Hyaluronan does not regulate human epidermal keratinocyte proliferation and differentiation

Research output: Contribution to journalArticle

Abstract

Hyaluronan (HA) is synthesized by three HA synthases (HAS1, 2, and 3) and secreted in the extracellular matrix. In human skin, large amounts of HA are found in the dermis. HA is also synthesized by keratinocytes in the epidermis though its epidermal functions are not clearly identified yet. To investigate HA functions, we studied the effects of HA depletion on human keratinocyte physiology within in vitro reconstructed human epidermis. Inhibition of HA synthesis with 4-methylumbelliferone (4MU) did not modify the expression profile of the epidermal differentiation markers involucrin, keratin 10, and filaggrin during tissue reconstruction. In contrast, when keratinocytes were incubated with 4MU, cell proliferation was decreased. In an attempt to rescue the proliferation function, HA samples of various mean molecular masses were added to keratinocyte cultures treated with 4MU. These samples were unable to rescue the initial proliferation rate. Furthermore, treatments with HA-specific hyaluronidase, while removing almost all HA in keratinocyte cultures, did not alter the differentiation or proliferation processes. The differences between 4MU and hyaluronidase effects did not result from differences in intracellular HA, sulfated glycosaminoglycan concentration, apoptosis, or levels of HA receptors, all of which remained unchanged. Similarly, knockdown of UDP-glucose 6-dehydrogenase (UGDH) using lentiviral shRNA effectively decreased HA production but did not affect proliferation rate. Overall, these data suggest that HA levels in the human epidermis are not directly correlated with keratinocyte proliferation and differentiation and that incubation of cells with 4MU cannot equate with HA removal.

Original languageEnglish
Pages (from-to)6347-6358
Number of pages18
JournalThe Journal of Biological Chemistry
Volume291
Issue number12
DOIs
Publication statusPublished - Jan 2016

Fingerprint

Hyaluronic Acid
Keratinocytes
Hymecromone
Epidermis
Hyaluronoglucosaminidase
A73025
Uridine Diphosphate Glucose Dehydrogenase
Keratin-10
CD44 Antigens
Uridine Diphosphate Glucose
Differentiation Antigens
Physiology
Cell proliferation
Molecular mass
Dermis
Small Interfering RNA
Extracellular Matrix
Cell Differentiation
Skin
Oxidoreductases

Cite this

@article{0545dffde8a7473587e772baf35c8a07,
title = "Hyaluronan does not regulate human epidermal keratinocyte proliferation and differentiation",
abstract = "Hyaluronan (HA) is synthesized by three HA synthases (HAS1, 2, and 3) and secreted in the extracellular matrix. In human skin, large amounts of HA are found in the dermis. HA is also synthesized by keratinocytes in the epidermis though its epidermal functions are not clearly identified yet. To investigate HA functions, we studied the effects of HA depletion on human keratinocyte physiology within in vitro reconstructed human epidermis. Inhibition of HA synthesis with 4-methylumbelliferone (4MU) did not modify the expression profile of the epidermal differentiation markers involucrin, keratin 10, and filaggrin during tissue reconstruction. In contrast, when keratinocytes were incubated with 4MU, cell proliferation was decreased. In an attempt to rescue the proliferation function, HA samples of various mean molecular masses were added to keratinocyte cultures treated with 4MU. These samples were unable to rescue the initial proliferation rate. Furthermore, treatments with HA-specific hyaluronidase, while removing almost all HA in keratinocyte cultures, did not alter the differentiation or proliferation processes. The differences between 4MU and hyaluronidase effects did not result from differences in intracellular HA, sulfated glycosaminoglycan concentration, apoptosis, or levels of HA receptors, all of which remained unchanged. Similarly, knockdown of UDP-glucose 6-dehydrogenase (UGDH) using lentiviral shRNA effectively decreased HA production but did not affect proliferation rate. Overall, these data suggest that HA levels in the human epidermis are not directly correlated with keratinocyte proliferation and differentiation and that incubation of cells with 4MU cannot equate with HA removal.",
author = "J{\'e}r{\'e}my Malaisse and Val{\'e}rie Pendaries and Fanny Hontoir and {De Glas}, Val{\'e}rie and {Van Vlaender}, Daniel and Michel Simon and {Lambert de Rouvroit}, Catherine and Yves Poumay and Bruno Flamion",
note = "Copyright {\circledC} 2015, The American Society for Biochemistry and Molecular Biology.",
year = "2016",
month = "1",
doi = "10.1074/jbc.M115.661348",
language = "English",
volume = "291",
pages = "6347--6358",
journal = "The Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "12",

}

TY - JOUR

T1 - Hyaluronan does not regulate human epidermal keratinocyte proliferation and differentiation

AU - Malaisse, Jérémy

AU - Pendaries, Valérie

AU - Hontoir, Fanny

AU - De Glas, Valérie

AU - Van Vlaender, Daniel

AU - Simon, Michel

AU - Lambert de Rouvroit, Catherine

AU - Poumay, Yves

AU - Flamion, Bruno

N1 - Copyright © 2015, The American Society for Biochemistry and Molecular Biology.

PY - 2016/1

Y1 - 2016/1

N2 - Hyaluronan (HA) is synthesized by three HA synthases (HAS1, 2, and 3) and secreted in the extracellular matrix. In human skin, large amounts of HA are found in the dermis. HA is also synthesized by keratinocytes in the epidermis though its epidermal functions are not clearly identified yet. To investigate HA functions, we studied the effects of HA depletion on human keratinocyte physiology within in vitro reconstructed human epidermis. Inhibition of HA synthesis with 4-methylumbelliferone (4MU) did not modify the expression profile of the epidermal differentiation markers involucrin, keratin 10, and filaggrin during tissue reconstruction. In contrast, when keratinocytes were incubated with 4MU, cell proliferation was decreased. In an attempt to rescue the proliferation function, HA samples of various mean molecular masses were added to keratinocyte cultures treated with 4MU. These samples were unable to rescue the initial proliferation rate. Furthermore, treatments with HA-specific hyaluronidase, while removing almost all HA in keratinocyte cultures, did not alter the differentiation or proliferation processes. The differences between 4MU and hyaluronidase effects did not result from differences in intracellular HA, sulfated glycosaminoglycan concentration, apoptosis, or levels of HA receptors, all of which remained unchanged. Similarly, knockdown of UDP-glucose 6-dehydrogenase (UGDH) using lentiviral shRNA effectively decreased HA production but did not affect proliferation rate. Overall, these data suggest that HA levels in the human epidermis are not directly correlated with keratinocyte proliferation and differentiation and that incubation of cells with 4MU cannot equate with HA removal.

AB - Hyaluronan (HA) is synthesized by three HA synthases (HAS1, 2, and 3) and secreted in the extracellular matrix. In human skin, large amounts of HA are found in the dermis. HA is also synthesized by keratinocytes in the epidermis though its epidermal functions are not clearly identified yet. To investigate HA functions, we studied the effects of HA depletion on human keratinocyte physiology within in vitro reconstructed human epidermis. Inhibition of HA synthesis with 4-methylumbelliferone (4MU) did not modify the expression profile of the epidermal differentiation markers involucrin, keratin 10, and filaggrin during tissue reconstruction. In contrast, when keratinocytes were incubated with 4MU, cell proliferation was decreased. In an attempt to rescue the proliferation function, HA samples of various mean molecular masses were added to keratinocyte cultures treated with 4MU. These samples were unable to rescue the initial proliferation rate. Furthermore, treatments with HA-specific hyaluronidase, while removing almost all HA in keratinocyte cultures, did not alter the differentiation or proliferation processes. The differences between 4MU and hyaluronidase effects did not result from differences in intracellular HA, sulfated glycosaminoglycan concentration, apoptosis, or levels of HA receptors, all of which remained unchanged. Similarly, knockdown of UDP-glucose 6-dehydrogenase (UGDH) using lentiviral shRNA effectively decreased HA production but did not affect proliferation rate. Overall, these data suggest that HA levels in the human epidermis are not directly correlated with keratinocyte proliferation and differentiation and that incubation of cells with 4MU cannot equate with HA removal.

UR - http://www.jbc.org/content/early/2015/12/01/jbc.M115.661348

U2 - 10.1074/jbc.M115.661348

DO - 10.1074/jbc.M115.661348

M3 - Article

C2 - 26627828

VL - 291

SP - 6347

EP - 6358

JO - The Journal of Biological Chemistry

JF - The Journal of Biological Chemistry

SN - 0021-9258

IS - 12

ER -