair: Androgens Induce CXCL12 in Human Dermal Papilla and Dermal Sheath Cup
Humanized CXCL12 antibody delays onset and modulates immune response in alopecia areata mice: insights from single-cell RNA sequencing

... 29 Under conditions of hypoxia, endothelial cells often rely on glycolysis rather than oxidative phosphorylation for energy production. 30 Previous studies demonstrated that hypoxia is a common pathologic factor for retinal neovascularization diseases. 31 By stimulating glycolytic pathways, FGF2 helps these cells adapt to lowoxygen environments, thus facilitating the formation of new blood vessels. ...

Stem cell factor and cKIT modulate endothelial glycolysis in hypoxia
  • Citing Article
  • March 2024

Cardiovascular Research

... In addition, the CXCL12 level was increased in the dermal papilla region of alopecia areata (AA) patients and is regarded as a stress-sentinel that activates immune cells [6]. We further demonstrated that the CXCL12-neutralizing antibody promotes hair growth in androgenic alopecia (AGA) and AA [7]. In that study, we found that CXCL12 is highly expressed in dermal fibroblasts (DFs) compared with hair follicles in mouse skin. ...

CXCL12 Neutralizing Antibody Promotes Hair Growth in Androgenic Alopecia and Alopecia Areata

International Journal of Molecular Sciences

... In any event, CD8 + and CD4 + T cells are of primary interest as the drivers of actual hair loss [15]. Other scientists have taken a different approach whereby HF keratinocytes [91], dermal papilla cells [92], adiposederived stem cells [93], or immune cells from non-AA affected volunteers [94] have been induced to express an inflammatory state and then candidate AA treatments were assessed for their ability to suppress the phenotype. Such cell cultures could provide a basic platform for initial screening of treatments that inhibit leukocyte activation, cell migration, cytokine secretion, or cytotoxic activity. ...

Involvement of DKK1 secreted from adipose-derived stem cells in alopecia areata
  • Citing Article
  • November 2023

... Under low intracellular ROS levels, p53 is activated by ATM and checkpoint kinase 2, CHK2, promoting the transcription of antioxidant enzymes that contribute to the detoxification, and upregulating the levels of reducing molecules, such as nicotinamide adenine dinucleotide phosphate (NADPH) and glutathione. Some examples of p53-activated ROSdetoxifying molecules reported in an UVR context are depicted in Figure 3, including the en-zymes manganese superoxide dismutase 2, SOD2 [49], glutathione peroxidase 1, GPx1 [50], peroxiredoxin, PRDX1 [51], and NAD(P)H quinone dehydrogenase 1 (NQO1) [52]. In addition, the p53 transcriptional targets sestrin SESN1 and SESN2 interact with Nrf2 antagonist Kelch Like ECH Associated Protein 1 (Keap1), promoting its degradation [53]. ...

Antioxidative and Anti-photooxidative Potential of Interruptins from the Edible Fern Cyclosorus terminans in Human Skin Cells
  • Citing Article
  • June 2023

Current Pharmaceutical Biotechnology

... However, when human DP spheres formed with the PRP scaffold were combined with human neonatal foreskin for a chamber assay, hair follicle induction did not occur. To date, there have been no reports of successful hair follicle regeneration using DP cells and epidermal cells derived from humans [128,129]. Therefore, to evaluate the hair follicle formation ability of human DP cells, they can be mixed with mouse-derived epidermal cells to assess their hair follicle regenerative potential. Similarly, to evaluate the hair follicle formation ability of human epidermal cells, their regenerative potential can be assessed by mixing them with mouse dermal cells (Figure 3). ...

Hair growth promoting effects of human dermal papilla cells in pig
  • Citing Article
  • April 2023

Experimental Dermatology

... Frontiers in Pharmacology frontiersin.org recognized as secreted proteins that induce hair growth (Cazzola and Perez-Moreno, 2022;Kim and Sung, 2023). Therefore, we investigated how araliadiol affects the expression of anagen follicle markers and hair growth-promoting factors. ...

The effects of GPR40 agonists on hair growth are mediated by ANGPTL4
  • Citing Article
  • March 2023

Biomedicine & Pharmacotherapy

... Adipose-derived stem cells (ADSCs) were the first MSCs shown to stimulate hair growth through the secretion of several growth factors, including hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), insulin-like growth factor (IGF), and platelet-derived growth factor (PDGF). As a result, adiposederived stem cell (ASC) conditioned medium and ASC exosomes have been developed as cosmetic ingredients and are commercially available (39). In summary, ADSCs hold significant promise for promoting hair follicle growth and treating alopecia. ...

Hypoxia enhances the hair growth-promoting effects of embryonic stem cell-derived mesenchymal stem cells via NADPH oxidase 4
  • Citing Article
  • January 2023

Biomedicine & Pharmacotherapy

... Challenges in spheroid culture include the time needed to form stable spheroids and methods for large-scale production to ensure an adequate number for injection. Recently, we showed that stable and uniform spheroids can be efficiently generated within 2 h using a highly viscous methylcellulose medium [7] . Future advancements could include combining microwell technology, methylcellulosebased incubation, and automated production systems to enable large-scale spheroid manufacturing [8][9][10] . ...

Engineering of hybrid spheroids of mesenchymal stem cells and drug depots for immunomodulating effect in islet xenotransplantation

Science Advances