Lipid Pathways Research Articles

Holistic transcriptional responses of Cordyceps militaris to different culture temperatures

Cordyceps militaris is a medicinal entomopathogenic fungus containing valuable biometabolites for pharmaceutical applications. Its genetic inheritance and environmental factors play a crucial role in the production of biomass enriched with cordycepin. While temperature is a crucial controlled parameter for fungal cultivation, its impacts on growth and metabolite biosynthesis remains poorly characterized. This study aimed to investigate the metabolic responses and cordycepin production of C. militaris strain TBRC6039 under various temperature conditions through transcriptome analysis. Among 9599 expressed genes, 576 genes were significantly differentially expressed at culture temperatures of 15 and 25 °C. The changes in the transcriptional responses induced by these temperatures were found in several metabolisms involved in nutrient assimilation and energy source, including amino acids metabolism (e.g., glycine, serine and threonine metabolism) and lipid metabolism (e.g., biosynthesis of unsaturated fatty acids and steroid biosynthesis). At the lower temperature (15 °C), the biosynthetic pathways of lipids, specifically ergosterol and squalene, were the target for maintaining membrane function by transcriptional upregulation. Our study revealed the responsive mechanisms of C. militaris in acclimatization to temperature conditions that provide an insight on physiological manipulation for the production of metabolites by C. militaris.

Research in revealing the effects on Cuscuta chinensis to diarrhea type irritable bowel syndrome based on network pharmacology and molecular docking potential mechanism.

To explore the potential mechanism in Cuscuta sinensis on diarrhea-type irritable bowel syndrome using network pharmacology and molecular docking techniques. First, the active components and related targets of Cuscuta were found setting oral utilization >30% and drug-like properties greater than or equal to 0.18 as filter information from TCMSP database. The targets of diarrheal irritable bowel syndrome were compiled by searching DrugBank, GeneCards, OMIM, PharmGkb, and TTD databases. The intersections of drugs and targets related to the disease were taken for gene ontology enrichment and Kyoto encyclopedia of genes and genomes enrichment analyses, to elucidate the potential molecular mechanisms and pathway information of Cuscuta sinensis for the treatment of diarrheal irritable bowel syndrome. The protein-protein interaction network was constructed by using the STRING database and visualized with Cytoscape_v3.10.0 software to find the protein-protein interaction network core At last, molecular docking was performed to validate the combination of active compounds with the core target. The target information of Cuscuta and diarrhea-type irritable bowel syndrome was compiled, which can be resulted in 11 active compounds such as quercetin, kaempferol, isorhamnetin, β-sitosterol, and another 17 core targets such as TP53, IL6, AKT1, IL1B, TNF, EGFR, etc, whose Kyoto encyclopedia of genes and genomes was enriched in the pathways of lipids and atherosclerosis, chemical carcinogenesis-receptor activation, PI3K-Akt signaling pathway, and fluid shear stress and atherosclerosis, etc. Docking demonstrated that the core targets and the active compounds were able to be better combined. Cuscuta chinensis may exert preventive effects on diarrhea-type irritable bowel syndrome by reducing intestinal inflammation, protecting intestinal mucosa, and playing an important role in antioxidant response through multi-targets and multi-pathways.

Integrative metabolomics and gut microbiota analyses reveal the protective effects of DHA-enriched phosphatidylserine on bisphenol A-induced intestinal damage

The protective effects of docosahexaenoic acid-enriched phosphatidylserine (DHA-PS, 50 or 100 mg/kg) on bisphenol A (BPA)-induced intestinal damage were assessed. The biochemical indices, pathological examination, non-targeted metabolomics integrated with gut microbiota analysis, and immunofluorescence analysis were used to investigate the protective effects of DHA-PS and its underlying regulatory mechanism. The DHA-PS treatment improved the pathology of intestinal tract and increased the expression levels of Claudin-1, Occludin, and ZO-1. In addition, the DHA-PS treatment also notably decreased the levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α, and increased the antioxidant enzymes activities. Moreover, DHA-PS treatment increased the relative abundances of Akkermansia, Alistipes, Butyricicoccus, Coriobacteriaceae_UCG-002, Enterorhabdus, and Lachnospiraceae_UCG-006. DHA-PS treatment alleviated BPA-induced intestinal damage by regulating the kynurenine pathway of tryptophan, lipid, and arachidonic acid metabolisms. Overall, this study suggested that DHA-PS could alleviate BPA-induced intestinal damage by enhancing the intestinal barrier integrity, improving gut microbial composition and metabolites, and inhibiting the TLR4/NF-κB pathways.

Lipidomic signatures in Colombian adults with metabolic syndrome

Background and AimsMetabolic syndrome (MetS) comprises a set of risk factors that contribute to the development of chronic and cardiovascular diseases, increasing the mortality rate. Altered lipid metabolism is associated with the development of metabolic disorders such as insulin resistance, obesity, atherosclerosis, and metabolic syndrome; however, there is a lack of knowledge about lipids compounds and the lipidic pathways associated with this condition, particularly in the Latin-American population. Innovative approaches, such as lipidomic analysis, facilitate the identification of lipid species related to these risk factors. This study aimed to assess the plasma lipidome in subjects with MetS.MethodsThis correlation study included healthy adults and adults with MetS. Blood samples were analyzed. The lipidomic profile was determined using an Agilent Technologies 1260 liquid chromatography system coupled to a Q-TOF 6545 quadrupole mass analyzer with electrospray ionization. The main differences were determined between the groups.ResultsThe analyses reveal a distinct lipidomic profile between healthy adults and those with MetS, including increased concentrations of most identified glycerolipids -both triglycerides and diglycerides- and decreased levels of ether lipids and sphingolipids, especially sphingomyelins, in MetS subjects. Association between high triglycerides, waist circumference, and most differentially expressed lipids were found.ConclusionOur results demonstrate dysregulation of lipid metabolism in subjects with Mets, supporting the potential utility of plasma lipidome analysis for a deeper understanding of MetS pathophysiology.

Serum and urine lipidomic profiles identify biomarkers diagnostic for seropositive and seronegative rheumatoid arthritis.

Seronegative rheumatoid arthritis (RA) is defined as RA without circulating autoantibodies such as rheumatoid factor and anti-citrullinated protein antibodies; thus, early diagnosis of seronegative RA can be challenging. Here, we aimed to identify diagnostic biomarkers for seronegative RA by performing lipidomic analyses of sera and urine samples from patients with RA. We performed untargeted lipidomic analysis of sera and urine samples from 111 RA patients, 45 osteoarthritis (OA) patients, and 25 healthy controls (HC). These samples were divided into a discovery cohort (n = 97) and a validation cohort (n = 84). Serum samples from 20 patients with systemic lupus erythematosus (SLE) were also used for validation. The serum lipidome profile of RA was distinguishable from that of OA and HC. We identified a panel of ten serum lipids and three urine lipids in the discovery cohort that showed the most significant differences. These were deemed potential lipid biomarker candidates for RA. The serum lipid panel was tested using a validation cohort; the results revealed an accuracy of 79%, a sensitivity of 71%, and a specificity of 86%. Both seropositive and seronegative RA patients were differentiated from patients with OA, SLE, and HC. Three urinary lipids showing differential expression between RA from HC were identified with an accuracy of 84%, but they failed to differentiate RA from OA. There were five lipid pathways that differed between seronegative and seropositive RA. Here, we identified a panel of ten serum lipids as potential biomarkers that can differentiate RA from OA and SLE, regardless of seropositivity. In addition, three urinary lipids had diagnostic utility for differentiating RA from HC.

Impact of the Oral Administration of Polystyrene Microplastics on Hepatic Lipid, Glucose, and Amino Acid Metabolism in C57BL/6Korl and C57BL/6-Lepem1hwl/Korl Mice.

The impact of microplastics (MPs) on the metabolic functions of the liver is currently unclear and not completely understood. To investigate the effects of the administration of MPs on the hepatic metabolism of normal and obese mice, alterations in the lipid, glucose (Glu), and amino acid regulation pathways were analyzed in the liver and adipose tissues of C57BL/6Korl (wild type, WT) or C57BL/6-Lepem1hwl/Korl mice (leptin knockout, Lep KO) orally administered polystyrene (PS) MPs for 9 weeks. Significant alterations in the lipid accumulation, adipogenesis, lipogenesis, and lipolysis pathways were detected in the liver tissue of MP-treated WT and Lep KO mice compared to the vehicle-treated group. These alterations in their liver tissues were accompanied by an upregulation of the serum lipid profile, as well as alterations in the adipogenesis, lipogenesis, and lipolysis pathways in the adipose tissues of MP-treated WT and Lep KO mice. Specifically, the level of leptin was increased in the adipose tissues of MP-treated WT mice without any change in their food intake. Also, MP-induced disruptions in the glycogenolysis, Glu transporter type 4 (GLUT4)-5' AMP-activated protein kinase (AMPK) signaling pathway, levels of lipid intermediates, and the insulin resistance of the liver tissues of WT and Lep KO mice were observed. Furthermore, the levels of seven endogenous metabolites were remarkably changed in the serum of WT and Lep KO mice after MP administrations. Finally, the impact of the MP administration observed in both types of mice was further verified in differentiated 3T3-L1 adipocytes and HepG2 cells. Thus, these results suggest that the oral administration of MPs for 9 weeks may be associated with the disruption of lipid, Glu, and amino acid metabolism in the liver tissue of obese WT and Lep KO mice.

Mechanisms of low nighttime temperature promote oil accumulation in Brassica napus L. based on in-depth transcriptome analysis.

Rape (Brassica napus L.; AACC) is an important oil-bearing crop worldwide. Temperature significantly affects the production of oil crops; however, the mechanisms underlying temperature-promoted oil biosynthesis remain largely unknown. In this study, we found that a temperature-sensitive cultivar (O) could accumulate higher seed oil content under low nighttime temperatures (LNT,13°C) compared with a temperature-insensitive cultivar (S). We performed an in-depth transcriptome analysis of seeds from both cultivars grown under different nighttime temperatures. We found that low nighttime temperatures induced significant changes in the transcription patterns in the seeds of both cultivars. In contrast, the expression of genes associated with fatty acid and lipid pathways was higher in the O cultivar than in the S cultivar under low nighttime temperatures. Among these genes, we identified 14 genes associated with oil production, especially BnLPP and ACAA1, which were remarkably upregulated in the O cultivar in response to low nighttime temperatures compared to S. Further, a WGCNA analysis and qRT-PCR verification revealed that these genes were mainly regulated by five transcription factors, WRKY20, MYB86, bHLH144, bHLH95, and NAC12, whose expression was also increased in O compared to S under LNT. These results allowed the elucidation of the probable molecular mechanism of oil accumulation under LNT conditions in the O cultivar. Subsequent biochemical assays verified that BnMYB86 transcriptionally activated BnLPP expression, contributing to oil accumulation. Meanwhile, at LNT, the expression levels of these genes in the O plants were higher than at high nighttime temperatures, DEGs (SUT, PGK, PK, GPDH, ACCase, SAD, KAS II, LACS, FAD2, FAD3, KCS, KAR, ECR, GPAT, LPAAT, PAP, DGAT, STERO) related to lipid biosynthesis were also upregulated, most of which are used in oil accumulation.

Using Metabolomics to Understand Selective Breeding of Goats that Enables Tolerance of Plant Secondary Compounds

The objective of this study was to determine aspects of liver function in goats with a divergent propensity to consume juniper ( Juniperus spp.), a terpene -containing woody plant. Goats represent a significant contribution to sustainable food production and local economies worldwide. Climate change in arid and semiarid regions has led to a transition of grasslands to woodlands. Goats ( Capra hircus) are adapted to utilize woody plants as a food source; however, herbivory can be limited by plant secondary compounds (e.g., monoterpenes). Estimated breeding values (EBV) have been determined for domestic goats based on their propensity to consume above (H: EBV = 13.1 ± 0.2) or below (L: EBV = −14.8 ± 0.5) the herd average for proportion of juniper in the diet of free-ranging animals. To investigate the physiological and metabolic mechanisms enabling increased intake and tolerance of juniper, H and L adult male Spanish goats within young (2-yr-old) and old (3-5-yr-old) groups that spent their life grazing pastures containing juniper were randomly selected from our research herd. Twenty goats (n = 5 of each age and juniper consumption combination) were weighed and harvested. Serum samples and multiple organs (i.e., liver, kidney, heart, spleen, adrenal, testis) were collected, weighed, and preserved for analyses. With an initial focus on the liver, samples (200 mg) were submitted to Metabolon, Inc. (Morrisville, NC) to identify metabolites by ultra-high-performance liquid chromatography/tandem high resolution/accurate mass spectrometry chromatography methods. Data were analyzed using the PROC MIXED procedure of Statistical Analysis System (SAS Institute Inc., Cary, NC). Goat body weight (BW), relative organ weight (g/kg BW), and serum constituents were analyzed using a model that included age (O, Y), treatment (H, L), and the age x treatment interaction. Animals were the experimental unit and error degrees of freedom = 16. Statistical significance was determined at P ≤ 0.05. Multiple differences in metabolites of the lipid and protein pathways were identified. For example, the relative intensity of 19 monoacylglycerol metabolites was greater (P < 0.05) in H than L goats. This report, however, focuses on the gamma-glutamyl amino acid (GGAA) pathway due to its role in diagnosis of liver dysfunction. Relative liver weight (g/kg BW) was numerically greater (P = 0.15) in H (15.80 ± 0.75) than L (14.58 ± 0.39) goats. Serum gamma glutamyl transferase (U/L) was within the clinically normal range but was greater (P < 0.0001) in H (59.2 ± 2.1) versus L (44.3 ± 2.2), and in Y (55.3 ± 2.9) versus O (48.2 ± 3.2) goats. Of the 18 GGAAs identified from the GGAA super- and sub-pathways, the relative intensities of 11 metabolites were greater (P < 0.05) in liver from H than L; 8 were greater in HY versus LY, and only 2 were greater in OH versus OL goats. The increased expression of serum and liver components of the GGAA pathway for the goats with a positive estimated breeding value for juniper consumption may be indicative of metabolic mechanisms and biomarkers associated with increased tolerance of plant secondary compounds. Texas A&M AgriLife Research. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Microglia as a Target of Immunomodulation in Pediatric Autoimmune Neuropsychiatric Disorders associated with Streptococcal Infections (PANDAS)

PANDAS, or Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcal Infections, is a neuropsychiatric condition characterized by the abrupt onset of diverse neuropsychiatric manifestations following streptococcal infections in children. It's believed to involve immune dysregulation and autoimmune targeting of the brain basal ganglia. However, the precise mechanisms remain unclear. Hypothesis: considering the role of microglia in neuroinflammation, evidence for their activation in PET imaging of PANDAS patients, and increased number of CD68+/Iba1+ activated microglia in mice inoculated with group A Streptococcus (GAS), we hypothesized that as a consequence of disrupted blood-brain barrier (BBB), circulating factors modulate microglial functions towards a pro-inflammatory phenotype. Methods: Untargeted metabolomics was performed with sera from 10 children with PANDAS and 12 healthy controls. Human iPSC-derived microglia were generated and exposed to 5% serum from PANDAS patients or healthy controls in the presence or absence of Polymyxin B (PMB; endotoxin scavenger) and assessed for nitric oxide (NOx) production, immunophenotyped with flow cytometry (TMEM119, TREM2, CD68, and Iba-1) and bulk RNA-seq analysis. Results: Significant differences in serum metabolomic profile of PANDAS cases were observed, particularly in pathways associated with amino acid, fatty acid, lipid, steroid, and oxidative stress-related pathways. Treatment of iPSC-derived or primary human microglia with PANDAS serum led to a significant increase in NOx production, which was blunted by approx. 30% by PMB. Immunophenotyping revealed a significant reduction in homeostatic markers TMEM119 and TREM2, and expansion of CD68+ and Iba-1+ microglia associated with inflammatory response. Regulation of TMEM119 and TREM2 was endotoxin-independent, while PMB partially blunted the increase in CD68 and Iba-1. RNAseq revealed dramatic effects of PANDAS sera on microglial gene expression pattern, with upregulated genes associated with C-type lectin receptor, TLR, TNF, and NFk-B signaling pathways among others. Conclusion: The results suggest that as a result of BBB disruption in PANDAS, associated systemic inflammatory mediators modulate the phenotype and function of the brain microglia, a mechanism that likely contributes to neuroinflammation and the observed symptoms. The observed contribution of endotoxin may also suggest a dysfunction of intestinal barrier which may compound the systemic immune responses in PANDAS. Arizona Department of Health Services RFGA2022-010-23 (PK) & ADHS17-00007403 (SR, MD); The Alex Manfull Fund (PK). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Effects of gut bacterial community on differential growth of sea cucumber (Apostichopus japonicus): Molecular mechanisms based on host-gut microbiome co-metabolism

Gut microbiome and their metabolites are increasingly recognized for their pivotal role in regulating the health and growth of the host. The mechanism by which the host-gut microbiome co-metabolism affects the growth rate of sea cucumber (Apostichopus japonicus) remains unclear. In this study, the intestinal wall tissues of fast-growing (FG) and slow-growing (SG) A. japonicus were collected from the full-sib family for 16S amplicon sequencing, metagenomics, metabolomics, and transcriptomics analyses. The results showed a marked variation in the gut bacterial community of A. japonicus with different growth rates. Compared with SG group, FG A. japonicus had a higher abundance of Bacteroidetes. Many of these bacteria were associated with the degradation of polysaccharides, which could potentially affect growth performance. The abundance of Proteobacteria, however, was found to be higher in SG A. japonicus, thereby increasing the potential risk of disease. The metabolome results were further supported by the predicated KEGG functions of the gut microbiomes, lots of metabolites related to the metabolism and biosynthesis of fructose 6-phosphate, arachidonic acid, oleic acid, and dihydroxyacetone phosphate were upregulated in the FG A. japonicus. Furthermore, the genes (IGF1, HK, PK, PCK1) were significantly upregulated in the FG A. japonicus, which were mainly enriched in the metabolic pathways of lipids, amino acids, and carbohydrates. Gut bacterial community via their unique metabolic pathways, which in turn affected the metabolic phenotypes of the organism and eventually led to differences in growth performance of A. japonicus. These findings might help understand the molecular mechanisms of growth gap in sea cucumber, thereby increasing the mariculture production.

Metabolism of Inflammatory T helper 17 cells from Type 2 Diabetic Patients

Abstract Around 90% of the 537 million global diabetic adults suffer from type 2 diabetes (T2D), a metabolic disorder marked by heightened blood glucose due to insulin resistance or inadequate insulin production. T2D onset involves chronic systemic inflammation, predominantly influenced by T-helper 17 (Th17) cells. In T2D patients, defective fatty acid metabolic pathways contribute to Th17-driven inflammation. While lipid metabolism's role is established in in vitro Th17 cells, in vivo differentiated Th17 cells remain underexplored due to their low abundance. We addressed this challenge with a novel Th17 cell enrichment assay, capturing CD4+ T cells and magnetically purifying them based on IL-17A/17F/21 secretion for scRNAseq. From this transcriptomic data we established that the subsets of Th17 cells found in mice - pathogenic and non-pathogenic, are also present in humans, and that in vivo differentiated Th17 cells have a unique lipid metabolic phenotype from in vitro differentiated Th17 cells. We also characterized the metabolic state of these Th17 cells by utilizing the metabolic modeling algorithm Compass. Overlaying scRNAseq data from diabetic and control patients, we identified key lipid pathways driving Th17 inflammation in T2D. This pioneering study on endogenous Th17 cells in healthy and T2D patients, coupled with functional validation, unveils potential targets for developing immunotherapy for T2D.

JAK/STAT Inhibition Normalizes Lipid Composition in 3D Human Epidermal Equivalents Challenged with Th2 Cytokines.

Derangement of the epidermal barrier lipids and dysregulated immune responses are key pathogenic features of atopic dermatitis (AD). The Th2-type cytokines interleukin IL-4 and IL-13 play a prominent role in AD by activating the Janus Kinase/Signal Transduction and Activator of Transcription (JAK/STAT) intracellular signaling axis. This study aimed to investigate the role of JAK/STAT in the lipid perturbations induced by Th2 signaling in 3D epidermal equivalents. Tofacitinib, a low-molecular-mass JAK inhibitor, was used to screen for JAK/STAT-mediated deregulation of lipid metabolism. Th2 cytokines decreased the expression of elongases 1, 3, and 4 and serine-palmitoyl-transferase and increased that of sphingolipid delta(4)-desaturase and carbonic anhydrase 2. Th2 cytokines inhibited the synthesis of palmitoleic acid and caused depletion of triglycerides, in association with altered phosphatidylcholine profiles and fatty acid (FA) metabolism. Overall, the ceramide profiles were minimally affected. Except for most sphingolipids and very-long-chain FAs, the effects of Th2 on lipid pathways were reversed by co-treatment with tofacitinib. An increase in the mRNA levels of CPT1A and ACAT1, reduced by tofacitinib, suggests that Th2 cytokines promote FA beta-oxidation. In conclusion, pharmacological inhibition of JAK/STAT activation prevents the lipid disruption caused by the halted homeostasis of FA metabolism.

Multi-tissue profiling of oxylipins reveal a conserved up-regulation of epoxide:diol ratio that associates with white adipose tissue inflammation and liver steatosis in obesity

BackgroundObesity drives maladaptive changes in the white adipose tissue (WAT) which can progressively cause insulin resistance, type 2 diabetes mellitus (T2DM) and metabolic dysfunction-associated liver disease (MASLD). Obesity-mediated loss of WAT homeostasis can trigger liver steatosis through dysregulated lipid pathways such as those related to polyunsaturated fatty acid (PUFA)-derived oxylipins. However, the exact relationship between oxylipins and metabolic syndrome remains elusive and cross-tissue dynamics of oxylipins are ill-defined. MethodsWe quantified PUFA-related oxylipin species in the omental WAT, liver biopsies and plasma of 88 patients undergoing bariatric surgery (female N=79) and 9 patients (female N=4) undergoing upper gastrointestinal surgery, using UPLC-MS/MS. We integrated oxylipin abundance with WAT phenotypes (adipogenesis, adipocyte hypertrophy, macrophage infiltration, type I and VI collagen remodelling) and the severity of MASLD (steatosis, inflammation, fibrosis) quantified in each biopsy. The integrative analysis was subjected to (i) adjustment for known risk factors and, (ii) control for potential drug-effects through UPLC-MS/MS analysis of metformin-treated fat explants ex vivo. FindingsWe reveal a generalized down-regulation of cytochrome P450 (CYP)-derived diols during obesity conserved between the WAT and plasma. Notably, epoxide:diol ratio, indicative of soluble epoxide hydrolyse (sEH) activity, increases with WAT inflammation/fibrosis and hepatic steatosis and T2DM. Increased 12,13-EpOME:DiHOME in WAT and liver is a marker of worsening metabolic syndrome in patients with obesity. InterpretationThese findings suggest a dampened sEH activity and a possible role of fatty acid diols during metabolic syndrome in major metabolic organs such as WAT and liver. They also have implications in view of the clinical trials based on sEH inhibition for metabolic syndrome. FundingWellcome Trust (PS3431_WMIH); Duke-NUS (Intramural Goh Cardiovascular Research Award (Duke-NUS-GCR/2022/0020); National Medical Research Council (OFLCG22may-0011); National Institute of Environmental Health Sciences (Z01 ES025034); NIHR-Imperial Biomedical Research Centre

Lipidomics Analysis Unravels Aberrant Lipid Species and Pathways Induced by Zinc Oxide Nanoparticles in Kidney Cells.

Zinc oxide nanoparticles (ZnO NPs) are widely used in versatile applications, from high technology to household products. While numerous studies have examined the toxic gene profile of ZnO NPs across various tissues, the specific lipid species associated with adverse effects and potential biomarkers remain elusive. In this study, we conducted a liquid chromatography-mass spectrometry based lipidomics analysis to uncover potential lipid biomarkers in human kidney cells following treatment with ZnO NPs. Furthermore, we employed lipid pathway enrichment analysis (LIPEA) to elucidate altered lipid-related signaling pathways. Our results demonstrate that ZnO NPs induce cytotoxicity in renal epithelial cells and modulate lipid species; we identified 64 lipids with a fold change (FC) > 2 and p < 0.01 with corrected p < 0.05 in HK2 cells post-treatment with ZnO NPs. Notably, the altered lipids between control HK2 cells and those treated with ZnO NPs were associated with the sphingolipid, autophagy, and glycerophospholipid pathways. This study unveils novel potential lipid biomarkers of ZnO NP nanotoxicity, representing the first lipidomic profiling of ZnO NPs in human renal epithelial cells.

Dynamics of the Trypanosoma cruzi infection in adipose tissue: Assessing gene expression of PNPLA2, FASN, and ACAT1 under Benzonidazole treatment and indirect mononuclear immune cells interaction

Trypanosoma cruzi is a parasite with a high capacity to adapt to the host. Animal models have already demonstrated that the tropism of this parasite occurs not only in cardiac/digestive tissues but also in adipose tissue (AT). That said, the consequences ofT. cruziinfection for AT and the implications of treatment with Benzonidazole in this tissue are under discussion. Here, we tested the hypothesis that T. cruzi infection in adipose tissue upon treatment with Benzonidazole (Bz) and the interaction of mononuclear immune cells (PBMC) influences the relative expression of ACAT1, FASN, and PNPLA2 genes. Thus, stem cells derived from adipose tissue (ADSC) after adipogenic differentiation were indirectly cultivated with PBMC after infection with the T. cruzi Y strain and treatment with Bz. We use the TcSAT-IAM system and RT-qPCR to evaluate the parasite load and the relative quantification (ΔCt) of the ACAT1, FASN, and PNPLA2 genes. Our results demonstrate that treatment with Bz did not reduce adipocyte infection in the presence (p-value: 0.5796) or absence (p-value: 0.1854) of cultivation with PBMC. In addition, even though there is no statistical difference when compared to the control group (AT), T. cruzi induces the FASN expression (Rq: 14.00). However, treatment with Bz in AT suggests the increases of PNPLA2 expression levels (Rq: 12.58), even in the absence of T. cruzi infection. During indirect cultivation with PBMC, T. cruzi smooths the expression of PNPLA2 (Rq: 0.824) and instigates the expression of ACAT1 (Rq: 1.632) and FASN (Rq: 1.394). Furthermore, the treatment with Bz during infection induces PNPLA2 expression (Rq: 1.871), maintaining FASN expression levels (Rq: 1.334). Given this, our results indicate that treatment with Benzonidazole did not decrease T. cruzi infection in adipose tissue. However, treating the adipocyte cells with Bz during the interaction with PBMC cells influences the lipid pathways scenario, inducing lipolytic metabolism through the expression of PNPLA2.

Relative expression of genes related to volatile organic compounds in non-climacteric and climacteric melons

Melon (Cucumis melo L.) is an important species in the cucurbit family with a large economic importance in the world. Two melon cultivars commercially important in Brazil are ‘Yellow’ and ‘Gaúcho’. In addition to their economic importance, these two cultivars display phenotypic differences in aroma, a major trait determining fruit quality. Volatile organic compounds (VOCs) impart the different aroma found in this fruit and their biosynthesis is associated with fatty acid and amino acid metabolism. Using RT-qPCR techniques, the expression of seven genes (CmLOX9, CmLOX18, CmBCAT1, CmArAT1, CmPDC1, CmADH1, and CmAAT1) was determined during ripening. The lipid pathway played a strong role in determining aroma composition in non-climacteric ‘Yellow’ melons. Most volatiles decreased during ripening, explaining the non-aromatic characteristic of this cultivar. In climacteric ‘Gaúcho’ melons, the amino acid pathway was the main one related to the biosynthesis of esters, which contribute to the aroma of this cultivar. Volatile products of the branched-chain amino acid pathway correlated with CmADH1 and CmAAT1 expression, demonstrating their role in volatile synthesis in this climacteric melon cultivar. In addition, CmPDC1 contributes to the formation of aldehydes at the beginning of this pathway.

Human Airway Organoids and Multimodal Imaging-Based Toxicity Evaluation of 1-Nitropyrene.

Despite significant advances in understanding the general health impacts of air pollution, the toxic effects of air pollution on cells in the human respiratory tract are still elusive. A robust, biologically relevant in vitro model for recapitulating the physiological response of the human airway is needed to obtain a thorough understanding of the molecular mechanisms of air pollutants. In this study, by using 1-nitropyrene (1-NP) as a proof-of-concept, we demonstrate the effectiveness and reliability of evaluating environmental pollutants in physiologically active human airway organoids. Multimodal imaging tools, including live cell imaging, fluorescence microscopy, and MALDI-mass spectrometry imaging (MSI), were implemented to evaluate the cytotoxicity of 1-NP for airway organoids. In addition, lipidomic alterations upon 1-NP treatment were quantitatively analyzed by nontargeted lipidomics. 1-NP exposure was found to be associated with the overproduction of reactive oxygen species (ROS), and dysregulation of lipid pathways, including the SM-Cer conversion, as well as cardiolipin in our organoids. Compared with that of cell lines, a higher tolerance of 1-NP toxicity was observed in the human airway organoids, which might reflect a more physiologically relevant response in the native airway epithelium. Collectively, we have established a novel system for evaluating and investigating molecular mechanisms of environmental pollutants in the human airways via the combinatory use of human airway organoids, multimodal imaging analysis, and MS-based analyses.

Identification of Plasma Exosomes hsa_circ_0001360 and hsa_circ_0000038 as Key Biomarkers of Coronary Heart Disease.

Coronary heart disease (CHD) is the leading cause of death and disability worldwide. Accumulating evidence reveals that atherosclerosis (AS), characterized by systemic, chronic, and multifocal disease, and is the primary pathological basis of cardiovascular diseases, including CHD. However, the molecular underpinnings of CHD are still far from well understood. Our study attempted to identify aberrant plasma exosome-derived circRNAs and key exosomal circRNA biomarkers for CHD. The expression profiles of mRNAs, circRNAs, and lncRNAs in the blood exosomes of CHD patients and healthy controls were obtained from the exoRBase database. The corresponding miRNAs of the differentially expressed mRNAs, circRNAs, and lncRNAs were predicted via ENCORI and the miRcode database. LncRNAs/circRNAs and mRNAs with the cotargeted miRNAs were selected to construct an interaction network. Multiple machine learning algorithms have been used to explore potential biomarkers, followed by verification in patients with CHD using real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR). Based on the cutoff criterion of P < 0.05, we identified 85 differentially expressed circRNAs (4 upregulated and 81 downregulated), 43 differentially expressed lncRNAs (24 upregulated and 19 downregulated), and 312 differentially expressed mRNAs (55 upregulated and 257 downregulated). Functional enrichment analysis revealed that the differentially expressed mRNAs were involved mainly in neutrophil extracellular trap (NET) formation and the nucleotide-binding oligomerization domain- (NOD-) like receptor signaling pathway. Further analysis revealed that the DEGs in the circRNA/lncRNA-miRNA-mRNA interaction network were closely related to lipid and atherosclerotic signaling pathways. Hsa_circ_0001360 and hsa_circ_0000038 were identified as potential biomarkers for CHD based on three machine learning algorithms. The relative expression levels of hsa_circ_0001360 and hsa_circ_0000038 were significantly altered in plasma exosomes from patients with CHD. ROC curve analysis revealed that the areas under the curve (AUCs) were 0.860, 0.870, and 0.940 for hsa_circ_0001360, hsa_circ_0000038, and the two-gene combination, respectively. The circRNA/lncRNA-miRNA-mRNA interaction network might help to elucidate the pathogenesis of CHD. Hsa_circ_0001360 combined with hsa_circ_0000038 might be an important diagnostic biomarker.

Metabolomic Profiles and Pathways in Osteoarthritic Human Cartilage: A Comparative Analysis with Healthy Cartilage.

Osteoarthritis (OA) is a chronic joint disease with heterogenous metabolic pathology. To gain insight into OA-related metabolism, metabolite extracts from healthy (n = 11) and end-stage osteoarthritic cartilage (n = 35) were analyzed using liquid chromatography-mass spectrometry metabolomic profiling. Specific metabolites and metabolic pathways, including lipid and amino acid pathways, were differentially regulated in osteoarthritis-derived and healthy cartilage. The detected alterations in amino acids and lipids highlighted key differences in bioenergetic resources, matrix homeostasis, and mitochondrial alterations in OA-derived cartilage compared to healthy cartilage. Moreover, the metabolomic profiles of osteoarthritic cartilage separated into four distinct endotypes, highlighting the heterogenous nature of OA metabolism and the diverse landscape within the joint in patients. The results of this study demonstrate that human cartilage has distinct metabolomic profiles in healthy and end-stage OA patients. By taking a comprehensive approach to assess metabolic differences between healthy and osteoarthritic cartilage and within osteoarthritic cartilage alone, several metabolic pathways with distinct regulation patterns were detected. Additional investigation may lead to the identification of metabolites that may serve as valuable indicators of disease status or potential therapeutic targets.

Abstract 4448: Effects of adagrasib on cholesterol, lipid and glucose gene expression regulation in tumor xenograft models and patient samples

Abstract Introduction: Clinically active KRASG12C inhibitors have represented a key research breakthrough and led to novel treatment options for lung, colorectal and other cancer patients harboring this mutation. While treatment with KRASG12C inhibitors are clinically active in the majority of patients, the depth and duration of response is variable and most patients ultimately progress. This variation in therapeutic response highlights the need for a better understanding of drug mechanism of action and the identification of rational combination strategies. Oncogenic KRAS mutations hyperactivate the MAPK pathway, create an immunosuppressive tumor microenvironment, and promote glycolysis even in the presence of oxygen commonly known as the Warburg effect. This altered metabolic state is marked by increased glucose uptake and lactate production, providing necessary substrates for rapid tumor growth. Results: Pharmacological inhibition of KRASG12C with a potent and selective KRASG12C inhibitor, adagrasib, revealed marked alterations of metabolic gene expression programs in tumor samples collected from mouse xenograft studies and repeat biopsies provided by patients enrolled on adagrasib clinical trials. Upregulation of genes involved in reverse cholesterol transport including ApoE, NR1H3 (LXRa) and ABCA1 was observed across both preclinical tumor xenografts and patient samples following adagrasib treatment. Additionally, decreased expression of Glut1 and low-density lipoprotein receptor (LDLR) genes were also observed in both tumor models and patients suggesting that adagrasib treatment results in an extensive shift in uptake and utilization of cholesterol, lipid, and glucose. The impact of adagrasib treatment on circulating metabolic parameters in tumor xenograft-bearing mice were investigated in follow up studies. Conclusions: Taken together, these data suggest adagrasib modulates cholesterol efflux, glucose and lipid pathways and alters tumor and systemic metabolic regulation. Additional research may yield a targetable collateral vulnerability and rational combinatorial strategy. Citation Format: Fabiola Shelton, Natalie Hoffman, David Trinh, Xousaen M. Helu, Andrew Calinisan, Allan Hebbert, Larry Yan, David M. Briere, Laura Hover, Julio Fernandez-Benet, Kenna Anderes, James G. Christensen, Jill Hallin, Peter Olson. Effects of adagrasib on cholesterol, lipid and glucose gene expression regulation in tumor xenograft models and patient samples [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4448.