Molecular profiling and feasibility using a comprehensive hybrid capture panel on a consecutive series of non-small-cell lung cancer patients from a single centre.

BACKGROUND: Targeted next-generation sequencing (NGS) is recommended to screen actionable genomic alterations (GAs) in patients with non-small-cell lung cancer (NSCLC). We determined the feasibility to detect actionable GAs using TruSight™ Oncology 500 (TSO500) in 200 consecutive patients with NSCLC. MATERIALS AND METHODS: DNA and RNA were sequenced on an Illumina® NextSeq 550 instrument and processed using the TSO500 Docker pipeline. Clinical actionability was defined within the molecular tumour board following European Society for Medical Oncology (ESMO) guidelines for oncogene-addicted NSCLC. Overall survival (OS) was estimated as per the presence of druggable GAs and treatment with targeted therapy. RESULTS: Most patients were males (69.5%) and former or current smokers (86.5%). Median age was 64 years. The most common histological type and tumour stage were lung adenocarcinoma (81%) and stage IV (64%), respectively. Sequencing was feasible in most patients (93.5%) and actionable GAs were found in 26.5% of patients. A high concordance was observed between single-gene testing and TSO500 NGS panel. Patients harbouring druggable GAs and receiving targeted therapy achieved longer OS compared to patients without druggable GAs. Conversely, patients with druggable GAs not receiving targeted therapy had a trend toward shorter OS compared with driver-negative patients. CONCLUSIONS: Hybrid capture sequencing using TSO500 panel is feasible to analyse clinical samples from patients with NSCLC and is an efficient tool for screening actionable GAs.

Lipidomic signature of stroke recurrence after transient ischemic attack.

While TIA patients have transient symptoms, they should not be underestimated, as they could have an underlying pathology that may lead to a subsequent stroke: stroke recurrence (SR). Previously, it has been described the involvement of lipids in different vascular diseases. The aim of the current study was to perform a lipidomic analysis to identify differences in the lipidomic profile between patients with SR and patients without. Untargeted lipidomic analysis was performed in plasma samples of 460 consecutive TIA patients recruited < 24 h after the onset of symptoms. 37 (8%) patients suffered SR at 90 days. Lipidomic profiling disclosed 7 lipid species differentially expressed between groups: 5 triacylglycerides (TG), 1 diacylglyceride (DG), and 1 alkenyl-PE (plasmalogen) [specifically, TG(56:1), TG(63:0), TG(58:2), TG(50:5), TG(53:7, DG(38:5)) and PE(P-18:0/18:2)]. 6 of these 7 lipid species belonged to the glycerolipid family and a plasmalogen, pointing to bioenergetics pathways, as well as oxidative stress response. In this context, it was proposed the PE(P-18:0/18:2) as potential biomarker of SR condition.The observed changes in lipid patterns suggest pathophysiological mechanisms associated with lipid droplets metabolism and antioxidant protection that is translated to plasma level as consequence of a more intensive or high-risk ischemic condition related to SR.

Metabolomics reveals that fittest trail runners show a better adaptation of bioenergetic pathways.

OBJECTIVES: To analyze the effect in the blood metabolome of trail running, a demanding sport that takes place in the natural environment, places considerable strain on both muscles and joints. While metabolic responses to aerobic exercise have been analyzed in-depth, few studies have focused on trail running. DESIGN: Observational study to analyze changes in 35 different metabolites - representative of aerobic exercise-induced by a simulated 21-km trail race with an uphill gradient of 1400 m. METHODS: We performed a semiquantitative metabolomics study consisting of capillary blood microsampling and targeted screening with liquid chromatography and mass spectrometry to analyze, in 33 licensed athletes, changes concerning 35 metabolites. RESULTS: We observed significant changes in many metabolites, including increased acetyl-carnitine and taurine concentrations (false discovery rate-corrected paired t-test P value 1.63 × 10-13, and P value 5.021 × 10-12, respectively) and decreased carnitine and proline concentrations (P value 6.33 × 10-10, and P value 1.21 × 10-9, respectively). Metabolic responses to trail running were largely independent of sex but were influenced by the level of training, with runners with a higher level showing resistance to exercise-induced changes in taurine, 1-methyl histidine, acetyl-carnitine, and hypoxanthine concentrations. Performance (measured as race time) was inversely correlated with changes in specific metabolites (including taurine, serotonin, and hypoxanthine) and directly correlated with increases in glutathione. CONCLUSIONS: Our findings demonstrate the usefulness of metabolomics studies for analyzing exercise-induced physiological changes and show individual differences associated with the level of training and performance.

Plasma methionine metabolic profile is associated with longevity in mammals.

Methionine metabolism arises as a key target to elucidate the molecular adaptations underlying animal longevity due to the negative association between longevity and methionine content. The present study follows a comparative approach to analyse plasma methionine metabolic profile using a LC-MS/MS platform from 11 mammalian species with a longevity ranging from 3.5 to 120 years. Our findings demonstrate the existence of a species-specific plasma profile for methionine metabolism associated with longevity characterised by: i) reduced methionine, cystathionine and choline; ii) increased non-polar amino acids; iii) reduced succinate and malate; and iv) increased carnitine. Our results support the existence of plasma longevity features that might respond to an optimised energetic metabolism and intracellular structures found in long-lived species.

Genome-wide profiling of non-smoking-related lung cancer cells reveals common RB1 rearrangements associated with histopathologic transformation in EGFR-mutant tumors.

BACKGROUND: The etiology and the molecular basis of lung adenocarcinomas (LuADs) in nonsmokers are currently unknown. Furthermore, the scarcity of available primary cultures continues to hamper our biological understanding of non-smoking-related lung adenocarcinomas (NSK-LuADs). PATIENTS AND METHODS: We established patient-derived cancer cell (PDC) cultures from metastatic NSK-LuADs, including two pairs of matched EGFR-mutant PDCs before and after resistance to tyrosine kinase inhibitors (TKIs), and then performed whole-exome and RNA sequencing to delineate their genomic architecture. For validation, we analyzed independent cohorts of primary LuADs. RESULTS: In addition to known non-smoker-associated alterations (e.g. RET, ALK, EGFR, and ERBB2), we discovered novel fusions and recurrently mutated genes, including ATF7IP, a regulator of gene expression, that was inactivated in 5% of primary LuAD cases. We also found germline mutations at dominant familiar-cancer genes, highlighting the importance of genetic predisposition in the origin of a subset of NSK-LuADs. Furthermore, there was an over-representation of inactivating alterations at RB1, mostly through complex intragenic rearrangements, in treatment-naive EGFR-mutant LuADs. Three EGFR-mutant and one EGFR-wild-type tumors acquired resistance to EGFR-TKIs and chemotherapy, respectively, and histology on re-biopsies revealed the development of small-cell lung cancer/squamous cell carcinoma (SCLC/LuSCC) transformation. These features were consistent with RB1 inactivation and acquired EGFR-T790M mutation or FGFR3-TACC3 fusion in EGFR-mutant tumors. CONCLUSIONS: We found recurrent alterations in LuADs that deserve further exploration. Our work also demonstrates that a subset of NSK-LuADs arises within cancer-predisposition syndromes. The preferential occurrence of RB1 inactivation, via complex rearrangements, found in EGFR-mutant tumors appears to favor SCLC/LuSCC transformation under growth-inhibition pressures. Thus RB1 inactivation may predict the risk of LuAD transformation to a more aggressive type of lung cancer, and may need to be considered as a part of the clinical management of NSK-LuADs patients.

Lipid profile of cerebrospinal fluid in multiple sclerosis patients: a potential tool for diagnosis.

Multiple sclerosis (MS) is a complex multifactorial neuropathology. Although its etiology remains unclear, it has been demonstrated that the immune system attacks myelin, leading to demyelination and axonal damage. The involvement of lipids as one of the main components of myelin sheaths in MS and other demyelinating diseases has been postulated. However, it is still a matter of debate whether specific alteration patterns exist over the disease course. Here, using a lipidomic approach, we demonstrated that, at the time of diagnosis, the cerebrospinal fluid of MS patients presented differences in 155 lipid species, 47 of which were identified. An initial hierarchical clusterization was used to classify MS patients based on the presence of 25 lipids. When a supervised method was applied in order to refine this classification, a lipidomic signature was obtained. This signature was composed of 15 molecules belonging to five different lipid families including fatty acids (FAs). An FA-targeted approach revealed differences in two members of this family: 18:3n3 and 20:0 (arachidic acid). These results reveal a CSF lipidomic signature in MS patients at the time of diagnosis that might be considered as a potential diagnostic tool.

Cellular Uptake and Efflux of Palbociclib In Vitro in Single Cell and Spheroid Models.

Adequate drug distribution through tumors is essential for treatment to be effective. Palbociclib is a cyclin-dependent kinase 4/6 inhibitor approved for use in patients with hormone receptor positive, human epidermal growth factor receptor 2 negative metastatic breast cancer. It has unusual physicochemical properties, which may significantly influence its distribution in tumor tissue. We studied the penetration and distribution of palbociclib in vitro, including the use of multicellular three-dimensional models and mathematical modeling. MCF-7 and DLD-1 cell lines were grown as single cell suspensions (SCS) and spheroids; palbociclib uptake and efflux were studied using liquid chromatography-tandem mass spectrometry. Intracellular concentrations of palbociclib for MCF-7 SCS (C max 3.22 µM) and spheroids (C max 2.91 µM) were 32- and 29-fold higher and in DLD-1, 13- and 7-fold higher, respectively, than the media concentration (0.1 µM). Total palbociclib uptake was lower in DLD-1 cells than MCF-7 cells in both SCS and spheroids. Both uptake and efflux of palbociclib were slower in spheroids than SCS. These data were used to develop a mathematical model of palbociclib transport that quantifies key parameters determining drug penetration and distribution. The model reproduced qualitatively most features of the experimental data and distinguished between SCS and spheroids, providing additional support for hypotheses derived from the experimental data. Mathematical modeling has the potential for translating in vitro data into clinically relevant estimates of tumor drug concentrations. SIGNIFICANCE STATEMENT: This study explores palbociclib uptake and efflux in single cell suspension and spheroid models of cancer. Large intracellular concentrations of palbociclib are found after drug exposure. The data from this study may aid understanding of the intratumoural pharmacokinetics of palbociclib, which is useful in understanding how drug distributes within tumor tissue and optimizing drug efficacy. Biomathematical modelling has the potential to derive intratumoural drug concentrations from plasma pharmacokinetics in patients.

Exceptional human longevity is associated with a specific plasma phenotype of ether lipids.

A lipid profile resistant to oxidative damage is an inherent trait associated with animal lifespan. However, there is a lack of lipidomic studies on human longevity. Here we use mass spectrometry based technologies to detect and quantify 137 ether lipids to define a phenotype of healthy humans with exceptional lifespan. Ether lipids were chosen because of their antioxidant properties and ability to modulate oxidative stress. Our results demonstrate that a specific ether lipid signature can be obtained to define the centenarian state. This profile comprises higher level of alkyl forms derived from phosphatidylcholine with shorter number of carbon atoms and double bonds; and decreased content in alkenyl forms from phosphatidylethanolamine with longer chain length and higher double bonds. This compositional pattern suggests that ether lipids from centenarians are more resistant to lipid peroxidation, and that ether lipid signature expresses an optimized feature associated with exceptional human longevity. These results are in keeping with the free radical theory of aging.

Drug delivery in a tumour cord model: a computational simulation.

The tumour vasculature and microenvironment is complex and heterogeneous, contributing to reduced delivery of cancer drugs to the tumour. We have developed an in silico model of drug transport in a tumour cord to explore the effect of different drug regimes over a 72 h period and how changes in pharmacokinetic parameters affect tumour exposure to the cytotoxic drug doxorubicin. We used the model to describe the radial and axial distribution of drug in the tumour cord as a function of changes in the transport rate across the cell membrane, blood vessel and intercellular permeability, flow rate, and the binding and unbinding ratio of drug within the cancer cells. We explored how changes in these parameters may affect cellular exposure to drug. The model demonstrates the extent to which distance from the supplying vessel influences drug levels and the effect of dosing schedule in relation to saturation of drug-binding sites. It also shows the likely impact on drug distribution of the aberrant vasculature seen within tumours. The model can be adapted for other drugs and extended to include other parameters. The analysis confirms that computational models can play a role in understanding novel cancer therapies to optimize drug administration and delivery.

Activation of sirtuin 1 as therapy for the peroxisomal disease adrenoleukodystrophy.

Oxidative stress and mitochondrial failure are prominent factors in the axonal degeneration process. In this study, we demonstrate that sirtuin 1 (SIRT1), a key regulator of the mitochondrial function, is impaired in the axonopathy and peroxisomal disease X-linked adrenoleukodystrophy (X-ALD). We have restored SIRT1 activity using a dual strategy of resveratrol treatment or by the moderate transgenic overexpression of SIRT1 in a X-ALD mouse model. Both strategies normalized redox homeostasis, mitochondrial respiration, bioenergetic failure, axonal degeneration and associated locomotor disabilities in the X-ALD mice. These results indicate that the reactivation of SIRT1 may be a valuable strategy to treat X-ALD and other axonopathies in which the control of redox and energetic homeostasis is impaired.

Concurrent systemic therapy with radiotherapy for the treatment of poor-risk patients with unresectable stage III non-small-cell lung cancer: a review of the literature.

BACKGROUND: There is no consensus on the therapeutic approach to poor-risk patients with unresectable stage III non-small-cell lung cancer (NSCLC), despite the increasing number of these patients in current clinical practice. In terms of survival, the combination of concurrent systemic therapy with standard radiotherapy might be advantageous over radiotherapy alone. The purpose of this review is to ascertain the feasibility, safety and efficacy of the combination of concurrent systemic therapy and standard radiotherapy in these patients. METHODS: A computer-based literature search was carried out using PubMed and Science Direct for relevant publications; data reported at major conferences in abstract form were also included. RESULTS: In unresectable stage III NSCLC, advanced age, poor performance status, weight loss and comorbidities are factors that influence treatment options and disease outcomes in clinical practice. Prospective studies including poor-risk patients have been reviewed. Trials specifically recruiting poor-risk patients have been separated into those using chemotherapy and those using targeted agents with or without chemotherapy. Only two phase III studies specifically including poor-risk patients have been published. Some recent studies suggested that tolerable radio-sensitizing therapy combined with radiotherapy can provide longer survival outcomes than those reported earlier with chemo-radiotherapy or with radiotherapy alone. CONCLUSIONS: There is an unmet need to develop well-designed clinical trials with tolerable combinations of systemic therapy and radiotherapy specifically tailored to this lung cancer population. Such trials should incorporate careful comorbidity measurement and, in older adults, a validated geriatric assessment.

Neurotoxicidad por baclofeno en paciente con insuficiencia renal crónica terminal / Baclofen neurotoxicity in a patient with end-stage chronic renal failure

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Atherosclerosis prevention by nutritional factors: a meta-analysis in small animal models.

BACKGROUND AND AIMS: Atherosclerosis prevention in small laboratory models has been used as a preclinical stage in the development of functional foods with claimed antiatherogenic properties. However, a high heterogeneity of experimental atherosclerosis models as well as species-specific differences in lipoprotein metabolism could limit the usefulness of these developments. To solve this, we have performed a meta-analysis on the effects of nutritional complements (i.e. less than 2% of diet) with potential antiatherogenic properties in mice, rabbits and hamsters, and compared the outcomes with those obtained in humans. METHODS AND RESULTS: A meta-analysis comprising works dealing with dietary prevention of experimental atherosclerosis (i.e. macroscopic and/or pathological evidences of atheromatosis in aorta) has been performed (n = 110 works). Quality criteria were applied resulting in selection of 16 works comprising 511 animals. Despite high heterogeneity, there is a significant effect of nutritional interventions reducing atheroma globally (mean effect 24.38% (95% CI: 13.24-35.51%) of prevention). In mouse studies (20.64% (95% CI: 8.38-32.90%)) and in rabbits (40.48% (95% CI: 6.73-74.23%)) this effect was significant, in contrast with hamster-based works (95% CI: 13.66-49.48%). Meta-regression showed that reduction of atheroma plaque formation was not linked to changes either in total circulating cholesterol or LDL cholesterol levels. CONCLUSION: Nutritional addition of selected compounds significantly prevents experimental atheromatosis, but the reproduction of positive effects observed in humans was very limited. These analyses reinforce the need for adequate standardization of atherosclerosis studies in preclinical models and for human intervention trials.

Compatibility of entomopathogenic nematodes with fipronil.

The survival and infectivity of infective juveniles (IJs) of three species of entomopathogenic nematodes, Steinernema carpocapsae Weiser, S. arenarium (Artyukhovsky) (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae), were determined after exposure to different concentrations (250, 500, 1000 and 2000 ppm) of fipronil, an insecticide acting on the GABA receptors to block the chloride channel. Heterorhabditis bacteriophora was very tolerant to all concentrations of fipronil, with the highest mortality of 17% being observed at 2000 ppm of fipronil after 72 h exposure. Steinernema carpocapsae showed a similar response, with the highest mortality of 11.25% of IJs being observed after 72 h exposure to 2000 ppm of fipronil. Steinernema arenarium was, however, more sensitive to fipronil, and at 2000 ppm mortality rates of 94.6% and 100% were observed after 24 and 72 h, respectively. Fipronil had negligible effects on the infectivity of the three nematode species tested. The IJs which survive exposure to all concentrations of fipronil tested can infect and reproduce in Galleria larvae. The moderate effects on entomopathogenic nematodes of a lower fipronil concentration (250 ppm) and the field rates (12-60 ppm) of fipronil used as insecticide, suggest that direct mixing of entomopathogenic nematodes and fipronil at field rates is a viable integrated pest management option.

El ácido palmítico estimula la producción de interleucina 6 en células musculares esqueléticas por activación de la proteincinasa C y el factor nuclear kB / Palmitate stimulates interleukin-6 production in skeletal muscle cells through activation of protein kinase C and nuclear factor kB

Introducción. Los mecanismos causantes de la aparición de resistencia a la insulina mediada por ácidos grasos libres en el músculo esquelético no son bien conocidos. Diversos datos sugieren que existe una relación entre inflamación y diabetes mellitus tipo 2 que podría estar implicada en la aparición de esta patología. Material, métodos y resultados. La incubación de células musculares esqueléticas C2C12 con palmitato 0,5 mM incrementó los niveles de ARN mensajero (ARNm) (inducción de 3,5 veces, p < 0,05) y la secreción (control 375 ± 57 frente a palmitato 1.129 ± 177 pg/ml, p < 0,001) de la citocina proinflamatoria interleucina 6 (IL-6). El tratamiento con palmitato aumentó la activación del factor nuclear (NF)-kB y la coincubación de las células con palmitato y el inhibidor de NF-kB pirrolidina ditiocarbamato evitó tanto la expresión como la secreción de IL-6. Por otra parte, la incubación de las células tratadas con palmitato con un potente inhibidor específico de la proteincinasa C (PKC), la calfostina C, o con PMA, que disminuye los valores de PKC en incubaciones largas, suprimió la inducción de la producción de IL-6. Finalmente, las células musculares esqueléticas expuestas a palmitato mostraron una caída de los valores de ARNm y de proteína del transportador de glucosa 4 (GLUT4), pero en presencia de un anticuerpo contra la IL-6, que neutraliza su actividad biológica, se evitaron estas reducciones. Conclusiones. Estos estudios sugieren que la IL-6 puede mediar parte de los efectos del palmitato sobre la sensibilidad a la insulina (AU)

Introduction. The mechanisms by which elevated levels of free fatty acids in skeletal muscle cause insulin resistance are not well understood. In addition, accumulating evidence suggests a link between inflammation and type 2 diabetes, which could be involved in the development of this disorder. Material, methods and results. Exposure of C2C12 skeletal muscle cells to 0.5 mM palmitate increased mRNA levels (3.5-fold induction, p<0.05) and secretion (control 375±57 vs palmitate 1129±177 pg/ml, p<0.001) of the proinflammatory cytokine interleukin (IL)-6. Palmitate increased nuclear factor (NF)-kB activation and coincubation of the cells with palmitate and the NF-kB inhibitor pyrrolidine dithiocarbamate prevented both IL-6 expression and secretion. Furthermore, incubation of palmitate-treated cells with calphostin C, a strong and specific inhibitor of protein kinase C (PKC), and PMA, which down-regulates PKC in long-term incubations, abolished induction of IL-6 production. Finally, exposure of skeletal muscle cells to palmitate decreased mRNA and protein levels of GLUT4. However, in the presence of anti-IL-6 antibody, which neutralizes the biological activity of mouse IL-6 in cell culture, these reductions were prevented. Conclusions. These findings suggest that IL-6 may mediate several of the prodiabetic effects of palmitate (AU)

D) ¿Modula el metabolismo lipídico del miocardio la hipertrofia cardíaca? / No disponible

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