Characterization of vitamin D production by human ocular barrier cells

Jawaher A Alsalem; Deepali Patel; Radhika Susarla; Miguel Coca-Prados; Rosemary Bland; Elizabeth A Walker; Saaeha Rauz; Graham R Wallace

doi:10.1167/iovs.13-13019

Characterization of vitamin D production by human ocular barrier cells

Jawaher A Alsalem, Deepali Patel, Radhika Susarla, Miguel Coca-Prados, Rosemary Bland, Elizabeth A Walker, Saaeha Rauz, Graham R Wallace

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Abstract

PURPOSE: Vitamin D3 is a secosteroid mainly synthesized from the conversion of the skin precursor 7-dehydrocholesterol (7DHC) to vitamin D3 by ultraviolet (UV) B sunlight. Extrarenal synthesis of vitamin D3 has been reported in many tissues and cells, including barrier sites. This study characterizes the expression of components of vitamin D3 signaling in human ocular barrier cells.

METHODS: Primary human scleral fibroblasts (HSF), human corneal endothelial (HCEC-12), nonpigmented ciliary body epithelial (ODM-2), and adult retinal pigment epithelial (ARPE-19) cell lines were analyzed for the expression of vitamin D receptor (VDR), the vitamin D3 activating enzymes 1α-hydroxylase (CYP27B1), 25-hydroxylases (CYP27A1 and CYP2R1), the vitamin D3 inactivating enzyme 24-hydroxylase (CYP24A1), and the endocytic receptors cubilin and megalin using a combination of RT-PCR, immunocytochemistry, and enzyme immunoassay (EIA).

RESULTS: The HSF, HCEC-12, ODM-2, and ARPE-19 express mRNA and protein for all vitamin D3 synthesizing and metabolizing components. The cell types tested, except HSF, are able to convert inactive 25-hydroxyvitamin D3 (25[OH]D3) into active 1,25-hydroxyvitamin D3 (1,25[OH]2D3).

CONCLUSIONS: This novel study demonstrated that ocular barrier epithelial cells express the machinery for vitamin D3 and can produce 1,25(OH)2D3. We suggest that vitamin D3 might have a role in immune regulation and barrier function in ocular barrier epithelial cells.

Original language	English
Pages (from-to)	2140-2147
Number of pages	8
Journal	Investigative Ophthalmology & Visual Science (IOVS)
Volume	55
Issue number	4
DOIs	https://doi.org/10.1167/iovs.13-13019
Publication status	Published - Apr 2014

Keywords

25-Hydroxyvitamin D3 1-alpha-Hydroxylase
Cell Line
Cholecalciferol
Endothelium, Corneal
Gene Expression Regulation
Humans
Immunoenzyme Techniques
Immunohistochemistry
RNA, Messenger
Retinal Pigment Epithelium
Reverse Transcriptase Polymerase Chain Reaction
Sclera

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AlSalem_et_al_Characterization_of_Vitamin_D_Investigative_Ophthalmology_&_Visual_Science_2014
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@article{40be2b004e474ffcafd75183bec1deba,

title = "Characterization of vitamin D production by human ocular barrier cells",

abstract = "PURPOSE: Vitamin D3 is a secosteroid mainly synthesized from the conversion of the skin precursor 7-dehydrocholesterol (7DHC) to vitamin D3 by ultraviolet (UV) B sunlight. Extrarenal synthesis of vitamin D3 has been reported in many tissues and cells, including barrier sites. This study characterizes the expression of components of vitamin D3 signaling in human ocular barrier cells.METHODS: Primary human scleral fibroblasts (HSF), human corneal endothelial (HCEC-12), nonpigmented ciliary body epithelial (ODM-2), and adult retinal pigment epithelial (ARPE-19) cell lines were analyzed for the expression of vitamin D receptor (VDR), the vitamin D3 activating enzymes 1α-hydroxylase (CYP27B1), 25-hydroxylases (CYP27A1 and CYP2R1), the vitamin D3 inactivating enzyme 24-hydroxylase (CYP24A1), and the endocytic receptors cubilin and megalin using a combination of RT-PCR, immunocytochemistry, and enzyme immunoassay (EIA).RESULTS: The HSF, HCEC-12, ODM-2, and ARPE-19 express mRNA and protein for all vitamin D3 synthesizing and metabolizing components. The cell types tested, except HSF, are able to convert inactive 25-hydroxyvitamin D3 (25[OH]D3) into active 1,25-hydroxyvitamin D3 (1,25[OH]2D3).CONCLUSIONS: This novel study demonstrated that ocular barrier epithelial cells express the machinery for vitamin D3 and can produce 1,25(OH)2D3. We suggest that vitamin D3 might have a role in immune regulation and barrier function in ocular barrier epithelial cells.",

keywords = "25-Hydroxyvitamin D3 1-alpha-Hydroxylase, Cell Line, Cholecalciferol, Endothelium, Corneal, Gene Expression Regulation, Humans, Immunoenzyme Techniques, Immunohistochemistry, RNA, Messenger, Retinal Pigment Epithelium, Reverse Transcriptase Polymerase Chain Reaction, Sclera",

author = "Alsalem, {Jawaher A} and Deepali Patel and Radhika Susarla and Miguel Coca-Prados and Rosemary Bland and Walker, {Elizabeth A} and Saaeha Rauz and Wallace, {Graham R}",

year = "2014",

month = apr,

doi = "10.1167/iovs.13-13019",

language = "English",

volume = "55",

pages = "2140--2147",

journal = "Investigative Ophthalmology & Visual Science (IOVS)",

issn = "0146-0404",

publisher = "Association for Research in Vision and Ophthalmology",

number = "4",

}

TY - JOUR

T1 - Characterization of vitamin D production by human ocular barrier cells

AU - Alsalem, Jawaher A

AU - Patel, Deepali

AU - Susarla, Radhika

AU - Coca-Prados, Miguel

AU - Bland, Rosemary

AU - Walker, Elizabeth A

AU - Rauz, Saaeha

AU - Wallace, Graham R

PY - 2014/4

Y1 - 2014/4

N2 - PURPOSE: Vitamin D3 is a secosteroid mainly synthesized from the conversion of the skin precursor 7-dehydrocholesterol (7DHC) to vitamin D3 by ultraviolet (UV) B sunlight. Extrarenal synthesis of vitamin D3 has been reported in many tissues and cells, including barrier sites. This study characterizes the expression of components of vitamin D3 signaling in human ocular barrier cells.METHODS: Primary human scleral fibroblasts (HSF), human corneal endothelial (HCEC-12), nonpigmented ciliary body epithelial (ODM-2), and adult retinal pigment epithelial (ARPE-19) cell lines were analyzed for the expression of vitamin D receptor (VDR), the vitamin D3 activating enzymes 1α-hydroxylase (CYP27B1), 25-hydroxylases (CYP27A1 and CYP2R1), the vitamin D3 inactivating enzyme 24-hydroxylase (CYP24A1), and the endocytic receptors cubilin and megalin using a combination of RT-PCR, immunocytochemistry, and enzyme immunoassay (EIA).RESULTS: The HSF, HCEC-12, ODM-2, and ARPE-19 express mRNA and protein for all vitamin D3 synthesizing and metabolizing components. The cell types tested, except HSF, are able to convert inactive 25-hydroxyvitamin D3 (25[OH]D3) into active 1,25-hydroxyvitamin D3 (1,25[OH]2D3).CONCLUSIONS: This novel study demonstrated that ocular barrier epithelial cells express the machinery for vitamin D3 and can produce 1,25(OH)2D3. We suggest that vitamin D3 might have a role in immune regulation and barrier function in ocular barrier epithelial cells.

AB - PURPOSE: Vitamin D3 is a secosteroid mainly synthesized from the conversion of the skin precursor 7-dehydrocholesterol (7DHC) to vitamin D3 by ultraviolet (UV) B sunlight. Extrarenal synthesis of vitamin D3 has been reported in many tissues and cells, including barrier sites. This study characterizes the expression of components of vitamin D3 signaling in human ocular barrier cells.METHODS: Primary human scleral fibroblasts (HSF), human corneal endothelial (HCEC-12), nonpigmented ciliary body epithelial (ODM-2), and adult retinal pigment epithelial (ARPE-19) cell lines were analyzed for the expression of vitamin D receptor (VDR), the vitamin D3 activating enzymes 1α-hydroxylase (CYP27B1), 25-hydroxylases (CYP27A1 and CYP2R1), the vitamin D3 inactivating enzyme 24-hydroxylase (CYP24A1), and the endocytic receptors cubilin and megalin using a combination of RT-PCR, immunocytochemistry, and enzyme immunoassay (EIA).RESULTS: The HSF, HCEC-12, ODM-2, and ARPE-19 express mRNA and protein for all vitamin D3 synthesizing and metabolizing components. The cell types tested, except HSF, are able to convert inactive 25-hydroxyvitamin D3 (25[OH]D3) into active 1,25-hydroxyvitamin D3 (1,25[OH]2D3).CONCLUSIONS: This novel study demonstrated that ocular barrier epithelial cells express the machinery for vitamin D3 and can produce 1,25(OH)2D3. We suggest that vitamin D3 might have a role in immune regulation and barrier function in ocular barrier epithelial cells.

KW - 25-Hydroxyvitamin D3 1-alpha-Hydroxylase

KW - Cell Line

KW - Cholecalciferol

KW - Endothelium, Corneal

KW - Gene Expression Regulation

KW - Humans

KW - Immunoenzyme Techniques

KW - Immunohistochemistry

KW - RNA, Messenger

KW - Retinal Pigment Epithelium

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Sclera

U2 - 10.1167/iovs.13-13019

DO - 10.1167/iovs.13-13019

M3 - Article

C2 - 24576880

SN - 0146-0404

VL - 55

SP - 2140

EP - 2147

JO - Investigative Ophthalmology & Visual Science (IOVS)

JF - Investigative Ophthalmology & Visual Science (IOVS)

IS - 4

ER -

Characterization of vitamin D production by human ocular barrier cells

Abstract

Keywords

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