Formalin-fixation and paraffin-embedding (FFPE) is the worldwide gold standard for tissue preserv... more Formalin-fixation and paraffin-embedding (FFPE) is the worldwide gold standard for tissue preservation and routine material for general diagnostics and cancer in particular. Despite the fact that histology is complemented by other modalities such as in situ hybridization (ISH) or immunohistochemistry (IHC), the pathology can fail to provide the right diagnosis in a certain number of cases. Overall, the standard process is time-consuming, labor intensive and doesn’t reach the right sensitivity and specificity. Lipids are central players in cancer. Unfortunately, to this date lipids aren’t used for diagnostics and lipid analysis from FFPE has never been reported due to tissue processing which limits their detection. Here, we report a new method for cancer diagnostics based on their unique tissue-lipidomic signatures through rapid molecular screening of FFPE sections without dewaxing using Water-Assisted Laser Desorption Ionization Mass Spectrometry (WALDI-MS) - SpiderMass. Advantageously, the method does not require human intervention. The annotated tissues are first analyzed to train the system and then blind samples can be diagnosed in less than a few minutes. The platform was demonstrated for the diagnostics of canine and human sarcoma, including cases which are known to be problematic in the gold standard workflow. Interrogation of blind samples against the built models revealed over 95% probability values to discriminate specific subtypes
F14512 is a new topoisomerase II inhibitor containing a spermine moiety that facilitates selectiv... more F14512 is a new topoisomerase II inhibitor containing a spermine moiety that facilitates selective uptake by tumor cells and increases topoisomerase II poisoning. F14512 is currently in Phase I/II clinical trials in patients with acute myeloid leukemia. The aim of this study is to investigate the potential of F14512 in a new clinical indication and to compare its clinical efficacy with the reference topoisomerase II inhibitor Etoposide. Because of the many similarities between human and dog lymphomas, we are seeking to determine the tolerance, efficacy, PK/PD relationship of F14512 in this indication and potential biomarkers that could be translated into human trials. Firstly, we successfully initiated two Phase 1 dose-escalation trials with 23 and 27 dogs with naturally occurring lymphomas using F14512 and Etoposide, respectively, with endpoints including safety, therapeutic efficacy and biomarker studies. Secondly, we initiated a randomized double blind study with two groups of 24 dogs with naturally occurring lymphomas in order to compare the clinical efficacy of F14512 and Etoposide in both newly diagnosed and relapsing cases. The Phase 1 trial demonstrated that F14512 could be safely administered to dogs with lymphoma resulting in strong therapeutic efficacy with a response rate of 91% (21/23) with 10 complete responses, 11 partial responses, one stable disease and one progressive disease. Phosphorylation of histone H2AX was studied as a potential pharmacodynamic biomarker of F14512. Etoposide displayed modest therapeutic efficacy with a response rate of 19% (5/27) with 1 complete response, 4 partial responses, and 6 stable diseases. The comparative clinical study was then initiated with both compounds administered at the recommended dose identified in the Phase 1 trial. Inclusion of all the dogs from the randomized double blind comparative study will be finalized at the beginning of 2016 and the results of the study will be disclosed at the AACR 2016 meeting. This work shows that naturally occurring cancers in dogs can be of great interest in translational research in order to support preclinical and clinical development of new compounds. Citation Format: Bruno Gomes, François Serres, Juliette Hordeaux, Zacharie Segaoula, Emmanuel Bouchaert, Séverine Wadoux, Laurent Marescaux, Franck Floch, Pierre Boyé, Kevin Geeraert, Ingrid Bemelmans, Thierry Marchal, Corinne Fournel, Grégoire Zorza, Pierre Ferré, Aurélie Pétain, Nicolas Guilbaud, Dominique Tierny. Randomized double-blind clinical study of F14512, a new polyamine-vectorized anticancer drug and Etoposide in naturally occurring canine lymphoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4185.
SummaryThe underlying mechanisms of the ketogenic diet (KD) remain unknown. Involvement of peroxi... more SummaryThe underlying mechanisms of the ketogenic diet (KD) remain unknown. Involvement of peroxisome proliferator–activated receptor‐α (PPARα) has been suggested. The aim of this study was to assess the anticonvulsant properties of fenofibrate, a PPARα agonist. Wistar rats were fed at libitum during 14 days by regular diet, KD, regular diet containing 0.2% fenofibrate (F), or KD containing 0.2% fenofibrate (KD + F). Pentylenetetrazol (PTZ) threshold and latencies to the onset of status epilepticus induced by lithium–pilocarpine were used to assess diet treatments with anticonvulsive effects. Myoclonic and generalized seizure PTZ thresholds were increased in F‐ and KD‐treated animals in comparison to control. No difference was observed between KD + F group and the others groups (control, F, KD). Latencies to the onset of status epilepticus were increased in F and KD groups compared to control. Fenofibrate exerts anticonvulsive properties comparable to KD in adult rats using PTZ and lithium–pilocarpine models. The underlying mechanisms such as PPARα activation and others should be investigated. These findings may provide insights into future directions to simplify KD protocols.
ABSTRACT Introduction Le diabète de type 2 (T2D) est un trouble métabolique de l’homéostasie du g... more ABSTRACT Introduction Le diabète de type 2 (T2D) est un trouble métabolique de l’homéostasie du glucose. Il est caractérisé par une hyperglycémie chronique qui résulte en partie d’une production excessive de glucose par le foie conséquence au développement d’une résistance à l’insuline. Le T2D est une pathologie multifactorielle à la fois génétique et environnementale. Récemment des études d’associations de gènes (GWAS) dans différentes cohortes ont mis en évidence une forte corrélation entre le locus CDKN2A et le risque de développement du T2D en se basant sur certains paramètres métaboliques tel que la glycémie à jeun. Le locus CDKN2A code pour des protéines régulatrices du cycle cellulaire dont la protéine p16INK4a. p16INK4a est largement décrite dans la littérature pour son rôle suppresseur de tumeurs et comme marqueur de sénescence, cependant son rôle dans le contrôle de l’homéostasie hépatique du glucose n’a jamais été rapporté Matériels et méthodes Afin de déterminer le rôle de p16INK4a dans le métabolisme hépatique du glucose, nous avons utilisé in vivo des souris sauvages (p16+/+) et déficientes pour p16INK4a (p16-/-) et in vitro des hépatocytes primaires ainsi que la lignée AML12. Résultats Nous avons montré qu’après un jeune, les souris p16-/- présentent une hypoglycémie moins prononcée qui se traduit par une expression hépatique plus élevée de gènes de la néoglucogenèse tels que PEPCK, G6Pase et PGC1a. De plus, les hépatocytes primaires de souris p16-/- présentent une meilleur réponse au glucagon que ceux des p16+/+. Enfin, nous avons montré que la diminution d’expression de p16INK4a par siRNA dans les AML12 suffit à induire l’expression des gènes de la néoglucogenèse et potentialise la réponse de ces cellules à différents stimuli gluconéogenique. L’effet observé dépend de l’activation de la voie PKA-CREB-PGC1A. Conclusion L’ensemble de ces données montrent pour la première fois que p16INK4a pourrait jouer un rôle un cours du développement du T2D.
Supplementary figures 1-3. Supplementary Figure 1: F14512 structure (A) and effects on Namalwa ly... more Supplementary figures 1-3. Supplementary Figure 1: F14512 structure (A) and effects on Namalwa lymphoma cell line (B-E). Supplementary Figure 2: DNA damage induced by F14512 in Namalwa cells. Supplementary Figure 3: Illustration of decreased cell count and cell viability in tumoral lymph nodes after F14512 infusion.
Purpose: F14512 is a new topoisomerase II inhibitor containing a spermine moiety that facilitates... more Purpose: F14512 is a new topoisomerase II inhibitor containing a spermine moiety that facilitates selective uptake by tumor cells and increases topoisomerase II poisoning. F14512 is currently in a phase I/II clinical trial in patients with acute myeloid leukemia. The aim of this study was to investigate F14512 potential in a new clinical indication. Because of the many similarities between human and dog lymphomas, we sought to determine the tolerance, efficacy, pharmacokinetic/pharmacodynamic (PK/PD) relationship of F14512 in this indication, and potential biomarkers that could be translated into human trials.Experimental Design: Twenty-three dogs with stage III–IV naturally occurring lymphomas were enrolled in the phase I dose-escalation trial, which consisted of three cycles of F14512 i.v. injections. Endpoints included safety and therapeutic efficacy. Serial blood samples and tumor biopsies were obtained for PK/PD and biomarker studies.Results: Five dose levels were evaluated to ...
Breast cancer is the most frequent cancer among women causing the greatest number of cancer-relat... more Breast cancer is the most frequent cancer among women causing the greatest number of cancer-related deaths. Cancer heterogeneity is a main obstacle to therapies. Around 96% of the drugs fail from discovery to the clinical trial phase probably because of the current unreliable preclinical models. New models emerge such as companion dogs who develop spontaneous mammary tumors resembling human breast cancer in many clinical and molecular aspects. The present work aimed at developing a robust canine mammary tumor model in the form of tumoroids which recapitulate the tumor diversity and heterogeneity. We conducted a complete characterization of these canine mammary tumoroids through histologic, molecular and proteomic analysis, demonstrating their strong similarity to the primary tumor. We demonstrated that these tumoroids can be used as a drug screening model. In fact, we showed that Paclitaxel, a human chemotherapeutic, could killed canine tumoroids with the same efficacy as human tumo...
Formalin-fixation and paraffin-embedding (FFPE) is the worldwide gold standard for tissue preserv... more Formalin-fixation and paraffin-embedding (FFPE) is the worldwide gold standard for tissue preservation and routine material for general diagnostics and cancer in particular. Despite the fact that histology is complemented by other modalities such as in situ hybridization (ISH) or immunohistochemistry (IHC), the pathology can fail to provide the right diagnosis in a certain number of cases. Overall, the standard process is time-consuming, labor intensive and doesn’t reach the right sensitivity and specificity. Lipids are central players in cancer. Unfortunately, to this date lipids aren’t used for diagnostics and lipid analysis from FFPE has never been reported due to tissue processing which limits their detection. Here, we report a new method for cancer diagnostics based on their unique tissue-lipidomic signatures through rapid molecular screening of FFPE sections without dewaxing using Water-Assisted Laser Desorption Ionization Mass Spectrometry (WALDI-MS) - SpiderMass. Advantageously, the method does not require human intervention. The annotated tissues are first analyzed to train the system and then blind samples can be diagnosed in less than a few minutes. The platform was demonstrated for the diagnostics of canine and human sarcoma, including cases which are known to be problematic in the gold standard workflow. Interrogation of blind samples against the built models revealed over 95% probability values to discriminate specific subtypes
F14512 is a new topoisomerase II inhibitor containing a spermine moiety that facilitates selectiv... more F14512 is a new topoisomerase II inhibitor containing a spermine moiety that facilitates selective uptake by tumor cells and increases topoisomerase II poisoning. F14512 is currently in Phase I/II clinical trials in patients with acute myeloid leukemia. The aim of this study is to investigate the potential of F14512 in a new clinical indication and to compare its clinical efficacy with the reference topoisomerase II inhibitor Etoposide. Because of the many similarities between human and dog lymphomas, we are seeking to determine the tolerance, efficacy, PK/PD relationship of F14512 in this indication and potential biomarkers that could be translated into human trials. Firstly, we successfully initiated two Phase 1 dose-escalation trials with 23 and 27 dogs with naturally occurring lymphomas using F14512 and Etoposide, respectively, with endpoints including safety, therapeutic efficacy and biomarker studies. Secondly, we initiated a randomized double blind study with two groups of 24 dogs with naturally occurring lymphomas in order to compare the clinical efficacy of F14512 and Etoposide in both newly diagnosed and relapsing cases. The Phase 1 trial demonstrated that F14512 could be safely administered to dogs with lymphoma resulting in strong therapeutic efficacy with a response rate of 91% (21/23) with 10 complete responses, 11 partial responses, one stable disease and one progressive disease. Phosphorylation of histone H2AX was studied as a potential pharmacodynamic biomarker of F14512. Etoposide displayed modest therapeutic efficacy with a response rate of 19% (5/27) with 1 complete response, 4 partial responses, and 6 stable diseases. The comparative clinical study was then initiated with both compounds administered at the recommended dose identified in the Phase 1 trial. Inclusion of all the dogs from the randomized double blind comparative study will be finalized at the beginning of 2016 and the results of the study will be disclosed at the AACR 2016 meeting. This work shows that naturally occurring cancers in dogs can be of great interest in translational research in order to support preclinical and clinical development of new compounds. Citation Format: Bruno Gomes, François Serres, Juliette Hordeaux, Zacharie Segaoula, Emmanuel Bouchaert, Séverine Wadoux, Laurent Marescaux, Franck Floch, Pierre Boyé, Kevin Geeraert, Ingrid Bemelmans, Thierry Marchal, Corinne Fournel, Grégoire Zorza, Pierre Ferré, Aurélie Pétain, Nicolas Guilbaud, Dominique Tierny. Randomized double-blind clinical study of F14512, a new polyamine-vectorized anticancer drug and Etoposide in naturally occurring canine lymphoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4185.
SummaryThe underlying mechanisms of the ketogenic diet (KD) remain unknown. Involvement of peroxi... more SummaryThe underlying mechanisms of the ketogenic diet (KD) remain unknown. Involvement of peroxisome proliferator–activated receptor‐α (PPARα) has been suggested. The aim of this study was to assess the anticonvulsant properties of fenofibrate, a PPARα agonist. Wistar rats were fed at libitum during 14 days by regular diet, KD, regular diet containing 0.2% fenofibrate (F), or KD containing 0.2% fenofibrate (KD + F). Pentylenetetrazol (PTZ) threshold and latencies to the onset of status epilepticus induced by lithium–pilocarpine were used to assess diet treatments with anticonvulsive effects. Myoclonic and generalized seizure PTZ thresholds were increased in F‐ and KD‐treated animals in comparison to control. No difference was observed between KD + F group and the others groups (control, F, KD). Latencies to the onset of status epilepticus were increased in F and KD groups compared to control. Fenofibrate exerts anticonvulsive properties comparable to KD in adult rats using PTZ and lithium–pilocarpine models. The underlying mechanisms such as PPARα activation and others should be investigated. These findings may provide insights into future directions to simplify KD protocols.
ABSTRACT Introduction Le diabète de type 2 (T2D) est un trouble métabolique de l’homéostasie du g... more ABSTRACT Introduction Le diabète de type 2 (T2D) est un trouble métabolique de l’homéostasie du glucose. Il est caractérisé par une hyperglycémie chronique qui résulte en partie d’une production excessive de glucose par le foie conséquence au développement d’une résistance à l’insuline. Le T2D est une pathologie multifactorielle à la fois génétique et environnementale. Récemment des études d’associations de gènes (GWAS) dans différentes cohortes ont mis en évidence une forte corrélation entre le locus CDKN2A et le risque de développement du T2D en se basant sur certains paramètres métaboliques tel que la glycémie à jeun. Le locus CDKN2A code pour des protéines régulatrices du cycle cellulaire dont la protéine p16INK4a. p16INK4a est largement décrite dans la littérature pour son rôle suppresseur de tumeurs et comme marqueur de sénescence, cependant son rôle dans le contrôle de l’homéostasie hépatique du glucose n’a jamais été rapporté Matériels et méthodes Afin de déterminer le rôle de p16INK4a dans le métabolisme hépatique du glucose, nous avons utilisé in vivo des souris sauvages (p16+/+) et déficientes pour p16INK4a (p16-/-) et in vitro des hépatocytes primaires ainsi que la lignée AML12. Résultats Nous avons montré qu’après un jeune, les souris p16-/- présentent une hypoglycémie moins prononcée qui se traduit par une expression hépatique plus élevée de gènes de la néoglucogenèse tels que PEPCK, G6Pase et PGC1a. De plus, les hépatocytes primaires de souris p16-/- présentent une meilleur réponse au glucagon que ceux des p16+/+. Enfin, nous avons montré que la diminution d’expression de p16INK4a par siRNA dans les AML12 suffit à induire l’expression des gènes de la néoglucogenèse et potentialise la réponse de ces cellules à différents stimuli gluconéogenique. L’effet observé dépend de l’activation de la voie PKA-CREB-PGC1A. Conclusion L’ensemble de ces données montrent pour la première fois que p16INK4a pourrait jouer un rôle un cours du développement du T2D.
Supplementary figures 1-3. Supplementary Figure 1: F14512 structure (A) and effects on Namalwa ly... more Supplementary figures 1-3. Supplementary Figure 1: F14512 structure (A) and effects on Namalwa lymphoma cell line (B-E). Supplementary Figure 2: DNA damage induced by F14512 in Namalwa cells. Supplementary Figure 3: Illustration of decreased cell count and cell viability in tumoral lymph nodes after F14512 infusion.
Purpose: F14512 is a new topoisomerase II inhibitor containing a spermine moiety that facilitates... more Purpose: F14512 is a new topoisomerase II inhibitor containing a spermine moiety that facilitates selective uptake by tumor cells and increases topoisomerase II poisoning. F14512 is currently in a phase I/II clinical trial in patients with acute myeloid leukemia. The aim of this study was to investigate F14512 potential in a new clinical indication. Because of the many similarities between human and dog lymphomas, we sought to determine the tolerance, efficacy, pharmacokinetic/pharmacodynamic (PK/PD) relationship of F14512 in this indication, and potential biomarkers that could be translated into human trials.Experimental Design: Twenty-three dogs with stage III–IV naturally occurring lymphomas were enrolled in the phase I dose-escalation trial, which consisted of three cycles of F14512 i.v. injections. Endpoints included safety and therapeutic efficacy. Serial blood samples and tumor biopsies were obtained for PK/PD and biomarker studies.Results: Five dose levels were evaluated to ...
Breast cancer is the most frequent cancer among women causing the greatest number of cancer-relat... more Breast cancer is the most frequent cancer among women causing the greatest number of cancer-related deaths. Cancer heterogeneity is a main obstacle to therapies. Around 96% of the drugs fail from discovery to the clinical trial phase probably because of the current unreliable preclinical models. New models emerge such as companion dogs who develop spontaneous mammary tumors resembling human breast cancer in many clinical and molecular aspects. The present work aimed at developing a robust canine mammary tumor model in the form of tumoroids which recapitulate the tumor diversity and heterogeneity. We conducted a complete characterization of these canine mammary tumoroids through histologic, molecular and proteomic analysis, demonstrating their strong similarity to the primary tumor. We demonstrated that these tumoroids can be used as a drug screening model. In fact, we showed that Paclitaxel, a human chemotherapeutic, could killed canine tumoroids with the same efficacy as human tumo...
Uploads
Papers by Emmanuel Bouchaert