Michel Lagarde

Docosahexaenoic acid (DHA, C22 : 6 n‐3) is a critical omega‐3 polyunsaturated fatty acid (PUFA) with essential roles in brain and retinal functions. Its selective uptake into these tissues is mediated by several pathways, including the transporter Mfsd2a, which preferentially utilizes lysophosphatidylcholine‐DHA (LPC‐DHA) as a substrate. Due to the low de novo synthesis of DHA, optimizing its dietary intake and exploring novel delivery strategies are important in pathologies that affect PUFA proportion. The present work focuses on developing an efficient chemo‐enzymatic methodology to access structured mixed DHA phosphatidylcholines (mixed DHA‐PCs), valuable biological tools for enhancing drug and/or DHA delivery to the brain and retina. Chemo‐enzymatic methods were established to synthesize mixed sn1‐ or sn2‐DHA‐PC isomers, to study the importance of DHA position, from cost‐effective glycerophosphorylcholine (GPC). The approach aligns with green chemistry principles by leveraging lipase‐catalyzed regioselective acylation and deacylation reactions, thus reducing the number of chemical steps compared to traditional methods. The resulting mixed PCs AceDoPC®, DoAcePC (stabilized forms of LPC‐DHA), and phenolic‐DHA‐PCs were produced at a scale suitable for future in vivo studies, enabling investigations into the positional effect of DHA on brain and retinal uptake.

Stroke is a devastating neurological disorder that is characterized by the sudden disruption of blood flow to the brain. Lipids are essential components of brain structure and function and play pivotal roles in stroke pathophysiology. Dysregulation of lipid signaling pathways modulates key cellular processes such as apoptosis, inflammation, and oxidative stress, exacerbating ischemic brain injury. In the present review, we summarize the roles of lipids in stroke pathology in different models (cell cultures, animal, and human studies). Additionally, the potential of lipids, especially polyunsaturated fatty acids, to promote neuroprotection and their use as biomarkers in stroke are discussed.

Docosahexaenoic acid-containing lysophosphatidylcholine (DHA-LysoPC) is presented as the main transporter of DHA from blood plasma to the brain. This is related to the major facilitator superfamily domain-containing protein 2A (Mfsd2a) symporter expression in the blood–brain barrier that recognizes the various lyso-phospholipids that have choline in their polar head. In order to stabilize the DHA moiety at the sn-2 position of LysoPC, the sn-1 position was esterified by the shortest acetyl chain, creating the structural phospholipid 1-acetyl,2-docosahexaenoyl-glycerophosphocholine (AceDoPC). This small structure modification allows the maintaining of the preferential brain uptake of DHA over non-esterified DHA. Additional properties were found for AceDoPC, such as antioxidant properties, especially due to the aspirin-like acetyl moiety, as well as the capacity to generate acetylcholine in response to the phospholipase D cleavage of the polar head. Esterification of DHA within DHA-LysoPC or AceDoPC could elicit more potent neuroprotective effects against neurological diseases.

We investigated the synthesis of N-docosahexaenoylethanolamine (synaptamide) in neuronal cells from unesterified docosahexaenoic acid (DHA) or DHA-lysophosphatidylcholine (DHA-lysoPC), the two major lipid forms that deliver DHA to the brain, in order to understand the formation of this neurotrophic and neuroprotective metabolite of DHA in the brain. Both substrates were taken up in Neuro2A cells and metabolized to N-docosahexaenoylphosphatidylethanolamine (NDoPE) and synaptamide in a time- and concentration-dependent manner, but unesterified DHA was 1.5 to 2.4 times more effective than DHA-lysoPC at equimolar concentrations. The plasmalogen NDoPE (pNDoPE) amounted more than 80% of NDoPE produced from DHA or DHA-lysoPC, with 16-carbon-pNDoPE being the most abundant species. Inhibition of N-acylphosphatidylethanolamine-phospholipase D (NAPE-PLD) by hexachlorophene or bithionol significantly decreased the synaptamide production, indicating that synaptamide synthesis is mediated at least in part via NDoPE hydrolysis. NDoPE formation occurred much more rapidly than synaptamide production, indicating a precursor–product relationship. Although NDoPE is an intermediate for synaptamide biosynthesis, only about 1% of newly synthesized NDoPE was converted to synaptamide, possibly suggesting additional biological function of NDoPE, particularly for pNDoPE, which is the major form of NDoPE produced.

AceDoPC^® is a structured glycerophospholipid that targets the brain with docosahexaenoic acid (DHA) and is neuroprotective in the experimental ischemic stroke. AceDoPC^® is a stabilized form of the physiological 2-DHA-LysoPC with an acetyl group at the sn1 position; preventing the migration of DHA from the sn2 to sn1 position. In this study we aimed to know the bioavailability of ¹³C-labeled DHA after oral intake of a single dose of ¹³C-AceDoPC^®, in comparison with ¹³C-DHA in triglycerides (TAG), using gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) to assess the ¹³C enrichment of DHA-containing lipids. ¹³C-DHA enrichment in plasma phospholipids was significantly higher after intake of AceDoPC^® compared with TAG-DHA, peaking after 24 h in both cases. In red cells, ¹³C-DHA enrichment in choline phospholipids was comparable from both sources of DHA, with a maximum after 72 h, whereas the ¹³C-DHA enrichment in ethanolamine phospholipids was higher from AceDoPC^® compared to TAG-DHA, and continued to increase after 144 h. Overall, our study indicates that DHA from AceDoPC^® is more efficient than from TAG-DHA for a sustained accumulation in red cell ethanolamine phospholipids, which has been associated with increased brain accretion.