K6PC-5, a direct activator of sphingosine kinase 1, promotes epidermal differentiation through intracellular CA 2+ signaling

Jeong Hee Hong, Jong Kyung Youm, Mi Jung Kwon, Byeong Deog Park, Yong Moon Lee, Syng Ill Lee, DongMin Shin, Seung Hun Lee

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Sphingosine-1-phosphate (S1P), a bioactive sphingolipid metabolite, regulates multiple cellular responses such as Ca 2+ signaling, growth, survival, and differentiation. Because sphingosine kinase (SphK) is the enzyme directly responsible for production of S1P, many factors have been identified that regulate its activity and subsequent S1P levels. Here we synthesized a previously unidentified SphK activator, K6PC-5, and have studied its effects on intracellular Ca 2+ signaling in HaCaT cells and epidermal differentiation in murine skin. K6PC-5, a hydrophobic compound chemically named N-(1,3-dihydroxyisopropyl)-2-hexyl-3-oxo-decanamide, activated SphK (obtained from C57BL/6 murine blood and F9-12 cell lysates) in a dose-dependent manner. K6PC-5 induced both intracellular Ca 2+ concentration ([Ca 2+ ] i ) oscillations in HaCaT cells and Ca 2+ mobilization in hairless mouse epidermis. Both dimethylsphingosine (DMS) and dihydroxysphingosine (DHS), SphK inhibitors, and transfection of SphK1-siRNA blocked K6PC-5-induced increases in [Ca 2+ ] i . The K6PC-5-induced [Ca 2+ ] i oscillations were dependent on thapsigargin-sensitive Ca 2+ stores and Ca 2+ entry, but independent of the classical phospholipase C-mediated pathway. In addition, K6PC-5 enhanced the expression of involucrin and filaggrin, specific differentiation-associated marker proteins in HaCaT cells, whereas transfection of SphK1-siRNA blocked the increase of involucrin. Topical K6PC-5 also enhanced the expression of involucrin, loricrin, filaggrin, and keratin 5 in intact murine epidermis. Finally, topical K6PC-5 inhibited epidermal hyperplasia by exerting antiproliferative effects on keratinocytes in murine epidermis. These results suggest that K6PC-5 acts to regulate both differentiation and proliferation of keratinocytes via [Ca 2+ ] i responses through S1P production. Thus, regulation of S1P levels may represent a novel approach for treatment of skin disorders characterized by abnormal differentiation and proliferation, such as atopic dermatitis and psoriasis.

Original languageEnglish
Pages (from-to)2166-2178
Number of pages13
JournalJournal of Investigative Dermatology
Volume128
Issue number9
DOIs
Publication statusPublished - 2008 Jan 1

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Epidermis
Keratinocytes
Small Interfering RNA
Transfection
Skin
Keratin-5
Hairless Mouse
Sphingolipids
Thapsigargin
Differentiation Antigens
Type C Phospholipases
Atopic Dermatitis
Metabolites
Psoriasis
Hyperplasia
Cell Differentiation
Blood
sphingosine 1-phosphate
sphingosine kinase
Enzymes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Dermatology
  • Cell Biology

Cite this

Hong, Jeong Hee ; Youm, Jong Kyung ; Kwon, Mi Jung ; Park, Byeong Deog ; Lee, Yong Moon ; Lee, Syng Ill ; Shin, DongMin ; Lee, Seung Hun. / K6PC-5, a direct activator of sphingosine kinase 1, promotes epidermal differentiation through intracellular CA 2+ signaling In: Journal of Investigative Dermatology. 2008 ; Vol. 128, No. 9. pp. 2166-2178.
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abstract = "Sphingosine-1-phosphate (S1P), a bioactive sphingolipid metabolite, regulates multiple cellular responses such as Ca 2+ signaling, growth, survival, and differentiation. Because sphingosine kinase (SphK) is the enzyme directly responsible for production of S1P, many factors have been identified that regulate its activity and subsequent S1P levels. Here we synthesized a previously unidentified SphK activator, K6PC-5, and have studied its effects on intracellular Ca 2+ signaling in HaCaT cells and epidermal differentiation in murine skin. K6PC-5, a hydrophobic compound chemically named N-(1,3-dihydroxyisopropyl)-2-hexyl-3-oxo-decanamide, activated SphK (obtained from C57BL/6 murine blood and F9-12 cell lysates) in a dose-dependent manner. K6PC-5 induced both intracellular Ca 2+ concentration ([Ca 2+ ] i ) oscillations in HaCaT cells and Ca 2+ mobilization in hairless mouse epidermis. Both dimethylsphingosine (DMS) and dihydroxysphingosine (DHS), SphK inhibitors, and transfection of SphK1-siRNA blocked K6PC-5-induced increases in [Ca 2+ ] i . The K6PC-5-induced [Ca 2+ ] i oscillations were dependent on thapsigargin-sensitive Ca 2+ stores and Ca 2+ entry, but independent of the classical phospholipase C-mediated pathway. In addition, K6PC-5 enhanced the expression of involucrin and filaggrin, specific differentiation-associated marker proteins in HaCaT cells, whereas transfection of SphK1-siRNA blocked the increase of involucrin. Topical K6PC-5 also enhanced the expression of involucrin, loricrin, filaggrin, and keratin 5 in intact murine epidermis. Finally, topical K6PC-5 inhibited epidermal hyperplasia by exerting antiproliferative effects on keratinocytes in murine epidermis. These results suggest that K6PC-5 acts to regulate both differentiation and proliferation of keratinocytes via [Ca 2+ ] i responses through S1P production. Thus, regulation of S1P levels may represent a novel approach for treatment of skin disorders characterized by abnormal differentiation and proliferation, such as atopic dermatitis and psoriasis.",
author = "Hong, {Jeong Hee} and Youm, {Jong Kyung} and Kwon, {Mi Jung} and Park, {Byeong Deog} and Lee, {Yong Moon} and Lee, {Syng Ill} and DongMin Shin and Lee, {Seung Hun}",
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K6PC-5, a direct activator of sphingosine kinase 1, promotes epidermal differentiation through intracellular CA 2+ signaling . / Hong, Jeong Hee; Youm, Jong Kyung; Kwon, Mi Jung; Park, Byeong Deog; Lee, Yong Moon; Lee, Syng Ill; Shin, DongMin; Lee, Seung Hun.

In: Journal of Investigative Dermatology, Vol. 128, No. 9, 01.01.2008, p. 2166-2178.

Research output: Contribution to journalArticle

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AU - Hong, Jeong Hee

AU - Youm, Jong Kyung

AU - Kwon, Mi Jung

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AU - Lee, Yong Moon

AU - Lee, Syng Ill

AU - Shin, DongMin

AU - Lee, Seung Hun

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N2 - Sphingosine-1-phosphate (S1P), a bioactive sphingolipid metabolite, regulates multiple cellular responses such as Ca 2+ signaling, growth, survival, and differentiation. Because sphingosine kinase (SphK) is the enzyme directly responsible for production of S1P, many factors have been identified that regulate its activity and subsequent S1P levels. Here we synthesized a previously unidentified SphK activator, K6PC-5, and have studied its effects on intracellular Ca 2+ signaling in HaCaT cells and epidermal differentiation in murine skin. K6PC-5, a hydrophobic compound chemically named N-(1,3-dihydroxyisopropyl)-2-hexyl-3-oxo-decanamide, activated SphK (obtained from C57BL/6 murine blood and F9-12 cell lysates) in a dose-dependent manner. K6PC-5 induced both intracellular Ca 2+ concentration ([Ca 2+ ] i ) oscillations in HaCaT cells and Ca 2+ mobilization in hairless mouse epidermis. Both dimethylsphingosine (DMS) and dihydroxysphingosine (DHS), SphK inhibitors, and transfection of SphK1-siRNA blocked K6PC-5-induced increases in [Ca 2+ ] i . The K6PC-5-induced [Ca 2+ ] i oscillations were dependent on thapsigargin-sensitive Ca 2+ stores and Ca 2+ entry, but independent of the classical phospholipase C-mediated pathway. In addition, K6PC-5 enhanced the expression of involucrin and filaggrin, specific differentiation-associated marker proteins in HaCaT cells, whereas transfection of SphK1-siRNA blocked the increase of involucrin. Topical K6PC-5 also enhanced the expression of involucrin, loricrin, filaggrin, and keratin 5 in intact murine epidermis. Finally, topical K6PC-5 inhibited epidermal hyperplasia by exerting antiproliferative effects on keratinocytes in murine epidermis. These results suggest that K6PC-5 acts to regulate both differentiation and proliferation of keratinocytes via [Ca 2+ ] i responses through S1P production. Thus, regulation of S1P levels may represent a novel approach for treatment of skin disorders characterized by abnormal differentiation and proliferation, such as atopic dermatitis and psoriasis.

AB - Sphingosine-1-phosphate (S1P), a bioactive sphingolipid metabolite, regulates multiple cellular responses such as Ca 2+ signaling, growth, survival, and differentiation. Because sphingosine kinase (SphK) is the enzyme directly responsible for production of S1P, many factors have been identified that regulate its activity and subsequent S1P levels. Here we synthesized a previously unidentified SphK activator, K6PC-5, and have studied its effects on intracellular Ca 2+ signaling in HaCaT cells and epidermal differentiation in murine skin. K6PC-5, a hydrophobic compound chemically named N-(1,3-dihydroxyisopropyl)-2-hexyl-3-oxo-decanamide, activated SphK (obtained from C57BL/6 murine blood and F9-12 cell lysates) in a dose-dependent manner. K6PC-5 induced both intracellular Ca 2+ concentration ([Ca 2+ ] i ) oscillations in HaCaT cells and Ca 2+ mobilization in hairless mouse epidermis. Both dimethylsphingosine (DMS) and dihydroxysphingosine (DHS), SphK inhibitors, and transfection of SphK1-siRNA blocked K6PC-5-induced increases in [Ca 2+ ] i . The K6PC-5-induced [Ca 2+ ] i oscillations were dependent on thapsigargin-sensitive Ca 2+ stores and Ca 2+ entry, but independent of the classical phospholipase C-mediated pathway. In addition, K6PC-5 enhanced the expression of involucrin and filaggrin, specific differentiation-associated marker proteins in HaCaT cells, whereas transfection of SphK1-siRNA blocked the increase of involucrin. Topical K6PC-5 also enhanced the expression of involucrin, loricrin, filaggrin, and keratin 5 in intact murine epidermis. Finally, topical K6PC-5 inhibited epidermal hyperplasia by exerting antiproliferative effects on keratinocytes in murine epidermis. These results suggest that K6PC-5 acts to regulate both differentiation and proliferation of keratinocytes via [Ca 2+ ] i responses through S1P production. Thus, regulation of S1P levels may represent a novel approach for treatment of skin disorders characterized by abnormal differentiation and proliferation, such as atopic dermatitis and psoriasis.

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