Desaturase Research Articles

Biosynthesis and Significance of Fatty Acids, Glycerophospholipids, and Triacylglycerol in the Processes of Glioblastoma Tumorigenesis

One area of glioblastoma research is the metabolism of tumor cells and detecting differences between tumor and healthy brain tissue metabolism. Here, we review differences in fatty acid metabolism, with a particular focus on the biosynthesis of saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA) by fatty acid synthase (FASN), elongases, and desaturases. We also describe the significance of individual fatty acids in glioblastoma tumorigenesis, as well as the importance of glycerophospholipid and triacylglycerol synthesis in this process. Specifically, we show the significance and function of various isoforms of glycerol-3-phosphate acyltransferases (GPAT), 1-acylglycerol-3-phosphate O-acyltransferases (AGPAT), lipins, as well as enzymes involved in the synthesis of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI), and cardiolipin (CL). This review also highlights the involvement of diacylglycerol O-acyltransferase (DGAT) in triacylglycerol biosynthesis. Due to significant gaps in knowledge, the GEPIA database was utilized to demonstrate the significance of individual enzymes in glioblastoma tumorigenesis. Finally, we also describe the significance of lipid droplets in glioblastoma and the impact of fatty acid synthesis, particularly docosahexaenoic acid (DHA), on cell membrane fluidity and signal transduction from the epidermal growth factor receptor (EGFR).

Abstract 1250: Aberrant polyunsaturated fatty acids desaturation impairs survival and anti-tumor immunity of tumor resident memory CD8+ T cell in hepatocellular carcinoma

Abstract Background & Aim: Aberrant polyunsaturated fatty acids (PUFAs) metabolism is closely associated with the immunosuppressive tumor microenvironment (TME). Tissue-resident memory T cell (Trm) plays a central role against solid tumors, including HCC, depending on the uptake of exogenous fatty acids to maintain survival and anti-tumor immunity. Here we aimed to explore the potential interaction in TME between HCC-derived PUFAs and Trm function in a series of integrated clinical, basic and translational studies. Methods: Tissue interstitial fluids (TIFs) were isolated and analyzed from fresh paired adjacent tissues and HCC for detecting the PUFAs alterations in the HCC TME. Integrated analyses were performed in multiple public databases and local cohorts. Fatty acid desaturase 1 (FADS1) knockdown (KD) HCC cell line was generated to observe the tumor biology and metabolic function. The effects of FADS1 knockout (KO) or overexpression were investigated in multiple spontaneous HCC mouse models via hydrodynamic tail vein injection (HTVI) in vivo. Mutation landscape analysis was used to explore the mechanism of FADS1 transcription. Therapeutic interventions by downregulating FADS1 were also evaluated in vivo. Results: Clinically, we found that the arachidonic acid (ARA) level was enriched in HCC TIFs. A rate-limiting enzyme controlling PUFA biosynthesis, especially ARA production from dihomo-γ-linolenic acid (DGLA), FADS1, was overexpressed in HCC patients. High expression of FADS1 predicted poor survival and was negatively correlated with CD8+CD103+Trm infiltration. Moreover, in the local clinical trial, we noticed that the expression of FADS1 was upregulated in non-response patients with anti-PD-1 mAb treatment. Functionally, we demonstrated that the ARA level was decreased in the conditioned medium (CM) from FADS1 KD cells. Applying CM prolonged the survival of CD8+CD103+Trm ex vivo. FADS1 KO inhibited tumor development by increasing intratumor infiltration and function of CD8+CD103+Trm. In contrast, overexpression of FADS1 promoted HCC progression by restraining immune surveillance in vivo. Mechanistically, we found that the expression of FADS1 was upregulated in the TP53 mutation group by analyzing the mutation landscape of HCC. Mutant P53 protein could bind to the promoter region of FADS1. Therapeutically, recombinant adeno-associated 8 (rAAV8)-shRNAs delivery and anti-PD-1 mAb administration inhibited the tumor growth of anti-PD-1 mAb treatment-resistant HCC model driven by CTNNB1N90 and MYC. Conclusions: We demonstrated that mutant P53 protein directly promoted FADS1 transcription, increasing ARA accumulation in HCC TME. FADS1 remodeling the metabolic patterns of PUFAs impaired CD8+CD103+Trm anti-tumor immunity, possibly leading to HCC progression. Citation Format: Tao Ding, Li Pang, Runying Long, Shinuan Zeng, Kevin TP Ng, Qingmei Zhang, Zhiwei Chen, Kwan Man. Aberrant polyunsaturated fatty acids desaturation impairs survival and anti-tumor immunity of tumor resident memory CD8+ T cell in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1250.

Dietary omega-3 fatty acid deficiency from pre-pregnancy to lactation affects expression of genes involved in hippocampal neurogenesis of the offspring

Maternal n-3 PUFA (omega-3) deficiency can affect brain development in utero and postnatally. Despite the evidence, the impacts of n-3 PUFA deficiency on the expression of neurogenesis genes in the postnatal hippocampus remained elusive. Since postnatal brain development requires PUFAs via breast milk, we examined the fatty acid composition of breast milk and hippocampal expression of neurogenesis genes in n-3 PUFA deficient 21d mice. In addition, the expression of fatty acid desaturases, elongases, free fatty acids signaling receptors, insulin and leptin, and glucose transporters were measured. Among the genes involved in neurogenesis, the expression of brain-specific tenascin-R (TNR) was downregulated to a greater extent (∼31 fold), followed by adenosine A2A receptor (A2AAR), dopamine receptor D2 (DRD2), glial cell line-derived neurotrophic factor (GDNF) expression in the n-3 PUFA deficient hippocampus. Increasing dietary LA to ALA (50:1) elevated the ARA to DHA ratio by ∼8 fold in the n-3 PUFA deficient breast milk, with an overall increase of total n-6/n-3 PUFAs by ∼15:1 (p<0.05) compared to n-3 PUFA sufficient (LA to ALA: 2:1) diet. The n-3 PUFA deficient mice exhibited upregulation of FADS1, FADS2, ELOVL2, ELOVL5, ELOVL6, GPR40, GPR120, LEPR, IGF1 and downregulation of GLUT1, GLUT3, and GLUT4 mRNA expression in hippocampus (p<0.05). Maternal n-3 PUFA deficiency affects the hippocampal expression of key neurogenesis genes in the offspring with concomitant expression of desaturase and elongase genes, suggesting the importance of dietary n-3 PUFA for neurodevelopment.

The chromosome-level genome of Eucommia ulmoides provides insights into sex differentiation and α-linolenic acid biosynthesis.

Eucommia ulmoides Oliver is a typical dioecious plant endemic to China that has great medicinal and economic value. Here, we report a high-quality chromosome-level female genome of E. ulmoides obtained by PacBio and Hi-C technologies. The size of the female genome assembly was 1.01 Gb with 17 pseudochromosomes and 31,665 protein coding genes. In addition, Hi-C technology was used to reassemble the male genome released in 2018. The reassembled male genome was 1.24 Gb with the superscaffold N50 (48.30 Mb), which was increased 25.69 times, and the number of predicted genes increased by 11,266. Genome evolution analysis indicated that E. ulmoides has undergone two whole-genome duplication events before the divergence of female and male, including core eudicot γ whole-genome triplication event (γ-WGT) and a recent whole genome duplication (WGD) at approximately 27.3 million years ago (Mya). Based on transcriptome analysis, EuAP3 and EuAG may be the key genes involved in regulating the sex differentiation of E. ulmoides. Pathway analysis showed that the high expression of ω-3 fatty acid desaturase coding gene EU0103017 was an important reason for the high α-linolenic acid content in E. ulmoides. The genome of female and male E. ulmoides presented here is a valuable resource for the molecular biological study of sex differentiation of E. ulmoides and also will provide assistance for the breeding of superior varieties.

G0S2 inhibits adipogenesis and unsaturated fatty acid biosynthesis by repressing the expression of SCD in chicken

SummaryFatty acids metabolic products and intramuscular fat (IMF) content determine meat quality in chickens. In this study, the mechanisms of G0/G1 switch gene 2 (G0S2) underlying modulation of chicken IMF content as well as fatty acid synthesis was investigated. Several advanced techniques including Gas Chromatography–Mass Spectrometer (GC–MS), RNA‐sequencing and lentiviral transfection were utilised. RNA‐sequencing showed that 2242 differentially expressed genes were significantly different, and many of these were related to lipid catabolism, such as stearoyl‐CoA desaturase 1 (SCD), acyl‐CoA synthetase bubblegum family member 2 (ACSBG2), and fatty acid desaturase 2 (FADS2), etc. Knocking down of G0S2 also affected the biological activities of signal transduction, catalytic activity, lipid metabolism, cell cycle and death, cell differentiation and immune response. GC–MS revealed that decreased lipid accumulation and lower unsaturated fatty acid percentage were observed in G0S2 knockdown adipocytes. Our results showed that lower expression of G0S2 decreased lipid accumulation mainly via down‐regulated SCD, then regulated mRNA levels of patatin‐like phospholipase domain containing 2 (PNPLA2), which was shown as a major triglyceride lipase in the liver and adipose tissue. Importantly, we further verified the regulation network of G0S2 in the thigh tissues of 12 Huangshan Black chickens, which was consistent with previous IMF results. Thus, our study highlights that G0S2 modulated chicken meat quality traits by the G0S2–SCD–PNPLA2 axis in lipid metabolism in vitro and in vivo. These findings are expected to provide a strategy from a genetic perspective for improving the meat quality.

Therapy-Resistant Acute Myeloid Leukemia Stem Cells Are Resensitized to Venetoclax + Azacitidine by Targeting Fatty Acid Desaturases 1 and 2.

Recent advances in targeting leukemic stem cells (LSCs) using venetoclax with azacitidine (ven + aza) has significantly improved outcomes for de novo acute myeloid leukemia (AML) patients. However, patients who relapse after traditional chemotherapy are often venetoclax-resistant and exhibit poor clinical outcomes. We previously described that fatty acid metabolism drives oxidative phosphorylation (OXPHOS) and acts as a mechanism of LSC survival in relapsed/refractory AML. Here, we report that chemotherapy-relapsed primary AML displays aberrant fatty acid and lipid metabolism, as well as increased fatty acid desaturation through the activity of fatty acid desaturases 1 and 2, and that fatty acid desaturases function as a mechanism of recycling NAD+ to drive relapsed LSC survival. When combined with ven + aza, the genetic and pharmacologic inhibition of fatty acid desaturation results in decreased primary AML viability in relapsed AML. This study includes the largest lipidomic profile of LSC-enriched primary AML patient cells to date and indicates that inhibition of fatty acid desaturation is a promising therapeutic target for relapsed AML.

Antidiabetic and anti-obesity properties of a polyphenol-rich flower extract from Tagetes erecta L. and its effects on Caenorhabditis elegans fat storages

Diabetes mellitus (DM) is a metabolic disease characterized by a high blood sugar level that can cause severe complications to the organism or even death when not treated. However, certain dietary habits and foods may have beneficial effects on this condition. A polyphenolic-rich extract (containing hyperoside, isoquercitrin, quercetin, ellagic acid, and vanillic acid) of Tageres erecta L. (T. erecta) was obtained from yellow and orange flowers using an ethanolic Soxhlet extraction. These extracts were screened for antidiabetic and anti-obesity properties using in vitro and in vivo procedures. The capacity to inhibit the enzymes lipase and α-glucosidase, as well as the inhibition of advance glycation end-products (AGEs) was tested in vitro. Caenorhabditis elegans (C. elegans) was used as an obesity in vivo model to assess extracts effects on fat accumulation using the wild-type strain N2 and a mutant with no N3 fatty acid desaturase activity BX24. Extracts from both cultivars (yellow and orange) T. erecta presented in vitro inhibitory activity against the enzymes lipase and α-glucosidase, showing lower IC50 values than acarbose (control). They also showed important activity in preventing AGEs formation. The polyphenol-rich matrices reduced the fat content of obese worms in the wild-type strain (N2) down to levels of untreated C. elegans, with no significant differences found between negative control (100% reduction) and both tested samples (p < 0.05). Meanwhile, the fat reduction was considerably lower in the BX24 mutants (fat-1(wa-9)), suggesting that N3 fatty acid desaturase activity could be partially involved in the T. erecta flower effect. Our findings suggested that polyphenols from T. erecta can be considered candidate bioactive compounds in the prevention and improvement of metabolic chronic diseases such as obesity and diabetes.

Contribution of the leaf and silique photosynthesis to the seeds yield and quality of oilseed rape (Brassica napus L.) in reproductive stage

Influences of photosynthesis of leaf and silique on seeds yield and quality of oilseed rape (Brassica napus L.) were explored in this study. A field comparing experiment with several rapeseed varieties was conducted and the results showed, that the leaf area index (LAI), silique surface area index (SAI), siliques number per plant, and biological yield were statistically classified as the first principal factors which greatly influenced seeds yield, the leaf net photosynthetic rate (Pn) and silique Pn were the second principal factors; the stomatal conductance (Gs) and chlorophyll a (Chl a) content were the first principal factors which influenced leaf Pn and silique Pn. A shading experiment was conducted and the results showed that, under treatments of the ZH1, ZH2, and ZH3 (shading rapeseed plants during flowering stage, during time from initial flowering until seeds ripening, and during time from flowering ending until seeds ripening, respectively), respectively the seeds yield per plant was reduced by 34.6%, 84.3%, and 86.1%, the seed protein content was significantly increased. The treatment ZH1 Not, but the ZH2 and ZH3 caused significant decrease in both seed oil content and oleic acid (C18:1) content in seed oil, and the contents of linoleic acid (C18:2), linolenic acid (C18:3) in oil were significantly increased, gene expression of the ACCase (Acetyl-CoA carboxylase), FAD2 (fatty acid desaturase), and FAD3 (ω-3 fatty acid dehydrogenase) in green seeds was restrained/changed. Thus the LAI, SAI, siliques number per plant, biological yield per plant, leaf Pn, silique Pn, and the Gs, Chl a content of leaf and silique formed an indexes system to be used in screening rapeseed variety with higher light efficiency and seeds yield; the silique photosynthesis inhibition and the photosynthates deficiency in rapeseed plant after flowering stage predominately influenced seeds yield and quality.

A near complete genome assembly of chia assists in identification of key fatty acid desaturases in developing seeds.

Chia is an annual crop whose seeds have the highest content of α-linolenic acid (ALA) of any plant known to date. We generated a high-quality assembly of the chia genome using circular consensus sequencing (CCS) of PacBio. The assembled six chromosomes are composed of 21 contigs and have a total length of 361.7 Mb. Genome annotation revealed a 53.5% repeat content and 35,850 protein-coding genes. Chia shared a common ancestor with Salvia splendens ~6.1 million years ago. Utilizing the reference genome and two transcriptome datasets, we identified candidate fatty acid desaturases responsible for ALA biosynthesis during chia seed development. Because the seed of S. splendens contains significantly lower proportion of ALA but similar total contents of unsaturated fatty acids, we suggest that strong expression of two ShFAD3 genes are critical for the high ALA content of chia seeds. This genome assembly will serve as a valuable resource for breeding, comparative genomics, and functional genomics studies of chia.

Combining a Mutant Allele of FAD2-1A with HD Improves the ω-6/ω-3 Ratio in Soybeans

The intake of foods with unbalanced ω-6/ω-3 ratios causes various health problems. Commodity soybeans generally have a ω-6/ω-3 ratio of 6–7:1. The recommended ratio in terms of health benefits is &lt;4:1. This study aimed to identify the appropriate combination of mutant alleles that can reduce the ω-6/ω-3 ratio using three segregating soybean populations. F2 individuals from each population were genotyped for three different alleles of microsomal delta-12 fatty acid desaturase 2 enzyme (FAD2-1A) and an allele of homeodomain-like transcriptional regulator (HD) genes, and their five major fatty acids were assessed. F2 seeds carrying both fad2-1a and hd had slightly different ω-6/ω-3 ratios according to the different fad2-1a alleles. The fad2-1aDEL, fad2-1aS117N, and fad2-1aW293STOP alleles combined with a hd allele resulted in ω-6/ω-3 ratios with a range of 1.9–2.7:1, 2.7–3.9:1, and 2.6–3.6:1 in soybean seeds, respectively. This study revealed that the induction of mutations in FAD2-1ADEL and HD was the most efficient strategy to improve the ω-6/ω-3 ratio and elevate the ω-3 fatty acid concentrations in soybean seeds. These results provide useful information in soybean breeding programs to release a new soybean cultivar with a lower ω-6/ω-3 ratio and elevated ω-3 fatty acids, which can be a beneficial ingredient for soybean-based foods.

A mutant cotton fatty acid desaturase 2-1d allele causes protein mistargeting and altered seed oil composition

BackgroundCotton (Gossypium sp.) has been cultivated for centuries for its spinnable fibers, but its seed oil also possesses untapped economic potential if, improvements could be made to its oleic acid content.ResultsPrevious studies, including those from our laboratory, identified pima accessions containing approximately doubled levels of seed oil oleic acid, compared to standard upland cottonseed oil. Here, the molecular properties of a fatty acid desaturase encoded by a mutant allele identified by genome sequencing in an earlier analysis were analyzed. The mutant sequence is predicted to encode a C-terminally truncated protein lacking nine residues, including a predicted endoplasmic reticulum membrane retrieval motif. We determined that the mutation was caused by a relatively recent movement of a Ty1/copia type retrotransposon that is not found associated with this desaturase gene in other sequenced cotton genomes. The mutant desaturase, along with its repaired isozyme and the wild-type A-subgenome homoeologous protein were expressed in transgenic yeast and stably transformed Arabidopsis plants. All full-length enzymes efficiently converted oleic acid to linoleic acid. The mutant desaturase protein produced only trace amounts of linoleic acid, and only when strongly overexpressed in yeast cells, indicating that the missing C-terminal amino acid residues are not strictly required for enzyme activity, yet are necessary for proper subcellular targeting to the endoplasmic reticulum membrane.ConclusionThese results provide the biochemical underpinning that links a genetic lesion present in a limited group of South American pima cotton accessions and their rare seed oil oleic acid traits. Markers developed to the mutant desaturase allele are currently being used in breeding programs designed to introduce this trait into agronomic upland cotton varieties.

Genome-wide comparative identification and analysis of membrane-FADS-like superfamily genes in freshwater economic fishes.

Membrane fatty acid desaturase (FADS)-like superfamily proteins (FADSs) are essential for the synthesis of unsaturated fatty acids (UFAs). Recently, studies on FADS in fishes have mostly focused on marine species, and a comprehensive analysis of the FADS superfamily, including the FADS, stearoyl-CoA desaturase (SCD), and sphingolipid delta 4-desaturase (DEGS) families, in freshwater economic fishes is urgently required. To this end, we conducted a thorough analysis of the number, gene/protein structure, chromosomal location, gene linkage map, phylogeny, and expression of the FADS superfamily. We identified 156 FADSs genes in the genome of 27 representative species. Notably, FADS1 and SCD5 were lost in most freshwater fish and other teleosts. All FADSs proteins contain 4 transmembrane helices and 2-3 amphipathic α-helices. FADSs in the same family are often linked on the same chromosome; moreover, FADS and SCD or DEGS are frequently collocated on the same chromosome. In addition, FADS, SCD, and DEGS family proteins share similar evolutionary patterns. Interestingly, FADS6, as a member of the FADS family, exhibits a similar gene structure and chromosome location to that of SCD family members, which may be the transitional form of FADS and SCD. This study shed light on the type, structure, and phylogenetic relationship of FADSs in freshwater fishes, offering a new perspective into the functional mechanism analysis of FADSs.

Zinc- and nickel-induced changes in fatty acid profiles in the zinc hyperaccumulator Arabidopsis halleri and non-accumulator Arabidopsis lyrata

This pilot study aimed at comparing zinc (Zn) and nickel (Ni) effects on the fatty acid (FA) profiles, oxidative stress and desaturase activity in the Zn hyperaccumulator Arabidopsis halleri and the excluder Arabidopsis lyrata to allow a better picture of the physiological mechanisms which may contribute to metal tolerance or acclimation. The most significant changes in the FA composition were observed in the shoots of the hyperaccumulator and in the roots of the excluder, and were not only metal-dependent, but also species-specific, since the most significant changes in the shoots of A. halleri were observed under Ni treatment, though Ni, in contrast to Zn, was accumulated mainly in its roots. Several FAs appeared in the roots and shoots of A. lyrata only upon metal exposure, whereas they were already found in control A. halleri. In both species, there was an increase in oleic acid under Ni treatment in both organs, whereas in Zn-treated plants the increase was shown only for the shoots. A rare conjugated α-parinaric acid was identified only in the shoots of metal-treated A. halleri. In the shoots of the hyperaccumulator, there was an increase in the content of saturated FAs and a decrease in the content of unsaturated FAs, while in the roots of the excluder, the opposite pattern was observed. These metal-induced changes in FA composition in the shoots of A. halleri can lead to a decrease in the fluidity of membranes, which could diminish the penetration of ROS into the membrane and thus maintain its stability.

Generation of Sesame Mutant Population by Mutagenesis and Identification of High Oleate Mutants by GC Analysis.

Sesame is one of the important oilseed crops in the world. Natural genetic variation exists in the sesame germplasm collection. Mining and utilizing the genetic allele variation from the germplasm collection is an important approach for seed quality improvement. The sesame germplasm accession, PI 263470, which has a significantly higher level of oleic acid (54.0%) than the average (39.5%), was identified by screening the entire USDA germplasm collection. The seeds from this accession were planted in a greenhouse. Leaf tissues and seeds were harvested from individual plants. DNA sequencing of the coding region of the fatty acid desaturase gene (FAD2) confirmed that this accession contained a natural mutation of G425A which may correspond to the deduced amino acid substitution of R142H leading to the high level of oleic acid, but it was a mixed accession with three genotypes (G/G, G/A, and A/A at the position). The genotype with A/A was selected and self-crossed for three generations. The purified seeds were used for EMS-induced mutagenesis to further enhance the level of oleic acid. A total of 635 M2 plants were generated from mutagenesis. Some mutant plants had significant morphological changes including leafy flat stems and others. M3 seeds were used for fatty acid composition analysis by gas chromatography (GC). Several mutant lines were identified with high oleic acid (70%). Six M3 mutant lines plus one control line were advanced to M7 or M8 generations. Their high oleate traits from M7 or M8 seeds harvested from M6 or M7 plants were further confirmed. The level of oleic acid from one mutant line (M7 915-2) was over 75%. The coding region of FAD2 was sequenced from these six mutants, but no mutation was identified. Additional loci may contribute to the high level of oleic acid. The mutants identified in this study can be used as breeding materials for sesame improvement and as genetic materials for forward genetic studies.

Membrane lipid metabolism and scavenging of reactive oxygen species are essential for cold resistance of Solanum habrochaites (LA1777): Lipidomics and transcriptomes analysis

Low temperature significantly limits the production of tomato (Solanum lycopersicum) in greenhouse. Solanum habrochaites is a wild species of tomato closely related to Solanum lycopersicum and is more tolerant to cold than S. lycopersicum. However, the mechanism by which S. habrochaites resists low temperature is unknown. This study aimed to analyze the physiology, transcriptomics, and lipidomics of LA1777 (S. habrochaites variety) after exposure to cold stress for 3 days to assess its specific mechanism to cold tolerance. A cold-sensitive cultivated tomato variety AC was used as controls. Physiological analysis showed that the two tomato varieties were significantly affected by cold stress. However, LA1777 had more stable physiological characteristics and maintained relatively good photosynthetic capacity than the AC variety. Transcriptome analysis suggested that the stability of cell membrane structure and function might promote cold tolerance of LA1777. Furthermore, 243 differentially expressed genes (DEGs) involved in lipid metabolism were identified. Particularly, the upregulation of temperature-induced lipocalin genes (TILs) and fatty acid desaturase genes (FADs) promoted cold tolerance by inhibiting the excessive accumulation of reactive oxygen species (ROS) and increasing membrane lipid unsaturation, respectively. Meanwhile, lipidomics data indicated that monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) levels were significantly reduced in both plants under cold stress. MGDG levels decreased by 28% and 54% in LA1777 and AC, respectively, while DGDG decreased by 7% and 29%, respectively. Further results showed that high phosphoinositide (PI) content may increase cold tolerance in LA1777 by maintaining phosphatidylinositol signaling system. Also, results showed that adenosine triphosphate (ATP) produced via β-oxidation of fatty acids in mitochondria can maintain a higher metabolic level of wild tomato under cold stress. A metabolic model of the mechanism of cold tolerance in tomato was also constructed. This study provides a basis for further analysis of the molecular mechanism of cold resistance in tomato, which is crucial for genetic improvement.

β-triketone herbicide exposure cause tyrosine and fat accumulation in Caenorhabditis elegans

β-triketone herbicides have been efficiently employed as an alternate to atrazine. Triketones are 4-hydroxyphenylpyruvate dioxygenase (HPPD) enzyme inhibitors and exposure is reported to cause significant increase in plasma tyrosine levels. In this study, we have employed a non-target organism Caenorhabditis elegans to determine the impact of β-triketone exposures at recommended field doses (RfD). Our results indicate sulcotrione and mesotrione, negatively influence the survival, behavior, and reproduction of the organism at RfD. Additionally, we have traced the parallels regarding the impact of triketones on the tyrosine metabolism pathway, in C. elegans to those in mammalian models, wherein the expression of the tyrosine metabolism pathway genes are altered, directly influencing tyrosine catabolism leading to significant tyrosine accumulation in exposed organism. Further, we investigated the impact of sulcotrione and mesotrione exposure on fat deposition (triglyceride levels, Oil-Red-O staining and lipidomics) and the fatty acid metabolism pathway. In the exposed worms, the expression of enlongases and fatty acid desaturases were up-regulated along with an increase in the levels of triglycerides. Thus, the data indicates a positive association of β-triketone exposure to mis-regulation of the fatty acid metabolism pathway genes leading to fat accumulation in worms. Therefore, β-triketone might be a potential obesogen.

Desaturation of sebaceous-type saturated fatty acids through the SCD1 and the FADS2 pathways impacts lipid neosynthesis and inflammatory response in sebocytes in culture.

Sebum is a lipid-rich mixture secreted by the sebaceous gland (SG) onto the skin surface. By penetrating through the epidermis, sebum may be involved in the regulation of epidermal and dermal cells in both healthy and diseased skin conditions. Saturated and monounsaturated fatty acids (FAs), found as free FAs (FFAs) and in bound form in neutral lipids, are essential constituents of sebum and key players of the inflammatory processes occurring in the pilosebaceous unit in acne-prone skin. Little is known on the interplay among uptake of saturated FFAs, their biotransformation, and induction of proinflammatory cytokines in sebocytes. In the human SG, palmitate (C16:0) is the precursor of sapienate (C16:1n-10) formed by insertion of a double bond (DB) at the Δ6 position catalysed by the fatty acid desaturase 2 (FADS2) enzyme. Conversely, palmitoleate (C16:1n-7) is formed by insertion of a DB at the Δ9 position catalysed by the stearoyl coenzyme A desaturase 1 (SCD1) enzyme. Other FFAs processed in the SG, also undergo these main desaturation pathways. We investigated lipogenesis and release of IL-6 and IL-8 pro-inflammatory cytokines in SZ95 sebocytes in vitro after treatment with saturated FFAs, that is, C16:0, margarate (C17:0), and stearate (C18:0) with or without specific inhibitors of SCD1 and FADS2 desaturase enzymes, and a drug with mixed inhibitory effects on FADS1 and FADS2 activities. C16:0 underwent extended desaturation through both SCD1 and FADS2 catalysed pathways and displayed the strongest lipoinflammatory effects. Inhibition of desaturation pathways proved to enhance lipoinflammation induced by SFAs in SZ95 sebocytes. Palmitate (C16:0), margarate (C17:0), and stearate (C18:0) are saturated fatty acids that induce different arrays of neutral lipids (triglycerides) and dissimilar grades of inflammation in sebocytes.

Fatty Acid Profile and Genetic Variants of Proteins Involved in Fatty Acid Metabolism Could Be Considered as Disease Predictor.

Circulating fatty acids (FA) have an endogenous or exogenous origin and are metabolized under the effect of many enzymes. They play crucial roles in many mechanisms: cell signaling, modulation of gene expression, etc., which leads to the hypothesis that their perturbation could be the cause of disease development. FA in erythrocytes and plasma rather than dietary FA could be used as a biomarker for many diseases. Cardiovascular disease was associated with elevated trans FA and decreased DHA and EPA. Increased arachidonic acid and decreased Docosahexaenoic Acids (DHA) were associated with Alzheimer's disease. Low Arachidonic acid and DHA are associated with neonatal morbidities and mortality. Decreased saturated fatty acids (SFA), increased monounsaturated FA (MUFA) and polyunsaturated FA (PUFA) (C18:2 n-6 and C20:3 n-6) are associated with cancer. Additionally, genetic polymorphisms in genes coding for enzymes implicated in FA metabolism are associated with disease development. FA desaturase (FADS1 and FADS2) polymorphisms are associated with Alzheimer's disease, Acute Coronary Syndrome, Autism spectrum disorder and obesity. Polymorphisms in FA elongase (ELOVL2) are associated with Alzheimer's disease, Autism spectrum disorder and obesity. FA-binding protein polymorphism is associated with dyslipidemia, type 2 diabetes, metabolic syndrome, obesity, hypertension, non-alcoholic fatty liver disease, peripheral atherosclerosis combined with type 2 diabetes and polycystic ovary syndrome. Acetyl-coenzyme A carboxylase polymorphisms are associated with diabetes, obesity and diabetic nephropathy. FA profile and genetic variants of proteins implicated in FA metabolism could be considered as disease biomarkers and may help with the prevention and management of diseases.

Identification of Key Lipogenesis Stages and Proteins Involved in Walnut Kernel Development.

Walnuts are abundant in oil content, especially for polyunsaturated fatty acids, but the understanding of their formation is limited. We collected walnut (Juglans regia L.) kernels at 60, 74, 88, 102, 116, 130, and 144 days after pollination (designated S1-S7). The ultrastructure and accumulation of oil bodies (OBs) were observed using transmission electron microscopy (TEM), and the oil content, fatty acid composition, and proteomic changes in walnut kernels were determined. The oil content and OB accumulation increased during the development and rose sharply from S1 to S3 stages, which are considered the key lipogenesis stage. A total of 5442 proteins were identified and determined as differentially expressed proteins (DEPs) using label-free proteomic analysis. Fatty acid desaturases (FAD) 2, FAD3, oleosin, and caleosin were essential and upregulated from the S1 to S3 stages. Furthermore, the highly expressed oleosin gene JrOLE14.7 from walnuts was cloned and overexpressed in transgenic Brassica napus. The overexpression of JrOLE14.7 increased the oil content, diameter, hundred weight of seeds and changed the fatty acid composition and OB size of Brassica napus seeds. These findings provide insights into the molecular mechanism of oil biosynthesis and the basis for the genetic improvement of walnuts.

Engineered human hepatocyte organoids enable CRISPR-based target discovery and drug screening for steatosis

The lack of registered drugs for nonalcoholic fatty liver disease (NAFLD) is partly due to the paucity of human-relevant models for target discovery and compound screening. Here we use human fetal hepatocyte organoids to model the first stage of NAFLD, steatosis, representing three different triggers: free fatty acid loading, interindividual genetic variability (PNPLA3 I148M) and monogenic lipid disorders (APOB and MTTP mutations). Screening of drug candidates revealed compounds effective at resolving steatosis. Mechanistic evaluation of effective drugs uncovered repression of de novo lipogenesis as the convergent molecular pathway. We present FatTracer, a CRISPR screening platform to identify steatosis modulators and putative targets using APOB−/− and MTTP−/− organoids. From a screen targeting 35 genes implicated in lipid metabolism and/or NAFLD risk, FADS2 (fatty acid desaturase 2) emerged as an important determinant of hepatic steatosis. Enhancement of FADS2 expression increases polyunsaturated fatty acid abundancy which, in turn, reduces de novo lipogenesis. These organoid models facilitate study of steatosis etiology and drug targets.