Intranet

Fyziologický ústav AV ČR

Špičková věda pro zdraví

Větvené estery mastných kyselin (FAHFA)


Bílá tuková tkáň (WAT) je komplexní endokrinní orgán a subklinický zánět této tkáně během obezity přispívá s rozvoji metabolických onemocnění. V roce 2014 byla objevena nová třída lipidových mediátorů pocházejících z WAT – větvené estery oxidovaných mastných kyselin (FAHFA). FAHFA jsou endogenní lipidy s protizánětlivými a antidiabetickými vlastnostmi, včetně zvýšení glukózové tolerance a sekrece inzulínu a glukagonu podobného peptidu 1 (GLP-1) při současném snížení zánětlivých odpovědí [1-5].

Jsou tvořeny mastnou kyselinou (například kyselinou palmitovou, PA), esterifikovanou na hydroxylovou skupinu hydroxy-mastné kyseliny (například kyseliny hydroxystearové, HSA), zkráceně PAHSA. Poloha větvení na uhlíku definuje regioizomer (například 5-PAHSA). Existuje několik skupin regioizomerů odvozených z palmitové, palmitoleové, stearové, olejové, linolové a docosahexaenové kyseliny doposud detekovaných ve tkáních [1-4, 6, 7]. Tuková tkáň představuje hlavní místo syntézy FAHFA [1, 2], ale příslušné biosyntetické enzymy nejsou známy. Serinová hydroláza [8] a threoninové hydrolázy [9] byly identifikovány jako enzymy metabolizující FAHFA. U lidí byly FAHFA detekovány v séru, mateřském mléce, mekoniu a tukové tkáni [1, 2, 10].

Grafická reprezentace FAHFA rodin rozdělené podle hydroxy mastné kyseliny a obarvená podle esterifikované kyseliny [11] (click for full image).

Dle naší hypotézy existují u myší i lidí nové FAHFA s protizánětlivými účinky odvozené od omega-3 PUFA , které mohou pozitivně ovlivnit metabolismus WAT při obezitě, obzvláště subklinický zánět. Díky experimentům na buněčných kulturách, myších a lidech budeme zkoumat struktury, efekty na zánět WAT, glukozovou toleranci WAT a molekulární mechanismu signalizace těchto nových lipidů. Naše výsledky přispějí k poznání mechanismů propojujících zánět, metabolismus a typ lipidů ve výživě.

Granty

S podporou:

  • GAČR projekt č. 20-00317S (2020-2022, PI: Ondřej Kuda)
  • MŠMT projekt č. LTAUSA18104 (2019-2022, PI: Ondřej Kuda)
  • GAČR projekt č. 17-10088Y (2017-2019, PI: Ondřej Kuda)
  • MŠMT projekt č. LTAUSA17173 (2017-2019, PI: Ondřej Kuda)
  • MŠMT projekt č.. LH14040 (2014-2016, PI: Ondřej Kuda)

Refs:

Naše publikace:

 

► 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

Branched esters of palmitic acid and hydroxy stearic acid are antiinflammatory and antidiabetic lipokines that belong to a family of fatty acid (FA) esters of hydroxy fatty acids (HFAs) called FAHFAs. FAHFAs themselves belong to oligomeric FA esters, known as estolides. Glycerol-bound FAHFAs in triacylglycerols (TAGs), named TAG estolides, serve as metabolite reservoir of FAHFAs mobilized by lipases upon demand. Here, we characterized the involvement of two major metabolic lipases, adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL), in TAG estolide and FAHFA degradation. We synthesized a library of 20 TAG estolide isomers with FAHFAs varying in branching position, chain length, saturation grade, and position on the glycerol backbone and developed an in silico mass spectra library of all predicted catabolic intermediates. We found that ATGL alone or coactivated by comparative gene identification-58 efficiently liberated FAHFAs from TAG estolides with a preference for more compact substrates where the estolide branching point is located near the glycerol ester bond. ATGL was further involved in transesterification and remodeling reactions leading to the formation of TAG estolides with alternative acyl compositions. HSL represented a much more potent estolide bond hydrolase for both TAG estolides and free FAHFAs. FAHFA and TAG estolide accumulation in white adipose tissue of mice lacking HSL argued for a functional role of HSL in estolide catabolism in vivo. Our data show that ATGL and HSL participate in the metabolism of estolides and TAG estolides in distinct manners and are likely to affect the lipokine function of FAHFAs.

  • 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.
  • Mass spectra of TAG estolide standards are available from MassBank of North America (MoNA)


 

► Kristyna Brejchova, Laurence Balas, Veronika Paluchova, Marie Brezinova, Thierry Durand, Ondrej Kuda
Understanding FAHFAs: From Structure to Metabolic Regulation
Progress in Lipid Research, Volume 79, July 2020, 101053. DOI https://doi.org/10.1016/j.plipres.2020.101053 fulltext PDF link

The discovery of branched fatty acid esters of hydroxy fatty acids (FAHFAs) in humans draw attention of many researches to their biological effects. Although FAHFAs were originally discovered in insects and plants, their introduction into the mammalian realm opened new horizons in bioactive lipid research. Hundreds of isomers from different families have been identified so far and their role in (patho) physiological processes is currently being explored. The family of palmitic acid esters of hydroxy stearic acids (PAHSAs), especially 5-PAHSA and 9-PAHSA regioisomers, stands out in the crowd of other FAHFAs for their anti-inflammatory and anti-diabetic effects. Beneficial effects of PAHSAs have been linked to metabolic disorders such as type 1 and type 2 diabetes, colitis, and chronic inflammation. Besides PAHSAs, a growing family of polyunsaturated FAHFAs exerts mainly immunomodulatory effects and biological roles of many other FAHFAs remains currently unknown. Therefore, FAHFAs represent unique lipid messengers capable of affecting many immunometabolic processes. The objective of this review is to summarize the knowledge concerning the diversity of FAHFAs, nomenclature, and their analysis and detection. Special attention is paid to the total syntheses of FAHFAs, optimal strategies, and to the formation of the stereocenter required for optically active molecules. Biosynthetic pathways of saturated and polyunsaturated FAHFAs in mammals and plants are reviewed together with their metabolism and degradation. Moreover, an overview of biological effects of branched FAHFAs is provided and many unanswered questions regarding FAHFAs are discussed.

  • Review of total syntheses of FAHFAs
  • FAHFA nomenclature & analytical procedures
  • Metabolism - synthesis and degradation
  • Biological effects on target organs
  • 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, in press DOI https://doi.org/10.1002/mnfr.201901238

Scope Docosahexaenoic acid ester of hydroxy linoleic acid (DHAHLA) is a bioactive lipids with anti‐inflammatory properties from the family of fatty acid esters of hydroxy fatty acids (FAHFA). Methods and results To explore the biosynthesis of 13‐DHAHLA from dietary oils, C57BL/6N mice were gavaged for 8 days with various corn oil / marine oil mixtures containing the same amount of DHA. Plasma levels of omega‐3 FAHFAs were influenced by the lipid composition of the mixtures but did not reflect the changes in bioavailability of polyunsaturated fatty acids in plasma. Triacylglycerol‐bound DHA and linoleic acid served as more effective precursors for 13‐DHAHLA synthesis than DHA bound in phospholipids or wax esters. Both 13(S)‐ and 13(R)‐DHAHLA inhibited antigen and PGE2‐induced chemotaxis and degranulation of mast cells to a comparable extent and 13(S)‐DHAHLA was identified as the predominant isomer in mouse adipose tissue. Conclusion Here we identified the optimal nutritional source of DHA, which supports production of anti‐inflammatory FAHFAs, as triacylglycerol‐based marine oil and also revealed a possible role of triacylglycerols in the synthesis of FAHFA lipokines.

  • 13-DHAHLA is an anti-inflammatory lipid mediator.
  • 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

Purpose Fatty acid esters of hydroxy fatty acids (FAHFAs) are a large family of endogenous bioactive lipids. To date, most of the studied FAHFAs are branched regioisomers of Palmitic Acid Hydroxyl Stearic Acid (PAHSA) that were reported to possess anti-diabetic and anti-inflammatory activity in humans and rodents. Recently, we have demonstrated that 9-PAHPA or 9-OAHPA intake increased basal metabolism and enhanced insulin sensitivity in healthy control diet-fed mice but induced liver damage in some mice. The present work aims to explore whether a long-term intake of 9-PAHPA or 9-OAHPA may have similar effects in obesogenic diet-fed mice. Methods C57Bl6 mice were fed with a control or high fat-high sugar (HFHS) diets for 12 weeks. The HFHS diet was supplemented or not with 9-PAHPA or 9-OAHPA. Whole-body metabolism was explored. Glucose and lipid metabolism as well as mitochondrial activity and oxidative stress status were analyzed. Results As expected, the intake of HFHS diet led to obesity and lower insulin sensitivity with minor effects on liver parameters. The long-term intake of 9-PAHPA or 9-OAHPA modulated favorably the basal metabolism and improved insulin sensitivity as measured by insulin tolerance test. On the contrary to what we have reported previously in healthy mice, no marked effect for these FAHFAs was observed on liver metabolism of obese diabetic mice. Conclusion This study indicates that both 9-PAHPA and 9-OAHPA may have interesting insulin-sensitizing effects in obese mice with lower insulin sensitivity.

  • 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

Branched fatty acid esters of hydroxy fatty acids (FAHFAs) are a new family of endogenous lipids recently discovered. Several studies reported that some FAHFAs have anti-diabetic and anti-inflammatory effects. The objective of this study was to explore the impact of two FAHFAs, 9-PAHPA or 9-OAHPA on the metabolism of mice. C57Bl/6 J male mice, 6-weeks old, were divided into 3 groups of 10 mice each. One group received a control diet and the two others groups received the control diet supplemented with 9-PAHPA or 9-OAHPA for 12 weeks. Mouse weight and body composition were monitored throughout the study. Some days before euthanasia, energy expenditure, glucose tolerance and insulin sensitivity were also determined. After sacrifice, blood and organs were collected for relevant molecular, biochemical and histological analyses. Although high intake of 9-PAHPA or 9-OAHPA increased basal metabolism, it had no direct effect on body weight. Interestingly, the 9-PAHPA or 9-OAHPA intake increased insulin sensitivity, but without modifying glucose tolerance. Nevertheless, 9-PAHPA intake induced a loss of glucose-stimulated insulin secretion. Surprisingly, both studied FAHFAs induced hepatic steatosis and fibrosis in some mice, which were more marked with 9-PAHPA. Finally, a slight remodeling of white adipose tissue was also observed with 9-PAHPA intake. In conclusion, the long-term high intake of 9-PAHPA or 9-OAHPA increased basal metabolism and insulin sensitivity in healthy mice. However, this effect, highly likely beneficial in a diabetic state, was accompanied by manifest liver damage in certain mice that should deserve special attention in both healthy and pathological studies.

  • 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

Branched esters of palmitic acid and hydroxy-stearic acid (PAHSA) are anti-inflammatory and anti-diabetic lipokines that connect glucose and lipid metabolism. We aimed to characterize involvement of the 5-PAHSA regioisomer in the adaptive metabolic response of white adipose tissue (WAT) to cold exposure (CE) in mice, exploring the crosstalk between glucose utilization and lipid metabolism. CE promoted local production of 5- and 9-PAHSAs in WAT. Metabolic labeling of de novo lipogenesis (DNL) using 2H2O revealed that 5-PAHSA potentiated the effects of CE and stimulated triacylglycerol/fatty acid (TAG/FA) cycling in WAT through impacting lipogenesis and lipolysis. Adipocyte lipolytic products were altered by 5-PAHSA through selective FA re-esterification. The impaired lipolysis in global adipose triglyceride lipase (ATGL) knockout mice reduced free PAHSA levels and uncovered a metabolite reservoir of TAG-bound PAHSAs (TAG-estolides) in WAT. Utilization of 13C isotope tracers and dynamic metabolomics documented that 5-PAHSA primed adipocytes for glucose metabolism in a different way from insulin, promoting DNL, and impeding TAG synthesis. In summary, our data revealed new cellular and physiological mechanisms underlying the beneficial effects of 5-PAHSA, its relation to insulin action in adipocytes, and independently confirmed a PAHSA metabolite reservoir linked to ATGL-mediated lipolysis.

 


 

 

► 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; 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 60 (2019), 1 November 2019, 151331; DOI https://doi.org/10.1016/j.tetlet.2019.151331

Organic synthesis of 13(S)-DHAHLA and 13(R)-DHAHLA

 


 

Kuda O, Brezinova M, Rombaldova M, Slavikova B, Posta M, Beier P, Janovska P, Veleba J, Kopecky J Jr, Kudova E, Pelikanova T, Kopecky J. Docosahexaenoic acid-derived fatty acid esters of hydroxy fatty acids (FAHFAs) with anti-inflammatory properties.

http://diabetes.diabetesjournals.org/content/65/9/2580

http://diabetes.diabetesjournals.org/content/65/11/3516.2 erratum - an incorrect version of the Supplementary Data was erroneously posted online and has been replaced with the correct version.

Chronický zánět přispívá ke vzniku cukrovky, stejně jako kardiovaskulárních, střevních i některých mozkových onemocnění. Tuky z mořských ryb napomáhají v prevenci zánětlivých onemocnění.

 

Anti-inflammatory effects  of novel lipokines of fatty acid esters of hydroxy fatty acids  family in obesity

Omega-3 polynenasycené mastné kyseliny (omega-3) z mořských ryb potlačují zánět a mají i některé příznivé metabolické účinky. Omega-3 snižují riziko vzniku kardiovaskulárních chorob, které jsou často spojené s obezitou a diabetem 2. typu, a také zlepšují lipidový metabolismus. Komplexní studium mechanismů působení omega-3 na myších modelech obezity ve Fyziologickém ústavu AV ČR, klinický výzkum na  obézních pacientech s diabetem 2. typu v Institutu klinické a experimentální medicíny a spolupráce s Ústavem organické chemie a biochemie AV ČR, vedly k objasnění struktury nových signálních molekul lipidové povahy - esterů mastných kyselin a hydroxylovaných mastných kyselin (FAHFA) - odvozených od dokosahexaenové kyseliny (DHA): 13-DHAHLA, 9-DHAHLA a 14-DHAHDHA. Tyto molekuly, které jsou syntetizovány tukovými buňkami a působí protizánětlivě, byly přítomny v séru a tukové tkáni obézních myší i diabetických pacientů, kterým byly podávány omega-3 v dietě. Tyto nově objevené látky, jejichž tvorbu lze stimulovat vhodnou dietou, se podílejí na zdraví prospěšných účincích omega-3 a mohou najít široké uplatnění v prevenci a léčbě řady závažných onemocnění.


 

Kuda O. Bioactive metabolites of docosahexaenoic acid. Biochimie. Jan 2017, DOI: 10.1016/j.biochi.2017.01.002

http://www.sciencedirect.com/science/article/pii/S0300908416302218

Přehled bioaktivních metabolitů DHA. Schema k tisku v JPEG DHA metabolites scheme

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Legend:
 13-DHAHLA, 13-(docosahexaenoyloxy)-hydroxylinoleic acid
 14-DHAHDHA, 14-(docosahexaenoyloxy)-hydroxydocosahexaenoic acid
 9-DHAHLA, 9-(docosahexaenoyloxy)-hydroxylinoleic acid
 AT-, aspirin-triggered-
 CEP, 2-(ω-carboxyethyl)pyrrole
 COX, cyclooxygenase
 DHEA, docosahexaenoyl ethanolamine
 DHG, docosahexaenoyl glycerol
 diHDHA, dihydroxydocosahexaenoic acid
 diHDHEA, dihydroxy-DHEA
 diHDPA, dihydroxydocosapentaenoic acid
 DPA, docosapentaenoic acid
 DPEP, dipeptidase
 eMar, 13,14-epoxy-maresin
 GGT, γ-glutamyl transferase
 GSH, glutathione
 GST, glutathione S-transferase
 GSTM4, glutathione S-transferase
 HEDPEA, hydroxy-epoxy-docosapentaenoyl ethanolamine
 HOHA, 4-hydroxy-7-oxohept-5-enoic acid 
 HpDHA, hydroperoxydocosahexaenoic acid
 HpDHEA, hydroperoxy-DHEA
 LOX, lipoxygenase
 MCTR, Maresin conjugates in tissue regeneration
 NAPE-PLD, N-acyl phosphatidylethanolamine-specific phospholipase D
 NAT, N-acyltransferase
 P450, cytochrome P450
 PCTR, Protectin conjugates in tissue regeneration
 PD, protectin D
 PE, phosphatidylethanolamine
 PGDH, hydroxyprostaglandin dehydrogenase
 RCTR, Resolvin conjugates in tissue regeneration
 ROS, reactive oxygen species
 RvD, resolvin D
 sEH, soluble epoxide hydrolase
 triHDHA, trihydroxydocosahexaenoic acid

 


Brezinova M, Kuda O, Hansikova J, Rombaldova M, Balsa L, Bardova K, Durand T, Rossmeisl M, Cerna M, Stranak Z, Kopecky J. 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; https://doi.org/10.1016/j.bbalip.2017.11.004

fulltext share link till 01/08/2018

 


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 Mar; db171087.; 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.

fulltext

 


► Ondrej Kuda On the Complexity of PAHSA Research. Cell Metabolism 2018, Sep 20; DOI https://doi.org/10.1016/j.cmet.2018.09.006

Comments on the methodological and conceptual problems when working with FAHFAs.

fulltext at https://www.cell.com/cell-metabolism/pdf/S1550-4131(18)30571-0.pdf

free fulltext link https://authors.elsevier.com/a/1Xq8i5WXUlA-Mk

 


Financováno GAČR projekt 17-10088Y (2017-2019, PI: Ondřej Kuda PhD., FGÚ), MŠMT projekt LH14040 (2014-2016, PI: Ondřej Kuda), MŠMT projekt LTAUSA17173 (2017-2019, PI: Ondřej Kuda).