Involvement of S1P1 receptor pathway in angiogenic effects of a novel adenosine-like nucleic acid analog COA-Cl in cultured human vascular endothelial cells

Junsuke Igarashi, Takeshi Hashimoto, Yasuo Kubota, Kazuyo Shoji, Tokumi Maruyama, Norikazu Sakakibara, Yoh Takuwa, Yoshihiro Ujihara, Yuki Katanosaka, Satoshi Mohri, Keiji Naruse, Tetsuo Yamashita, Ryuji Okamoto, Katsuya Hirano, Hiroaki Kosaka, Maki Takata, Ryoji Konishi, Ikuko Tsukamoto

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

COA-Cl (2Cl-C.OXT-A) is a recently developed adenosine-like nucleic acid analog that promotes angiogenesis via the mitogen-activated protein (MAP) kinases ERK1/2. Endothelial S1P1 receptor plays indispensable roles in developmental angiogenesis. In this study, we examined the functions of S1P1 in COA-Cl-induced angiogenic responses. Antagonists for S1P1, W146, and VPC23019, substantially but still partly inhibited the effects of COA-Cl with regard to ERK1/2 activation and tube formation in cultured human umbilical vein endothelial cells (HUVEC). Antagonists for adenosine A1 receptor and purinergic P2Y1 receptor were without effect. Genetic knockdown of S1P1 with siRNA, but not that of S1P3, attenuated COA-Cl-elicited ERK1/2 responses. The signaling properties of COA-Cl showed significant similarities to those of sphingosine 1-phosphate, an endogenous S1P1 ligand, in that both induced responses sensitive to pertussis toxin (Gα i/o inhibitor), 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester) (BAPTA-AM), (calcium chelator), and PP2 (c-Src tyrosine kinase inhibitor). COA-Cl elevated intracellular Ca2+ concentration and induced tyrosine phosphorylation of p130Cas, a substrate of c-Src, in HUVEC. COA-Cl displaced [3H]S1P in a radioligand-binding competition assay in chem-1 cells overexpressing S1P1. However, COA-Cl activated ERK1/2 in CHO-K1 cells that lack functional S1P1 receptor, suggesting the presence of additional yet-to-be-defined COA-Cl target in these cells. The results thus suggest the major contribution of S1P1 in the angiogenic effects of COA-Cl. However, other mechanism such as that seen in CHO-K1 cells may also be partly involved. Collectively, these findings may lead to refinement of the design of this nucleic acid analog and ultimately to development of small molecule-based therapeutic angiogenesis.

Original languageEnglish
Article numbere00068
JournalPharmacology Research and Perspectives
Volume2
Issue number5
DOIs
Publication statusPublished - Oct 1 2014

Fingerprint

Lysosphingolipid Receptors
CHO Cells
Human Umbilical Vein Endothelial Cells
Adenosine
Nucleic Acids
Endothelial Cells
Purinergic P2Y1 Receptors
Adenosine A1 Receptor Antagonists
Mitogen-Activated Protein Kinase 1
Pertussis Toxin
Small Interfering RNA
Tyrosine
Phosphorylation
Ligands
1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester
Therapeutics
Calcium Chelating Agents
sphingosine 1-phosphate
VPC23019
CSK tyrosine-protein kinase

Keywords

  • Angiogenesis
  • endothelial cells
  • nucleic acid
  • receptors
  • S1P receptors
  • signal transduction
  • sphingosine 1-phosphate

ASJC Scopus subject areas

  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Neurology

Cite this

Involvement of S1P1 receptor pathway in angiogenic effects of a novel adenosine-like nucleic acid analog COA-Cl in cultured human vascular endothelial cells. / Igarashi, Junsuke; Hashimoto, Takeshi; Kubota, Yasuo; Shoji, Kazuyo; Maruyama, Tokumi; Sakakibara, Norikazu; Takuwa, Yoh; Ujihara, Yoshihiro; Katanosaka, Yuki; Mohri, Satoshi; Naruse, Keiji; Yamashita, Tetsuo; Okamoto, Ryuji; Hirano, Katsuya; Kosaka, Hiroaki; Takata, Maki; Konishi, Ryoji; Tsukamoto, Ikuko.

In: Pharmacology Research and Perspectives, Vol. 2, No. 5, e00068, 01.10.2014.

Research output: Contribution to journalArticle

Igarashi, J, Hashimoto, T, Kubota, Y, Shoji, K, Maruyama, T, Sakakibara, N, Takuwa, Y, Ujihara, Y, Katanosaka, Y, Mohri, S, Naruse, K, Yamashita, T, Okamoto, R, Hirano, K, Kosaka, H, Takata, M, Konishi, R & Tsukamoto, I 2014, 'Involvement of S1P1 receptor pathway in angiogenic effects of a novel adenosine-like nucleic acid analog COA-Cl in cultured human vascular endothelial cells', Pharmacology Research and Perspectives, vol. 2, no. 5, e00068. https://doi.org/10.1002/prp2.68
Igarashi, Junsuke ; Hashimoto, Takeshi ; Kubota, Yasuo ; Shoji, Kazuyo ; Maruyama, Tokumi ; Sakakibara, Norikazu ; Takuwa, Yoh ; Ujihara, Yoshihiro ; Katanosaka, Yuki ; Mohri, Satoshi ; Naruse, Keiji ; Yamashita, Tetsuo ; Okamoto, Ryuji ; Hirano, Katsuya ; Kosaka, Hiroaki ; Takata, Maki ; Konishi, Ryoji ; Tsukamoto, Ikuko. / Involvement of S1P1 receptor pathway in angiogenic effects of a novel adenosine-like nucleic acid analog COA-Cl in cultured human vascular endothelial cells. In: Pharmacology Research and Perspectives. 2014 ; Vol. 2, No. 5.
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AU - Igarashi, Junsuke

AU - Hashimoto, Takeshi

AU - Kubota, Yasuo

AU - Shoji, Kazuyo

AU - Maruyama, Tokumi

AU - Sakakibara, Norikazu

AU - Takuwa, Yoh

AU - Ujihara, Yoshihiro

AU - Katanosaka, Yuki

AU - Mohri, Satoshi

AU - Naruse, Keiji

AU - Yamashita, Tetsuo

AU - Okamoto, Ryuji

AU - Hirano, Katsuya

AU - Kosaka, Hiroaki

AU - Takata, Maki

AU - Konishi, Ryoji

AU - Tsukamoto, Ikuko

PY - 2014/10/1

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N2 - COA-Cl (2Cl-C.OXT-A) is a recently developed adenosine-like nucleic acid analog that promotes angiogenesis via the mitogen-activated protein (MAP) kinases ERK1/2. Endothelial S1P1 receptor plays indispensable roles in developmental angiogenesis. In this study, we examined the functions of S1P1 in COA-Cl-induced angiogenic responses. Antagonists for S1P1, W146, and VPC23019, substantially but still partly inhibited the effects of COA-Cl with regard to ERK1/2 activation and tube formation in cultured human umbilical vein endothelial cells (HUVEC). Antagonists for adenosine A1 receptor and purinergic P2Y1 receptor were without effect. Genetic knockdown of S1P1 with siRNA, but not that of S1P3, attenuated COA-Cl-elicited ERK1/2 responses. The signaling properties of COA-Cl showed significant similarities to those of sphingosine 1-phosphate, an endogenous S1P1 ligand, in that both induced responses sensitive to pertussis toxin (Gα i/o inhibitor), 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester) (BAPTA-AM), (calcium chelator), and PP2 (c-Src tyrosine kinase inhibitor). COA-Cl elevated intracellular Ca2+ concentration and induced tyrosine phosphorylation of p130Cas, a substrate of c-Src, in HUVEC. COA-Cl displaced [3H]S1P in a radioligand-binding competition assay in chem-1 cells overexpressing S1P1. However, COA-Cl activated ERK1/2 in CHO-K1 cells that lack functional S1P1 receptor, suggesting the presence of additional yet-to-be-defined COA-Cl target in these cells. The results thus suggest the major contribution of S1P1 in the angiogenic effects of COA-Cl. However, other mechanism such as that seen in CHO-K1 cells may also be partly involved. Collectively, these findings may lead to refinement of the design of this nucleic acid analog and ultimately to development of small molecule-based therapeutic angiogenesis.

AB - COA-Cl (2Cl-C.OXT-A) is a recently developed adenosine-like nucleic acid analog that promotes angiogenesis via the mitogen-activated protein (MAP) kinases ERK1/2. Endothelial S1P1 receptor plays indispensable roles in developmental angiogenesis. In this study, we examined the functions of S1P1 in COA-Cl-induced angiogenic responses. Antagonists for S1P1, W146, and VPC23019, substantially but still partly inhibited the effects of COA-Cl with regard to ERK1/2 activation and tube formation in cultured human umbilical vein endothelial cells (HUVEC). Antagonists for adenosine A1 receptor and purinergic P2Y1 receptor were without effect. Genetic knockdown of S1P1 with siRNA, but not that of S1P3, attenuated COA-Cl-elicited ERK1/2 responses. The signaling properties of COA-Cl showed significant similarities to those of sphingosine 1-phosphate, an endogenous S1P1 ligand, in that both induced responses sensitive to pertussis toxin (Gα i/o inhibitor), 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester) (BAPTA-AM), (calcium chelator), and PP2 (c-Src tyrosine kinase inhibitor). COA-Cl elevated intracellular Ca2+ concentration and induced tyrosine phosphorylation of p130Cas, a substrate of c-Src, in HUVEC. COA-Cl displaced [3H]S1P in a radioligand-binding competition assay in chem-1 cells overexpressing S1P1. However, COA-Cl activated ERK1/2 in CHO-K1 cells that lack functional S1P1 receptor, suggesting the presence of additional yet-to-be-defined COA-Cl target in these cells. The results thus suggest the major contribution of S1P1 in the angiogenic effects of COA-Cl. However, other mechanism such as that seen in CHO-K1 cells may also be partly involved. Collectively, these findings may lead to refinement of the design of this nucleic acid analog and ultimately to development of small molecule-based therapeutic angiogenesis.

KW - Angiogenesis

KW - endothelial cells

KW - nucleic acid

KW - receptors

KW - S1P receptors

KW - signal transduction

KW - sphingosine 1-phosphate

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