Roles of Growth Factors on Corneal Morphogenesis

Summary

Principal Investigator: WINSTON KAO
Affiliation: University of Cincinnati
Country: USA
Abstract: Transforming growth factor ? (TGF-?) has a pivotal role in corneal wound healing. Previous studies revealed that activation of p38MAPK and Smad signaling pathway have distinct roles in mediating TGF-? signaling of corneal epithelium debridement and keratectomy, respectively. Such differences can be explained by the fact that healing epithelium of debridement migrates on basement membrane, whereas that of keratectomy migrates on collagenous matrix of denuded stroma, resulting in distinct integrin expression patterns in migrating epithelium within 1 hour of injuries. Thus, we hypothesize that interaction of different integrins with TGF-? receptors accounts for the difference in TGF-? signaling pathways, i.e., activation of p38MAPK in epithelium debridement and Smads cascades in keratectomy (Hypothesis 1). It has also been found that suppression of cell proliferation and activation of Activating Transcription Factor 2 (ATF2) are independent of TGF-? signaling following epithelium debridement. Thus, the activation of ATF2 by an alternative pathway, i.e., JNK, and its subsequent formation of Activating Protein-1 transcription factor (AP-1) complex plays a key role in the suppression of epithelial cell proliferation in the early healing phase of corneal injury (Hypotheisis 2). Specific Aim 1 will identify and characterize roles of integrins in TGF-? signaling pathways during the healing of corneal epithelium debridement and keratectomy using tritransgenic Cre-LoxP mouse models, i.e., Krt12rtTA/rtTA/tet-O-Cre/Tbr2f/f and Krt12rtTA/rtTA/tet-O-Cre/Smad4f/f in which floxed Tbr2 and Smad4 genes are ablated specifically in corneal epithelium upon doxycycline induction so that one can determine potential variations in signaling pathways in the absence and presence of Tbr2 and Smad4 (Aim 1A), to examine roles of integrins in mediating TGF-? receptor signaling (Aim 1B) and to examine efficacy of p38MAPK1 and Smad7 on modulation of cell migration and proliferation during wound healing (Aim 1C). Specific Aim 2 will elucidate roles of ATF2 and AP-1 in suppression of cell proliferation during corneal wound healing by identification of ATF2 and/or AP-1 complexes in healing epithelium of corneal epithelium debridement and keratectomy using immunoprecipitation and western blot analysis (Aim 2A), determine involvement of ATF2 in suppression of cell proliferation during healing of epithelium debridement by overexpression of dominant negative ATF2 and ?N-ATF2 mutant proteins (Aim 2B), and to determine effects of JNK and p38MAPK inhibitors on activation of ATF2 during corneal wound healing (Aim2C). These experiments will yield useful information for restoration of normal vision by intervening TBR2 and ATF2 signaling pathways of injured corneas. PUBLIC HEALTH RELEVANCE The proposed studies will examine the roles of TGF-? in modulating corneal functions using experimental animals that conditionally over express dominant negative mutant and/or wild type signal transduction molecules of TGF-? receptor mediated pathways, e.g., ATF2, p38MAPK, Smad7 by transgenes delivered with Adenoviral vectors, and conditional ablation of genes, i.e., Tbr2, Smad4 and cJun in corneal epithelium of tritransgenic mice, i.e., Krt12rtTA/rtTA/tet-O-Cre/Tbr2f/f, Krt12rtTA/rtTA/tet-O-Cre/Smad4f/f and Krt12rtTA/rtTA/tet-O-Cre/cJunf/f mice upon doxycycline induction. The proposed studies will fill gaps of our understanding of TGF-? signaling on corneal morphogenesis during wound healing as well as homeostasis in adults. Data obtained will yield useful information for a better understanding of corneal diseases at molecular and cellular levels in vivo and serve as basis for designing treatment regiments for corneal wound healing.
Funding Period: -------------------- - --------------------
more information: NIH RePORT

Top Publications

  1. pmc Loss of tumor necrosis factor alpha potentiates transforming growth factor beta-mediated pathogenic tissue response during wound healing
    Shizuya Saika
    Department of Ophthalmology, Wakayama Medical University, 811 1 Kimiidera, Wakayama, 641 0012, Japan
    Am J Pathol 168:1848-60. 2006
  2. pmc Dexamethasone induces cross-linked actin networks in trabecular meshwork cells through noncanonical wnt signaling
    Yong Yuan
    Crawley Vision Research Laboratory, Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, Ohio
    Invest Ophthalmol Vis Sci 54:6502-9. 2013
  3. pmc Targeted overexpression of TGF-α in the corneal epithelium of adult transgenic mice induces changes in anterior segment morphology and activates noncanonical Wnt signaling
    Yong Yuan
    Crawley Vision Research Laboratory, Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USA
    Invest Ophthalmol Vis Sci 54:1829-37. 2013
  4. pmc Wakayama symposium: challenges of future research in ocular surface cell biology
    Winston W Y Kao
    Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, Ohio 45267 0838, USA
    Ocul Surf 11:19-24. 2013
  5. pmc Mastermind-like transcriptional co-activator-mediated Notch signaling is indispensable for maintaining conjunctival epithelial identity
    Yujin Zhang
    Edith J Crawley Vision Research Center Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USA
    Development 140:594-605. 2013
  6. pmc Role of SH2-containing tyrosine phosphatase Shp2 in mouse corneal epithelial stratification
    Gracia Y Ng
    Edith J Crawley Vision Research Center Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, Ohio
    Invest Ophthalmol Vis Sci 54:7933-42. 2013
  7. pmc Promiscuous recombination of LoxP alleles during gametogenesis in cornea Cre driver mice
    Daniel Y Weng
    Department of Ophthalmology, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Mol Vis 14:562-71. 2008
  8. ncbi Signaling pathways in morphogenesis of cornea and eyelid
    Winston W Y Kao
    Department of Ophthalmology, University of Cincinnati, Cincinnati, Ohio 45267 0838, USA
    Ocul Surf 6:9-23. 2008
  9. pmc A role for the mitogen-activated protein kinase kinase kinase 1 in epithelial wound healing
    Maoxian Deng
    Department of Environmental Health and Center for Environmental Genetics, University of Cincinnati Medical Center, Cincinnati, OH 45267 0056, USA
    Mol Biol Cell 17:3446-55. 2006
  10. ncbi Ocular surface tissue morphogenesis in normal and disease states revealed by genetically modified mice
    Winston W Y Kao
    Department of Ophthalmology and Cell Biology, University of Cincinnati, 3225 Eden Avenue, Cincinnati, OH 45267, USA
    Cornea 25:S7-S19. 2006

Scientific Experts

  • Yuka Okada
  • WINSTON KAO
  • Ewa Anna Meyer-Blazejewska
  • Chia Yang Liu
  • Yujin Zhang
  • Hongshan Liu
  • Shizuya Saika
  • Yong Yuan
  • Mindy K Call
  • Osamu Yamanaka
  • Lung Kun Yeh
  • Yasuhito Hayashi
  • Kathleen C Flanders
  • Yoshitaka Ohnishi
  • Ai Kitano
  • Gracia Y Ng
  • Jianhua Zhang
  • I Jong Wang
  • Camila C Simões
  • Kazuto Terai
  • Jinsong Hao
  • Daniel Y Weng
  • Yuji Nakajima
  • Kazuo Ikeda
  • Takeshi Miyamoto
  • Maoxian Deng
  • Oliver Lam
  • Gen Sheng Feng
  • Minh Thanh T Nguyen
  • William D Hardie
  • Warren S Pear
  • Yan Yuan
  • Gracia Ng
  • Walden Ai
  • Katy Fischesser
  • James J Augsburger
  • David Schlessinger
  • Zelia M Correa
  • Noriko Terai
  • Abbot G Spaulding
  • Tai ichiro Chikama
  • Emanuele Pelosi
  • Candace W C Kao
  • James V Jester
  • S Kevin Li
  • Tyler Kochel
  • Pao Hsien Chu
  • Makoto M Taketo
  • Wei Lan Weng
  • George Babcock
  • Sandy Schwemberger
  • Chia Yi Kuan
  • Shigeyuki Kon
  • Susan R Rittling
  • Kumi Shirai
  • Toshimitsu Uede
  • David T Denhardt
  • Ken ichi Miyazaki
  • Maureen Mongan
  • Jianhua Yang
  • Maureen Sartor
  • Bing Su
  • Atsushi Takatori
  • Zhimin Peng
  • Marian Miller
  • Ying Xia
  • Wei Li Chen
  • Ranjani Parthasarathy
  • Lin Zhang
  • Akira Ooshima

Detail Information

Publications23

  1. pmc Loss of tumor necrosis factor alpha potentiates transforming growth factor beta-mediated pathogenic tissue response during wound healing
    Shizuya Saika
    Department of Ophthalmology, Wakayama Medical University, 811 1 Kimiidera, Wakayama, 641 0012, Japan
    Am J Pathol 168:1848-60. 2006
    ....
  2. pmc Dexamethasone induces cross-linked actin networks in trabecular meshwork cells through noncanonical wnt signaling
    Yong Yuan
    Crawley Vision Research Laboratory, Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, Ohio
    Invest Ophthalmol Vis Sci 54:6502-9. 2013
    ..The purpose of the study is to demonstrate that noncanonical Wnt signaling mediates the DEX-induced CLAN formation in TM cells...
  3. pmc Targeted overexpression of TGF-α in the corneal epithelium of adult transgenic mice induces changes in anterior segment morphology and activates noncanonical Wnt signaling
    Yong Yuan
    Crawley Vision Research Laboratory, Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USA
    Invest Ophthalmol Vis Sci 54:1829-37. 2013
    ..However, the underlying mechanisms remain unclear. Here we examined the effects of TGF-α overexpression on adult ocular surface homeostasis...
  4. pmc Wakayama symposium: challenges of future research in ocular surface cell biology
    Winston W Y Kao
    Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, Ohio 45267 0838, USA
    Ocul Surf 11:19-24. 2013
    ..To investigate possible mechanisms, we have developed mouse models in which the gene functions of ocular surface epithelia and stromas can be altered by Doxycycline induction in spatial and temporal specific manners...
  5. pmc Mastermind-like transcriptional co-activator-mediated Notch signaling is indispensable for maintaining conjunctival epithelial identity
    Yujin Zhang
    Edith J Crawley Vision Research Center Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USA
    Development 140:594-605. 2013
    ....
  6. pmc Role of SH2-containing tyrosine phosphatase Shp2 in mouse corneal epithelial stratification
    Gracia Y Ng
    Edith J Crawley Vision Research Center Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, Ohio
    Invest Ophthalmol Vis Sci 54:7933-42. 2013
    ..Shp2 protein tyrosine phosphatase mediates a wide variety of receptor tyrosine kinases (RTK) cell signaling. Herein, we investigate the role of Shp2 in corneal morphogenesis and homeostasis...
  7. pmc Promiscuous recombination of LoxP alleles during gametogenesis in cornea Cre driver mice
    Daniel Y Weng
    Department of Ophthalmology, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Mol Vis 14:562-71. 2008
    ..To examine whether promiscuous Cre/LoxP recombination happens during gametogenesis in double transgenic mice carrying LoxP modified alleles and Cre transgene driven by tissue-specific promoter outside the gonads of adult mice...
  8. ncbi Signaling pathways in morphogenesis of cornea and eyelid
    Winston W Y Kao
    Department of Ophthalmology, University of Cincinnati, Cincinnati, Ohio 45267 0838, USA
    Ocul Surf 6:9-23. 2008
    ..This review discusses known signaling transduction pathways involved in the morphogenesis of the cornea and eyelids during embryonic development and homeostasis in adults, using genetically modified experimental animal models...
  9. pmc A role for the mitogen-activated protein kinase kinase kinase 1 in epithelial wound healing
    Maoxian Deng
    Department of Environmental Health and Center for Environmental Genetics, University of Cincinnati Medical Center, Cincinnati, OH 45267 0056, USA
    Mol Biol Cell 17:3446-55. 2006
    ..Our data suggest that MEKK1 transmits wound signals, leading to the transcriptional activation of genes involved in ECM homeostasis, epithelial cell migration, and wound reepithelialization...
  10. ncbi Ocular surface tissue morphogenesis in normal and disease states revealed by genetically modified mice
    Winston W Y Kao
    Department of Ophthalmology and Cell Biology, University of Cincinnati, 3225 Eden Avenue, Cincinnati, OH 45267, USA
    Cornea 25:S7-S19. 2006
    ..These mouse lines can also be used as models for development of therapeutic treatment regimens of ocular surface diseases using gene therapy and stem cell strategies...
  11. ncbi Loss of osteopontin perturbs the epithelial-mesenchymal transition in an injured mouse lens epithelium
    Shizuya Saika
    Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan
    Lab Invest 87:130-8. 2007
    ..An in vitro experiment shows OPN facilitates cell adhesion of lens epithelial cell line. OPN is required for activation of Smad2/3 signal in an injured lens epithelium and lens cell EMT...
  12. ncbi TNFalpha suppression of corneal epithelium migration
    Yuka Okada
    Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan
    Mol Vis 13:1428-35. 2007
    ..To evaluate the role of tumor necrosis factor alpha (TNFalpha) in regulation of corneal epithelial cell migration...
  13. ncbi Effect of overexpression of PPARgamma on the healing process of corneal alkali burn in mice
    Shizuya Saika
    Dept of Ophthalmology, Wakayama Medical University, 811 1 Kimiidera, Wakayama, 641 0012, Japan
    Am J Physiol Cell Physiol 293:C75-86. 2007
    ..Together, these data suggest that overexpression of PPARgamma may represent an effective new strategy for treatment of ocular surface burns...
  14. pmc Crosstalk between TGF-beta and MAPK signaling during corneal wound healing
    Kazuto Terai
    Department of Ophthalmology, University of Cincinnati, Cincinnati, Ohio 45267 0838, USA
    Invest Ophthalmol Vis Sci 52:8208-15. 2011
    ..The aim of this study was to elucidate the mechanisms governing epithelial cell migration and proliferation during wound healing...
  15. doi Bone marrow mesenchymal stem cells can differentiate and assume corneal keratocyte phenotype
    Hongshan Liu
    Department of Ophthalmology, Edith Crawley Vision Research Center, University of Cincinnati, Cincinnati, OH, USA
    J Cell Mol Med 16:1114-24. 2012
    ..These observations suggest that corneal intrastromal transplantation of BM-MSC may be an effective treatment regimen for corneal diseases involving dysfunction of keratocytes...
  16. doi Clinical and histopathological features and immunoreactivity of human choroidal and ciliary melanomas as prognostic factors for metastasis and death
    Camila C Simões
    Department of Ophthalmology, University of Cincinnati College of Medicine, Stetson Building Suite 5300, 260 Stetson Street, Cincinnati, OH 45267 0527, USA
    Graefes Arch Clin Exp Ophthalmol 249:1795-803. 2011
    ....
  17. pmc Notch gain of function in mouse periocular mesenchyme downregulates FoxL2 and impairs eyelid levator muscle formation, leading to congenital blepharophimosis
    Yujin Zhang
    Edith J Crawley Vision Research Center Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USA
    J Cell Sci 124:2561-72. 2011
    ..These data strongly imply that a physiologically low level of Notch1 is crucial for proper FoxL2 expression in POMCs, which is, in turn, essential for Müeller muscle formation and normal eyelid development...
  18. pmc TRPV1 involvement in inflammatory tissue fibrosis in mice
    Yuka Okada
    Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan
    Am J Pathol 178:2654-64. 2011
    ..In conclusion, absence or blocking of TRPV1 suppressed inflammation and fibrosis/scarring during healing of alkali-burned mouse cornea. TRPV1 is a potential drug target for improving the outcome of inflammatory/fibrogenic wound healing...
  19. pmc From hair to cornea: toward the therapeutic use of hair follicle-derived stem cells in the treatment of limbal stem cell deficiency
    Ewa Anna Meyer-Blazejewska
    Department of Ophthalmology, University of Erlangen Nurnberg, Erlangen, Germany
    Stem Cells 29:57-66. 2011
    ..These data highlight the therapeutic properties of using HFSC to treat LSCD in a mouse model while demonstrating a strong translational potential and points to the niche as a key factor for determining stem cell differentiation...
  20. pmc Aberrant expression of a beta-catenin gain-of-function mutant induces hyperplastic transformation in the mouse cornea
    Yujin Zhang
    Department of Ophthalmology, College of Medicine, University of Cincinnati, Crawley Vision Research Center, Cincinnati, OH 45267 0838, USA
    J Cell Sci 123:1285-94. 2010
    ..Taken together, these results argue that beta-catenin activation is a crucial step during OSSN pathogenesis. Thus, inhibition of beta-catenin might be beneficial for treating this disease...
  21. pmc Gene delivery to cornea
    Jinsong Hao
    Division of Pharmaceutical Sciences, College of Pharmacy, University of Cincinnati, Cincinnati, OH 45267, USA
    Brain Res Bull 81:256-61. 2010
    ..The combination of iontophoresis and electroporation was found to be effective in delivering siRNA but not plasmid DNA into the corneal epithelium. Nanocarriers such as polymeric micelles are promising methods of corneal gene delivery...
  22. pmc A novel protocol of whole mount electro-immunofluorescence staining
    Hongshan Liu
    Department of Ophthalmology, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Mol Vis 15:505-17. 2009
    ..To develop a new method of whole mount immunostaining that improves the penetration of staining reagents into the cornea and decreases non-specific binding and background...
  23. ncbi Corneal morphogenesis during development and diseases
    Winston W Y Kao
    Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH 45267 0838, USA
    Eye Contact Lens 36:265-8. 2010
    ..To review the use of genetically modified mouse lines for elucidating corneal morphogenesis during embryonic development and diseases...