Metabolic Drug Research Articles

Repurposing metabolic drugs for COVID-19.

Repurposing metabolic drugs for COVID-19.

Integrated analysis identifies oxidative stress-related lncRNAs associated with progression and prognosis in colorectal cancer

BackgroundColorectal cancer (CRC) is one of the most common cancers in the world. Oxidative stress reactions have been reportedly associated with oncogenesis and tumor progression. By analyzing mRNA expression data and clinical information from The Cancer Genome Atlas (TCGA), we aimed to construct an oxidative stress-related long noncoding RNA (lncRNA) risk model and identify oxidative stress-related biomarkers to improve the prognosis and treatment of CRC.ResultsDifferentially expressed oxidative stress-related genes (DEOSGs) and oxidative stress-related lncRNAs were identified by using bioinformatics tools. An oxidative stress-related lncRNA risk model was constructed based on 9 lncRNAs (AC034213.1, AC008124.1, LINC01836, USP30-AS1, AP003555.1, AC083906.3, AC008494.3, AC009549.1, and AP006621.3) by least absolute shrinkage and selection operator (LASSO) analysis. The patients were then divided into high- and low-risk groups based on the median risk score. The high-risk group had a significantly worse overall survival (OS) (p < 0.001). Receiver operating characteristic (ROC) and calibration curves displayed the favorable predictive performance of the risk model. The nomogram successfully quantified the contribution of each metric to survival, and the concordance index and calibration plots demonstrated its excellent predictive capacity. Notably, different risk subgroups showed significant differences in terms of their metabolic activity, mutation landscape, immune microenvironment and drug sensitivity. Specifically, differences in the immune microenvironment implied that CRC patients in certain subgroups might be more responsive to immune checkpoint inhibitors.ConclusionsOxidative stress-related lncRNAs can predict the prognosis of CRC patients, which provides new insight for future immunotherapies based on potential oxidative stress targets.

Pharmacovigilance-based drug repurposing: searching for putative drugs with hypohidrosis or anhidrosis adverse events for use against hyperhidrosis

BackgroundDrug repurposing refers to the application of existing drugs to new therapeutic indications. As phenotypic indicators of human drug response, drug side effects may provide direct signals and unique opportunities for drug repurposing.ObjectivesWe aimed to identify drugs frequently associated with hypohidrosis or anhidrosis adverse reactions (that is, the opposite condition of hyperhidrosis) from the pharmacovigilance database, which could be potential candidates as anti-hyperhidrosis treatment agents.MethodsIn this observational, retrospective, pharmacovigilance study, adverse event reports of hypohidrosis or anhidrosis in the US Food and Drug Administration (FDA) Adverse Event Reporting System (FAERS) were assessed between January 2004 and December 2021 using reporting odds ratio (ROR) estimates and categorized by the World Health Organization Anatomical Therapeutic Chemical (ATC) classification code. The onset time of drug-associated hypohidrosis or anhidrosis was also examined.ResultsThere were 540 reports of 192 drugs with suspected drug-associated hypohidrosis or anhidrosis in the FAERS database, of which 39 drugs were found to have statistically significant signals. Nervous system drugs were most frequently reported (187 cases, 55.82%), followed by alimentary tract and metabolism drugs (35 cases, 10.45%), genitourinary system and sex hormones (28 cases, 8.36%), and dermatologicals (22 cases, 6.57%). The top 3 drug subclasses were antiepileptics, drugs for urinary frequency and incontinence, and antidepressants. Taking disproportionality signals, pharmacological characteristics of drugs and appropriate onset time into consideration, the main putative drugs for hyperhidrosis were glycopyrronium, solifenacin, oxybutynin, and botulinum toxin type A. Other drugs, such as topiramate, zonisamide, agalsidase beta, finasteride, metformin, lamotrigine, citalopram, ciprofloxacin, bupropion, duloxetine, aripiprazole, prednisolone, and risperidone need more investigation.ConclusionsSeveral candidate agents among hypohidrosis or anhidrosis-related drugs were identified that may be redirected for diminishing sweat production. There are affirmative data for some candidate drugs, and the remaining proposed candidate drugs without already known sweat reduction mechanisms of action should be further explored.

Metabolic Control, Diabetic Complications and Drug Therapy in a Cohort of Patients with Type 1 and Type 2 Diabetes in Secondary and Tertiary Care between 2004 and 2019.

Retrospective cross-sectional analysis of four single-centre cohorts between 2004 and 2019. All patients with T1DM or T2DM in secondary care (n = 5571) or tertiary care (n = 2001) were included. Statistical analyses were performed using linear mixed models. Diabetes duration increased in both patients with T1DM and T2DM in secondary care and tertiary care (p < 0.001). Patients in secondary care consistently showed good glycaemic control, while patients in tertiary care showed inadequate glycaemic control. All four cross-sectional cohorts showed a significant increase in the prevalence of nephropathy over time and three out of four cohorts (T1DM and T2DM in secondary care and T2DM in tertiary care) showed an increase in the prevalence of neuropathy (all p < 0.001). The incidence of severe hypoglycaemia was consistently low. The use of insulin pumps and insulin analogues in the therapy of T1DM increased significantly. The increased prevalence of complications is likely due to older age and longer diabetes duration. Low rates of hypoglycaemia, lower limb amputations and good glycaemic control in secondary care patients indicate a good structure of patient care.

Activity-balanced GLP-1/GDF15 dual agonist reduces body weight and metabolic disorder in mice and non-human primates

Obesity is a considerable health concern with limited pharmacotherapy options of low efficacy. Here, we develop a GLP-1/GDF15 fusion protein and explore its weight-lowering potential in animals. The molecule, QL1005, is engineered via fusing GLP-1 and GDF15 analogs by a peptide linker and conjugating it to a fatty acid for time-action extension. Invitro, the potency of QL1005 is superior to the GLP-1 analog semaglutide. In obese mice, QL1005 induces reductions in body weight, food intake, insulin, fasting glucose, and triglycerides. Notably, these metabolic effects come as a result of activities emanating from both GLP-1 and GDF15, in an individual pathway-balanced fashion. In a cynomolgus monkey model of obesity, QL1005 reduces body weight, food intake, insulin, and glucose in a dose-dependent manner with limited incidence of GI side effects. Altogether, this long-acting, dual GLP-1/GDF15 molecule demonstrates the promise of poly-pharmaceutical approaches in metabolic drug discovery and development.

CircPVT1 and PVT1/AKT3 show a role in cell proliferation, apoptosis, and tumor subtype-definition in small cell lung cancer.

Small cell lung cancer (SCLC) is treated as a homogeneous disease, although the expression of NEUROD1, ASCL1, POU2F3, and YAP1 identifies distinct molecular subtypes. The MYC oncogene, amplified in SCLC, was recently shown to act as a lineage-specific factor to associate subtypes with histological classes. Indeed, MYC-driven SCLCs show a distinct metabolic profile and drug sensitivity. To disentangle their molecular features, we focused on the co-amplified PVT1, frequently overexpressed and originating circular (circRNA) and chimeric RNAs. We analyzed hsa_circ_0001821 (circPVT1) and PVT1/AKT3 (chimPVT1) as examples of such transcripts, respectively, to unveil their tumorigenic contribution to SCLC. In detail, circPVT1 activated a pro-proliferative and anti-apoptotic program when over-expressed in lung cells, and knockdown of chimPVT1 induced a decrease in cell growth and an increase of apoptosis in SCLC in vitro. Moreover, the investigated PVT1 transcripts underlined a functional connection between MYC and YAP1/POU2F3, suggesting that they contribute to the transcriptional landscape associated with MYC amplification. In conclusion, we have uncovered a functional role of circular and chimeric PVT1 transcripts in SCLC; these entities may prove useful as novel biomarkers in MYC-amplified tumors.

DR5 Up-Regulation Induced by Dichloroacetate Sensitizes Tumor Cells to Lipid Nanoparticles Decorated with TRAIL.

Cancer resistance to treatments is a challenge that researchers constantly seek to overcome. For instance, TNF-related apoptosis-inducing ligand (TRAIL) is a potential good prospect as an anti-cancer therapy, as it attacks tumor cells but not normal cells. However, treatments based in soluble TRAIL provided incomplete clinical results and diverse formulations have been developed to improve its bioactivity. In previous works, we generated a new TRAIL formulation based in its attachment to the surface of unilamellar nanoliposomes (LUV-TRAIL). This formulation greatly increased apoptosis in a wide selection of tumor cell types, albeit a few of them remained resistant. On the other hand, it has been described that a metabolic shift in cancer cells can also alter its sensitivity to other treatments. In this work, we sought to increase the sensitivity of several tumor cell types resistant to LUV-TRAIL by previous exposure to the metabolic drug dichloroacetate (DCA), which forces oxidative phosphorylation. Results showed that DCA + LUV-TRAIL had a synergistic effect on both lung adenocarcinoma A549, colorectal HT29, and breast cancer MCF7 cells. Despite DCA inducing intracellular changes in a cell-type specific way, the increase in cell death by apoptosis was clearly correlated with an increase in death receptor 5 (DR5) surface expression in all cell lines. Therefore, DCA-induced metabolic shift emerges as a suitable option to overcome TRAIL resistance in cancer cells.

Evaluation of Renal Impairment Influence on Metabolic Drug Clearance using a Modelling Approach.

Chronic kidney disease (CKD) may alter drug renal elimination but is also known for interacting with hepatic metabolism via multiple uremic components. However, few global models, considering the five major cytochromes, have been published, and none specifically address the decrease in cytochrome P450 (CYP450) activity. The aim of our study was to estimate the possibility of quantifying residual cytochrome activity as a function of filtration rate, according to the data available in the literature. For each drug in the DDI-predictor database, we collected available pharmacokinetic data comparing drug exposition in the healthy patient and in various stages of CKD, before building a model capable of predicting the variation of exposure according to the degree of renal damage. We followed an In vivo Mechanistic Static Model (IMSM) approach, previously validated for predicting change in liver clearance. We estimated the remaining fraction parameters at glomerular filtration rate (GFR) = 0 and the alpha value of GFR to 50% impairment for the 5 major cytochromes using a non-linear constrained regression using Matlab software. Thirty-one compounds had usable pharmacokinetic data, with 51 AUC ratios between healthy and renal impaired patients. The remaining CYP3A4 activity was estimated to be 0.4 when CYP2D6, 2C9, 2C19 and 1A2 activity was estimated to be 0.43; 1; 0.73 and 0.7, respectively. The alpha value was estimated to be at 6.62; 25; 9.8; 1.38 and 11.04 for each cytochrome. In comparison with published data, all estimates but one were correctly predicted in the range of 0.5-2. Our approach was able to describe the impact of CKD on metabolic elimination. Modelling this process makes it possible to anticipate changes in clearance and drug exposure in CKD patients, with the advantage of greater simplicity than approaches based on physiologically-based pharmacokinetic modelling. However, a precise estimation of the impact of renal failure is not possible with an IMSM approach due to the large variability of the published data, and thus should rely on specific pharmacokinetic modelling for narrow therapeutic margin drugs.

Novel Muscle-Homing Peptide FGF1 Conjugate Based on AlphaFold for Type 2 Diabetes Mellitus

Drugs for metabolic diseases usually require systemic administration and act on multiple tissues, which may produce some unpredictable side effects. There have been many successful studies on targeted drugs, especially antitumor drugs. However, there is still little research on metabolic disease drugs targeting specific tissues. Fibroblast growth factor 1 (FGF1) is a potential therapy for type 2 diabetes (T2D) without the risk of hypoglycemia. However, the major impediment to the clinical application of FGF1 is its mitogenic potential. We previously engineered an FGF1 variant (named FGF1ΔHBS) to tune down its mitogenic activity via reducing the heparin-binding ability. However, other notable side effects still remained, including severe appetite inhibition, pathogenic loss of body weight, and increase in fatality rate. In this study, we used AlphaFold2 and PyMOL visualization tools to construct a novel FGF1ΔHBS conjugate fused with skeletal muscle-targeted (MT) peptide through a flexible peptide linker termed MT-FGF1ΔHBS. We found that MT-FGF1ΔHBS specifically homed to skeletal muscle tissue after systemic administration and induced a potent glucose-lowering effect in T2D mice without hypoglycemia. Mechanistically, MT-FGF1ΔHBS elicits the glucose-lowering effect via AMPK activation to promote the GLUT4 expression and translocation in skeletal muscle cells. Notably, compared with native FGF1ΔHBS, MT-FGF1ΔHBS had minimal effects on food intake and body weight and did not induce any hyperplasia in major tissues of both T2D and normal mice, indicating that this muscle-homing protein may be a promising candidate for T2D treatment. Our targeted peptide strategy based on computer-aided structure prediction in this study could be effectively applied for delivering agents to functional tissues to treat metabolic or other diseases, offering enhanced efficacy and reducing systemic off-target side effects.

Применение Кокарнита у пациенток с ишемической болезнью плаценты в послеродовом периоде

schemic placental disease, which includes such nosological entities as preeclampsia, fetal growth retardation and placental abruption, is commonly the cause of premature labor, which makes it an urgent medical and biological problem. To increase the efficacy of preventive measures, it seems promising to use metabolic therapy, which implies a direct effect on metabolic processes at the tissue and cellular levels. The antioxidant effect of metabolic drugs can reduce the formation of reactive oxygen species and peroxyl radicals in the blood and tissues, and prevent cellular damage. The authors present their experience concerning the use of Cocarnit® metabolic drug, which includes triphosadenine, cocarboxylase, cyanocobalamin and nicotinamide, in complex therapy in the late postpartum period in patients with ischemic placental disease. The use of this drug improves the hemodynamic parameter control, normalizes carbohydrate metabolism, contributes to the rapid restoration of hemoglobin levels in combination with iron preparations, as well as to a softer and faster rehabilitation of female patients in this group, increasing the probability of a favorable outcome in future pregnancy. It should be noted that the preliminary results obtained by the authors require further randomized comparative clinical trials on large samples. KEYWORDS: pregnancy, ischemic placental disease, postpartum period, fetal death, Cocarnit, triphosadenine, cocarboxylase, nicotinamide, cyanocobalamin, metabolic therapy. FOR CITATION: Dobrokhotova Yu.E., Dzhokhadze L.S. Cocarnit in female patients with ischemic placental disease in the postpartum period. Russian Journal of Woman and Child Health. 2023;6(4):385–390 (in Russ.). DOI: 10.32364/2618-8430-2023-6-4-10.

Metabolic adverse drug reactions in geriatric patients in Odisha

Background: Geriatric patients are at a greater risk of developing adverse drug reaction (ADR) because of changed metabolic activity and renal status associated with aging. As most of the elderly patients suffer from multiple chronic diseases (Diabetes, Hypertension, Arthritis, constipation, peptic ulcer disease, etc.), they are under polypharmacy and the risk of ADRs is exacerbated by multiple medications. Metabolic ADRs include weight gain, hypoglycemia, hyponatremia, hypokalemia, and the endocrinal abnormality are the frequent cause of hospital admission in geriatric patients. Most metabolic ADRs due to drugs are treatable. There are few such studies conducted in India regarding metabolic ADR in geriatric patients. Hence, this study aims to evaluate the metabolic ADRs encountered in geriatric patients with the following objectives in Odisha. Aim and Objectives: The prevalence and spectrum of metabolic ADRs in geriatric population were studied. The causality assessment, severity assessment, and preventability were done using standard scales. Materials and Method: This was a prospective and observational study conducted from September 2016 to September 2018 in Department of Pharmacology in collaboration with Department of Geriatric Medicine and Department of Medicine of SCB Medical College and Hospital. All geriatric patients (≥60 years) with ADR, reporting to the above departments, were included in the study. The detailed information of type of ADRs and its characteristics was filled up in Suspected ADR Reporting Form designed by Indian pharmacopeia commission version 1.3. The incidence and spectrum of metabolic ADRs were observed. Their causality, severity, and preventability were evaluated by the WHO-UMC System, Hartwig’s Severity Scale, and Schumock Thornton Preventability Scale, respectively. Result: A total of 236 ADRs were reported in 2 years, out of which 28.8% were metabolic ADRs. Metabolic ADRs were more reported in young old patients and often encountered ADR was hypoglycemia 73.5% due to use of oral anti diabetics. About 29% of the metabolic ADRs had a causal relation of probable association. About 94.2% were moderate in severity and 89.7% metabolic ADRs could be probably preventable. Conclusion: There was a probable causal association of metabolic ADR with the drug used and they were moderate verity. The metabolic ADRs found in our study can be preventable with appropriate means.

SGLT-2 Inhibitors in Heart Failure: A Review of Current Evidence.

Sodium-glucose co-transporter 2 (SGLT-2) inhibitors are the latest addition to guideline-directed medical therapy in heart failure (HF) with reduced ejection fraction with recent trials suggesting a significant reduction in adverse cardiovascular outcomes in patients with HF with mildly reduced and preserved ejection fraction. SGLT-2 inhibitors have evolved as metabolic drugs due to their multi-system effects and are indicated for the management of HF across the ejection fraction spectrum, type 2 diabetes, and chronic kidney disease. There is ongoing research to explore the mechanistic effects of SGLT-2 inhibitors in HF and to evaluate their use in worsening HF and after myocardial infarction. This review focuses on the evidence for SGLT-2 inhibitors from type 2 diabetes cardiovascular outcome and primary HF trials and discusses ongoing research related to their use in cardiovascular disease.

Antiproliferative effects of 13α/β-steroids on triple-negative MDA-MB-231 breast cancer cells: unraveling intracellular signaling without ERα

This study aimed to investigate the activities of novel 20(R)-3,20-dihydroxy-19-norpregn-1,3,5(10)-trienes (kuz7 and kuz8b) of natural 13β- and epimeric 13α-series against triple-negative MDA-MB-231 breast cancer cells. High antiproliferative activity of synthesized compounds kuz8b and kuz7 against MDA-MB-231 triple-negative cancer cells was revealed. The steroid kuz7 of natural 13β-configuration was more active against MDA-MB-231 cells than the 13α-steroid kuz8b. Cell cycle analysis revealed common patterns for the action of both tested compounds. The number of cells in the subG1 phase increased in a dose-dependent manner, indicating induction of apoptosis, which was also verified by PARP cleavage. In contrast, the number of cells in the G0/G1 phase decreases with increasing compound concentration. Steroid kuz7 at micromolar concentrations reduced the expression of GLUT1, a glucose transporter. High efficacy of the combination of kuz7 with biguanide metformin was shown, and synergistic effects on MDA-MB-231 cell growth and expression of the anti-apoptotic protein Bcl-2 were revealed. According to the obtained results, including the high activity of kuz7 against triple-negative cancer cells, the detected induction of apoptosis, and the decrease in GLUT1 expression, 13β-steroid kuz7 is of interest for further preclinical studies both alone and in combination with the metabolic drug metformin.

Effect of Vitamin E on Serum Creatinine level on Gentamicin Induced Nephrotoxicity in Long Evans rats

Background: The kidneys are one of the vital organs of our body to excrete metabolic waste products, drugs and chemicals in the form of urine. Objectives: This study was carried out to observe the effect of Vitamin E on gentamicin-induced nephrotoxicity by assessing serum creatinine level in Long Evans rats. Materials and Methods: The experimental study was carried out on 40 Long Evans rats of both sex with the weight ranges from 172-255 gm and the age ranges from 7 to 10 weeks. The rats were divided into four groups- Group A (control) received normal saline, group B, C and D received gentamicin for 6 days, rats of group C received vitamin E capsule for 9 days with gentamicin whereas, group D received vitamin E capsule with gentamicin for total 10 days. Serum creatinine level was measured at the end of experiment. Results: The mean (mean+ SD) serum creatinine levels in group A, B, C and D were 0.98+0.34, 2.36+ 0.44, 1.39+0.18 and 1.30+0.18 respectively. The differences between the groups were highly significant (p&lt;0.001) in group A&amp;B, B&amp;C and B&amp;D whereas the difference between C&amp;D was not significant (p&gt;0.50). Serum creatinine level on the normal saline control (group A) was within normal limit (0.98mg/dl). Serum creatinine level in gentamicin treated (group B) rats were more in comparison to gentamicin and vitamin E treated rats (group C&amp;D) and pretreatment with longer duration group (group D) showed lower serum creatinine level than the shorter one (group C) though there was no significant difference. Conclusion: Vitamin E treatment showed some protective effects against gentamicin induced nephrotoxicity. The results also indicated that effectiveness of vitamin E depends on a suitable duration of pretreatment for better protection against gentamicin-induced nephrotoxicity. KYAMC Journal Vol. 13, No. 03, October 2022: 149-152

The eighth International Forum on Stem Cells: virtual meeting, October 20-21, 2022.

The eighth International Forum on Stem Cells: virtual meeting, October 20-21, 2022.

Study of vinpocetine adsorption on the surface of liposomes obtained from soya lecitine

Liposomes are one of the best studied types of nanoparticles that are considered as contemporary and efficient means for delivery of various medicinal agents widely applied in the clinical practice. Taking into account the features of transportation, translocation through histological barriers, cellular membranes and metabolic transformations liposomal pharmaceutical drugs show unique properties, first of all connected with the distinctions in their pharmacokinetics. Objective of the study: determination of adsorption characteristics for vinpocetine applied on the surface of liposomes obtained from soya lecitine. Liposomes from soya lecitine were obtained by hydration/rehydration technique. In order to study characteristics of vinpocetine adsorption on the surface of liposomes technique of equilibrium dialysis was utilized. Sol of iron hydroxide was used as a comparison adsorbent. Separation of the dispersive medium from sol colloid particles was performed with the use of column chromatography. Comparative investigations of vinpocetine adsorption were performed both on comparison adsorbent and on the liposomes obtained from soya lecitine. Rather efficient adsorption of vinpocetine on the liposomes at their low concentrations was observed.

Effects of metabolic cancer therapy on tumor microenvironment.

Targeting tumor metabolism for cancer therapy is an old strategy. In fact, historically the first effective cancer therapeutics were directed at nucleotide metabolism. The spectrum of metabolic drugs considered in cancer increases rapidly - clinical trials are in progress for agents directed at glycolysis, oxidative phosphorylation, glutaminolysis and several others. These pathways are essential for cancer cell proliferation and redox homeostasis, but are also required, to various degrees, in other cell types present in the tumor microenvironment, including immune cells, endothelial cells and fibroblasts. How metabolism-targeted treatments impact these tumor-associated cell types is not fully understood, even though their response may co-determine the overall effectivity of therapy. Indeed, the metabolic dependencies of stromal cells have been overlooked for a long time. Therefore, it is important that metabolic therapy is considered in the context of tumor microenvironment, as understanding the metabolic vulnerabilities of both cancer and stromal cells can guide new treatment concepts and help better understand treatment resistance. In this review we discuss recent findings covering the impact of metabolic interventions on cellular components of the tumor microenvironment and their implications for metabolic cancer therapy.

CONVENTIONAL APPROACHES TO THE THERAPY OF HEREDITARY MYOPATHIES

The aim of the work was to analyze the available therapeutic options for the conventional therapy of hereditary myopathies.Materials and methods. When searching for the material for writing a review article, such abstract databases as PubMed and Google Scholar were used. The search was carried out on the publications during the period from 1980 to September 2022. The following words and their combinations were selected as parameters for the literature selection: “myopathy”, “Duchenne”, “myodystrophy”, “metabolic”, “mitochondrial”, “congenital”, “symptoms”, “replacement”, “recombinant”, “corticosteroids”, “vitamins”, “tirasemtiv”, “therapy”, “treatment”, “evidence”, “clinical trials”, “patients”, “dichloracetate”.Results. Congenital myopathies are a heterogeneous group of pathologies that are caused by atrophy and degeneration of muscle fibers due to mutations in genes. Based on a number of clinical and pathogenetic features, hereditary myopathies are divided into: 1) congenital myopathies; 2) muscular dystrophy; 3) mitochondrial and 4) metabolic myopathies. At the same time, treatment approaches vary significantly depending on the type of myopathy and can be based on 1) substitution of the mutant protein; 2) an increase in its expression; 3) stimulation of the internal compensatory pathways expression; 4) restoration of the compounds balance associated with the mutant protein function (for enzymes); 5) impact on the mitochondrial function (with metabolic and mitochondrial myopathies); 6) reduction of inflammation and fibrosis (with muscular dystrophies); as well as 7) an increase in muscle mass and strength. The current review presents current data on each of the listed approaches, as well as specific pharmacological agents with a description of their action mechanisms.Conclusion. Currently, the following pharmacological groups are used or undergoing clinical trials for the treatment of various myopathies types: inotropic, anti-inflammatory and antifibrotic drugs, antimyostatin therapy and the drugs that promote translation through stop codons (applicable for nonsense mutations). In addition, metabolic drugs, metabolic enzyme cofactors, mitochondrial biogenesis stimulators, and antioxidants can be used to treat myopathies. Finally, the recombinant drugs alglucosidase and avalglucosidase have been clinically approved for the replacement therapy of metabolic myopathies (Pompe’s disease).

Functional noninvasive detection of glycolytic pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) lacks effective treatment options beyond chemotherapy. Although molecular subtypes such as classical and QM (quasi-mesenchymal)/basal-like with transcriptome-based distinct signatures have been identified, deduced therapeutic strategies and targets remain elusive. Gene expression data show enrichment of glycolytic genes in the more aggressive and therapy-resistant QM subtype. However, whether the glycolytic transcripts are translated into functional glycolysis that could further be explored for metabolic targeting in QM subtype is still not known. We used different patient-derived PDAC model systems (conventional and primary patient-derived cells, patient-derived xenografts (PDX), and patient samples) and performed transcriptional and functional metabolic analysis. These included RNAseq and Illumina HT12 bead array, in vitro Seahorse metabolic flux assays and metabolic drug targeting, and in vivo hyperpolarized [1-13C]pyruvate and [1-13C]lactate magnetic resonance spectroscopy (HP-MRS) in PDAC xenografts. We found that glycolytic metabolic dependencies are not unambiguously functionally exposed in all QM PDACs. Metabolic analysis demonstrated functional metabolic heterogeneity in patient-derived primary cells and less so in conventional cell lines independent of molecular subtype. Importantly, we observed that the glycolytic product lactate is actively imported into the PDAC cells and used in mitochondrial oxidation in both classical and QM PDAC cells, although more actively in the QM cell lines. By using HP-MRS, we were able to noninvasively identify highly glycolytic PDAC xenografts by detecting the last glycolytic enzymatic step and prominent intra-tumoral [1-13C]pyruvate and [1-13C]lactate interconversion in vivo. Our study adds functional metabolic phenotyping to transcriptome-based analysis and proposes a functional approach to identify highly glycolytic PDACs as candidates for antimetabolic therapeutic avenues.

Clean-catching urine from pigs: A method for collecting quality specimens for urinalysis and microbiological culturing in a laboratory environment.

Urine is an important biological specimen for assessing various metabolic functions and drug clearance. In urinary tract infection research, urine is particularly important as urinary bacterial titres constitute the main diagnostic outcome for assessing the course of infection. Collecting uncontaminated urine samples from pigs can be done by bladder catheterization or suprapubic bladder aspiration, which are both laborious and invasive procedures that require the need for anaesthesia. To improve animal welfare and optimize urine sampling protocols, we developed a method of clean-catching midstream urine specimens from pigs during spontaneous micturition. The quality of urine specimens collected by clean-catch, bladder catheter and suprapubic bladder aspiration were compared using microbiological culturing. We show that urine specimens collected by clean-catch are only minimally contaminated by skin- and vaginal-commensals with no influence on urinary bacterial titres during ongoing infection. In conclusion, we describe a method in which spontaneous micturition can be prompted in pigs, facilitating fast and reliable collection of quality specimens suitable for microbiological culturing. The method supersedes the need for anaesthesia, which not only represents a considerable refinement in terms of animal welfare but also facilitates more frequent collection of specimens that can enhance the scientific outcome of experimental animal studies in pigs.