Branched fatty acid esters of hydroxy fatty acids (FAHFA)
White adipose tissue (WAT) is a complex endocrine organ and its low-grade inflammation in obesity contributes to the development of metabolic disorders. In 2014, a class of WAT-born lipid mediators - fatty acid esters of hydroxy fatty acids (FAHFA) was discovered. FAHFAs are endogenous lipids with anti-inflammatory and anti-diabetic properties, including the enhancement of glucose tolerance, and insulin and glucagon-like peptide 1 (GLP-1) secretion while reducing inflammatory responses [1-5].
They consist of a fatty acid (e.g. palmitic acid, PA) esterified to the hydroxyl group of a hydroxy fatty acid (e.g. hydroxystearic acid, HSA), abbreviated as PAHSA. The position of the branching carbon defines a regioisomer (e.g. 5-PAHSA). There are several regioisomer families derived from palmitic, palmitoleic, stearic, oleic, linoleic, and docosahexaenoic acid with tissue-specific distribution documented so far [1-4, 6, 7]. Adipose tissue represents a major site of FAHFAs synthesis [1, 2], but the biosynthetic enzymes involved are unknown . Serine hydrolase carboxyl ester lipase  and threonine hydrolases  were identified as FAHFA-metabolizing enzymes. In humans, FAHFAs were detected in the serum, breast milk, meconium, and adipose tissues [1, 2, 10].
Network representation of FAHFA families linked according to the hydroxy-backbone and colored according to the esterified fatty acid .
Our hypothesis is that novel FAHFAs derived from omega-3 PUFA, with anti-inflammatory properties, could be found in mice and humans and that they can beneficially affect adipose tissue metabolism in obesity, especially low-grade inflammation. We are also interested in FAHFA metabolic pathways, which seem to be as complex as eicosanoid-related pathways. Using experiments in cell cultures, mice and humans we explore the structures, effects on WAT inflammation, WAT glucose tolerance and molecular mechanisms of signaling of these new lipokines. Our results present a significant advance in research of the mechanisms connecting inflammation, metabolism, and nutritional lipids.
► Veronika Paluchova, Tomas Cajka, Thierry Durand, Claire Vigor, Chandra Dodia, Shampa Chatterjee, Aron B. Fisher, Ondrej Kuda✉ The role of peroxiredoxin 6 in biosynthesis of FAHFAs
Free Radical Biology and Medicine, Volume 193, Part 2, 20 November 2022, Pages 787-794. DOI https://doi.org/10.1016/j.freeradbiomed.2022.11.015
► Kristyna Brejchova, Veronika Paluchova, Marie Brezinova, Tomas Cajka, Laurence Balas, Thierry Durand, Marcela Krizova, Zbynek Stranak, Ondrej Kuda✉ Triacylglycerols containing branched palmitic acid ester of hydroxystearic acid (PAHSA) are present in the breast milk and hydrolyzed by carboxyl ester lipase
Food Chemistry, Volume 388, 15 September 2022, 132983. DOI https://doi.org/10.1016/j.foodchem.2022.132983
Effect of preterm birth (PB) or caesarean section (CS) on milk metabolome.
Colostrum levels of 5-PAHSA were negatively affected by PB and CS.
FAHFA-containing triacylglycerols are substrates of carboxyl ester lipase in milk.
5-PAHSA-containing lipids are resistant to carboxyl ester lipase hydrolysis.
► Martin Riecan, Veronika Paluchova, Magno Lopes, Kristyna Brejchova and Ondrej Kuda✉ Branched and linear fatty acid esters of hydroxy fatty acids (FAHFAs) relevant to human health
Pharmacology & Therapeutics, 2022 Mar;231:107972. DOI https://doi.org/10.1016/j.pharmthera.2021.107972
Two structurally and functionally distinct FAHFA superfamilies are recognized based on the position of the estolide bond: omega-FAHFAs and in-chain branched FAHFAs.
We propose a theoretical involvement of cytochrome P450 enzymes in the generation and degradation of saturated HFA backbones.
And present an overview of small-molecule inhibitors used in FAHFA research
► Kristyna Brejchova, Franz Peter Walter Radner, Laurence Balas, Veronika Paluchova, Tomas Cajka, Hana Chodounska, Eva Kudova, Margarita Schratter, Renate Schreiber, Thierry Durand, Rudolf Zechner, and Ondrej Kuda✉ Distinct roles of adipose triglyceride lipase and hormone-sensitive lipase in the catabolism of triacylglycerol estolides
PNAS January 12, 2021 118 (2) e2020999118. DOI https://doi.org/10.1073/pnas.2020999118
Fat mass is controlled by the balance of triacylglycerol (TAG) degradation and synthesis. Adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) are key players in TAG catabolism providing fatty acids (FAs) as energy substrates and metabolic intermediates.
Here, we show that ATGL and HSL metabolize TAGs containing antidiabetic lipid mediators (FA esters of hydroxy FAs), distinctly controlling the release of bioactive lipids.
Our paper connects lipolysis-mediated TAG metabolism with the regulation of antidiabetic signaling lipids.
FAHFA-ome in human adipose tissue - network of 583 FAHFAs from 21 families
► Veronika Paluchova, Anders Vik, Tomas Cajka, Marie Brezinova, Kristyna Brejchova, Viktor Bugajev, Lubica Draberova, Petr Draber, Jana Buresova, Petra Kroupova, Kristina Bardova, Martin Rossmeisl, Jan Kopecky, Trond Vidar Hansen, Ondrej Kuda✉ Triacylglycerol-rich oils of marine origin are optimal nutrients for induction of polyunsaturated docosahexaenoic acid ester of hydroxy linoleic acid (13-DHAHLA) with anti-inflammatory properties in mice.
Molecular Nutrition and Food Research, 2020 Jun;64(11):e1901238 DOI https://doi.org/10.1002/mnfr.201901238
The authors investigate DHA-rich marine oils as potential nutritional sources of 13-DHAHLA precursors and explore anti-inflammatory properties of the bioactive lipid.
The results suggest that triacylglycerol-based marine oils are superior to marine phospholipids and wax esters in the ability to increase levels of 13-DHAHLA in circulation.
Both 13(S)- and 13(R)-DHAHLA inhibited antigen and PGE2-induced chemotaxis and degranulation of mast cells.
► Melha Benlebna, Laurence Balas, Béatrice Bonafos, Laurence Pessemesse, Gilles Fouret, Claire Vigor, Sylvie Gaillet, Jacques Grober, Florence Bernex, Jean François Landrier, Ondrej Kuda, Thierry Durand, Charles Coudray, François Casas, Christine Feillet-Coudray Long-term intake of 9-PAHPA or 9-OAHPA modulates favorably the basal metabolism and exerts an insulin sensitizing effect in obesogenic diet-fed mice.
European Journal of Nutrition, 2020 in press DOI https://doi.org/10.1007/s00394-020-02391-1
9-PAHPA and 9-OAHPA increased insulin sensitivity in C57Bl/6J obese mice.
9-PAHPA and 9-OAHPA did not affect liver metabolism.
► Melha Benlebna, Laurence Balas, Beatrice Bonafos, Laurence Pessemesse, Claire Vigor, Jacques Grober, Florence Bernex, Gilles Fouret, Veronika Paluchova, Sylvie Gaillet, Jean Francois Landrier, Ondrej Kuda, Thierry Durand, Charles Coudray, François Casas, Christine Feillet-Coudray Long-term high dietary intake of 9-PAHPA or 9-OAHPA increases basal metabolism and insulin sensitivity but disrupts liver homeostasis in healthy mice.
Journal of Nutritional Biochemistry, Volume 79, May 2020, 108361, DOI https://doi.org/10.1016/j.jnutbio.2020.108361
9-PAHPA and 9-OAHPA are two branched fatty acid esters of hydroxy fatty acids (FAHFAs)
These both FAHFAs increased basal metabolism in C57Bl/6J healthy mice
9-PAHPA and 9-OAHPA increased insulin sensitivity in C57Bl/6J healthy mice
9-PAHPA and 9-OAHPA induced hepatic steatosis and fibrosis in some C57Bl/6J mice
► Veronika Paluchova, Marina Oseeva, Marie Brezinova, Tomas Cajka, Kristina Bardova, Katerina Adamcova, Petr Zacek, Kristyna Brejchova, Laurence Balas, Hana Chodounska, Eva Kudova, Renate Schreiber, Rudolf Zechner, Thierry Durand, Martin Rossmeisl, Nada A. Abumrad, Jan Kopecky, Ondrej Kuda✉ Lipokine 5-PAHSA is Regulated by Adipose Triglyceride Lipase and Primes Adipocytes for de novo Lipogenesis in Mice.
Diabetes. 2020 Mar;69(3):300-312. DOI https://doi.org/10.2337/db19-0494
► Marie Brezinova, Tomas Cajka, Marina Oseeva, Marek Stepan, Klara Dadova, Lenka Rossmeislova, Milos Matous, Michaela Siklova, Martin Rossmeisl, Ondrej Kuda✉ Exercise training induces insulin-sensitizing PAHSAs in adipose tissue of elderly women.
Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 1865 (2020) 158576; online 16 November 2019, 158576; DOI https://doi.org/10.1016/j.bbalip.2019.158576
Exercise training stimulates beneficial changes in adipose tissue of elderly women.
Exercise stimulates production of insulin-sensitizing lipid mediators PAHSAs.
Insulin sensitivity is associated with short chain TAGs in adipose tissue.
Ether lipids and TAG estolides were detected in serum and adipose tissue samples.
► Anders Vik, Trond Vidar Hansen, Ondrej Kuda Synthesis of both enantiomers of the docosahexaenoic acid ester of 13-hydroxyoctadecadienoic acid (13-DHAHLA).
Tetrahedron Letters Volume 60, Issue 52, 26 December 2019, 151331; DOI https://doi.org/10.1016/j.tetlet.2019.151331
► Ondrej Kuda✉, Marie Brezinova, Jan Silhavy, Vladimir Landa, Vaclav Zidek, Chandra Dodia, Franziska Kreuchwig, Marek Vrbacky, Laurence Balas, Thierry Durand, Norbert Hübner, Aron B. Fisher, Jan Kopecky and Michal Pravenec Nrf2-mediated Antioxidant Defense and Peroxiredoxin 6 are Linked to Biosynthesis of Palmitic Acid Ester of 9-Hydroxystearic Acid.
Diabetes 2018 Jun; 67(6): 1190-1199; DOI https://doi.org/10.2337/db17-1087
Comprehensive lipidomic analysis of rat white adipose tissue samples identified ~160 FAHFA regioisomers and QTL analysis highlighted several positional candidate genes in PAHSA metabolism. The results indicate that the synthesis of PAHSAs via carbohydrate-responsive element-binding protein (ChREBP)-driven de novo lipogenesis is linked to the adaptive antioxidant system and the remodelling of phospholipid hydroperoxides.
► Marie Brezinova, Ondrej Kuda✉, Jana Hansikova, Martina Rombaldova, Laurence Balas, Kristina Bardova, Thierry Durand, Martin Rossmeisl, Marcela Cerna, Zbynek Stranak, Jan Kopecky. Levels of palmitic acid ester of hydroxystearic acid (PAHSA) are reduced in the breast milk of obese mothers.
BBA - Molecular and Cell Biology of Lipids 1863 (2018) 126–131; DOI https://doi.org/10.1016/j.bbalip.2017.11.004
FAHFAs are present in human and murine breast milk.
Levels of 5-PAHSA are negatively affected by obesity.
Stereochemistry of PAHSA regioisomers is explored
The majority of the 5-PAHSA signal is of R configuration
► Ondrej Kuda, Marie Brezinova, Martina Rombaldova, Barbora Slavikova, Martin Posta, Petr Beier, Petra Janovska, Jiri Veleba, Jan Kopecky, Jr., Eva Kudova, Terezie Pelikanova and Jan Kopecky✉ Docosahexaenoic acid-derived fatty acid esters of hydroxy fatty acids (FAHFAs) with anti-inflammatory properties.
Diabetes 2016 Sep; 65 (9): 2580-2590. https://doi.org/10.2337/db16-0385
Omega-3 polyunsaturated fatty acids (omega-3) of marine origin alleviate inflammation, while having favorable metabolic effects. Omega-3 reduce the risk of development of cardiovascular disorders that are linked to obesity and type 2 diabetes, and also improve lipid metabolism. A complex research of omega-3-related mechanisms of action in mouse models of obesity at the Institute of Physiology CAS, clinical research on obese patients with type 2 diabetes in the Institute for Clinical and Experimental Medicine, and a collaboration with the Institute of Organic Chemistry and Biochemistry CAS led to the identification of structures of novel signaling molecules of lipid origin - esters of fatty acids and hydroxyl-fatty acids (FAHFA) - derived from docosahexaenoic acid (DHA): 13-DHAHLA, 9-DHAHLA a 14-DHAHDHA. These molecules, which are synthesized by adipose cells and exert anti-inflammatory effects, were detected in the serum and adipose tissue of both obese mice and diabetic patients following dietary intervention with omega-3. These newly discovered molecules, which can be endogenously synthesized when eating an appropriate diet, are involved in the beneficial health effects of omega-3 and have the potential for their wide use in the prevention and treatment of severe diseases.
Chronic low-grade inflammation contributes to the development of diabetes, as well as cardiovascular, gastrointestinal and certain brain disorders. Lipids of marine origin help to prevent inflammatory diseases.
Purification of 13-DHAHLA: Organic synthesis of docosahexaenoic acid-13-hydroxylinoleic acid (13-DHAHLA) was performed according to Steglich esterification from docosahexaenoic acid (DHA) and 13-hydroxylinoleic acid (13-HODE). The product was purified using silica-based Ag+ flash chromatography (low pressure silver-ion chromatography, Discovery Ag-Ion SPE sorbent, Sigma-Aldrich) and a combination of acetonitrile and acetone ).
► Ondrej Kuda✉ Bioactive metabolites of docosahexaenoic acid.
Biochimie. Jan 2017, DOI: 10.1016/j.biochi.2017.01.002