Caveolin-1 Controls Hyperresponsiveness to Mechanical Stimuli and Fibrogenesis- Associated RUNX2 Activation in Keloid Fibroblasts  Chao-Kai Hsu, Hsi-Hui.

Slides:



Advertisements
Similar presentations
IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway  Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,
Advertisements

SiRNA Knockdown of Tissue Inhibitor of Metalloproteinase-1 in Keloid Fibroblasts Leads to Degradation of Collagen Type I  Masayo Aoki, Koichi Miyake,
Caveolin-1 Controls Hyperresponsiveness to Mechanical Stimuli and Fibrogenesis- Associated RUNX2 Activation in Keloid Fibroblasts  Chao-Kai Hsu, Hsi-Hui.
PFKFB3, a Direct Target of p63, Is Required for Proliferation and Inhibits Differentiation in Epidermal Keratinocytes  Robert B. Hamanaka, Gökhan M. Mutlu 
TWEAK/Fn14 Activation Contributes to the Pathogenesis of Bullous Pemphigoid  Yale Liu, Lingling Peng, Liang Li, Chengfei Liu, Xiao Hu, Shengxiang Xiao,
S100A12 Induced in the Epidermis by Reduced Hydration Activates Dermal Fibroblasts and Causes Dermal Fibrosis  Jingling Zhao, Aimei Zhong, Emily E. Friedrich,
Development of Cell-Penetrating Asymmetric Interfering RNA Targeting Connective Tissue Growth Factor  Jihye Hwang, Chanil Chang, Ji Hyun Kim, Chang Taek.
Myocardin-Related Transcription Factors A and B Are Key Regulators of TGF-β1- Induced Fibroblast to Myofibroblast Differentiation  Beverly J. Crider, George.
Volume 138, Issue 3, Pages e2 (March 2010)
Activation of TGF-β1 by AQP3-Mediated H2O2 Transport into Fibroblasts of a Bleomycin-Induced Mouse Model of Scleroderma  Jingying Luo, Xin Liu, Jie Liu,
Hyaluronic Acid Decreases Lipid Synthesis in Sebaceous Glands
Tetsuo Toyama, Agnieszka P. Looney, Brendon M
Requirement of Zinc Transporter SLC39A7/ZIP7 for Dermal Development to Fine-Tune Endoplasmic Reticulum Function by Regulating Protein Disulfide Isomerase 
Sun A. Ham, Eun S. Kang, Hanna Lee, Jung S. Hwang, Taesik Yoo, Kyung S
TIEG1 Represses Smad7-Mediated Activation of TGF-β1/Smad Signaling in Keloid Pathogenesis  Zhi-Cheng Hu, Fen Shi, Peng Liu, Jian Zhang, Dong Guo, Xiao-Ling.
Caveolin-1 Controls Hyperresponsiveness to Mechanical Stimuli and Fibrogenesis- Associated RUNX2 Activation in Keloid Fibroblasts  Chao-Kai Hsu, Hsi-Hui.
The Antifibrotic Effect of α2AP Neutralization in Systemic Sclerosis Dermal Fibroblasts and Mouse Models of Systemic Sclerosis  Yosuke Kanno, En Shu,
Thrombomodulin Promotes Diabetic Wound Healing by Regulating Toll-Like Receptor 4 Expression  Tsung-Lin Cheng, Chao-Han Lai, Po-Ku Chen, Chia-Fong Cho,
Galectin-1 Accelerates Wound Healing by Regulating the Neuropilin-1/Smad3/NOX4 Pathway and ROS Production in Myofibroblasts  Yueh-Te Lin, Jhih-Sian Chen,
Interferon-γ Protects from Staphylococcal Alpha Toxin-Induced Keratinocyte Death through Apolipoprotein L1  Anne M. Brauweiler, Elena Goleva, Donald Y.M.
Heat Increases the Editing Efficiency of Human Papillomavirus E2 Gene by Inducing Upregulation of APOBEC3A and 3G  Yang Yang, Hexiao Wang, Xinrui Zhang,
Increased Lipocalin-2 Contributes to the Pathogenesis of Psoriasis by Modulating Neutrophil Chemotaxis and Cytokine Secretion  Shuai Shao, Tianyu Cao,
Green Tea Extract and (−)-Epigallocatechin-3-Gallate Inhibit Mast Cell-Stimulated Type I Collagen Expression in Keloid Fibroblasts via Blocking PI-3K/Akt.
Inhibition of DNA Methylation in the COL1A2 Promoter by Anacardic Acid Prevents UV- Induced Decrease of Type I Procollagen Expression  Min-Kyoung Kim,
Protease-Activated Receptor-2 (PAR-2) Expression in Human Fibroblasts is Regulated by Growth Factors and Extracellular Matrix  Barry L. Gruber, Mary J.
Fibronectin-Containing Extracellular Vesicles Protect Melanocytes against Ultraviolet Radiation-Induced Cytotoxicity  Bum-Ho Bin, Dae-Kyum Kim, Nan-Hyung.
Hypoxia Impairs Skin Myofibroblast Differentiation and Function
A Protective Mechanism of Visible Red Light in Normal Human Dermal Fibroblasts: Enhancement of GADD45A-Mediated DNA Repair Activity  Yeo Jin Kim, Hyoung-June.
Spleen Tyrosine Kinase Mediates EGFR Signaling to Regulate Keratinocyte Terminal Differentiation  Nan-Lin Wu, Duen-Yi Huang, Li-Fang Wang, Reiji Kannagi,
Estrogen Upregulates Slug to Enhance the Migration of Keratinocytes
Simvastatin Protects Human Melanocytes from H2O2-Induced Oxidative Stress by Activating Nrf2  Yuqian Chang, Shuli Li, Weinan Guo, Yuqi Yang, Weigang Zhang,
SiRNA-Targeting Transforming Growth Factor-β Type I Receptor Reduces Wound Scarring and Extracellular Matrix Deposition of Scar Tissue  Yi-Wen Wang, Nien-Hsien.
Histone H3K27 Demethylase JMJD3 in Cooperation with NF-κB Regulates Keratinocyte Wound Healing  Jungtae Na, Kwanghyun Lee, Wonho Na, Jee-Yoon Shin, Min-Jung.
Differential Expression of Matrix Metalloproteinases During Impaired Wound Healing of the Diabetes Mouse  Steven J. Wall, Dr, Damon Bevan, David W. Thomas,
Regeneration of Human Dermis by a Multi-Headed Peptide
Georgios Theocharidis, Zoe Drymoussi, Alexander P. Kao, Asa H
TWEAK/Fn14 Signals Mediate Burn Wound Repair
Aldosterone and Mineralocorticoid Receptor Antagonists Modulate Elastin and Collagen Deposition in Human Skin  Thomas F. Mitts, Severa Bunda, Yanting.
Prolonged Activation of ERK Contributes to the Photorejuvenation Effect in Photodynamic Therapy in Human Dermal Fibroblasts  Yong Hyun Jang, Gi-Bang Koo,
Heat Modulation of Tropoelastin, Fibrillin-1, and Matrix Metalloproteinase-12 in Human Skin In Vivo  Zhou Chen, Jin Young Seo, Yeon Kyung Kim, Se Rah.
Opposing Roles of Epidermal Integrins α3β1 and α9β1 in Regulation of mTLD/BMP-1– Mediated Laminin-γ2 Processing during Wound Healing  Whitney M. Longmate,
Collagen XVII Participates in Keratinocyte Adhesion to Collagen IV, and in p38MAPK- Dependent Migration and Cell Signaling  Hongjiang Qiao, Akihiko Shibaki,
Lack of Collagen VI Promotes Wound-Induced Hair Growth
Overexpression of CD109 in the Epidermis Differentially Regulates ALK1 Versus ALK5 Signaling and Modulates Extracellular Matrix Synthesis in the Skin 
Amadeus S. Zhu, Ang Li, Tabetha S. Ratliff, Martha Melsom, Luis A
Functional Implications of the IL-6 Signaling Pathway in Keloid Pathogenesis  Mohammad Ghazizadeh, Mamiko Tosa, Hajime Shimizu, Hiko Hyakusoku, Oichi Kawanami 
Electrical Stimulation Enhances Epidermal Proliferation in Human Cutaneous Wounds by Modulating p53–SIVA1 Interaction  Anil Sebastian, Syed A. Iqbal,
Collagen VII Half-Life at the Dermal-Epidermal Junction Zone: Implications for Mechanisms and Therapy of Genodermatoses  Tobias Kühl, Markus Mezger, Ingrid.
GSK3β Inhibition Blocks Melanoma Cell/Host Interactions by Downregulating N- Cadherin Expression and Decreasing FAK Phosphorylation  Jobin K. John, Kim.
Yabin Cheng, Guangdi Chen, Magdalena Martinka, Vincent Ho, Gang Li 
Promotion Effects of miR-375 on the Osteogenic Differentiation of Human Adipose- Derived Mesenchymal Stem Cells  Si Chen, Yunfei Zheng, Shan Zhang, Lingfei.
Integrin α4β1 and TLR4 Cooperate to Induce Fibrotic Gene Expression in Response to Fibronectin’s EDA Domain  Rhiannon M. Kelsh-Lasher, Anthony Ambesi,
Nrf2 Promotes Keratinocyte Proliferation in Psoriasis through Up-Regulation of Keratin 6, Keratin 16, and Keratin 17  Luting Yang, Xueli Fan, Tingting.
Xiamenmycin Attenuates Hypertrophic Scars by Suppressing Local Inflammation and the Effects of Mechanical Stress  Xiao-Jin Liu, Min-Juan Xu, Si-Teng Fan,
Reduced Expression of Connective Tissue Growth Factor (CTGF/CCN2) Mediates Collagen Loss in Chronologically Aged Human Skin  TaiHao Quan, Yuan Shao, Tianyuan.
The IL-6 Trans-Signaling-STAT3 Pathway Mediates ECM and Cellular Proliferation in Fibroblasts from Hypertrophic Scar  Sutapa Ray, Xiaoxi Ju, Hong Sun,
TNF-α Suppresses α-Smooth Muscle Actin Expression in Human Dermal Fibroblasts: An Implication for Abnormal Wound Healing  Mytien T. Goldberg, Yuan-Ping.
SIRT1, a Class III Histone Deacetylase, Regulates LPS-Induced Inflammation in Human Keratinocytes and Mediates the Anti-Inflammatory Effects of Hinokitiol 
YAP and TAZ Regulate Skin Wound Healing
Dermatopontin Expression is Decreased in Hypertrophic Scar and Systemic Sclerosis Skin Fibroblasts and is Regulated by Transforming Growth Factor-β1,
Critical Role of Paxillin in Aging of Human Skin
Loss of PTEN Expression by Dermal Fibroblasts Causes Skin Fibrosis
IL-18 Downregulates Collagen Production in Human Dermal Fibroblasts via the ERK Pathway  Hee Jung Kim, Seok Bean Song, Jung Min Choi, Kyung Moon Kim,
TWEAK/Fn14 Activation Contributes to the Pathogenesis of Bullous Pemphigoid  Yale Liu, Lingling Peng, Liang Li, Chengfei Liu, Xiao Hu, Shengxiang Xiao,
Cornulin Is Induced in Psoriasis Lesions and Promotes Keratinocyte Proliferation via Phosphoinositide 3-Kinase/Akt Pathways  Changji Li, Lei Xiao, Jinjing.
TSLP Is a Potential Initiator of Collagen Synthesis and an Activator of CXCR4/SDF-1 Axis in Keloid Pathogenesis  Jung U Shin, Seo Hyeong Kim, Hyeran Kim,
Collagen Synthesis Is Suppressed in Dermal Fibroblasts by the Human Antimicrobial Peptide LL-37  Hyun Jeong Park, Dae Ho Cho, Hee Jung Kim, Jun Young.
TAK1 Is Required for Dermal Wound Healing and Homeostasis
Aldo-Keto Reductase 1C3 Is Expressed in Differentiated Human Epidermis, Affects Keratinocyte Differentiation, and Is Upregulated in Atopic Dermatitis 
Hui Deng, Jing Liu, Yu Deng, Gangwen Han, Yiqun G. Shellman, Steven E
Presentation transcript:

Caveolin-1 Controls Hyperresponsiveness to Mechanical Stimuli and Fibrogenesis- Associated RUNX2 Activation in Keloid Fibroblasts  Chao-Kai Hsu, Hsi-Hui Lin, Hans I Harn, Rei Ogawa, Yang-Kao Wang, Yen-Ting Ho, Wan-Rung Chen, Yi-Chao Lee, Julia Yu-Yun Lee, Shyh-Jou Shieh, Chao-Min Cheng, John A. McGrath, Ming-Jer Tang  Journal of Investigative Dermatology  Volume 138, Issue 1, Pages 208-218 (January 2018) DOI: 10.1016/j.jid.2017.05.041 Copyright © 2017 The Authors Terms and Conditions

Figure 1 Keloid tissues are stiffer than normal tissue, whereas keloid fibroblasts (KFs) are softer than normal fibroblasts (NFs). (a) Western blot results of skin tissue from control subjects (n = 6) and keloid patients (n = 6). The protein levels of fibronectin (FN), collagen (COL) 1A1, COL3A1, COL11A1, and GAPDH (internal control) were analyzed. (b) Quantification results of FN, COL1A1, COL3A1, and COL11A1 from a. (c) Atomic force microscopy indentation results of skin tissue dissected from control subjects (n = 6) and keloid patients (n = 6). (d) Western blot results of NFs and KFs derived from control subjects (n = 6) and keloid patients (n = 6), respectively. The protein levels of FN, COL1A1, COL3A1, COL11A1, and GAPDH were analyzed. (e) Quantification results of FN, COL1A1, COL3A1, and COL11A1 from d. (f) Stiffness distribution histograms of NFs (n = 8) and KFs (n = 8). (g) Atomic force microscopy indentation results of NFs (n = 8) and KFs (n = 8). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. Journal of Investigative Dermatology 2018 138, 208-218DOI: (10.1016/j.jid.2017.05.041) Copyright © 2017 The Authors Terms and Conditions

Figure 2 RUNX2 is highly up-regulated in keloid tissues/fibroblasts. (a) Ingenuity pathway analysis results showing the network of RUNX2 and their close interactions with several extracellular matrix proteins and matrix metalloproteinases, which were related to wound healing and scar formation. (b) Immunohistochemical staining results for RUNX2 expression in normal skin tissue, normal scar tissue, hypertrophic scar, and keloid tissue (n = 4 in each). Scale bars = 100 μm. (c) Percentage of fibroblasts with nuclear RUNX2 in tissues from b. (d) Representative immunoblots of skin tissue from control subjects and keloid patients. The protein levels of phosphorylated RUNX2, RUNX2, and GAPDH were analyzed. (e) Quantification results of phosphorylated RUNX2 and RUNX2 from skin tissue of control subjects (n = 6) and keloid patients (n = 6). (f) Representative immunoblots of NFs and KFs. The protein levels of p-RUNX2, RUNX2, and GAPDH were analyzed. (g) Quantification results of p-RUNX2 and RUNX2 from NFs (n = 6) and KFs (n = 6). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. KF, keloid fibroblast; NF, normal fibroblast; p-, phosphorylated. Journal of Investigative Dermatology 2018 138, 208-218DOI: (10.1016/j.jid.2017.05.041) Copyright © 2017 The Authors Terms and Conditions

Figure 3 Keloid dermal fibroblasts show hyperresponsiveness to dermal tissue-equivalent matrix stiffness, which causes increased expression of RUNX2 and COL11A1. (a) Immunofluorescence study of NFs and KFs cultured on matrices of varying stiffness for 24 hours. Cells were stained for RUNX2 (green), the nucleus (blue), and F-actin (red). Scale bars = 20 μm. (b) Percentage of cells with nuclear RUNX2 in NFs (n = 5) and KFs (n = 5) from a. (c) Western blot results of NFs (n = 4) and KFs (n = 4) cultured on matrices of varying stiffness for 24 hours. (d) Representative reverse transcription-PCR results of NFs (n = 3) and KFs (n = 3) derived from explants on matrices of varying stiffness at day 3. The mRNA expressions of RUNX2, COL11A1, fibronectin (FN), and COL3A1 were analyzed. (e) Quantification results of RUNX2, COL11A1, FN, and COL3A1 mRNA from d. GAPDH-normalized data were compared with those of NFs cultured on 2 kPa PA gel. ∗P < 0.05, ∗∗∗P < 0.001. COL, collagen; FN, fibronectin; KF, keloid fibroblast; NF, normal fibroblast; PA, polyacrylamide. Journal of Investigative Dermatology 2018 138, 208-218DOI: (10.1016/j.jid.2017.05.041) Copyright © 2017 The Authors Terms and Conditions

Figure 4 Suppression of CAV1 in NFs causes cell softening, loss of stiffness-sensing ability, and increased expression of RUNX2 and FN. (a) Western blot results of NFs transfected with nontargeting control siRNA (siNC) or siCAV1 for 48 hours and KFs. (b) Quantification results of CAV1, RUNX2, and FN from a. GAPDH-normalized data were compared with those of NFs (n = 3) transfected with siNC. CAV1 knockdown in NFs causes (c) cell softening and (d) inability to change cell stiffness on matrices of varying stiffness. (e) Confocal immunofluorescence images obtained with anti-RUNX2 (green) in NFs and KFs cultured on dishes and treated with siNC or siCAV1 for 24 hours. Scale bars = 50 μm. (f) Percentage of cells with nuclear RUNX2 in dermal fibroblasts from e. (g) Percentage of cells with nuclear RUNX2 in siNC- or siCAV1-transfected NFs and KFs grown on PA gels of 20 kPa and 2 kPa for 24 hours. (h) Confocal immunofluorescence images obtained with anti-CAV1 (red), anti-RUNX2 (green), and nucleus (blue) in skin tissues from control subjects (n = 2) and keloid patients (n = 3). Scale bar = 50 μm. (i) Pearson correlation comparing CAV1 and RUNX2 intensity in skin tissues from h. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. FN, fibronectin; KF, keloid fibroblast; NC, nontargeting control; NF, normal fibroblast; PA, polyacrylamide; siRNA, small interfering RNA. Journal of Investigative Dermatology 2018 138, 208-218DOI: (10.1016/j.jid.2017.05.041) Copyright © 2017 The Authors Terms and Conditions

Figure 5 CAV1 knockdown promotes migratory ability of NFs. NFs (n = 3) and KFs (n = 3) were transfected with siNC or siCAV1 for 24 hours. After reaching a confluent monolayer, wound healing experiments were performed. (a) Quantification results of wound scratch (0 hours) and recovery (8 hours and 24 hours) from Supplementary Figure S5. Wound recovery (%) = [(wound area at 0 hours – wound area at 24 hours)/wound area at 0 hours] × 100%. (b) The representative force maps of fibroblasts under indicated conditions plated on polydimethylsiloxane micropost arrays. The color scale indicates the magnitude of traction force (nN). (c) Quantification results of total force and force per post generated in cells from b. (d) Western blot results of NFs or KFs transfected with siNC and siCAV1 for 48 hours. The protein levels of CAV1, pMLC2, MLC2, and GAPDH were analyzed. (e) Quantification results of CAV1, pMLC2, and MLC2 from d. ∗P < 0.05, ∗∗P < 0.01. KF, keloid fibroblast; NC, nontargeting control; NF, normal fibroblast; p, phosphorylated; siRNA, small interfering RNA. Journal of Investigative Dermatology 2018 138, 208-218DOI: (10.1016/j.jid.2017.05.041) Copyright © 2017 The Authors Terms and Conditions

Figure 6 Epigenetic control of CAV1 affects cell mechanics and RUNX2 expression in KFs. (a) Confocal immunofluorescence images of KFs plated onto culture dishes and treated with various doses of TSA for 24 hours. Scale bars = 20 μm. (b) Western blot results of KFs (n = 3) under indicated conditions. The protein levels of CAV1, RUNX2, FN, histone H3 acetyl K9 (histone H3AK9), and GAPDH were analyzed. Quantification results of (c) histone H3AK9, (d) CAV1, (e) RUNX2, and (f) FN from b. GAPDH-normalized data were compared with those of KFs treated with DMSO (TSA = 0 μmol/L). Treatment of KFs with 200 nmol/L TSA (g) hindered wound recovery, (h) increased cell stiffness, and (i) decreased the percentage of cells with nuclear RUNX2 on 2 kPa. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. C, control; FN, fibronectin; hr, hour; KF, keloid fibroblast; M, mol/L; TSA, trichostatin A. Journal of Investigative Dermatology 2018 138, 208-218DOI: (10.1016/j.jid.2017.05.041) Copyright © 2017 The Authors Terms and Conditions