Acylcarnitines Research Articles

Mitochondrial dysfunction: The pathological link between psoriasis and insulin resistance?

Psoriasis is strongly associated with insulin resistance (IR). Lipid profile disturbances and upregulation of enzymes crucial for fatty acid oxidation have been reported in patients with psoriasis. Mitochondrial ß-oxidation is altered in patients with IR. Common mitochondrial dysfunction may be involved in the origin of both diseases. This study aimed to evaluate mitochondrial ß-oxidation, intermediary metabolism, and mitochondrial content in psoriatic patients with or without IR and compare them to healthy controls. The participants were divided into three groups: (1) psoriasis and IR (n=26); (2) psoriasis without IR (n=17); and (3) healthy controls (n=17). Quantification of amino acids and acylcarnitines (AC) by tandem mass spectrometry, determination of urinary organic acids by gas chromatography/mass spectrometry (GC/MS), and mitochondrial DNA quantification were performed in all groups. When comparisons were made between the two psoriatic groups, no differences were found between: C5DC + C6OH, C16:1, Met/Leu, Met/Phe, C16:1/C16, and C5DC + C6OH/C4DC + C5OH ratios. Nine analytes were different: phenylalanine, Cit/Phe, and Cit/Tyr ratios, C0, C3, C5, C6DC, C16, and C18:1OH. There were no correlations between psoriasis area and severity index (PASI), body mass index (BMI) and duration of disease with ACs. A higher proportion of patients with psoriasis showed increased urine levels of uric acid and hippuric acid (p=0.01). The mtDNA content was significantly higher in cases than in controls, with no differences between IR and non-IR psoriatic patients. Psoriasis patients with and without IR have a different acylcarnitine profile reflecting impaired ß-oxidation. A distinctive profile of acylcarnitines suggests an involvement of mitochondrial function associated with an increase in stearoyl CoA desaturase (SCD) activity in psoriatic patients with and without IR.

A Marked Low-Grade Inflammation and a Significant Deterioration in Metabolic Status in First-Episode Schizophrenia: A Five-Year Follow-Up Study.

The objective of this study was to evaluate how schizophrenia spectrum disorders and applied long-term (5.1 years) antipsychotic (AP) treatment affect the serum level of acylcarnitines (ACs), cytokines and metabolic biomarkers and to characterize the dynamics of inflammatory and metabolic changes in the early course of the disorder. A total of 112 adults participated in the study (54 patients with first-episode psychosis (FEP) and 58 control subjects). Biomolecule profiles were measured at the onset of first-episode psychosis and 0.6 years and 5.1 years after the initiation of APs. The results of the present study confirmed that specific metabolic–inflammatory imbalance characterizes AP-naïve patients. Short-term (0.6-years) AP treatment has a favourable effect on psychotic symptoms, as well as the recovery of metabolic flexibility and resolution of low-level inflammation. However, 5.1 years of AP treatment resulted in weight gain and increased serum levels of interleukin (IL)-2, IL-4, IL-6, IL-10, interferon-γ, hexoses, acetylcarnitine, short-chain ACs (C3, C4) and long-chain ACs (C16:2, C18:1, C18:2). In conclusion, despite the improvement in psychotic symptoms, 5.1 years of AP treatment was accompanied by a pronounced metabolic–inflammatory imbalance, which was confirmed by the presence of enhanced pro-inflammatory activity and increased obesity with changes in the metabolism of carbohydrates, lipids, and their metabolites.

Examination of the regulation of energy metabolism, antioxidant response, and ammonia detoxification in hard clam, Mercenaria mercenaria, under hypersalinity stress

With global warming on the rise, hypersalinity events are occurring with increasing frequency due to accelerated evaporation rates of seawater. Hypersalinity affects the distribution, growth, and physiological processes of aquatic organisms, especially for osmoconforming bivalves that lack complete means of osmoregulation. In this way, knowledge concerning comprehensive metabolic regulation is limited in hypersalinity-stressed bivalves. The hard clam, Mercenaria mercenaria, is an important pond-farmed bivalve in China, with an annual output value of 150 million USD. This species can be farmed in salt pans because it is highly resistant to hypersalinity. In this study, we investigated the comprehensive pattern of metabolic regulation in hard clams under hypersalinity stress. We identified 979 metabolites in the gills of hard clams, 295 of which were differentially accumulated after different periods of exposure to hypersalinity (8 h, 1 day, and 5 days), compared with the control. Critical hypersalinity-responsive metabolites, including acyl carnitine, oxidized lipids, and several antioxidants were identified using K-means analysis. During hypersalinity exposure, the activities of hexokinase and pyruvate kinase, and the pyruvic acid content significantly increased, suggesting that glycolysis was enhanced. Alanine and glycine were identified as key free amino acids involved in osmoregulaion. After 5 days of hypersalinity exposure, the accumulation of alanine and lactic acid marked the initiation of anaerobic glycolysis. The up-regulation of amino acid catabolism and inhibition of ammonia excretion led to ammonia accumulation in the gills. Glutamine synthetase and glutamate dehydrogenase activities significantly increased to reduce ammonia toxicity. Many antioxidant metabolites, such as allantoin, cinnamic acid, and trigonelline, were dramatically up-regulated to relieve hypersalinity-induced oxidative stress. The accumulation of lysophosphatides suggested that cell membranes damaged by prolonged oxidative stress and/or hyperosmosis were dissolved by phospholipase A1. This process helped to maintain overall cellular homeostasis. Potential biomarkers of hypersalinity stress in hard clams were identified. These results will facilitate the assessment of the physiological status of hard clam in aquaculture and provide novel insights into the metabolic regulatory mechanisms of bivalves under hypersalinity stress.

Metabolic modulation as a common adaptive mechanism in patients with different subtypes of aortic valve stenosis

Abstract Background Aortic valve stenosis (AS) is a rapidly growing global health problem due to aging and population growth. If treatment by aortic valve replacement (AVR) is delayed, this leads to poor recovery of cardiac function and marked increase of mortality. In conditions of AS or heart failure a cardiac remodelling occurs including a change in cardiac substrate utilization. However, it is difficult to determine the stage of AS-induced cardiac remodelling and therefore the reversibility of this changes after AVR. As cardiac metabolism might be therapeutic target for patients not recovering after valve replacement a better understanding of the metabolic remodelling in different AS subtypes is necessary. Metabolomics constitute a potent screening tool for defining changes in global and cardiac-specific metabolism in cardiovascular disease. Methods/Results Serum metabolomic profiling for 1293 metabolites was performed in serum samples of 40 AS-patients and 10 healthy controls using Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectroscopy. AS-patients were divided in four groups (10 patients/group) with respect to ejection fraction (EF, normal/reduced) and aortic valve gradient (> or <40 mmHg). Principal component analysis revealed a clear difference of metabolic profiles in AS-patients vs controls. However, less separation between AS-subtypes could be detected. A Random Forest Analysis comparing all AS-patients to healthy controls resulted in a predictive accuracy of 94%. In AS-patients we observed an increase of acyl carnitines compared to control indicating an alteration of myocardial free fatty acid oxidation. Moreover, we detected a higher ketone body concentrations in AS-patients that could serve as an alternative fuel source for the heart. Interestingly, no differences between the AS subgroups could be detected indicating that changes occur independently from EF and AS-gradient. Additionally, several compounds involved in nitrogen balance were increased in AS-patients while numerous amino acids were decreased, possibly also due to alternative energy production. Increased circulating heme catabolites have been associated with heart failure. Here, we found increased heme catabolites only in patients with low EF and high-gradient with unclear significance. Conclusion In summary, our study indicates a shift of cardiac substrate utilization in patients with AS from free fatty acid oxidation to alternative substrates like ketone bodies and amino acids. As no differences between AS subtypes were observed, this metabolic remodelling may occur as an adaptive mechanism independent from EF and AS-gradient in patients with severe AS. Since our study was conducted by an unspecific approach with a small sample size, further investigation is necessary for a better understanding of the specific myocardial metabolic changes in AS patients. Funding Acknowledgement Type of funding sources: None.

Rare Cause of Arrhythmia and Seizures in a Late-Preterm Newborn

Rare Cause of Arrhythmia and Seizures in a Late-Preterm Newborn

Establishment and Validation of Reference Values for Amino Acids and Acylcarnitines in Dried Blood Spots for Omani Newborns Using Tandem Mass Spectrometry.

ObjectivesTo establish a reference range for acylcarnitines (ACs) and amino acids (AAs) concentrations in dried blood spot (DBS) samples of Omani neonates to detect inborn errors of metabolism (IEM), and to evaluate the effect of age and sex on ACs and AAs.MethodsElectrospray-ionization tandem mass spectrometry (+ESI-MS/MS) was used to determine ACs and AAs concentrations in DBS samples collected from 1302 healthy newborns (0–7 days) delivered at Sultan Qaboos University Hospital between August 2008 and May 2009.ResultsMore than fifty biomarkers that allow diagnosis of various IEMs were measured, their 1stand 99thpercentile values determined, and compared with published international data. Our results were comparable with the corresponding figures from Collaborative Laboratory Integrated Report, despite a much smaller sample size. We found that age had a significant effect on most ACs and AAs except decadienoylcarnitin, decenoylcarnitine, adipylcarnitine, palmitoylcarnitine, steatoylcarnitine, tyrosin, phenylalanine, and valine. Sex of the neonate had insignificant effect on most ACs and AAs except free-carnitine, acetylcarnitine, hexanoylcarnitine, octanoylcarnitine, malonylcarnitine, decanoylcarnitine, dodecenoylcarnitine, dodecanoylcarnitine, and tetradecanoylcarnitine.ConclusionsTandem mass spectrometer is a highly effective tool for high throughput screening of IEM. This study is the first to publish reference intervals for ACs and AAs from DBS samples of Omani newborns. The results may prove to be of significance when determining cut-off values for newborn screening in the near future.

Multi-omics analysis reveals the host-microbe interactions in aged rhesus macaques.

Aging is a complex multifactorial process that greatly affects animal health. Multi-omics analysis is widely applied in evolutionary biology and biomedical research. However, whether multi-omics can provide sufficient information to reveal comprehensive changes in aged non-human primates remains unclear. Here, we explored changes in host–microbe interactions with aging in Chinese rhesus macaques (Macaca mulatta lasiota, CRs) using multi-omics analysis. Results showed marked changes in the oral and gut microbiomes between young and aged CRs, including significantly reduced probiotic abundance and increased pathogenic bacterial abundance in aged CRs. Notably, the abundance of Lactobacillus, which can metabolize tryptophan to produce aryl hydrocarbon receptor (AhR) ligands, was decreased in aged CRs. Consistently, metabolomics detected a decrease in the plasma levels of AhR ligands. In addition, free fatty acid, acyl carnitine, heparin, 2-(4-hydroxyphenyl) propionic acid, and docosahexaenoic acid ethyl ester levels were increased in aged CRs, which may contribute to abnormal fatty acid metabolism and cardiovascular disease. Transcriptome analysis identified changes in the expression of genes associated with tryptophan metabolism and inflammation. In conclusion, many potential links among different omics were found, suggesting that aged CRs face multiple metabolic problems, immunological disorders, and oral and gut diseases. We determined that tryptophan metabolism is critical for the physiological health of aged CRs. Our findings demonstrate the value of multi-omics analyses in revealing host–microbe interactions in non-human primates and suggest that similar approaches could be applied in evolutionary and ecological research of other species.

Longitudinal Metabolomics and Lipidomics Analyses Reveal Alterations Associated with Envenoming by Bothrops asper and Daboia russelii in an Experimental Murine Model.

Longitudinal metabolomics and lipidomics analyses were carried out on the blood plasma of mice injected intramuscularly with venoms of the viperid species Bothrops asper or Daboia russelii. Blood samples were collected 1, 3, 6, and 24 h after venom injection, and a control group of non-envenomed mice was included. Significant perturbations in metabolomics and lipidomics were observed at 1, 3, and 6 h, while values returned close to those of control mice by 24 h, hence reflecting a transient pattern of metabolic disturbance. Both venoms induced significant changes in amino acids, as well as in several purines and pyrimidines, and in some metabolites of the tricarboxylic acid cycle. KEGG analysis of metabolic pathways that showed those with the greatest change included aminoacyl tRNA synthesis and amino acid biosynthesis and metabolism pathways. With regard to lipid metabolism, there was an increase in triglycerides and some acyl carnitines and a concomitant drop in the levels of some phospholipids. In addition, envenomed mice had higher levels of cortisol, heme, and some oxidative stress markers. The overall pattern of metabolic changes in envenomed mice bears similarities with the patterns described in several traumatic injuries, thus underscoring a metabolic response/adaptation to the injurious action of the venoms.

Using an integrated feature-based molecular network and lipidomics approach to reveal the differential lipids in yak shanks and flanks

Yak shanks and flanks are often used as food ingredients, but the lipid composition of these two parts may differ significantly. These meat parts were subjected to a lipidomics analysis using UHPLC-Q-Obitrap. Several computational tools, including feature-based molecular networks, ms-dial, and lipidone, were used to perform deep mining on the entire dataset. The analysis annotated 355 lipid species from 20 subclasses. Lipid chains have a length distribution of 16 to 20 carbons, with unsaturation ranging from 0 to 5. The results revealed that 71 lipids were significantly different in these muscles, including phosphatidylethanolamines (PEs) (16:0/20:4), PEs (18:0/19:1), PEs (18:1/22:5), sphingomyelins (SMs) (36:2; 3O), and carnitines (CARs) (22:0). Furthermore, the metabolic pathways of glycerophospholipids and sphingolipids act as important roles in the differences of these lipid components. This study obtained a comprehensive lipid profile, which is critical for understanding the precise nutritional differences in different yak meat sections.

Dose- and time-dependent manners of moxifloxacin induced liver injury by targeted metabolomics study.

Moxifloxacin is the most widely prescribed antibiotics due to its excellent oral bioavailability and broad-spectrum antibacterial effect. Despite of its popularity, the rare and severe liver injury induced by moxifloxacin is a big concern that cannot be ignored in clinical practice. However, the early warning and related metabolic disturbances of moxifloxacin induced hepatoxicity were rarely reported. In this study, the dose- and time-dependent manners of moxifloxacin induced liver injury were investigated by a targeted metabolomics method. In dose-dependent experiment, three different dosages of moxifloxacin were administered to the rats, including 36 mg kg−1 d−1, 72 mg kg−1 d−1, and 108 mg kg−1 d−1. In time-dependent experiment, moxifloxacin was orally administered to the rats for 3, 7 or 14 consecutive days. Pathological analysis showed that moxifloxacin caused obvious transient hepatotoxicity, with the most serious liver injury occurred in the 7 days continuous administration group. The transient liver injury can be automatically restored over time. Serum levels of liver function related biochemical indicators, including ALT, AST, TBIL, alkaline phosphatase, superoxide dismutase, and malondialdehyde, were also measured for the evaluation of liver injury. However, these indicators can hardly be used for the early warning of hepatotoxicity caused by moxifloxacin due to their limited sensitivity and significant hysteresis. Targeted metabolomics study demonstrated that serum concentrations of fatty acyl carnitines, fatty acids and dehydroepiandrosterone can change dynamically with the severity of moxifloxacin related liver injury. The elevated serum levels of fatty acyl carnitine, fatty acid and dehydroepiandrosterone were promising in predicting the hepatotoxicity induced by moxifloxacin.

Targeted Metabolomics Revealed a Sex-Dependent Signature for Metabolic Syndrome in the Mexican Population.

Metabolic syndrome (MetS) is a group of several metabolic conditions predisposing to chronic diseases. Individuals diagnosed with MetS are physiologically heterogeneous, with significant sex-specific differences. Therefore, we aimed to investigate the potential sex-specific serum modifications of amino acids and acylcarnitines (ACs) and their relationship with MetS in the Mexican population. This study included 602 participants from the Health Workers Cohort Study. Forty serum metabolites were analyzed using a targeted metabolomics approach. Multivariate regression models were used to test associations of clinical and biochemical parameters with metabolomic profiles. Our findings showed a serum amino acid signature (citrulline and glycine) and medium-chain ACs (AC14:1, AC10, and AC18:10H) associated with MetS. Glycine and AC10 were specific metabolites representative of discrimination according to sex-dependent MetS. In addition, we found that glycine and short-chain ACs (AC2, AC3, and AC8:1) are associated with age-dependent MetS. We also reported a significant correlation between body fat and metabolites associated with sex-age-dependent MetS. In conclusion, the metabolic profile varies by MetS status, and these differences are sex-age-dependent in the Mexican population.

Progressive Metabolic Abnormalities Associated with the Development of Neonatal Bronchopulmonary Dysplasia.

Objective: To assess the longitudinal metabolic patterns during the evolution of bronchopulmonary dysplasia (BPD) development. Methods: A case-control dataset of preterm infants (<32-week gestation) was obtained from a multicenter database, including 355 BPD cases and 395 controls. A total of 72 amino acid (AA) and acylcarnitine (AC) variables, along with infants’ calorie intake and growth outcomes, were measured on day of life 1, 7, 28, and 42. Logistic regression, clustering methods, and random forest statistical modeling were utilized to identify metabolic variables significantly associated with BPD development and to investigate their longitudinal patterns that are associated with BPD development. Results: A panel of 27 metabolic variables were observed to be longitudinally associated with BPD development. The involved metabolites increased from 1 predominant different AC by day 7 to 19 associated AA and AC compounds by day 28 and 16 metabolic features by day 42. Citrulline, alanine, glutamate, tyrosine, propionylcarnitine, free carnitine, acetylcarnitine, hydroxybutyrylcarnitine, and most median-chain ACs (C5:C10) were the most associated metabolites down-regulated in BPD babies over the early days of life, whereas phenylalanine, methionine, and hydroxypalmitoylcarnitine were observed to be up-regulated in BPD babies. Most calorie intake and growth outcomes revealed similar longitudinal patterns between BPD cases and controls over the first 6 weeks of life, after gestational adjustment. When combining with birth weight, the derived metabolic-based discriminative model observed some differences between those with and without BPD development, with c-statistics of 0.869 and 0.841 at day 7 and 28 of life on the test data. Conclusions: The metabolic panel we describe identified some metabolic differences in the blood associated with BPD pathogenesis. Further work is needed to determine whether these compounds could facilitate the monitoring and/or investigation of early-life metabolic status in the lung and other tissues for the prevention and management of BPD.

Dairy Milk Casein and Whey Proteins Differentially Alter the Postprandial Lipidome in Persons with Prediabetes: A Comparative Lipidomics Study.

Dairy milk, likely through its bioactive proteins, has been reported to attenuate postprandial hyperglycemia-induced oxidative stress responses implicated in cardiovascular diseases (CVDs). However, how its major proteins, whey and casein, alter metabolic excursions of the lipidome in persons with prediabetes is unclear. Therefore, the objective of this study was to examine whey or casein protein ingestion on glucose-induced alternations in lipidomic responses in adults (17 males and 6 females) with prediabetes. In this clinical study, participants consumed glucose alone, glucose + nonfat milk (NFM), or glucose with either whey (WHEY) or casein (CASEIN) protein, and plasma samples were collected at multiple time points. Lipidomics data from plasma samples was acquired using an ultra-high-performance liquid chromatography-high-resolution mass spectrometry-based platform. Our results indicated that glucose ingestion alone induced the largest number of changes in plasma lipids. WHEY showed an earlier and stronger impact to maintain the stability of the lipidome compared with CASEIN. WHEY protected against glucose-induced changes in glycerophospholipid and sphingolipid (SP) metabolism, while ether lipid metabolism and SP metabolism were the pathways most greatly impacted in CASEIN. Meanwhile, the decreased acyl carnitines and fatty acid (FA) 16:0 levels could attenuate lipid peroxidation after protein intervention to protect insulin secretory capacity. Diabetes-associated lipids, the increased phosphatidylethanolamine (PE) 34:2 and decreased phosphatidylcholine (PC) 34:3 in the NFM-T90min, elevated PC 35:4 and decreased CE 18:1 to a CE 18:2 ratio in the WHEY-T180min, decreased lysophosphatidylcholine (LPC) 22:6 and LPC 22:0/0:0 in the CASEIN-T90min, and decreased PE 36:1 in the CASEIN-T180min, indicating a decreased risk for prediabetes. Collectively, our study suggested that dairy milk proteins are responsible for the protective effect of non-fat milk on glucose-induced changes in the lipidome, which may potentially influence long-term CVD risk.

Combining data acquisition modes in liquid-chromatography–tandem mass spectrometry for comprehensive determination of acylcarnitines in human serum

Acylcarnitines (ACs) are metabolites involved in fatty acid β-oxidation and organic acid metabolism. Metabolic disorders associated to these two processes can be evaluated by determining the complete profile of ACs. In this research, we present an overall strategy for identification, confirmation, and quantitative determination of acylcarnitines in human serum. By this strategy we identified the presence of 47 ACs from C2 to C24 with detection of the unsaturation degree by application of a data-independent acquisition (DIA) liquid chromatography–tandem mass spectrometry (LC–MS/MS) method. Complementary, quantitative determination of ACs is based on a high-throughput and fully automated method consisting of solid-phase extraction on-line coupled to LC–MS/MS in data-dependent acquisition (DDA) to improve analytical features avoiding the errors associated to sample processing. Quantitation limits were at pg mL–1 level, the intra-day and between-day variability were below 15–20%, respectively; and the accuracy, expressed as bias, was always within ± 25%. The proposed method was tested with 40 human volunteers to determine the relative concentration of ACs in serum and identify predominant forms. Significant differences were detected by comparing the ACs profile of obese versus non-obese individuals.

Genomic analysis of 9 infants with hypermethioninemia by whole-exome sequencing among in Henan, China

BackgroundHypermethioninemia is an inborn error of metabolism with elevated plasma methionine (Met) caused by methionine adenosyltransferase deficiency. Methionine adenosyltransferase (MAT) I/III deficiency is the most common cause of hypermethioninemia. Except for increased blood Met, most patients have no symptoms, but a small number have nervous system complications, including cognitive impairment and mental retardation. ObjectiveTo investigate the gene variation of patients with hypermethioninemia in newborns in Henan province. Methods9 cases of hypermethioninemia were screened for amino acids profile and acyl carnitine by tandem mass spectrometric (MS/MS) among 245 054 newborns. We performed whole-exome sequencing on 9 families of infants with hypermethioninemia. We identified mutated genes under different models of inheritance and further assessed these mutations through Sanger sequencing and association analysis. ResultsThe incidence of neonatal hypermethioninemia was 1:27 228 in Henan province. A total of ten mutations in the MAT1A gene in the 9 patients were identified, including nine reported mutations (c.1070C > T, c.895C > T, c.100 T > A, c.315C > A, c.529C > T, c.623A > C, c.407G > T, c.1066C > T, 867G > T) and one novel mutations (c.772G > C). c.772G > C was detected in 2 families and is the most common variant. 7 infants (7/9) with hypermethioninemia were genetically autosomal dominant, and 2 infants (2/9) with hypermethioninemia were genetically autosomal recessive. ConclusionOur findings expand the mutational spectrum of hypermethioninemia, with the description of one new mutation. They improve the understanding of the genetic background and clinical manifestation of MAT1A in Chinese patients.

Abstract 3056: Gut microbial co-metabolism influences the tumor microenvironment

Abstract Right sided colorectal cancers (RCRCs) have distinct molecular phenotypes and clinical presentations to those on the left. Gut microbial co-metabolic interactions with the RCRC microenvironment are poorly defined and present a novel therapeutic target. Here, we present a multi-omics analysis of RCRCs. A prospective observational pilot study was performed at Imperial College London NHS Trust, UK. Treatment naive patients undergoing elective resectional surgery for primary RCRCs were recruited. Fresh intra-luminal fecal samples were obtained at three anatomical sites: 5 cm pre-, tumor, and 5 cm post-tumor. Microsatellite instability (MSI) and CDX2 expression were immunohistochemically evaluated. Metataxonomic analysis was carried out on a MiSeq Platform (Illumina, USA). Metabolomics analysis was performed using liquid chromatography mass spectrometry (Acquity UPLC, Xevo G2-S Q-TOF, Waters). After pre-processing, univariate and multivariate statistics were carried out in MetaboAnalyst and RStudio. 20 patients (8 females, mean age 69.9(53-83)) were included. Of the n=20 RCRCs, n=2 were ileo-cecal valve, n=4 were cecal, n=4 were cecum/proximal ascending colon, n=4 were mid-ascending colon, n=2 were ascending/hepatic flexure, n=2 were hepatic flexure, and n=2 were transverse colon cancers. Tumor classification according to TNM 8 indicated n=4 T1, n=1 T2, n=11 T3, n=4 T4; n=12 as N0, n=6 as N1, and n=2 as N2; n=2 as M0, n=1 as M1c, and n=17 as MX. All n=20 were CDX2 positive and n=16 had normal expression of mismatch repair proteins, n=4 showed MSI. Triglyceride (TG) species were significantly associated with T3-4 rather than T1-2 (p&amp;lt;0.01). Ceramide and diacylglycerol species were significantly associated with T1-2 rather than T3-4 (p&amp;lt;0.01). Multiple TG, an acyl carnitine, and a lysophosphatidic acid (LPA) species were associated with MSI status rather than no MSI status (p&amp;lt;0.01). An integrative analysis pipeline was developed to investigate statistically significant covariance between specific metabolites and taxa (p&amp;lt;0.01). Specifically, Procrustes similarity analysis of the multi-omics dataset revealed a negative correlation at pre- (p=0.606) and tumor level (p=0.738), but significant similarity at post-tumor level (p=0.045). This pattern in post-tumor may indicate cancer specific co-metabolism. At the post-tumor site, the LPA species increased in MSI, showed a negative correlation with members of the Oscillospiraceae family. Ceramide species, significantly increased in early-stage CRC, demonstrate negative correlation with the Lachnospiraceae family. Ongoing work is examining site-matched tissue samples utilizing imaging mass cytometry to define immune interactions. Mapping the intra-luminal fecal metabolome and metataxonome in RCRC reveals potential bacterial-host co-metabolic targets that could be leveraged for stratifying immunotherapy treatment. Citation Format: Petra Paizs, Duncan Roberts, Nathan Danckert, James Lewis, Maria Sani, Marta Jamroziak, Robert Goldin, Julian Marchesi, Lauren Ford, James Alexander, Zoltan Takats, James Kinross. Gut microbial co-metabolism influences the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3056.

Evaluating immune response and metabolic related biomarkers pre-allogenic hematopoietic stem cell transplant in acute myeloid leukemia.

Although hematopoietic stem cell transplantation (HCT) is the only curative treatment for acute myeloid leukemia (AML), it is associated with significant treatment related morbidity and mortality. There is great need for predictive biomarkers associated with overall survival (OS) and clinical outcomes. We hypothesized that circulating metabolic, inflammatory, and immune molecules have potential as predictive biomarkers for AML patients who receive HCT treatment. This retrospective study was designed with an exploratory approach to comprehensively characterize immune, inflammatory, and metabolomic biomarkers. We identified patients with AML who underwent HCT and had existing baseline plasma samples. Using those samples (n = 34), we studied 65 blood based metabolomic and 61 immune/inflammatory related biomarkers, comparing patients with either long-term OS (≥ 3 years) or short-term OS (OS ≤ 1 years). We also compared the immune/inflammatory response and metabolomic biomarkers in younger vs. older AML patients (≤30 years vs. ≥ 55 years old). In addition, the biomarker profiles were analyzed for their association with clinical outcomes, namely OS, chronic graft versus host disease (cGVHD), acute graft versus host disease (aGVHD), infection and relapse. Several baseline biomarkers were elevated in older versus younger patients, and baseline levels were lower for three markers (IL13, SAA, CRP) in patients with OS ≥ 3 years. We also identified immune/inflammatory response markers associated with aGVHD (IL-9, Eotaxin-3), cGVHD (Flt-1), infection (D-dimer), or relapse (IL-17D, bFGF, Eotaxin-3). Evaluation of metabolic markers demonstrated higher baseline levels of medium- and long-chain acylcarnitines (AC) in older patients, association with aGVHD (lactate, long-chain AC), and cGVHD (medium-chain AC). These differentially expressed profiles merit further evaluation as predictive biomarkers.

Neurotoxicity assessment of QoI strobilurin fungicides azoxystrobin and trifloxystrobin in human SH-SY5Y neuroblastoma cells: Insights from lipidomics and mitochondrial bioenergetics

Strobilurin fungicides are quinone outside inhibitors (QoI) used to treat fungal pathogens for agricultural and residential use. Here, we compared the potential for neurotoxicity of the widely used strobilurins, azoxystrobin (AZS) and trifloxystrobin (TFS), in differentiated human SH-SY5Y cells. Fungicides did not include cytotoxicity up to 200 µM but both induced loss of cell viability at 48 h, with TFS showing slightly higher toxicity that AZS. Caspase 3/7 activity was induced in SH-SY5Y cells by both fungicides at 48 h (50 µM for AZS and 25 µM for TFS). ATP levels were reduced following a 24-hour exposure to > 25 µM AZS and > 6.25 µM TFS and both fungicides rapidly impaired oxidative respiration (~12.5 µM for AZS and ~3.125 µM TFS) and decreased oligomycin-induced ATP production, maximal respiration, and mitochondrial spare capacity. AZS at 100 µM showed a continual impairment of mitochondrial membrane potential (MMP) between 4 and 48 h while TFS at > 50 µM decreased MMP at 24 h. Taken together, TFS exerted higher mitochondrial toxicity at lower concentrations compared to AZS in SH-SY5Y cells. To discern toxicity mechanisms of strobilurin fungicides, lipidomics was conducted in SH-SY5Y cells following exposure to 6.25 µM and 25 µM AZS, and a total of 1595 lipids were detected, representing 49 different lipid classes. Lipid classes with the largest proportion of lipids detected in SH-SY5Y cells included triglycerides (17%), phosphatidylethanolamines (8%), ether-linked triglycerides (8%), phosphatidylcholines (7%), ether-linked phosphatidylethanolamines (6%), and diacylglycerols (5%). Together, these 5 lipid classes accounted for over 50% of the total lipids measured in SH-SY5Y cells. Lipids that were increased by AZS included acyl carnitine, which plays a role in long chain fatty acid utilization for mitochondrial β-oxidation, as well as non-modified, ether linked, and oxidized triacylglycerols, suggesting compensatory upregulation of triglyceride biosynthesis. The ceramide HexCer-NS, linked to neurodegenerative diseases, was decreased in abundance following AZS exposure. In summary, strobilurin fungicides rapidly inhibit mitochondrial oxidative respiration and alter the abundance of several lipids in neuronal cells, relevant for understanding environmental exposure risks related to their neurotoxicity.

Determinants of blood acylcarnitine concentrations in healthy individuals of the European Prospective Investigation into Cancer and Nutrition

SummaryBackground & aimsCirculating levels of acylcarnitines (ACs) have been associated with the risk of various diseases such as cancer and type 2 diabetes. Diet and lifestyle factors have been shown to influence AC concentrations but a better understanding of their biological, lifestyle and metabolic determinants is needed.MethodsCirculating ACs were measured in blood by targeted (15 ACs) and untargeted metabolomics (50 ACs) in 7770 and 395 healthy participants of the European Prospective Investigation into Cancer and Nutrition (EPIC), respectively. Associations with biological and lifestyle characteristics, dietary patterns, self-reported intake of individual foods, estimated intake of carnitine and fatty acids, and fatty acids in plasma phospholipid fraction and amino acids in blood were assessed.ResultsAge, sex and fasting status were associated with the largest proportion of AC variability (partial-r up to 0.19, 0.18 and 0.16, respectively). Some AC species of medium or long-chain fatty acid moiety were associated with the corresponding fatty acids in plasma (partial-r = 0.24) or with intake of specific foods such as dairy foods containing the same fatty acid. ACs of short-chain fatty acid moiety (propionylcarnitine and valerylcarnitine) were moderately associated with concentrations of branched-chain amino acids (partial-r = 0.5). Intake of most other foods and of carnitine showed little association with AC levels.ConclusionsOur results show that determinants of ACs in blood vary according to their fatty acid moiety, and that their concentrations are related to age, sex, diet, and fasting status. Knowledge on their potential determinants may help interpret associations of ACs with disease risk and inform on potential dietary and lifestyle factors that might be modified for disease prevention.

Plasma Acylcarnitines as Metabolic Signatures of Declining Health-Related Quality of Life Measure in Community-Dwelling Older Adults: A Combined Cross-sectional and Longitudinal Pilot Study.

Health-related quality of life (HRQoL) measures are predictors of adverse health outcomes in older adults. Studies have demonstrated cross-sectional associations between HRQoL measures and blood-based biochemical markers. Acylcarnitines (ACs) are a class of metabolites generated in the mitochondria and are predictive of multiple geriatric syndromes. Changes in ACs reflect alterations in central carbon metabolic pathways. However, the prospective relationship between plasma ACs and declining HRQoL has not been examined. This study aimed to investigate both cross-sectional and longitudinal associations of baseline ACs with baseline and declining EuroQol-5 Dimension/EuroQol Visual Analogue Scale (EQ-5D/EQ-VAS) in community-dwelling older adults. One hundred and twenty community-dwelling older adults with EQ-5D/EQ-VAS measurements at baseline and follow-up were included. We quantified ACs at baseline using targeted plasma metabolomics profiling. Multivariate regressions were performed to examine cross-sectional and longitudinal associations between the measures. Cross-sectionally, ACs showed no significant associations with either EQ-5D index or EQ-VAS scores. Longitudinally, multiple baseline short-chain ACs were significantly and inversely associated with declining EQ-5D index score, explaining up to 8.5% of variance in the decline. Within a cohort of community-dwelling older adults who had high HRQoL at baseline, we showed that higher levels of short-chain ACs are longitudinally associated with declining HRQoL. These findings reveal a novel association between central carbon metabolic pathways and declining HRQoL. Notably, dysregulation in mitochondrial central carbon metabolism could be detected prior to clinically important decline in HRQoL, providing the first evidence of objective biomarkers as novel predictors to monitor HRQoL in nonpharmacological interventions and epidemiology.