Metabolic Screening Research Articles

Newly discovered variants in unexplained neonatal encephalopathy.

The genetic background of neonatal encephalopathy (NE) is complicated and early diagnosis is beneficial to optimizing therapeutic strategy for patients. NE Patients with unclear etiology received regular clinical tests including ammonia test, metabolic screening test, amplitude-integrated electroencephalographic (aEEG) monitoring, brain Magnetic Resonance Imaging (MRI) scanning, and genetic test. The protein structure change was predicted using Dynamut2 and RoseTTAFold. 15 out of a total of 113 NE Patients were detected with newly reported pathogenic variants. In this sub-cohort, (1) seizure was the primary initial symptoms; (2) four patients had abnormal metabolic screening results, and two of them were also diagnosed with excessive blood ammonia concentration; (3) the brain MRI results were irregular in three infants and the brain waves were of moderate-severe abnormality in about a half of the patients. The novel pathogenic variants discovered in this study belonged to 12 genes, and seven of them were predicted to introduce a premature translation termination. In-silicon predictions showed that four variants were destructive to the protein structure of KCNQ2. Our study expands the mutation spectrum of genes associated with NE and introduces new evidence for molecular diagnosis in this newborn illness.

Validation of the nutritrauma concept for the detection of potential harmful effects of medical nutritional treatment in critically ill patients in real life.

Introduction: medical nutritional treatment (MNT) can be complex and may be associated with potential metabolic complications, which has been recently described as nutritrauma. Objective: the aim of our work is to describe whether the application of the nutritrauma concept in real life is feasible and useful to detect the metabolic complications associated with the prescription of MNT. Material and methods: in this descriptive, prospective study at a single center we enrolled 30 consecutive critically ill patients in a 14-bed medical-surgical intensive care unit. The nutritrauma strategy implementation was based in four "M" steps: Metabolic screening, MNT prescription, biochemical Monitoring, and nutritional Management. Results: we analyzed 28 patients (mean age, 69.7 ± 11.3 years; APACHE II, 18.1 ± 8.1; SOFA, 7.5 ± 3.7; Nutric Score, modified, 4.3 ± 2.01, and mean BMI, 27.2 ± 3.8). The main cause of admission was sepsis (46.4 %). Length of ICU stay was 20.6 ± 15.1 days; 39.3 % of subjects died during their ICU stay. Enteral nutrition (82.1 %) was more frequent than parenteral nutrition (17.9 %). During nutritional monitoring, 54 specific laboratory determinations were made. Hyperglycemia was the most frequent metabolic alteration (83.3 % of measurements). Electrolyte disturbances included hypocalcemia (50 %), hypophosphatemia (29.6 %) and hypokalemia (27.8 %). The most frequent lipid profile abnormalities were hypocholesterolemia (64.8 %) and hypertriglyceridemia (27.8 %). Furthermore, nutritional prescription was modified for 53.6 % of patients: increased protein dosage (25 %), increased calorie dosage (21.4 %) and change to organ-specific diet (17.8 %). Conclusions: in conclusion, the application of the nutritrauma approach facilitates detection of metabolic complications and an evaluation of the appropriate prescription of MNT.

Characterization of PKCα-rutin interactions and their application as a treatment strategy for pulmonary arterial hypertension by inhibiting ferroptosis.

As a common plant-derived dietary flavonoid, rutin receives widespread attention because of its good antioxidant bioactivities. Protein kinase Cα (PKCα) is a serine/threonine kinase that is involved in uncountable cellular processes, among which ferroptosis, a novel form of cell death, is triggered by lipid peroxidation and has been reported to be associated with pulmonary arterial hypertension (PAH). But it is still not well appreciated how rutin inhibits ferroptosis in PAH and what function PKCα has in this process. In this study, we first observed whether rutin could prevent PAH by attenuating ferroptosis with a PAH animal model and pulmonary artery smooth muscle cells (PASMCs) under hypoxia. Mitochondrial metabolomics and network pharmacology were employed to clarify the metabolic alterations and screen target proteins, and the results showed that PKCα was a vital node in rutin regulating mitochondrial metabolism related to ferroptosis in PAH. Based on molecular docking and multispectral analysis, we found that rutin could directly interact with PKCα through hydrogen bonds, which could induce static quenching, and then influence the secondary structure of PKCα. In conclusion, these findings mainly point to a novel mechanism that rutin protects PAH rats by modifying the structure and altering the activity of PKCα, and thus suppressing ferroptosis. This work reveals that the interaction behaviors between small molecules and bio-macromolecules are a critical factor to develop natural biological active ingredients and gives an insight into the potential applications of flavonoids in health and disease.

Prevalence and outcome of inborn errors of metabolism among high-risk neonates: an Egyptian single-center study

Background Inborn errors of metabolism (IEM) are genetic disorders caused by incompetence in enzymatic reactions of intermediary metabolism due to an insufficient amount or unstable form of the involved enzyme. Aim To assess the prevalence and outcome of IEM among high-risk neonates for early diagnosis and detection of treatable causes to minimize morbidity and mortality rate in neonates. Methods This cross-sectional study was carried out on 684 neonates at the Neonatal Intensive Care Unit in the Pediatrics Department at Zagazig University Children’s Hospital from February 2020 to February 2023. Extended metabolic screen (EMS) was done for full-term neonates and preterm of more than 32 weeks of gestation. Further investigations such as plasma ammonia, plasma lactate, urine organic acid analysis, blood gas, serum anion gap, and the blood glucose level were also measured. Results The prevalence of IEM was 4.4%. The most common presentations among the cases of IEM were not doing well, respiratory distress, encephalopathy, and convulsion. As regards provisional diagnosis, 11 cases were diagnosed as maple syrup urine disease, 5 cases with glycine encephalopathy, 5 cases with medium-chain acyl CoA dehydrogenase deficiency,4 cases with methylmalonic acidemia, 2 cases with isovaleric acidemia, 2 cases with citrullinemia type 1, and one case with propionic academia; 12 (40%) of IEM cases died. Conclusion IEM disorders are not rare diseases in high-risk neonates with attention to consanguinity, which is a common tradition in our country. The delay in diagnosis of IEM leads to high morbidity and mortality.

Adherence to the monitoring of metabolic syndrome in patients receiving antipsychotics in outpatient clinics in Saudi Arabia.

Monitoring protocols have been developed because patients taking atypical antipsychotics are more prone to developing metabolic syndrome, which leads to possible increased mortality and morbidity. The aim of this study was to assess the degree of adherence to the recommendations of metabolic syndrome monitoring. This study was conducted in two large psychiatric facilities in the Eastern Province of Saudi Arabia. A retrospective analysis of the medical records of 350 patients taking antipsychotic medications was done, and an assessment was made of the frequency of metabolic monitoring at each of the intervals as suggested by the American Diabetes Association. Data was analyzed using SPSS; descriptive statistics. were computed and Chi-square test was used to determine statistical significance for association between categorical variable. The mean age of the patients was 34.9 ± 18 years; 64.6% were males. Olanzapine was the most prescribed medication (43.7%, n = 153), followed by quetiapine (17.4%, n = 61). Only one-third of the patients (29.6%) completed all the baseline parameters. Documentation of baseline parameters was low for glucose level (38.9%), lipid panel (17.3%), weight (25.2%), and waist circumference (1.4%). Adherence to yearly monitoring was much lower than at baseline (mean percentage: 29.6% vs. 1.7%). Furthermore, 45% of the patients were classified as obese and 10% had metabolic comorbidity. Individuals with mental illness who were taking antipsychotics did not undergo proper metabolic screening during antipsychotic treatment. Barriers to adherence to the monitoring guidelines should be examined and addressed. Giving assistance to practitioners to recall the required laboratory tests and vitals at certain intervals could help improve metabolic monitoring practices.

Selective Screening for Inborn Errors of Metabolism Using Tandem Mass Spectrometry in Newborns of the West Kazakhstan: Pilot Study

Tandem mass spectrometry can detect and quantify many metabolites in a single blood spot to diagnose amino acid disorders, organic acids, fatty acid oxidation, and urea cycle disorders. The use of tandem mass spectrometry (MS/MS) is expanding for the implementation of newborn screening programs for inborn errors of metabolism and for selective screening of children of different ages. In Kazakhstan, the use of MS/MS in metabolic screening programs is not yet developed due to the high cost of equipment and consumables and the lack of special screening centers and specialists. Data on the prevalence of most inborn errors of metabolism in Kazakhstan are not presented in the literature. Aim: to perform selective screening for hereditary metabolic diseases among newborns in western Kazakhstan using the liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Methods: Selective screening was performed among 250 newborns with suspected hereditary metabolic disorders using tandem mass spectrometry. Results: The results of selective newborn screening were interpreted by comparison with reference values established for this group. Diagnosis was based on clinical signs, blood levels of amino acids, acylcarnitines, succinylacetone, urine organic acids, and gene mutation tests. An assessment of 37 inborn errors of metabolism frequencies in high-risk newborns was performed. Conclusion: The research will further develop the national as selective as expanded newborn screening programs.

Clinical Characteristics of Pediatric Patients with Inborn Errors of Metabolism Admitted to Namazi Hospital, a Tertiary Referral Center in Shiraz, Southern Iran

Background: Inborn errors of metabolism (IEMs) are rare genetic disorders that usually cause disease by blocking a metabolic pathway. Delayed diagnosis often occurs due to nonspecific symptoms. This study assesses the demographic and clinical characteristics of pediatric patients with IEMs admitted to Namazi hospital, a referral center in southern Iran. Methods: All 1-month-old to 18-year-old patients who were admitted to the pediatric ward at Namazi hospital, a referral center in southern Iran, and diagnosed with IEMs were enrolled. Patients with incomplete information were excluded. Comprehensive demographic and clinical data, including age, sex, parental consanguinity, family history of IEM, presenting symptoms, number of hospitalizations, age of onset, and diagnosis of the disease, were collected. All the data were entered into SPSS version 22 and analyzed. Results: Two hundred patients, 53.5% male and 46.5% female, were enrolled in the study. The predominant clinical symptoms observed were lethargy and vomiting. The most prevalent IEMs were aminoacidopathies, organic acidemia, and mitochondrial disease. Consanguinity was found in 57.5% of patients, while a family history of metabolic diseases was observed in 16.5%. A statistically significant association was identified between consanguinity and the type of disease. Conclusion: The clinical symptoms of IEMs are nonspecific and may be misdiagnosed as other diseases, such as septicemia. We should always keep IEMs in mind to detect the disease earlier and prevent significant morbidities and mortality through appropriate and timely treatment. Due to the prevalence of family marriages in Iran, we should also consider metabolic screening.

Association between cholesterol and blood pressure examination in Sampangan, Semarang City: a preliminary study

Purpose: This study aims to determine the baseline prevalence and association between blood pressure and cholesterol examination. 
 Method: This preliminary study used a cross-sectional design to analyze the secondary blood pressure and cholesterol levels data in the Sampangan sub-district, Semarang City. The total sampling technique covered those enrolled in the metabolic syndrome screening from January to April 2023. The logistic regression test was used in data analysis. 
 Results: 58 respondents took blood pressure checks, while only 40 had cholesterol checks. The probability of males getting changes in cholesterol levels was higher than that of females (OR = 8.69; p-value = 0.01). However, neither (female nor male) had a significant relationship, and there was no difference in the chances of changes in blood pressure (OR = 1.43; p = 0.72). Respondents above 60 years had a significant association with changes in blood pressure compared to those under 60 years (OR = 0.09; p = 0.007).
 Conclusion: This study revealed males of a certain age experience higher changes in cholesterol levels compared to females. People over 60 years old tend to have higher blood pressure compared to those under 60 years old. These findings propose an outreach strategy for health screening for patients with limited mobility, transportation access, and finances for routine checkups at public or government healthcare facilities.

Novel Techniques and Models for Studying the Role of the Gut Microbiota in Drug Metabolism.

The gut microbiota, known as the second human genome, plays a vital role in modulating drug metabolism, significantly impacting therapeutic outcomes and adverse effects. Emerging research has elucidated that the microbiota mediates a range of modifications of drugs, leading to their activation, inactivation, or even toxication. In diverse individuals, variations in the gut microbiota can result in differences in microbe-drug interactions, underscoring the importance of personalized approaches in pharmacotherapy. However, previous studies on drug metabolism in the gut microbiota have been hampered by technical limitations. Nowadays, advances in biotechnological tools, such as microbially derived metabolism screening and microbial gene editing, have provided a deeper insight into the mechanism of drug metabolism by gut microbiota, moving us toward personalized therapeutic interventions. Given this situation, our review summarizes recent advances in the study of gut-microbiota-mediated drug metabolism and showcases techniques and models developed to navigate the challenges posed by the microbial involvement in drug action. Therefore, we not only aim at understanding the complex interaction between the gut microbiota and drugs and outline the development of research techniques and models, but we also summarize the specific applications of new techniques and models in researching gut-microbiota-mediated drug metabolism, with the expectation of providing new insights on how to study drug metabolism by gut microbiota.

Long-Term Management of Patients with Mild Urea Cycle Disorders Identified through the Newborn Screening: An Expert Opinion for Clinical Practice.

Urea cycle disorders (UCDs) are a group of rare inborn errors of metabolism caused by a deficiency in one of the six enzymes or one of the two transporters involved in the urea cycle. Current guidelines suggest that early diagnosis and treatment of mild UCDs may improve survival and prevent decompensation and neurocognitive impairment. Nevertheless, clinical studies are very difficult to carry out in this setting due to the rarity of the diseases, and high-level evidence is scant and insufficient to draw conclusions and provide clinical guidelines. With the early introduction of newborn screening, the Italian healthcare organization fostered an advancement in expertise in metabolic disease management and screening programs, by allocating resources, and favoring the expansion of newborn screening. A group of experts operating in Italian centers decided to share their experience and provide advice for the management of mild UCDs in clinical practice. A consensus was reached by the Estimate-Talk-Estimate (ETE) method. Five items were identified, and statements for each item were agreed. Briefly, the panel advised completing the diagnosis by expanded newborn screening (ENS) with biochemical and genetic confirmation and by following up with the patient during the first year of life, with a routine laboratory and metabolic profile as well as with clinical observation. Early initiation of therapy is advised and should be followed by therapy adjustment once the diagnostic profile is completed. The therapy should be based on a low-protein diet and nitrogen scavengers. The long-term follow-up is based on growth and nutritional assessment, clinical and neurocognitive evaluation, and laboratory and instrumental parameter monitoring.

A protocol for a turbidimetric assay using a Saccharomyces cerevisiae thiamin biosynthesis mutant to estimate total vitamin B1 content in plant tissue samples

BackgroundUnderstanding thiamin (thiamine; vitamin B1) metabolism in plants is crucial, as it impacts plant nutritional value as well as stress tolerance. Studies aimed at elucidating novel aspects of thiamin in plants rely on adequate assessment of thiamin content. Mass spectrometry-based methods provide reliable quantification of thiamin as well as closely related biomolecules. However, these techniques require expensive equipment and expertise. Microbiological turbidimetric assays can evaluate the presence of thiamin in a given sample, only requiring low-cost, standard lab equipment. Although these microbiological assays do not reach the accuracy provided by mass spectrometry-based methods, the ease with which they can be deployed in an inexpensive and high-throughput manner, makes them a favorable method in many circumstances. However, the thiamin research field could benefit from a detailed step-by-step protocol to perform such assays as well as a further assessment of its potential and limitations.ResultsHere, we show that the Saccharomyces cerevisiae thiamin biosynthesis mutant thi6 is an ideal candidate to be implemented in a turbidimetric assay aimed at assessing the content of thiamin and its phosphorylated equivalents (total vitamer B1). An optimized protocol was generated, adapted from a previously established microbiological assay using the thi4 mutant. A step-by-step guidance for this protocol is presented. Furthermore, the applicability of the assay is illustrated by assessment of different samples, including plant as well as non-plant materials. In doing so, our work provides an extension of the applicability of the microbiological assay on top of providing important considerations upon implementing the protocol.ConclusionsAn inexpensive, user-friendly protocol, including step-by-step guidance, which allows adequate estimation of vitamer B1 content of samples, is provided. The method is well-suited to screen materials to identify altered vitamer B1 content, such as in metabolic engineering or screening of germplasm.

The Prevalence of PCOS in South Asia

Polycystic ovary syndrome (PCOS) is a common endocrine disorder in reproductive-aged women, with ethnic diversity in its manifestation. South Asian patients have a much higher prevalence of PCOS than the controls. A greater PCOS burden among South Asian than White women receiving infertility care has been reported. South Asian patients showed an increased degree of hirsutism, early onset of symptoms, severe insulin resistance, metabolic risks (impaired glucose tolerance (IGT), insulin resistance (IR), diabetes mellitus (DM), hyperlipidemia) and endometrial pathology as compared with Caucasians. South Asians with PCOS also have greater odds of being centrally obese, with one third having the metabolic syndrome that bears no relationship to the androgenic phenotype. South Asians with PCOS manifest early and seek health-care pre-maritally, with irregular menses and hirsutism predominating. They have greater central obesity in proportion to a ‘lower’ BMI, which is linked to their metabolic risk. MetS affects one-third, with age [Formula: see text]35 years, Acantosis Nigricans and BMI [Formula: see text]25 kg/m2 (Asian cut off) having significant association. In a study when Chinese, Filipina and South Asians were compared, the prevalence of PCOS and obesity was significantly higher in South Asians whereas that of diabetes was higher among Filipinas. Genetic components play important roles in the pathogenesis of PCOS, and genome-wide association studies of PCOS have also suggested that similar genetic risk factors exist between Asian and Caucasian patients. Among women with PCOS, Asians have been reported to have lower insulin (vs. Black and Hispanic women) and higher glucose (vs. non-Hispanic White women) levels in response to an oral glucose load. Greater awareness in the clinicians about the differences in PCOS and diabetes among south Asian subgroups is important for targeting screening and prevention efforts. Due to the baseline ethnic predisposition to metabolic syndrome, comprehensive metabolic screening, and counselling regarding future risks of chronic disease should be provided.

Section 15 - Inborn Errors of Metabolism and Newborn Screening

Section 15 - Inborn Errors of Metabolism and Newborn Screening

Prenatal air pollution, fetal β-cell dysfunction and neurodevelopmental delay

BackgroundEmerging evidence has reported significant associations of prenatal air pollution exposure with neurodevelopmental delay in offspring. Sensitive exposure windows and the modifiable factor remain elusive. ObjectiveWe aim to identify sensitive windows of air pollution during pregnancy on neurodevelopmental delay, and examine whether cord blood C-peptide mediates the relationship. MethodsThis study included 7438 mother-newborn pairs in Hefei, China, from 2015 to 2021. Weekly exposure to particulate matter of aerodynamic diameter <2.5 µm, 10 µm (PM2.5, PM10), nitrogen dioxide (NO2) and carbon monoxide (CO) was estimated at regulatory air monitoring stations in Hefei. Denver Developmental Screening Test-II and the Gesell Developmental Schedules were applied to assess the neurodevelopmental delay in children 6–36 mon of age. Distributed lag nonlinear models examined sensitive time windows of prenatal air pollutants exposure. Mediation analysis estimated the mediating role of cord blood C-peptide. ResultsThe sensitive PM2.5, PM10, NO2, and CO exposure windows associated with neurodevelopmental delay were throughout pregnancy. Weekly air pollutants exposure was related to higher neurodevelopmental delay risks [cumulative odds ratio (OR): 1.40(1.29,1.53) in PM2.5 (per 10 μg/m3), 1.40(1.28,1.53) in PM10 (per 10 μg/m3), 1.41(1.30,1.52) in CO (per 0.1 mg/m3), and 1.49(1.29,1.72) in NO2 (per 5 μg/m3)]. Mediation analysis indicated 18.3 % contributions of cord C-peptide to the relationship [average mediation effect: 0.04(0.01.0.06); average direct effect: 0.15(0.07.0.25)]. Conclusions: Exposure to air pollution throughout pregnancy is linked to neurodevelopmental delay mediated by poorer fetal β-cell function. Screening and treatment of abnormal glucose metabolism in infants could benefit the prevention of air pollution-associated neurodevelopment delay.

Multiomic screening of invasive GBM cells reveals targetable transsulfuration pathway alterations.

While the poor prognosis of glioblastoma arises from the invasion of a subset of tumor cells, little is known of the metabolic alterations within these cells that fuel invasion. We integrated spatially addressable hydrogel biomaterial platforms, patient site-directed biopsies, and multiomics analyses to define metabolic drivers of invasive glioblastoma cells. Metabolomics and lipidomics revealed elevations in the redox buffers cystathionine, hexosylceramides, and glucosyl ceramides in the invasive front of both hydrogel-cultured tumors and patient site-directed biopsies, with immunofluorescence indicating elevated reactive oxygen species (ROS) markers in invasive cells. Transcriptomics confirmed upregulation of ROS-producing and response genes at the invasive front in both hydrogel models and patient tumors. Among oncologic ROS, H2O2 specifically promoted glioblastoma invasion in 3D hydrogel spheroid cultures. A CRISPR metabolic gene screen revealed cystathionine γ-lyase (CTH), which converts cystathionine to the nonessential amino acid cysteine in the transsulfuration pathway, to be essential for glioblastoma invasion. Correspondingly, supplementing CTH knockdown cells with exogenous cysteine rescued invasion. Pharmacologic CTH inhibition suppressed glioblastoma invasion, while CTH knockdown slowed glioblastoma invasion in vivo. Our studies highlight the importance of ROS metabolism in invasive glioblastoma cells and support further exploration of the transsulfuration pathway as a mechanistic and therapeutic target.

Melaleuca leucadendra Pharmacological Network for Identifying Potential Target of Alcohol Dependence

Melaleuca leucadendra (ML) contains a compound that is potentially a candidate for alcohol dependence. Alcohol dependence is defined by desire, tolerance, anxiety with alcohol, and continuing to drink even though the consequences are dangerous. The study aims to analyze the potential of the ML compound content for alcohol dependence therapy within silico-based pharmacological chain analysis. ML compound data is obtained from the Knapsack database, screening of absorption, distribution, metabolism, and excretion (ADME) of the compounds ML with SwissADME, prediction of the protein of the target compounder ML with the Swiss TargetpPrediction, Gene cards, venny, analysis of the pharmacological network with String-DB and its visualization with Cytoscape version 3.10.0. The pathways correlated with therapy are dopamine receptors, dopamine carriers, serotonin, gamma-aminobutyric acid receptors, and toll-like receptors for known therapeutic target proteins: OPRM1, DRD2, ALDH2, ADH1B, ADH1A, ADH1C, ADH4, SLC6A3, CNR1, POMC, ARRB2, and NCS1. Alcohol-dependent therapies include alpha-Campholenal, Benzaldehyde, trans-Pinocarveol, Borneol, linalool, alfa-Terpineol, (-)-alpha-Bisabolol oxide B, alphaterpine acetate, and Caryophylla-4(148),15-dien-5alphaol.

Network Pharmacology Analysis of Cananga odorata as a Treatment for Anxiety Disorders

Anxiety disorder is a psychological disorder associated with the existence of mental disorders and experiencing constant anxiety. In C. odorata, part of the flower has the potential as a sedative for the nervous system and for dealing with anxiety. The study aims to analyze the potential compound content in Cananga odorata for treating anxiety disorders with In silico-based pharmacological network analysis. CO compound data from the KNApSAck database, Absorption, Distribution, Metabolism, and Excretion (ADME) screening using SwissADME, predicted target proteins using Swiss Target Prediction, GeneCards, Venny, String DB pharmacological network analysis, Visualization with Cytoscape version 3.10.0, and Way2drug. The results of the pharmacological tissue analysis of the compound content in C. odorata obtained 45 compounds, and 18 known active components meet the criteria of Absorption, Distribution, Metabolism, and Excretion (ADME) that correspond to the drug compounder (Drug Likeness/DL). Based on the pathway that correlates with anxiety disorder therapy are the neurotransmitter systems like serotonin receptors and dopamine receptors. The known therapeutic target proteins are HTR1A, HTR2A, SLC6A4, NR3C1, MAOA, DRD4, HSP90AA1, JUN, ten active compounds associated with C. odorata namely Anonaine, (+)-Reticuline, linalool, (-)-Coreximine, Micheline A, (-)-Ushinsunine beta-N-oxide, 4-Terpineol, alpha-Terpeneol, Sampangine, Anaxagoreine. Based on the results of research, C. odorata is potentially a treatment for anxiety disorder.

The Therapeutic Role of Olea europaea in Alcohol Dependence Base in Network Pharmacology Analysis

Alcohol dependence is a state of alcohol becoming a vital part of the life of a person who consumes it; when discontinued, it can lead to a wide range of physical and mental health disorders as well as a decrease in life productivity in people with alcohol dependence. Olea Europaea (OE) is a plant capable of treating alcohol dependence. The method used in silico-based pharmacological grid analysis to determine the ability of the OE compound to treat alcohol dependence. EO compound data is obtained from the KnapSack database, absorption, distribution, metabolism, and excretion (ADME) screening using SwissADME, target protein prediction using SwissTargetPrediction, Gene cards, venny, pharmacological grid analysis with String-DB, visualization with Cytoscape 3.10.0. Results are obtained from 63 OE compounds, and 17 have ADME criteria matching the drug compounder (Drug Likeness/DL). The pathways that correlate with therapy are dopamine receptors, dopamine transporter, serotonin receptor, gamma-aminobutyric acid receptor, and toll-like receptors for known therapeutic target proteins: OPRM1, DRD2, ALDH2, ADH1B, ADH1A, ADH1C, ADH4, ADH7, SLC6A3, CNR1, POMC, ARRB2, and NCS1. Compounds associated with alcohol dependency therapy include Hexanal, Nonadienal, Octanal, 3-Hexenal, 3-Methyl-butanal, Methyl nominate, Cinchonidine, cinchonine, (9S)-10,11-Dihydrocinchonan-9-ol, Oleuropeic acid, Butyl acetate, cis-3-hexenyl acetate, and (S)-2,3-Epoxysqualene. Based on the findings, OE is a potential drug candidate for alcohol dependence.

The dark side of orange: Multiorganismic continuum dynamics within a lichen of the Atacama Desert

ABSTRACT Over the decades our understanding of lichens has shifted to the fact that they are multiorganismic, symbiotic microecosystems, with their complex interactions coming to the fore due to recent advances in microbiomics. Here, we present a mutualistic-parasitic continuum dynamics scenario between an orange lichen and a lichenicolous fungus from the Atacama Desert leading to the decay of the lichen’s photobiont and leaving behind a black lichen thallus. Based on isolation, sequencing, and ecophysiological approaches including metabolic screenings of the symbionts, we depict consequences upon infection with the lichenicolous fungus. This spans from a loss of the lichen’s photosynthetic activity and an increased roughness of its surface to an inhibition of the parietin synthesis as a shared pathway between the photobiont and the mycobiont, including a shift of secondary metabolism products. This degree of relations has rarely been documented before, although lichenicolous fungi have been studied for over 200 years, adding an additional level to the view of interactions within lichens.

TMET-22. MULTI-OMIC SCREENING OF INVASIVE GBM CELLS IN ENGINEERED BIOMATERIALS AND PATIENT BIOPSIES REVEALS TARGETABLE TRANSSULFURATION PATHWAY ALTERATIONS

Abstract While the poor prognosis of glioblastoma arises from the invasion of a subset of tumor cells, little is known of the metabolic alterations within these cells that fuel invasion. We integrated spatially addressable hydrogel biomaterial platforms, patient site-directed biopsies, and multi-omics analyses to define metabolic drivers of invasive glioblastoma cells. Metabolomics and lipidomics revealed elevations in the redox buffers cystathionine, hexosylceramides, and glucosyl ceramides in the invasive front of both hydrogel-cultured tumors and patient site-directed biopsies, with immunofluorescence indicating elevated reactive oxygen species (ROS) markers in invasive cells. Transcriptomics confirmed upregulation of ROS-producing and response genes at the invasive front in both hydrogel models and patient tumors. Amongst oncologic ROS, hydrogen peroxide specifically promoted glioblastoma invasion in 3D hydrogel spheroid cultures. A CRISPR metabolic gene screen revealed cystathionine gamma lyase (CTH), which converts cystathionine to the non-essential amino acid cysteine in the transsulfuration pathway, to be essential for glioblastoma invasion. Correspondingly, supplementing CTH knockdown cells with exogenous cysteine rescued invasion. Pharmacologic CTH inhibition suppressed glioblastoma invasion, while CTH knockdown slowed glioblastoma invasion in vivo. Our studies highlight the importance of ROS metabolism in invasive glioblastoma cells and support further exploration of the transsulfuration pathway as a mechanistic and therapeutic target.