Anticancerous antibody development targeting death receptors

Le développement d'anticorps thérapeutiques a suscité beaucoup d'intérêt au cours des dernières décennies. Plus de 30 d'entre eux ont été approuvés et sont utilisés pour traiter des patients atteints de cancer. Les récepteurs agonistes de TRAIL (DR4 ou DR5) sont surexprimés par les cellules tumorales et capables de déclencher leur mort. Ils représentent donc des cibles idéales. Malgré des résultats encourageants, la plupart des essais cliniques basés sur des anticorps monoclonaux ciblants DR4 ou DR5 ont été interrompus. Cependant, les connaissances actuelles ouvrent des perspectives thérapeutiques de choix pour l'utilisation de tels anticorps en oncologie. Afin de développer de nouveaux anticorps anti-DR4 et anti-DR5 reconnaissant sélectivement les protéines natives d’intérêt, et doués de propriétés antitumorales, nous avons opté pour une approche d'immunisation génétique basée sur des injections hydrodynamiques d'ADN complémentaire. Cette approche nous a permis d'obtenir des réponses humorales significatives, et après fusions des rates correspondantes, de générer 21 anticorps monoclonaux capables de reconnaître spécifiquement et avec une très grande affinité les récepteurs DR4 ou DR5, sous leurs formes natives. Parmi ces anticorps monoclonaux, deux sont doués de propriétés pro-apoptotiques, et quatre sont capables d'accroitre le potentiel pro-apoptotique du ligand TRAIL. Les propriétés antitumorales de l'anticorps anti-DR4 le plus puissant, l’AcM-C#16, ont également été validées in-vivo dans des modèles de xénogreffes.L'ensemble de ce travail démontre, et ce pour la première fois, que la méthode d'immunisation ADN par injection hydrodynamique peut être utilisée pour générer des anticorps monoclonaux thérapeutiques efficaces ciblant des récepteurs de la superfamille du TNF. Au-delà du système TRAIL, cette approche d'immunisation, peu exploitée, pourrait ouvrir de nouvelles perspectives thérapeutiques en l'adaptant à de nouvelles cibles.Development of therapeutic antibodies has attracted many interests in recent decades. More than 30 of them have been approved and are used to treat cancer patients. TRAIL agonist receptors (DR4 or DR5) are overexpressed by the tumour cells and are able to trigger their death. Therefore, they represent ideal targets. Despite encouraging results, most clinical trials based on monoclonal antibodies targeting DR4 or DR5 have been discontinued. However, current knowledge opens therapeutic perspectives of choice for the use of such antibodies in oncology. In order to develop new anti-DR4 and anti-DR5 antibodies recognizing selectively the native form of the proteins of interest, and endowed with antitumor properties, we have chosen to perform a genetic immunization approach based on hydrodynamic injections of complementary DNA. This approach allowed us to obtain significant humoral responses, and after fusions of the corresponding spleens, to generate 21 monoclonal antibodies capable of recognizing specifically and with very high affinity DR4 or DR5 receptors, in their native forms. Of these monoclonal antibodies, two are display pro-apoptotic properties, and four are capable of enhancing TRAIL pro-apoptotic potential. The antitumor properties of the most potent anti-DR4 antibody, mAb-C16, have also been validated using in-vivo xenografts models.Altogether this work demonstrates, for the first time, that the DNA immunization hydrodynamic injection method can be used to generate therapeutically effective monoclonal antibodies targeting TNF superfamily receptors. Beyond the TRAIL system, this immunization approach, scarcely exploited, could open new therapeutic perspectives by adapting it to new targets

Génération et caractérisation de nouveaux anticorps anti-DR4 et anti-DR5

Development of therapeutic antibodies has attracted many interests in recent decades. More than 30 of them have been approved and are used to treat cancer patients. TRAIL agonist receptors (DR4 or DR5) are overexpressed by the tumour cells and are able to trigger their death. Therefore, they represent ideal targets. Despite encouraging results, most clinical trials based on monoclonal antibodies targeting DR4 or DR5 have been discontinued. However, current knowledge opens therapeutic perspectives of choice for the use of such antibodies in oncology. In order to develop new anti-DR4 and anti-DR5 antibodies recognizing selectively the native form of the proteins of interest, and endowed with antitumor properties, we have chosen to perform a genetic immunization approach based on hydrodynamic injections of complementary DNA. This approach allowed us to obtain significant humoral responses, and after fusions of the corresponding spleens, to generate 21 monoclonal antibodies capable of recognizing specifically and with very high affinity DR4 or DR5 receptors, in their native forms. Of these monoclonal antibodies, two are display pro-apoptotic properties, and four are capable of enhancing TRAIL pro-apoptotic potential. The antitumor properties of the most potent anti-DR4 antibody, mAb-C16, have also been validated using in-vivo xenografts models.Altogether this work demonstrates, for the first time, that the DNA immunization hydrodynamic injection method can be used to generate therapeutically effective monoclonal antibodies targeting TNF superfamily receptors. Beyond the TRAIL system, this immunization approach, scarcely exploited, could open new therapeutic perspectives by adapting it to new targets.Le développement d'anticorps thérapeutiques a suscité beaucoup d'intérêt au cours des dernières décennies. Plus de 30 d'entre eux ont été approuvés et sont utilisés pour traiter des patients atteints de cancer. Les récepteurs agonistes de TRAIL (DR4 ou DR5) sont surexprimés par les cellules tumorales et capables de déclencher leur mort. Ils représentent donc des cibles idéales. Malgré des résultats encourageants, la plupart des essais cliniques basés sur des anticorps monoclonaux ciblants DR4 ou DR5 ont été interrompus. Cependant, les connaissances actuelles ouvrent des perspectives thérapeutiques de choix pour l'utilisation de tels anticorps en oncologie. Afin de développer de nouveaux anticorps anti-DR4 et anti-DR5 reconnaissant sélectivement les protéines natives d’intérêt, et doués de propriétés antitumorales, nous avons opté pour une approche d'immunisation génétique basée sur des injections hydrodynamiques d'ADN complémentaire. Cette approche nous a permis d'obtenir des réponses humorales significatives, et après fusions des rates correspondantes, de générer 21 anticorps monoclonaux capables de reconnaître spécifiquement et avec une très grande affinité les récepteurs DR4 ou DR5, sous leurs formes natives. Parmi ces anticorps monoclonaux, deux sont doués de propriétés pro-apoptotiques, et quatre sont capables d'accroitre le potentiel pro-apoptotique du ligand TRAIL. Les propriétés antitumorales de l'anticorps anti-DR4 le plus puissant, l’AcM-C#16, ont également été validées in-vivo dans des modèles de xénogreffes.L'ensemble de ce travail démontre, et ce pour la première fois, que la méthode d'immunisation ADN par injection hydrodynamique peut être utilisée pour générer des anticorps monoclonaux thérapeutiques efficaces ciblant des récepteurs de la superfamille du TNF. Au-delà du système TRAIL, cette approche d'immunisation, peu exploitée, pourrait ouvrir de nouvelles perspectives thérapeutiques en l'adaptant à de nouvelles cibles

Génération et caractérisation de nouveaux anticorps anti-DR4 et anti-DR5

Development of therapeutic antibodies has attracted many interests in recent decades. More than 30 of them have been approved and are used to treat cancer patients. TRAIL agonist receptors (DR4 or DR5) are overexpressed by the tumour cells and are able to trigger their death. Therefore, they represent ideal targets. Despite encouraging results, most clinical trials based on monoclonal antibodies targeting DR4 or DR5 have been discontinued. However, current knowledge opens therapeutic perspectives of choice for the use of such antibodies in oncology. In order to develop new anti-DR4 and anti-DR5 antibodies recognizing selectively the native form of the proteins of interest, and endowed with antitumor properties, we have chosen to perform a genetic immunization approach based on hydrodynamic injections of complementary DNA. This approach allowed us to obtain significant humoral responses, and after fusions of the corresponding spleens, to generate 21 monoclonal antibodies capable of recognizing specifically and with very high affinity DR4 or DR5 receptors, in their native forms. Of these monoclonal antibodies, two are display pro-apoptotic properties, and four are capable of enhancing TRAIL pro-apoptotic potential. The antitumor properties of the most potent anti-DR4 antibody, mAb-C16, have also been validated using in-vivo xenografts models.Altogether this work demonstrates, for the first time, that the DNA immunization hydrodynamic injection method can be used to generate therapeutically effective monoclonal antibodies targeting TNF superfamily receptors. Beyond the TRAIL system, this immunization approach, scarcely exploited, could open new therapeutic perspectives by adapting it to new targets.Le développement d'anticorps thérapeutiques a suscité beaucoup d'intérêt au cours des dernières décennies. Plus de 30 d'entre eux ont été approuvés et sont utilisés pour traiter des patients atteints de cancer. Les récepteurs agonistes de TRAIL (DR4 ou DR5) sont surexprimés par les cellules tumorales et capables de déclencher leur mort. Ils représentent donc des cibles idéales. Malgré des résultats encourageants, la plupart des essais cliniques basés sur des anticorps monoclonaux ciblants DR4 ou DR5 ont été interrompus. Cependant, les connaissances actuelles ouvrent des perspectives thérapeutiques de choix pour l'utilisation de tels anticorps en oncologie. Afin de développer de nouveaux anticorps anti-DR4 et anti-DR5 reconnaissant sélectivement les protéines natives d’intérêt, et doués de propriétés antitumorales, nous avons opté pour une approche d'immunisation génétique basée sur des injections hydrodynamiques d'ADN complémentaire. Cette approche nous a permis d'obtenir des réponses humorales significatives, et après fusions des rates correspondantes, de générer 21 anticorps monoclonaux capables de reconnaître spécifiquement et avec une très grande affinité les récepteurs DR4 ou DR5, sous leurs formes natives. Parmi ces anticorps monoclonaux, deux sont doués de propriétés pro-apoptotiques, et quatre sont capables d'accroitre le potentiel pro-apoptotique du ligand TRAIL. Les propriétés antitumorales de l'anticorps anti-DR4 le plus puissant, l’AcM-C#16, ont également été validées in-vivo dans des modèles de xénogreffes.L'ensemble de ce travail démontre, et ce pour la première fois, que la méthode d'immunisation ADN par injection hydrodynamique peut être utilisée pour générer des anticorps monoclonaux thérapeutiques efficaces ciblant des récepteurs de la superfamille du TNF. Au-delà du système TRAIL, cette approche d'immunisation, peu exploitée, pourrait ouvrir de nouvelles perspectives thérapeutiques en l'adaptant à de nouvelles cibles

Dombeya formosa (Malvaceae s. l.): A New Species Endemic to La Réunion (Indian Ocean) Based on Morphological and Molecular Evidence

International audienceDombeya formosa (Malvaceae s. l. or Dombeyaceae), a new species endemic to La Réunion, is described and illustrated. The species is distinguishable by its ovate leaves with round or slightly cordate bases, round to lightly acute apices and glabrescent abaxial surfaces, subulate and persistent stipules, umbellate inflorescences, and acute apices of floral bracts. Dombeya formosa is morphologically close to Dombeya punctata Cav. but differs from it in having ovate leaves and acute apices on floral bracts. Dombeya formosa is also morphologically similar to D. ficulnea Baill. but can be distinguished from it by its persistent and subulate stipules as well as the glabrescent adaxial surface of its leaves. Our study, focusing on 16 quantitative floral characters, demonstrates that there is a clear morphological distinction between D. formosa, D. punctata, and D. ficulnea. In comparison to the other two species, Dombeya formosa possesses smaller flowers and additionally shows cryptic dioecy along with flower-size dimorphism between sexes. Amongst the Mascarene Dombeyoideae, it is the only species distributed up to an altitude of 2,100 m. The phylogenetic position of Dombeya formosa is consistent with the morphological features and locates the species within the clade endemic to La Réunion characterized by umbellate inflorescences

Circulating acetylated polyamines correlate with Covid-19 severity in cancer patients

International audienceCancer patients are particularly susceptible to the development of severe Covid-19, prompting us to investigate the serum metabolome of 204 cancer patients enrolled in the ONCOVID trial. We previously described that the immunosuppressive tryptophan/kynurenine metabolite anthranilic acid correlates with poor prognosis in non-cancer patients. In cancer patients, we observed an elevation of anthranilic acid at baseline (without Covid-19 diagnosis) and no further increase with mild or severe Covid-19. We found that, in cancer patients, Covid-19 severity was associated with the depletion of two bacterial metabolites, indole-3-proprionate and 3-phenylproprionate, that both positively correlated with the levels of several inflammatory cytokines. Most importantly, we observed that the levels of acetylated polyamines (in particular N1-acetylspermidine, N1,N8-diacetylspermidine and N1,N12-diacetylspermine), alone or in aggregate, were elevated in severe Covid-19 cancer patients requiring hospitalization as compared to uninfected cancer patients or cancer patients with mild Covid-19. N1-acetylspermidine and N1,N8-diacetylspermidine were also increased in patients exhibiting prolonged viral shedding (>40 days). An abundant literature indicates that such acetylated polyamines increase in the serum from patients with cancer, cardiovascular disease or neurodegeneration, associated with poor prognosis. Our present work supports the contention that acetylated polyamines are associated with severe Covid-19, both in the general population and in patients with malignant disease. Severe Covid-19 is characterized by a specific metabolomic signature suggestive of the overactivation of spermine/spermidine N1-acetyl transferase-1 (SAT1), which catalyzes the first step of polyamine catabolism

Intestinal Akkermansia muciniphila predicts clinical response to PD-1 blockade in patients with advanced non-small-cell lung cancer

43siAside from PD-L1 expression, biomarkers of response to immune checkpoint inhibitors (ICIs) in non-small-cell lung cancer (NSCLC) are needed. In a previous retrospective analysis, we documented that fecal Akkermansia muciniphila (Akk) was associated with clinical benefit of ICI in patients with NSCLC or kidney cancer. In the current study, we performed shotgun-metagenomics-based microbiome profiling in a large cohort of patients with advanced NSCLC (n = 338) treated with first- or second-line ICIs to prospectively validate the predictive value of fecal Akk. Baseline stool Akk was associated with increased objective response rates and overall survival in multivariate analyses, independent of PD-L1 expression, antibiotics, and performance status. Intestinal Akk was accompanied by a richer commensalism, including Eubacterium hallii and Bifidobacterium adolescentis, and a more inflamed tumor microenvironment in a subset of patients. However, antibiotic use (20% of cases) coincided with a relative dominance of Akk above 4.8% accompanied with the genus Clostridium, both associated with resistance to ICI. Our study shows significant differences in relative abundance of Akk that may represent potential biomarkers to refine patient stratification in future studies.reservedopenDerosa, Lisa; Routy, Bertrand; Thomas, Andrew Maltez; Iebba, Valerio; Zalcman, Gerard; Friard, Sylvie; Mazieres, Julien; Audigier-Valette, Clarisse; Moro-Sibilot, Denis; Goldwasser, François; Silva, Carolina Alves Costa; Terrisse, Safae; Bonvalet, Melodie; Scherpereel, Arnaud; Pegliasco, Hervé; Richard, Corentin; Ghiringhelli, François; Elkrief, Arielle; Desilets, Antoine; Blanc-Durand, Felix; Cumbo, Fabio; Blanco, Aitor; Boidot, Romain; Chevrier, Sandy; Daillère, Romain; Kroemer, Guido; Alla, Laurie; Pons, Nicolas; Le Chatelier, Emmanuelle; Galleron, Nathalie; Roume, Hugo; Dubuisson, Agathe; Bouchard, Nicole; Messaoudene, Meriem; Drubay, Damien; Deutsch, Eric; Barlesi, Fabrice; Planchard, David; Segata, Nicola; Martinez, Stéphanie; Zitvogel, Laurence; Soria, Jean-Charles; Besse, BenjaminDerosa, Lisa; Routy, Bertrand; Thomas, Andrew Maltez; Iebba, Valerio; Zalcman, Gerard; Friard, Sylvie; Mazieres, Julien; Audigier-Valette, Clarisse; Moro-Sibilot, Denis; Goldwasser, François; Silva, Carolina Alves Costa; Terrisse, Safae; Bonvalet, Melodie; Scherpereel, Arnaud; Pegliasco, Hervé; Richard, Corentin; Ghiringhelli, François; Elkrief, Arielle; Desilets, Antoine; Blanc-Durand, Felix; Cumbo, Fabio; Blanco, Aitor; Boidot, Romain; Chevrier, Sandy; Daillère, Romain; Kroemer, Guido; Alla, Laurie; Pons, Nicolas; Le Chatelier, Emmanuelle; Galleron, Nathalie; Roume, Hugo; Dubuisson, Agathe; Bouchard, Nicole; Messaoudene, Meriem; Drubay, Damien; Deutsch, Eric; Barlesi, Fabrice; Planchard, David; Segata, Nicola; Martinez, Stéphanie; Zitvogel, Laurence; Soria, Jean-Charles; Besse, Benjami

Immune responses during COVID-19 infection

International audienceOver the past 16 years, three coronaviruses (CoVs), severe acute respiratory syndrome CoV (SARS-CoV) in 2002, Middle East respiratory syndrome CoV (MERS-CoV) in 2012 and 2015, and SARS-CoV-2 in 2020, have been causing severe and fatal human epidemics. The unpredictability of coronavirus disease-19 (COVID-19) poses a major burden on health care and economic systems across the world. This is caused by the paucity of in-depth knowledge of the risk factors for severe COVID-19, insufficient diagnostic tools for the detection of SARS-CoV-2, as well as the absence of specific and effective drug treatments. While protective humoral and cellular immune responses are usually mounted against these betacoronaviruses, immune responses to SARS-CoV2 sometimes derail towards inflammatory tissue damage, leading to rapid admissions to intensive care units. The lack of knowledge on mechanisms that tilt the balance between these two opposite outcomes poses major threats to many ongoing clinical trials dealing with immunostimulatory or immunoregulatory therapeutics. This review will discuss innate and cognate immune responses underlying protective or deleterious immune reactions against these pathogenic coronaviruses

Biosynthetic proteins targeting the SARS-CoV-2 spike as anti-virals

International audienceThe binding of the SARS-CoV-2 spike to angiotensin-converting enzyme 2 (ACE2) promotes virus entry into the cell. Targeting this interaction represents a promising strategy to generate antivirals. By screening a phage-display library of biosynthetic protein sequences build on a rigid alpha-helicoidal HEAT-like scaffold (named αReps), we selected candidates recognizing the spike receptor binding domain (RBD). Two of them (F9 and C2) bind the RBD with affinities in the nM range, displaying neutralisation activity in vitro and recognizing distinct sites, F9 overlapping the ACE2 binding motif. The F9-C2 fusion protein and a trivalent αRep form (C2-foldon) display 0.1 nM affinities and EC 50 of 8–18 nM for neutralization of SARS-CoV-2. In hamsters, F9-C2 instillation in the nasal cavity before or during infections effectively reduced the replication of a SARS-CoV-2 strain harbouring the D614G mutation in the nasal epithelium. Furthermore, F9-C2 and/or C2-foldon effectively neutralized SARS-CoV-2 variants (including delta and omicron variants) with EC 50 values ranging from 13 to 32 nM. With their high stability and their high potency against SARS-CoV-2 variants, αReps provide a promising tool for SARS-CoV-2 therapeutics to target the nasal cavity and mitigate virus dissemination in the proximal environment

Cross-tissue single-cell landscape of human monocytes and macrophages in health and disease

10.1016/j.immuni.2021.07.007IMMUNITY5481883-

Metabolomic analyses of COVID-19 patients unravel stage-dependent and prognostic biomarkers

Abstract The circulating metabolome provides a snapshot of the physiological state of the organism responding to pathogenic challenges. Here we report alterations in the plasma metabolome reflecting the clinical presentation of COVID-19 patients with mild (ambulatory) diseases, moderate disease (radiologically confirmed pneumonitis, hospitalization and oxygen therapy), and critical disease (in intensive care). This analysis revealed major disease- and stage-associated shifts in the metabolome, meaning that at least 77 metabolites including amino acids, lipids, polyamines and sugars, as well as their derivatives, were altered in critical COVID-19 patient’s plasma as compared to mild COVID-19 patients. Among a uniformly moderate cohort of patients who received tocilizumab, only 10 metabolites were different among individuals with a favorable evolution as compared to those who required transfer into the intensive care unit. The elevation of one single metabolite, anthranilic acid, had a poor prognostic value, correlating with the maintenance of high interleukin-10 and -18 levels. Given that products of the kynurenine pathway including anthranilic acid have immunosuppressive properties, we speculate on the therapeutic utility to inhibit the rate-limiting enzymes of this pathway including indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase