Drug sensitivity profiling of 3D tumor tissue cultures in the pediatric precision oncology program INFORM

The international precision oncology program INFORM enrolls relapsed/refractory pediatric cancer patients for comprehensive molecular analysis. We report a two-year pilot study implementing ex vivo drug sensitivity profiling (DSP) using a library of 75-78 clinically relevant drugs. We included 132 viable tumor samples from 35 pediatric oncology centers in seven countries. DSP was conducted on multicellular fresh tumor tissue spheroid cultures in 384-well plates with an overall mean processing time of three weeks. In 89 cases (67%), sufficient viable tissue was received; 69 (78%) passed internal quality controls. The DSP results matched the identified molecular targets, including BRAF, ALK, MET, and TP53 status. Drug vulnerabilities were identified in 80% of cases lacking actionable (very) high-evidence molecular events, adding value to the molecular data. Striking parallels between clinical courses and the DSP results were observed in selected patients. Overall, DSP in clinical real-time is feasible in international multicenter precision oncology programs

Drug sensitivity profiling of 3D tumor tissue cultures in the pediatric precision oncology program INFORM

The international precision oncology program INFORM enrolls relapsed/refractory pediatric cancer patients for comprehensive molecular analysis. We report a two-year pilot study implementing ex vivo drug sensitivity profiling (DSP) using a library of 75–78 clinically relevant drugs. We included 132 viable tumor samples from 35 pediatric oncology centers in seven countries. DSP was conducted on multicellular fresh tumor tissue spheroid cultures in 384-well plates with an overall mean processing time of three weeks. In 89 cases (67%), sufficient viable tissue was received; 69 (78%) passed internal quality controls. The DSP results matched the identified molecular targets, including BRAF, ALK, MET, and TP53 status. Drug vulnerabilities were identified in 80% of cases lacking actionable (very) high-evidence molecular events, adding value to the molecular data. Striking parallels between clinical courses and the DSP results were observed in selected patients. Overall, DSP in clinical real-time is feasible in international multicenter precision oncology programs

Drug sensitivity profiling of 3D tumor tissue cultures in the pediatric precision oncology program INFORM

The international precision oncology program INFORM enrolls relapsed/refractory pediatric cancer patients for comprehensive molecular analysis. We report a two-year pilot study implementing ex vivo drug sensitivity profiling (DSP) using a library of 75-78 clinically relevant drugs. We included 132 viable tumor samples from 35 pediatric oncology centers in seven countries. DSP was conducted on multicellular fresh tumor tissue spheroid cultures in 384-well plates with an overall mean processing time of three weeks. In 89 cases (67%), sufficient viable tissue was received; 69 (78%) passed internal quality controls. The DSP results matched the identified molecular targets, including BRAF, ALK, MET, and TP53 status. Drug vulnerabilities were identified in 80% of cases lacking actionable (very) high-evidence molecular events, adding value to the molecular data. Striking parallels between clinical courses and the DSP results were observed in selected patients. Overall, DSP in clinical real-time is feasible in international multicenter precision oncology programs.Peer reviewe

Physiological and pharmaceutical approaches in a mouse model of amyotrophic lateral sclerosis

La sclérose latérale amyotrophique (SLA) est une maladie neurodégénérative caractérisée par une mort sélective des motoneurones. Les mécanismes impliqués dans cette pathologie sont encore mal connus. Il apparait néanmoins que la SLA est une maladie multifactorielle impliquant différents partenaires tels que les motoneurones, les cellules gliales et musculaires. Le modèle murin SOD1G93A développe un phénotype semblable à celui observé chez les patients SLA. L'étude précise du patron locomoteur des SOD1G93A nous a permis de re-définir la date d'apparition des symptômes à 2 mois d'âge soit 1 un mois avant la date communément admise. Nous corrélons cette donnée fonctionnelle à des modifications histologiques au niveau médullaire, en particulier sur la composante gliale, et musculaire. Sur cette base, nous avons développé deux approches in vivo, l'une physiologique et l'autre pharmacologique. D'une part nous avons caractérisé les effets d'exercices physiques de différentes intensités chez les souris SOD1G93A. Cette étude à mis en évidence le rôle primordial de l'environnement et ne démontre pas d'effet de l'exercice sur la survie des souris SOD1G93A. D'autre part, nous rapportons une augmentation de survie des souris SOD1G93A après traitement chronique à faible dose d'une molécule anti-glutamatergique, la gacyclidine. A forte dose cette molécule semble avoir un effet néfaste.En parallèle, nous avons décrit la répartition anatomique de la sérotonine et d'un de ses récepteurs dans la moelle épinière humaine. Nous observons de grandes similarités topographiques avec les murins et primates, et validons ainsi l'utilisation future de ces modèles animaux dans les pathologies affectant la locomotion tel que la SLA.Amytrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by a selective death of motoneurons. Pathogenesis and mechanisms of selective vulnerability are not yet fully understood although there is growing evidences that ALS is a complex multi-factorial disease that involves several partners such as neuron, glial and muscle cells. Transgenic mice over-expressing a human mutated form of the gene coding for SOD1 develop a dominantly inherited adult-onset paralytic disorder that mimics human ALS symptoms. The precise description of the SOD1G93A mice locomotor pattern using a gait analysis method allowed us to refine symptoms onset at two months of age. This is one month earlier than described in the literature. We correlate these functional modifications to histological alterations of (1) the glial component of the spinal cord, and (2) muscles. From this referent study, we have then developed and evaluated a physiological and pharmacological in vivo approaches. In our first study, we have characterized the effect of different intensities of physical exercise on SOD1G93A mice. Our study not only demonstrates the crucial role of the environment but also that exercise does not have an impact on the survival of SOD1G93A mice. In the second part of our work, we report an increase in SOD1G93A mice survival when they had been chronically treated with a low dose of gacyclidine, an anti-glutamatergic molecule. At higher dose, this molecule seems to be detrimental.In a parallel study, we have carried out the anatomical description of serotonin and one of its receptor in the adult human spinal cord. We observe topographic similarities with rodents and primates, thus validating their further use as animal models, to study motor pathologies such as ALS

Population Density and Temperature Influence the Return on Maternal Investment in Wild House Mice

In mammals, reproduction is influenced by sexual competition, temperature and food availability and these factors might be crucial already during early life. Favorable early life environment and high maternal investment are expected to improve survival and reproduction. For example, in mammals, maternal investment via lactation predicts offspring growth. As body mass is often associated with fitness consequences, females have the potential to influence offspring fitness through their level of investment, which might interact with effects of population density and temperature. Here, we investigate the relationship between house mouse (Mus musculus domesticus) pup body mass at day 13 (used as approximation for weaning mass) and individual reproductive parameters, as well as longevity, under natural variation in population density and temperature (as approximation for season). Further, we assessed the extent to which mothers influence the body mass of their offspring until weaning. To do so, we analyzed life data of 384 house mice from a free-living wild commensal population that was not food limited. The mother’s contribution accounted for 49% of the variance in pup body mass. Further, we found a complex effect of population density, temperature and maternal investment on life-history traits related to fitness: shorter longevity with increasing pup body mass at day 13, delayed first reproduction of heavier pups when raised at warmer temperatures, and increased lifetime reproductive success for heavier pups at high densities. Our study shows that the effects of maternal investment are not independent of the effects of the environment. It thus highlights the importance of considering ecological conditions in combination with maternal effects to unravel the complexity of pup body mass on fitness measures

Approche physiologiques et pharmacologiques dans un modèle murin de la sclérose latérale amytrophique

La sclérose latérale amyotrophique (SLA) est une maladie neurodégénérative caractérisée par une mort sélective des motoneurones. Les mécanismes impliqués dans cette pathologie sont encore mal connus. Il apparait néanmoins que la SLA est une maladie multifactorielle impliquant différents partenaires tels que les motoneurones, les cellules gliales et musculaires. Le modèle murin SOD1G93A développe un phénotype semblable à celui observé chez les patients SLA. L'étude précise du patron locomoteur des SOD1G93A nous a permis de re-définir la date d'apparition des symptômes à 2 mois d'âge soit 1 un mois avant la date communément admise. Nous corrélons cette donnée fonctionnelle à des modifications histologiques au niveau médullaire, en particulier sur la composante gliale, et musculaire. Sur cette base, nous avons développé deux approches in vivo, l'une physiologique et l'autre pharmacologique. D'une part nous avons caractérisé les effets d'exercices physiques de différentes intensités chez les souris SOD1G93A. Cette étude à mis en évidence le rôle primordial de l'environnement et ne démontre pas d'effet de l'exercice sur la survie des souris SOD1G93A. D'autre part, nous rapportons une augmentation de survie des souris SOD1G93A après traitement chronique à faible dose d'une molécule anti-glutamatergique, la gacyclidine. A forte dose cette molécule semble avoir un effet néfaste.En parallèle, nous avons décrit la répartition anatomique de la sérotonine et d'un de ses récepteurs dans la moelle épinière humaine. Nous observons de grandes similarités topographiques avec les murins et primates, et validons ainsi l'utilisation future de ces modèles animaux dans les pathologies affectant la locomotion tel que la SLA.Amytrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by a selective death of motoneurons. Pathogenesis and mechanisms of selective vulnerability are not yet fully understood although there is growing evidences that ALS is a complex multi-factorial disease that involves several partners such as neuron, glial and muscle cells. Transgenic mice over-expressing a human mutated form of the gene coding for SOD1 develop a dominantly inherited adult-onset paralytic disorder that mimics human ALS symptoms. The precise description of the SOD1G93A mice locomotor pattern using a gait analysis method allowed us to refine symptoms onset at two months of age. This is one month earlier than described in the literature. We correlate these functional modifications to histological alterations of (1) the glial component of the spinal cord, and (2) muscles. From this referent study, we have then developed and evaluated a physiological and pharmacological in vivo approaches. In our first study, we have characterized the effect of different intensities of physical exercise on SOD1G93A mice. Our study not only demonstrates the crucial role of the environment but also that exercise does not have an impact on the survival of SOD1G93A mice. In the second part of our work, we report an increase in SOD1G93A mice survival when they had been chronically treated with a low dose of gacyclidine, an anti-glutamatergic molecule. At higher dose, this molecule seems to be detrimental.In a parallel study, we have carried out the anatomical description of serotonin and one of its receptor in the adult human spinal cord. We observe topographic similarities with rodents and primates, thus validating their further use as animal models, to study motor pathologies such as ALS.MONTPELLIER-BU Pharmacie (341722105) / SudocSudocFranceF

Dynamic Diversity of Glial Response Among Species in Spinal Cord Injury

International audienceThe glial scar that forms after traumatic spinal cord injury (SCI) is mostly composed of microglia, NG2 glia, and astrocytes and plays dual roles in pathophysiological processes induced by the injury. On one hand, the glial scar acts as a chemical and physical obstacle to spontaneous axonal regeneration, thus preventing functional recovery, and, on the other hand, it partly limits lesion extension. The complex activation pattern of glial cells is associated with cellular and molecular crosstalk and interactions with immune cells. Interestingly, response to SCI is diverse among species: from amphibians and fishes that display rather limited (if any) glial scarring to mammals that exhibit a well-identifiable scar. Additionally, kinetics of glial activation varies among species. In rodents, microglia become activated before astrocytes, and both glial cell populations undergo activation processes reflected amongst others by proliferation and migration toward the injury site. In primates, glial cell activation is delayed as compared to rodents. Here, we compare the spatial and temporal diversity of the glial response, following SCI amongst species. A better understanding of mechanisms underlying glial activation and scar formation is a prerequisite to develop timely glial cell-specific therapeutic strategies that aim to increase functional recovery