Quiescence of Limbal Epithelial Stem Cells Summary The stem cells (SCs) of the corneal epithelium located in the limbal ?palisades of Vogt? are the ultimate source of maintaining corneal epithelial homeostasis. Clinically, loss of limbal SCs or dysfunction of the limbal niche leads to corneal blindness due to limbal SC deficiency (LSCD). During the last funding period, we successfully isolated and expanded limbal niche cells (NC) and established in vitro niches by SC-NC reunion in 3D Matrigel or HC-HA/PTX3, i.e., a novel matrix purified from amniotic membrane. Using these in vitro models, we have proven the hypothesis that maintenance of close SC-NC contact is crucial for preventing SCs from adopting corneal fate decision and maintaining SC self-renewal and quiescence through a balancing act between Wnt and BMP signaling. Herein, we propose to explore the signaling mechanism wherein SC quiescence is controlled by delineating how HC-HA/PTX3 transmits CD44-mediated signaling to maintain the NC phenotype and activate BMP signaling (Aim 1) and how PCP signaling reinforces BMP signaling in NCs to augment SC quiescence (Aim 2). Furthermore, we will determine how Notch signaling is established between SCs and NCs through the polar expression of Notch ligands and receptors under the influence of PCP signaling so as to transmit BMP signaling from NCs to SCs to achieve SC quiescence (Aim 3). In 3D Matrigel, noggin suppresses BMP signaling to allow full activation of Wnt signaling in order to promote SC self-renewal. In HC-HA/PTX3, we will determine whether noggin acts differently from in 3D Matrigel by inducing an ?alert? stage of SC self-renewal without affecting pre-existing BMP and PCP signaling by upregulating NKD1 that continues to suppress Wnt signaling (Aim 4). Successful completion of the above Aims will shed new light on the signaling mechanism wherein SC quiescence and self-renewal are maintained by close contact with NCs. Consequently, we will learn a better strategy to maintain the SC pool so as to ensure homeostasis of the corneal epithelium. This new knowledge will also help us devise a new tissue engineering strategy for the corneal epithelium by successful recapitulation of the regulatory mechanism executed by close interaction with NCs. Future studies of how such signaling might be altered by pathogenic factors derived from non-resolving inflammation will help us unravel the pathogenesis and potential new therapies of LSCD. Collectively, these studies may one day help us realize the considerable promise held by adult SCs in treating a number of diseases in the body.

Public Health Relevance

The ultimate goal of our proposed research is to better understand the pathogenesis of limbal stem cell deficiency caused by a number of corneal blinding diseases and to develop a novel and effective tissue engineering strategy to maximize the ex vivo expansion of adult somatic lineage-committed epithelial stem cells. Our progress made in the last funding period have led us to recapitulate in vitro limbal niches exhibiting stem cell self-renewal and quiescence upon close contact with limbal niche cells. Hence, we propose to characterize the signaling pathways transmitted from the novel matrix, HC-HA/PTX3 (derived from amniotic membrane), to limbal niche cells and finally to limbal epithelial stem cells in controlling quiescence and self- renewal, of which both are critical for maintaining the stem cell pool. We believe that these studies will make a significant advance in the field of regenerative medicine by generating important knowledge that will unravel new therapeutic paradigms through restoration of the limbal niche health. Such novel approaches may one day help us realize the considerable promise held by adult SCs in treating a number of diseases in the body.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY006819-31
Application #
9431215
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Mckie, George Ann
Project Start
1986-05-01
Project End
2021-02-28
Budget Start
2018-03-01
Budget End
2019-02-28
Support Year
31
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Tissuetech, Inc.
Department
Type
DUNS #
167232888
City
Miami
State
FL
Country
United States
Zip Code
33173
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He, Hua; Kuriyan, Ajay E; Su, Chen-Wei et al. (2017) Inhibition of Proliferation and Epithelial Mesenchymal Transition in Retinal Pigment Epithelial Cells by Heavy Chain-Hyaluronan/Pentraxin 3. Sci Rep 7:43736
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Tseng, Scheffer C G (2016) HC-HA/PTX3 Purified From Amniotic Membrane as Novel Regenerative Matrix: Insight Into Relationship Between Inflammation and Regeneration. Invest Ophthalmol Vis Sci 57:ORSFh1-8
Chen, Szu-Yu; Han, Bo; Zhu, Ying-Ting et al. (2015) HC-HA/PTX3 Purified From Amniotic Membrane Promotes BMP Signaling in Limbal Niche Cells to Maintain Quiescence of Limbal Epithelial Progenitor/Stem Cells. Stem Cells 33:3341-55
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Han, Bo; Chen, Szu-Yu; Zhu, Ying-Ting et al. (2014) Integration of BMP/Wnt signaling to control clonal growth of limbal epithelial progenitor cells by niche cells. Stem Cell Res 12:562-73
Zhang, Suzhen; Zhu, Ying-Ting; Chen, Szu-Yu et al. (2014) Constitutive expression of pentraxin 3 (PTX3) protein by human amniotic membrane cells leads to formation of the heavy chain (HC)-hyaluronan (HA)-PTX3 complex. J Biol Chem 289:13531-42
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