Laminin N-terminus α31 regulates keratinocyte adhesion and migration through modifying the organisation and proteolytic processing of laminin 332

Published on Jul 28, 2020in bioRxiv
· DOI :10.1101/617597
Lee D. Troughton5
Estimated H-index: 5
(University of Liverpool),
Valentina Iorio4
Estimated H-index: 4
(University of Liverpool)
+ 3 AuthorsKevin J. Hamill17
Estimated H-index: 17
(University of Liverpool)
Sources
Abstract
Abstract Laminin N-terminus α31 (LaNt α31), a member of the laminin superfamily, expressed at low levels in intact epithelium but upregulated during wound repair. Increased expression of LaNt α31 reduced migration rate of corneal keratinocytes through an unknown mechanism. Here, we investigated whether LaNt α31 influences cell behaviour through modulating laminin-mediated processes. Adenoviral delivery of LaNt α31 into corneal epithelial cells led to reduced migration speed and increased cell spreading and changed laminin 332 organisation from diffuse arcs to tight clusters. Enhanced recruitment of collagen XVII and bullous pemphigoid antigen 1e to β4 integrin, indicating early maturation of hemidesmosomes, and changed focal adhesion distribution were also identified. LaNt α31 and laminin β3 co-immunoprecipitated from doubly transduced cells and were deposited together in live imaging experiment. Moreover, LaNt α31 expression led to increased matrix metalloproteinase (MMP) activity and proteolytic processing of laminin α3, and the inhibition of MMP activity rescued the laminin and hemidesmosome phenotypes. Provision of cell-derived extracellular matrix rescued the cell spreading and motility effects. These findings reveal LaNt α31 as a new player in regulating cell-to-matrix adhesion through its ability to influence laminin organisation and proteolytic processing.
References68
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Abstract Re-epithelialization describes the resurfacing of a skin wound with new epithelium. In response to various stimuli including that of growth factors, cytokines and extracellular matrix (ECM), wound edge epidermal keratinocytes undergo cytoskeleton rearrangements compatible with their motile behaviour and develop protrusive adhesion contacts. Matrix metalloproteinases (MMP) expression is crucial for proper cell movement and ECM remodelling; however, their deposition mechanism is unknown i...
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#1Lee D. Troughton (University of Liverpool)H-Index: 5
#2Tobias Zech (University of Liverpool)H-Index: 20
Last. Kevin J. Hamill (University of Liverpool)H-Index: 17
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Laminin N-terminus α31 (LaNt α31) is an alternative splice isoform derived from the laminin α3 gene. The LaNt α31 protein is enriched around the terminal duct lobular units in normal breast tissue. In the skin and cornea the protein influences epithelial cell migration and tissue remodelling. However, LaNt α31 has never been investigated in a tumour environment. Here we analysed LaNt α31 in invasive ductal carcinoma and determined its contribution to breast carcinoma invasion. LaNt α31 expressio...
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#2Raphael Reuten (UCPH: University of Copenhagen)H-Index: 10
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Laminin N terminus α31 (LaNt α31) is a netrin-like protein derived from alternative splicing of the laminin α3 gene. Although LaNt α31 has been demonstrated to influence corneal and skin epithelial cell function, its expression has not been investigated beyond these tissues. In this study, we used immunohistochemistry to characterise the distribution of this protein in a wide-array of human tissue sections in comparison to laminin α3. These data revealed widespread LaNt α31 expression. In epithe...
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Purpose: Laminin N-terminus (LaNt) α31 is a relatively unstudied protein derived from the laminin α3 gene but structurally similar to netrins. LaNt α31 has, to date, been investigated only in two-dimensional (2D) keratinocyte culture where it influences cell migration and adhesion, processes integral to wound repair. Here we investigated LaNt α31 distribution in ocular surface epithelium, during limbal stem cell activation, and corneal wound healing. Methods: Human, mouse, and pig eyes, ex vivo ...
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Cited By4
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The laminins (LM) are a family of basement membranes glycoproteins with essential structural roles in supporting epithelia, endothelia, nerves and muscle adhesion, and signaling roles in regulating cell migration, proliferation, stem cell maintenance and differentiation. Laminins are obligate heterotrimers comprised of ,  and chains that assemble intracellularly. However, extracellularly these heterotrimers then assemble into higher-order networks via interaction between their laminin N-term...
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#1Conor James Sugden (University of Liverpool)H-Index: 2
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Abstract Laminins are essential components of all basement membranes where they regulate an extensive array of tissue functions. Alternative splicing from the laminin α3 gene produces a non-laminin but netrin-like protein, Laminin N terminus α31 (LaNt α31). LaNt α31 is widely expressed in intact tissue and is upregulated in epithelial cancers and during wound healing. In vitro functional studies have shown that LaNt α31 can influence numerous aspects of epithelial cell behaviour via modifying ma...
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#1Conor James Sugden (University of Liverpool)H-Index: 2
#2Valentina Iorio (University of Liverpool)H-Index: 4
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Alternative splicing of the laminin α3 gene gives rise to a netrin-like protein termed LaNt α31, the major structural feature of which is a laminin N-terminal domain. LaNt α31 is expressed across a wide range of tissues, is upregulated in cancers, and ex vivo and in vitro functional studies have indicated that this relatively unstudied protein influences wound repair, stem cell activity, and tumour progression via modifying matrix organisation. However, LaNt α31 functionality has never been inve...
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Laminin N-terminus α31 (LaNt α31) is an alternative splice isoform derived from the laminin α3 gene. The LaNt α31 protein is enriched around the terminal duct lobular units in normal breast tissue. In the skin and cornea the protein influences epithelial cell migration and tissue remodelling. However, LaNt α31 has never been investigated in a tumour environment. Here we analysed LaNt α31 in invasive ductal carcinoma and determined its contribution to breast carcinoma invasion. LaNt α31 expressio...
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Laminin N terminus α31 (LaNt α31) is a netrin-like protein derived from alternative splicing of the laminin α3 gene. Although LaNt α31 has been demonstrated to influence corneal and skin epithelial cell function, its expression has not been investigated beyond these tissues. In this study, we used immunohistochemistry to characterise the distribution of this protein in a wide-array of human tissue sections in comparison to laminin α3. These data revealed widespread LaNt α31 expression. In epithe...
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