Abdi Ghaffari, Ruhangiz T. Kilani, Aziz Ghahary

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,

Oxysterols Induce Differentiation in Human Keratinocytes and Increase Ap-1-Dependent Involucrin Transcription Karen Hanley, Dean C. Ng, ShanShan He,

Enhanced Vascularization of Cultured Skin Substitutes Genetically Modified to Overexpress Vascular Endothelial Growth Factor1 Dorothy M. Supp, Andrew.

PKC-δ and -η, MEKK-1, MEK-6, MEK-3, and p38-δ Are Essential Mediators of the Response of Normal Human Epidermal Keratinocytes to Differentiating Agents

NF-κB Accumulation Associated with COL1A1 Transactivators Defects during Chronological Aging Represses Type I Collagen Expression through a –112/–61-bp.

Matrix Metalloproteinase 9 Expression is Coordinately Modulated by the KRE-M9 and 12-O-Tetradecanoyl-Phorbol-13-Acetate Responsive Elements Takashi Kobayashi,

Kynurenine Increases Matrix Metalloproteinase-1 and -3 Expression in Cultured Dermal Fibroblasts and Improves Scarring In Vivo Yunyuan Li, Ruhangiz T.

Topical Application of 17β-Estradiol Increases Extracellular Matrix Protein Synthesis by Stimulating TGF-β Signaling in Aged Human Skin In Vivo Eui Dong.

Complement Factor H: A Biomarker for Progression of Cutaneous Squamous Cell Carcinoma Pilvi M. Riihilä, Liisa M. Nissinen, Risto Ala-aho, Markku Kallajoki,

The Calcium-Sensing Receptor-Dependent Regulation of Cell–Cell Adhesion and Keratinocyte Differentiation Requires Rho and Filamin A Chia-Ling Tu, Wenhan.

BM-40(Osteonectin, SPARC) Is Expressed Both in the Epidermal and in the Dermal Compartment of Adult Human Skin Nicholas Hunzelmann, Martin Hafner, Sabine.

Expression of Type XVI Collagen in Human Skin Fibroblasts: Enhanced Expression in Fibrotic Skin Diseases Atsushi Akagi, Shingo Tajima, Yutaka Nagai

IFNλ Stimulates MxA Production in Human Dermal Fibroblasts via a MAPK-Dependent STAT1-Independent Mechanism Adewonuola A. Alase, Yasser M. El-Sherbiny,

Wnt5a/β-Catenin Signaling Drives Calcium-Induced Differentiation of Human Primary Keratinocytes Tanja Popp, Dirk Steinritz, Andreas Breit, Janina Deppe,

Age-Associated Increase in Skin Fibroblast–Derived Prostaglandin E2 Contributes to Reduced Collagen Levels in Elderly Human Skin Yong Li, Dan Lei, William.

The Transcriptional Coactivator DRIP/Mediator Complex Is Involved in Vitamin D Receptor Function and Regulates Keratinocyte Proliferation and Differentiation

Xu Shi-wen, Christopher P. Denton, Alan M. Holmes, Carol M

Andreea M. Bujor, Jaspreet Pannu, Shizhong Bu, Edwin A. Smith, Robin C

Hypoxia Impairs Skin Myofibroblast Differentiation and Function

Th2 Cytokines Increase Staphylococcus aureus Alpha Toxin–Induced Keratinocyte Death through the Signal Transducer and Activator of Transcription 6 (STAT6)

Histamine Contributes to Tissue Remodeling via Periostin Expression

Olivier Le Saux, Severa Bunda, Christopher M

Keratinocyte-Releasable Stratifin Functions as a Potent Collagenase-Stimulating Factor in Fibroblasts Aziz Ghahary, Feridoun Karimi-Busheri, Yvonne Marcoux,

Membrane Type 1 Matrix Metalloproteinase Regulates Cellular Invasiveness and Survival in Cutaneous Epidermal Cells Usha Nagavarapu, Kenneth Relloma,

Transcriptional Control of the Mouse Col7a1 Gene in Keratinocytes: Basal and Transforming Growth Factor-β Regulated Expression Michael Naso, Jouni Uitto,

Regulation of IL-33 Expression by IFN-γ and Tumor Necrosis Factor-α in Normal Human Epidermal Keratinocytes Jitlada Meephansan, Hidetoshi Tsuda, Mayumi.

Tomoyasu Hattori, Lukasz Stawski, Sashidhar S

Isolation (From a Basal Cell Carcinoma) of a Functionally Distinct Fibroblast-Like Cell Type that Overexpresses Ptch Anthony J. Dicker, Magdalena M.

Cytokine Profiling and Stat3 Phosphorylation in Epithelial–Mesenchymal Interactions between Keloid Keratinocytes and Fibroblasts Cheh P. Lim, Toan T.

Aquaporin-3 Re-Expression Induces Differentiation in a Phospholipase D2-Dependent Manner in Aquaporin-3-Knockout Mouse Keratinocytes Vivek Choudhary,

MiR-196a Downregulation Increases the Expression of Type I and III Collagens in Keloid Fibroblasts Kazuya Kashiyama, Norisato Mitsutake, Michiko Matsuse,

Human Keratinocytes Constitutively Express Interleukin-18 and Secrete Biologically Active Interleukin-18 After Treatment with Pro-Inflammatory Mediators.

Regulation of Fibulin-2 Gene Expression by Integrin α3β1 Contributes to the Invasive Phenotype of Transformed Keratinocytes Dara S. Missan, Sridar V.

Sema4D, the Ligand for Plexin B1, Suppresses c-Met Activation and Migration and Promotes Melanocyte Survival and Growth Joanne Soong, Yulin Chen, Elina.

Profiling Motility Signal-Specific Genes in Primary Human Keratinocytes Chieh-Fang Cheng, Jianhua Fan, Balaji Bandyopahdhay, Dennis Mock, Shengxi Guan,

Stratifin-Induced Matrix Metalloproteinase-1 in Fibroblast Is Mediated by c-fos and p38 Mitogen-Activated Protein Kinase Activation Eugene Lam, Runhangiz.

Inhibitory Effect of β-Thujaplicin on Ultraviolet B-Induced Apoptosis in Mouse Keratinocytes Takako Baba, Hajime Nakano, Katsuto Tamai, Daisuke Sawamura,

Epithelial Cells in the Hair Follicle Bulge do not Contribute to Epidermal Regeneration after Glucocorticoid-Induced Cutaneous Atrophy Dmitry V. Chebotaev,

Glucosamine sulfate modulates the levels of aggrecan and matrix metalloproteinase-3 synthesized by cultured human osteoarthritis articular chondrocytes

Post-Transcriptional Regulation of Melanin Biosynthetic Enzymes by cAMP and Resveratrol in Human Melanocytes Richard A. Newton, Anthony L. Cook, Donald.

Reduced Expression of Connective Tissue Growth Factor (CTGF/CCN2) Mediates Collagen Loss in Chronologically Aged Human Skin TaiHao Quan, Yuan Shao, Tianyuan.

Differentiated Keratinocyte-Releasable Stratifin ( Sigma) Stimulates MMP-1 Expression in Dermal Fibroblasts Aziz Ghahary, Yvonne Marcoux, Feridoun.

TNF-α Suppresses α-Smooth Muscle Actin Expression in Human Dermal Fibroblasts: An Implication for Abnormal Wound Healing Mytien T. Goldberg, Yuan-Ping.

Human Keratinocytes Respond to Osmotic Stress by p38 Map Kinase Regulated Induction of HSP70 and HSP27 M. Garmyn, A. Pupe Journal of Investigative Dermatology

A Role for Estrogen Receptor-α and Estrogen Receptor-β in Collagen Biosynthesis in Mouse Skin Margaret Markiewicz, Sergey Znoyko, Lukasz Stawski, Angela.

Critical Role of Paxillin in Aging of Human Skin

Insulin-Like Growth Factor-Binding Protein 7 Regulates Keratinocyte Proliferation, Differentiation and Apoptosis Janna Nousbeck, Ofer Sarig, Nili Avidan,

Hyaluronan Synthase 3 Regulates Hyaluronan Synthesis in Cultured Human Keratinocytes Tetsuya Sayo, Yoshinori Sugiyama, Yoshito Takahashi, Naoko Ozawa,

Green Tea Polyphenol Epigallocatechin-3-Gallate Suppresses Collagen Production and Proliferation in Keloid Fibroblasts via Inhibition of the STAT3-Signaling.

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,

Increased Expression of Wnt2 and SFRP4 in Tsk Mouse Skin: Role of Wnt Signaling in Altered Dermal Fibrillin Deposition and Systemic Sclerosis Julie Bayle,

Differential Roles of Insulin Receptor and Insulin-Like Growth Factor-1 Receptor in Differentiation of Murine Skin Keratinocytes Efrat Wertheimer, Meirav.

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.

Society for Investigative Dermatology 2010 Meeting Minutes

Anna Flammiger, Robert Besch, Anthony L. Cook, Tanja Maier, Richard A

The Effect of LXR Activators on AP-1 Proteins in Keratinocytes

Volume 56, Issue 5, Pages (November 1999)

Post-Transcriptional Regulation of UV Induced TNF-α Expression

Yunyuan Li, Edward E. Tredget, Abdi Ghaffari, Xiaoyue Lin, Ruhangiz T

Keratinocytes Inhibit Expression of Connective Tissue Growth Factor in Fibroblasts In Vitro by an Interleukin-1α-Dependent Mechanism Daniel Nowinski,

John M. Lamar, Vandana Iyer, C. Michael DiPersio

The Relaxin Gene Knockout Mouse: A Model of Progressive Scleroderma

Kristiina Airola, Norbert E. Fusenig

Effects of Hepatocyte Growth Factor on the Expression of Type I Collagen and Matrix Metalloproteinase-1 in Normal and Scleroderma Dermal Fibroblasts

Hepatocyte Growth Factor/Scatter Factor (HGF/SF) Induces Vascular Permeability Factor (VPF/VEGF) Expression by Cultured Keratinocytes Jens Gille, Mona.

Ultraviolet-B-Induced G1 Arrest is Mediated by Downregulation of Cyclin-Dependent Kinase 4 in Transformed Keratinocytes Lacking Functional p53 Arianna.

The Activity of Caspase-1 Is Increased in Lesional Psoriatic Epidermis

Matrix Metalloproteinase Inhibitor BB-3103 Unlike the Serine Proteinase Inhibitor Aprotinin Abrogates Epidermal Healing of Human Skin Wounds Ex Vivo1

Heparin-Binding EGF-Like Growth Factor Is Induced by Disruption of Lipid Rafts and Oxidative Stress in Keratinocytes and Participates in the Epidermal.

Presentation transcript:

Keratinocyte-Conditioned Media Regulate Collagen Expression in Dermal Fibroblasts Abdi Ghaffari, Ruhangiz T. Kilani, Aziz Ghahary Journal of Investigative Dermatology Volume 129, Issue 2, Pages 340-347 (February 2009) DOI: 10.1038/jid.2008.253 Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Keratinocyte-releasable factor(s) reduces the expression of type I collagen in dermal fibroblasts. (a) The levels of 4.8 and 5.8kb transcripts of pro-α1(I) collagen mRNA (COL1A1) in fibroblasts alone (F), fibroblasts co-cultured with keratinocytes (K/F), and fibroblasts co-cultured with fibroblasts (F/F) were evaluated by northern analysis. The same blot was re-hybridized for 18S ribosomal RNA and used as a control for RNA loading. The signals were then quantified by densitometry and the ratio of COL1A1/18S was determined. (b) The means of COL1A1/18S in K/F and F/F groups from two separate experiments were calculated and are depicted as a histogram. (c) COL1A1 mRNA expression in fibroblast-populated collagen gel was evaluated in the presence of a keratinocyte (K/F) or a fibroblast (F/F) top layer. Journal of Investigative Dermatology 2009 129, 340-347DOI: (10.1038/jid.2008.253) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Collagen-inhibitory effect of KCM in dermal fibroblasts is time dependent. (a) Fibroblasts were treated with KCM for 1, 3, 6, 12, 24, 36, or 48hours. Total RNA was then extracted and subjected to northern analysis. The blots were initially hybridized with cDNA specific for COL1A1 and subsequently with cDNA for 18S ribosomal RNA used as a loading control. (b) Similar time-dependent experiments in which fibroblasts were treated with KCM for 12, 24, 36, and 48hours, with control for 24 (C-24) and 48 (C-48) hours. The level of intracellular type I procollagen protein was evaluated by western blot using β-actin expression as a loading control. (c) For quantitative analysis, this experiment was repeated four times, and the ratio of type I collagen/β-actin (percentage of control, C-24) was calculated and the mean±SD is depicted and shown here. *P<0.01 and **P<0.001 between the control (C-24) and the cells treated with KCM for 36 and 48hours, respectively. Journal of Investigative Dermatology 2009 129, 340-347DOI: (10.1038/jid.2008.253) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Crude determination of KD-CIF molecular weight. (a) KCM was passed through a 50kDa cutoff centrifugal filter, and corresponding filtrate and retentate were used to treat dermal fibroblasts. Total KCM and non-conditioned medium (NCM) were also used as positive and negative controls, respectively. COL1A1 expression level was determined by northern blot analysis. Total KCM (lane 1), retentate (lane 2, >50kDa), and filtrate (lane 3, <50kDa) of 50kDa cutoff filter, as well as the same fractions for NCM, are shown here. (b) The same experimental protocol as above but with a 30kDa cutoff filter. Total KCM (lane 1), retentate (lane 2, >30kDa), and filtrate (lane 3, <30kDa) of 30kDa cutoff filter and corresponding fractions of NCM are presented. Expression of 18S ribosomal RNA was used as a control for loading. (c) Graph represents the mean ratio of COL1A1/18S RNA (percentage of control, which is NCM-treated cells)±SD of four separate experiments. *P<0.01 and **P<0.001 between control and KCM or KCM fraction of 30–50kDa treated cells, respectively. Journal of Investigative Dermatology 2009 129, 340-347DOI: (10.1038/jid.2008.253) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Stability of the keratinocyte-derived collagen-inhibitory factor(s). (a) Dermal fibroblasts were treated with medium alone (lane C) or KCM that had been collected and subjected to nothing (lane 1), acidification/neutralization (lane 2), or heat at 56°C for 30minutes (lane 3). Total RNA was extracted after 24hours and subjected to northern blot analysis for COL1A1 mRNA. The corresponding blots were subsequently re-hybridized with 18S ribosomal RNA and used as a loading control. (b) In a similar set of experiments, the levels of pro-α1(I) collagen protein and β-actin were evaluated after 48hours by western blot analysis. For quantitative analysis, the mean±SD of pro-α1(I) collagen/β-actin obtained from three separate experiments was then determined and the corresponding histogram is depicted in the lower panel. *P<0.05, the significant difference between test and control groups. (c) Cells were treated with nothing (C), KCM, or ammonium sulfate-precipitated proteins from KCM (PTT). Total proteins were extracted and evaluated for pro-α1(I) collagen protein and β-actin by western blot analysis. The ratio of mean±SD of pro-α1(I) collagen/β-actin was then determined from three different experiments, and the corresponding data are depicted in the lower panel. *P<0.05, significant difference between test (KCM) and control (C). No significant difference was observed between control and PPT (P>0.05). Journal of Investigative Dermatology 2009 129, 340-347DOI: (10.1038/jid.2008.253) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Keratinocyte differentiation has no effect on KD-CIF activity. (a) The multilayer formation by differentiated keratinocytes on day 14 (right panel) as compared with a monolayer of cells on day 0 (left panel). (b) Total proteins from keratinocyte lysate after 0, 2, 4, 6, 8, 10, and 12 days of culture were extracted and subjected to western blot analysis for involucrin protein used as a marker of keratinocyte differentiation. The level of β-actin was tested as a loading control. (c) Dermal fibroblasts were treated with KCM collected on days 0, 4, 8, 10, and 12 of keratinocyte culture. Non-conditioned medium was used as control (lane C). Total cell extracts were prepared and intracellular type I procollagen and MMP-1 protein levels were determined by western blot analysis. β-Actin is shown as loading control. Journal of Investigative Dermatology 2009 129, 340-347DOI: (10.1038/jid.2008.253) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions