TY - JOUR
T1 - NAPE-PLD controls OEA synthesis and fat absorption by regulating lipoprotein synthesis in an in vitro model of intestinal epithelial cells
AU - Igarashi, Miki
AU - Watanabe, Kazuhide
AU - Tsuduki, Tsuyoshi
AU - Kimura, Ikuo
AU - Kubota, Naoto
N1 - Funding Information:
The authors thank Dr. Tatsuya Sugawara (Kyoto University, Kyoto, Japan) for help in drafting the manuscript. The authors also thank Ms. Kumiko Tachikawa (Tokyo University of Agriculture and Technology, Tokyo, Japan) for excellent technical assistance. This research was supported by Grants-in-Aid for Scientific Research (KAKENHI), the Japan Society for the Promotion of Science 17K07809 (to M.L), the Kao Research Council for the Study of Healthcare Science (2016) (to M.L), and the Project of the National Agriculture and Food Research Organization (NARO) Bio-oriented Technology Research Advancement Institution (advanced integration research for agriculture and interdisciplinary fields) (to M.L and T.T.). The authors declare no conflicts of interest.
Publisher Copyright:
© FASEB
PY - 2019/3/1
Y1 - 2019/3/1
N2 - Oleoylethanolamide (OEA), a fatty acid ethanolamide (FAE), is a lipid mediator that controls food intake and lipid metabolism. Accumulating data imply the importance of intestinal OEA in controlling satiety in addition to gastrointestinal peptide hormones. Although the biochemical pathway of FAE production has been illustrated, the enzymes responsible for the cleavage of OEA from its precursor N-acyl-phosphatidylethanolamine (NAPE) must be identified among reported candidates in the gut. In this study, we assessed the involvement of NAPE-specific phospholipase D (NAPE-PLD), which can directly release FAEs from NAPE, in intestinal OEA synthesis and lipid metabolism. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPER-associated protein 9 (Cas9)-mediated deletion of the NAPE-PLD gene in intestinal epithelial-like Caco-2 cells reduced OEA levels, regardless of their differentiation states. Transcriptome analysis revealed that deletion of NAPE-PLD activates a transcriptional program for nutrient transportation, including lipids and lipoproteins, and inactivates cell-cycle or mitosis-related genes in Caco-2 cells. In addition, the basolateral secretion of lipoproteins was increased in NAPE-PLD-deleted cells although lipoprotein size was not affected. By contrast, cellular lipid levels were reduced in NAPE-PLD-deleted cells. Overall, these results indicate that NAPE-PLD plays important roles in OEA synthesis and fat absorption by regulating lipoprotein production in the intestinal epithelial cells.—Igarashi, M., Watanabe, K., Tsuduki, T., Kimura, I., Kubota, N. NAPE-PLD controls OEA synthesis and fat absorption by regulating lipoprotein synthesis in an in vitro model of intestinal epithelial cells. FASEB J. 33, 3167–3179 (2019). www.fasebj.org.
AB - Oleoylethanolamide (OEA), a fatty acid ethanolamide (FAE), is a lipid mediator that controls food intake and lipid metabolism. Accumulating data imply the importance of intestinal OEA in controlling satiety in addition to gastrointestinal peptide hormones. Although the biochemical pathway of FAE production has been illustrated, the enzymes responsible for the cleavage of OEA from its precursor N-acyl-phosphatidylethanolamine (NAPE) must be identified among reported candidates in the gut. In this study, we assessed the involvement of NAPE-specific phospholipase D (NAPE-PLD), which can directly release FAEs from NAPE, in intestinal OEA synthesis and lipid metabolism. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPER-associated protein 9 (Cas9)-mediated deletion of the NAPE-PLD gene in intestinal epithelial-like Caco-2 cells reduced OEA levels, regardless of their differentiation states. Transcriptome analysis revealed that deletion of NAPE-PLD activates a transcriptional program for nutrient transportation, including lipids and lipoproteins, and inactivates cell-cycle or mitosis-related genes in Caco-2 cells. In addition, the basolateral secretion of lipoproteins was increased in NAPE-PLD-deleted cells although lipoprotein size was not affected. By contrast, cellular lipid levels were reduced in NAPE-PLD-deleted cells. Overall, these results indicate that NAPE-PLD plays important roles in OEA synthesis and fat absorption by regulating lipoprotein production in the intestinal epithelial cells.—Igarashi, M., Watanabe, K., Tsuduki, T., Kimura, I., Kubota, N. NAPE-PLD controls OEA synthesis and fat absorption by regulating lipoprotein synthesis in an in vitro model of intestinal epithelial cells. FASEB J. 33, 3167–3179 (2019). www.fasebj.org.
KW - Caco-2
KW - NAPE-PLD
KW - fatty acid ethanolamide
UR - http://www.scopus.com/inward/record.url?scp=85075114023&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85075114023&partnerID=8YFLogxK
U2 - 10.1096/fj.201801408R
DO - 10.1096/fj.201801408R
M3 - Article
C2 - 30399323
AN - SCOPUS:85075114023
VL - 33
SP - 3167
EP - 3179
JO - FASEB Journal
JF - FASEB Journal
SN - 0892-6638
IS - 3
ER -