Efficacy and safety data for checkpoint inhibitors in advanced melanoma under real-life conditions: A monocentric study conducted in Nice from 2010 to 2016.

BACKGROUND: Immunotherapies using anti-CTLA4 and anti-PD1 antibodies have revolutionised the management of patients with advanced melanoma. The aim of our study was to analyse the efficacy and safety of immunotherapies in patients with advanced melanoma under real-life conditions. METHODS: We conducted a monocentric, retrospective, observational study that included all patients treated with immunotherapies (ipilimumab, i.e. ipi; nivolumab, i.e. niv and pembrolizumab, i.e. pbr) for advanced melanoma with exclusion of primary mucosal or ocular melanoma. The primary endpoint was progression-free survival (PFS). RESULTS: A total of 110 patients were included. Median PFS was better in the anti-PD1 group than in the anti-CTLA4 group (3.9 months vs. 2.9 months, P=0.025). The frequency of adverse events of any grade was 53.4% with ipi, 66.7% with niv and 75% with pbr. DISCUSSION: Our study shows slightly inferior efficacy data vs. clinical trials of ipi and niv because patients were presenting more severe illness at inclusion. Nevertheless, the study argues in favour of the superiority of anti-PD1 antibodies vs. anti-CTLA4 antibodies in terms of PFS and best overall response. Moreover, our study exhibits safety data comparable to those from clinical trials except for a lower frequency with ipi. CONCLUSION: Our efficacy and safety data obtained under real-life conditions are reassuring since they are consistent with data already published.

Pharmacological modulation of LMNA SRSF1-dependent splicing abrogates diet-induced obesity in mice.

Bakground/Objectives:Intense drug discovery efforts in the metabolic field highlight the need for novel strategies for the treatment of obesity. Alternative splicing (AS) and/or polyadenylation enable the LMNA gene to express distinct protein isoforms that exert opposing effects on energy metabolism and lifespan. Here we aimed to use the splicing factor SRSF1 that contribute to the production of these different isoforms as a target to uncover new anti-obesity drug. SUBJECTS/METHODS: Small molecules modulating SR protein activity and splicing were tested for their abilities to interact with SRSF1 and to modulate LMNA (AS). Using an LMNA luciferase reporter we selected molecules that were tested in diet-induced obese (DIO) mice. Transcriptomic analyses were performed in the white adipose tissues from untreated and treated DIO mice and mice fed a chow diet. RESULTS: We identified a small molecule that specifically interacted with the RS domain of SRSF1. ABX300 abolished DIO in mice, leading to restoration of adipose tissue homeostasis. In contrast, ABX300 had no effect on mice fed a standard chow diet. A global transcriptomic analysis revealed similar profiles of white adipose tissue from DIO mice treated with ABX300 and from untreated mice fed a chow diet. Mice treated with ABX300 exhibited an increase in O2 consumption and a switch in fuel preference toward lipids. CONCLUSIONS: Targeting SRSF1 with ABX300 compensates for changes in RNA biogenesis induced by fat accumulation and consequently represents a novel unexplored approach for the treatment of obesity.

White-to-brite conversion in human adipocytes promotes metabolic reprogramming towards fatty acid anabolic and catabolic pathways.

OBJECTIVE: Fat depots with thermogenic activity have been identified in humans. In mice, the appearance of thermogenic adipocytes within white adipose depots (so-called brown-in-white i.e., brite or beige adipocytes) protects from obesity and insulin resistance. Brite adipocytes may originate from direct conversion of white adipocytes. The purpose of this work was to characterize the metabolism of human brite adipocytes. METHODS: Human multipotent adipose-derived stem cells were differentiated into white adipocytes and then treated with peroxisome proliferator-activated receptor (PPAR)γ or PPARα agonists between day 14 and day 18. Gene expression profiling was determined using DNA microarrays and RT-qPCR. Variations of mRNA levels were confirmed in differentiated human preadipocytes from primary cultures. Fatty acid and glucose metabolism was investigated using radiolabelled tracers, Western blot analyses and assessment of oxygen consumption. Pyruvate dehydrogenase kinase 4 (PDK4) knockdown was achieved using siRNA. In vivo, wild type and PPARα-null mice were treated with a ß3-adrenergic receptor agonist (CL316,243) to induce appearance of brite adipocytes in white fat depot. Determination of mRNA and protein levels was performed on inguinal white adipose tissue. RESULTS: PPAR agonists promote a conversion of white adipocytes into cells displaying a brite molecular pattern. This conversion is associated with transcriptional changes leading to major metabolic adaptations. Fatty acid anabolism i.e., fatty acid esterification into triglycerides, and catabolism i.e., lipolysis and fatty acid oxidation, are increased. Glucose utilization is redirected from oxidation towards glycerol-3-phophate production for triglyceride synthesis. This metabolic shift is dependent on the activation of PDK4 through inactivation of the pyruvate dehydrogenase complex. In vivo, PDK4 expression is markedly induced in wild-type mice in response to CL316,243, while this increase is blunted in PPARα-null mice displaying an impaired britening response. CONCLUSIONS: Conversion of human white fat cells into brite adipocytes results in a major metabolic reprogramming inducing fatty acid anabolic and catabolic pathways. PDK4 redirects glucose from oxidation towards triglyceride synthesis and favors the use of fatty acids as energy source for uncoupling mitochondria.

In vivo 31P nuclear magnetic resonance (NMR) study of cerebral metabolism during histotoxic hypoxia in mice.

The alterations of cerebral energetic metabolism and intracellular brain pH that occur during histotoxic hypoxia were estimated in mice by in vivo 31P nuclear magnetic resonance (NMR) spectrometry. The brain spectra obtained by means of chronically implanted surface coils connected to a special designed probe were recorded sequentially and continuously before, during, and after histotoxic hypoxia induced by an injection of potassium cyanide. The levels of PCr, ATP, inorganic phosphate, and intracellular pH estimated from the records of the 31P cerebral spectra and the cerebral cortical activity allow noninvasive monitoring of both the energetic metabolism and the functional state of the brain in unanesthetized animals. The time courses of these different parameters are largely the same as those obtained previously by invasive methods, however, the simultaneous and continuous monitoring performed in this study exhibits several unexpected dissociations between, respectively, onset of coma, decrease in PCr level and intracellular pHi, and recovery of normal levels of PCr and intracellular pH. These dissociations indicate that tissue acidosis plays a minor role in the changes in PCr levels, compared with ATP, and they confirm that the thresholds of oxidative metabolism required for functional tissue activity and a normal rate of ATP are clearly different.

Potency test of killed polio vaccines. Statistical analysis of test on chicken and preliminary results in combination with ELISA D-antigen determinations.

The authors resume a several years' experience of potency testing inactivated polio vaccines with an in vivo-vitro assay based on inoculation of 3 week-old chicken and measure of neutralizing antibodies at 5 days on Hep 2 cells in a microplate system. Critical points, specificity and reproducibility are successively outlined. Preliminary combining results with determination of D-antigen by ELISA method and human immunization are submitted in view of a future standardization.

Mouse tumour tests for quality control of C. parvum preparations.

Description of two mouse tumor tests selected to control the quality of C. parvum pilot productions: transplanted Ehrlich's ascitic tumor in inbred Swiss mice and transplanted YC8 ascitic tumor in isogenic Balb/c mice. The results obtained during three years of control are analyzed, particularly accuracy, long range adequacy and reproducibility of tumor challenges and C, parvum stimulations. The effects of some parameters of the tumor tests, e.g. dose-responses, treatment schedules, tumor-host relationships and standard preparations are discussed.