Potentiel thérapeutique de la modulation de la transition epithélio-mésenchymateuse via le ciblage de nétrine 1 dans le cancer

Netrin 1 (NTN1) is a secreted molecule initially discovered in the 1940s for its implication in the development of the neural system through the guidance of commissural axons. The laboratory in which I conducted my thesis work under the direction of Dr. Patrick Mehlen showed that the reactivation of NTN1 expression in adulthood and its overexpression is associated with the development of numerous aggressive cancers. Due to its induction of manifold signalling pathways implicated in the modulation of inflammation, proliferation, survival and cell migration, its therapeutic targeting offers great potential in the treatment of cancer. In 2016, the French biotech start-up NETRIS Pharma that arose from the laboratory, developed a monoclonal humanized antibody directed against NTN1. This antibody inhibits the interaction of NTN1 with its dependence receptor Uncoordinated-5B (UNC5B), triggering the death of cells through the induction of apoptosis. Tested in phase I clinical trials on multiple advanced and metastatic cancer indications, the antibody has proven its absence of toxicity and its efficacy in humans. Furthermore, our work on metastatic endometrial cancer patients’ biopsies carried out over the course of the clinical trial, along with preclinical in vitro and in vivo models of this disease, have highlighted the effect of the treatment on the epithelial and mesenchymal profile of cancer cells. The work conducted over the course of my thesis and presented in this manuscript demonstrates the effect of NTN1 targeting by this antibody on the death of tumour cells as well as the epithelialization of tumours, leading to the inhibition of mesenchymal cellular properties, associated with a modulation of the immune infiltrate and of stromal cells. Altogether, these effects are also linked to a sensitization of cancer tissues to standard chemotherapy treatment options. My thesis was financed by an industrial convention of formation by research (Cifre) from the Ministry of Higher Education and Research, in close collaboration between the laboratory of Dr. Mehlen and the NETRIS Pharma start-up.La nétrine 1 (NTN1) est une molécule sécrétée initialement découverte dans les années 1940 pour son implication dans le développement du système nerveux via le guidage des axones commissuraux. Le laboratoire dans lequel j'ai effectué mon travail de thèse dirigé par le Dr. Patrick Mehlen a montré que la réactivation, voire surexpression, de l'expression de NTN1 à l'âge adulte est associée au développement de nombreux cancers agressifs. Liée à l'activation de nombreuses voies de signalisation impliquées dans la modulation de l'inflammation, la prolifération, la survie, et la migration cellulaire, son ciblage thérapeutique présente un potentiel dans le traitement cancéreux. En 2016, la start-up française NETRIS Pharma, issue du laboratoire, a développé un anticorps monoclonal humanisé dirigé contre NTN1. Cet anticorps inhibe l'interaction de NTN1 avec son récepteur à dépendance Uncoordinated-5B (UNC5B), induisant la mort des cellules par apoptose. Testé en essai clinique de phase I sur de multiples indications cancéreuses avancées à métastatiques, l'anticorps a prouvé sa non-toxicité et montré une efficacité chez l'Homme. De plus, nos travaux sur des biopsies métastatiques de patientes atteintes de cancer de l'endomètre réalisées au cours de cet essai clinique, complétés par l'analyse des modèles précliniques in vitro et in vivo de cette pathologie, ont mis en évidence un nouvel effet du traitement sur le profil épithélial et mésenchymateux des cellules cancéreuses. Les travaux réalisés au cours de ma thèse et présentés dans ce manuscrit démontrent l'effet du ciblage de NTN1 par un nouvel anticorps humanisé sur la mort des cellules tumourales ainsi que sur l'épithélialisation des tumeurs, conduisant à l'inhibition de la transition des cellules tumourales vers des phénotypes et fonctions cellulaires mésenchymateuses. Cet effet est également associé à une modification de l'infiltrat immunitaire et des cellules stromales, tout cela couplé à une sensibilisation des tissus cancéreux à des traitements standards de type chimio-thérapeutiques. Ces résultats ont permis la publication de deux articles dans la revue Nature. Ma thèse a été financée par une Convention industrielle de formation par la recherche (Cifre) du ministère de l'Enseignement Supérieur de la Recherche et de l'Innovation, en étroite collaboration entre le laboratoire du Dr. Mehlen au CRCL et de la start-up NETRIS Pharma

Potentiel thérapeutique de la modulation de la transition epithélio-mésenchymateuse via le ciblage de nétrine 1 dans le cancer

Netrin 1 (NTN1) is a secreted molecule initially discovered in the 1940s for its implication in the development of the neural system through the guidance of commissural axons. The laboratory in which I conducted my thesis work under the direction of Dr. Patrick Mehlen showed that the reactivation of NTN1 expression in adulthood and its overexpression is associated with the development of numerous aggressive cancers. Due to its induction of manifold signalling pathways implicated in the modulation of inflammation, proliferation, survival and cell migration, its therapeutic targeting offers great potential in the treatment of cancer. In 2016, the French biotech start-up NETRIS Pharma that arose from the laboratory, developed a monoclonal humanized antibody directed against NTN1. This antibody inhibits the interaction of NTN1 with its dependence receptor Uncoordinated-5B (UNC5B), triggering the death of cells through the induction of apoptosis. Tested in phase I clinical trials on multiple advanced and metastatic cancer indications, the antibody has proven its absence of toxicity and its efficacy in humans. Furthermore, our work on metastatic endometrial cancer patients’ biopsies carried out over the course of the clinical trial, along with preclinical in vitro and in vivo models of this disease, have highlighted the effect of the treatment on the epithelial and mesenchymal profile of cancer cells. The work conducted over the course of my thesis and presented in this manuscript demonstrates the effect of NTN1 targeting by this antibody on the death of tumour cells as well as the epithelialization of tumours, leading to the inhibition of mesenchymal cellular properties, associated with a modulation of the immune infiltrate and of stromal cells. Altogether, these effects are also linked to a sensitization of cancer tissues to standard chemotherapy treatment options. My thesis was financed by an industrial convention of formation by research (Cifre) from the Ministry of Higher Education and Research, in close collaboration between the laboratory of Dr. Mehlen and the NETRIS Pharma start-up.La nétrine 1 (NTN1) est une molécule sécrétée initialement découverte dans les années 1940 pour son implication dans le développement du système nerveux via le guidage des axones commissuraux. Le laboratoire dans lequel j'ai effectué mon travail de thèse dirigé par le Dr. Patrick Mehlen a montré que la réactivation, voire surexpression, de l'expression de NTN1 à l'âge adulte est associée au développement de nombreux cancers agressifs. Liée à l'activation de nombreuses voies de signalisation impliquées dans la modulation de l'inflammation, la prolifération, la survie, et la migration cellulaire, son ciblage thérapeutique présente un potentiel dans le traitement cancéreux. En 2016, la start-up française NETRIS Pharma, issue du laboratoire, a développé un anticorps monoclonal humanisé dirigé contre NTN1. Cet anticorps inhibe l'interaction de NTN1 avec son récepteur à dépendance Uncoordinated-5B (UNC5B), induisant la mort des cellules par apoptose. Testé en essai clinique de phase I sur de multiples indications cancéreuses avancées à métastatiques, l'anticorps a prouvé sa non-toxicité et montré une efficacité chez l'Homme. De plus, nos travaux sur des biopsies métastatiques de patientes atteintes de cancer de l'endomètre réalisées au cours de cet essai clinique, complétés par l'analyse des modèles précliniques in vitro et in vivo de cette pathologie, ont mis en évidence un nouvel effet du traitement sur le profil épithélial et mésenchymateux des cellules cancéreuses. Les travaux réalisés au cours de ma thèse et présentés dans ce manuscrit démontrent l'effet du ciblage de NTN1 par un nouvel anticorps humanisé sur la mort des cellules tumourales ainsi que sur l'épithélialisation des tumeurs, conduisant à l'inhibition de la transition des cellules tumourales vers des phénotypes et fonctions cellulaires mésenchymateuses. Cet effet est également associé à une modification de l'infiltrat immunitaire et des cellules stromales, tout cela couplé à une sensibilisation des tissus cancéreux à des traitements standards de type chimio-thérapeutiques. Ces résultats ont permis la publication de deux articles dans la revue Nature. Ma thèse a été financée par une Convention industrielle de formation par la recherche (Cifre) du ministère de l'Enseignement Supérieur de la Recherche et de l'Innovation, en étroite collaboration entre le laboratoire du Dr. Mehlen au CRCL et de la start-up NETRIS Pharma

Pharmacological targeting of netrin-1 inhibits EMT in cancer.

Epithelial-to-mesenchymal transition (EMT) regulates tumour initiation, progression, metastasis and resistance to anti-cancer therapy1-7. Although great progress has been made in understanding the role of EMT and its regulatory mechanisms in cancer, no therapeutic strategy to pharmacologically target EMT has been identified. Here we found that netrin-1 is upregulated in a primary mouse model of skin squamous cell carcinoma (SCC) exhibiting spontaneous EMT. Pharmacological inhibition of netrin-1 by administration of NP137, a netrin-1-blocking monoclonal antibody currently used in clinical trials in human cancer (ClinicalTrials.gov identifier NCT02977195 ), decreased the proportion of EMT tumour cells in skin SCC, decreased the number of metastases and increased the sensitivity of tumour cells to chemotherapy. Single-cell RNA sequencing revealed the presence of different EMT states, including epithelial, early and late hybrid EMT, and full EMT states, in control SCC. By contrast, administration of NP137 prevented the progression of cancer cells towards a late EMT state and sustained tumour epithelial states. Short hairpin RNA knockdown of netrin-1 and its receptor UNC5B in EPCAM+ tumour cells inhibited EMT in vitro in the absence of stromal cells and regulated a common gene signature that promotes tumour epithelial state and restricts EMT. To assess the relevance of these findings to human cancers, we treated mice transplanted with the A549 human cancer cell line-which undergoes EMT following TGFβ1 administration8,9-with NP137. Netrin-1 inhibition decreased EMT in these transplanted A549 cells. Together, our results identify a pharmacological strategy for targeting EMT in cancer, opening up novel therapeutic interventions for anti-cancer therapy.info:eu-repo/semantics/publishe

Netrin-1 blockade inhibits tumour growth and EMT features in endometrial cancer

Netrin-1 is upregulated in cancers as a protumoural mechanism1. Here we describe netrin-1 upregulation in a majority of human endometrial carcinomas (ECs) and demonstrate that netrin-1 blockade, using an anti-netrin-1 antibody (NP137), is effective in reduction of tumour progression in an EC mouse model. We next examined the efficacy of NP137, as a first-in-class single agent, in a Phase I trial comprising 14 patients with advanced EC. As best response we observed 8 stable disease (8 out of 14, 57.1%) and 1 objective response as RECIST v.1.1 (partial response, 1 out of 14 (7.1%), 51.16% reduction in target lesions at 6 weeks and up to 54.65% reduction during the following 6 months). To evaluate the NP137 mechanism of action, mouse tumour gene profiling was performed, and we observed, in addition to cell death induction, that NP137 inhibited epithelial-to-mesenchymal transition (EMT). By performing bulk RNA sequencing (RNA-seq), spatial transcriptomics and single-cell RNA-seq on paired pre- and on-treatment biopsies from patients with EC from the NP137 trial, we noted a net reduction in tumour EMT. This was associated with changes in immune infiltrate and increased interactions between cancer cells and the tumour microenvironment. Given the importance of EMT in resistance to current standards of care2, we show in the EC mouse model that a combination of NP137 with carboplatin-paclitaxel outperformed carboplatin-paclitaxel alone. Our results identify netrin-1 blockade as a clinical strategy triggering both tumour debulking and EMT inhibition, thus potentially alleviating resistance to standard treatments

Epidemiology and outcomes of hospital-acquired bloodstream infections in intensive care unit patients: the EUROBACT-2 international cohort study

Purpose In the critically ill, hospital-acquired bloodstream infections (HA-BSI) are associated with significant mortality. Granular data are required for optimizing management, and developing guidelines and clinical trials. Methods We carried out a prospective international cohort study of adult patients (≥ 18 years of age) with HA-BSI treated in intensive care units (ICUs) between June 2019 and February 2021. Results 2600 patients from 333 ICUs in 52 countries were included. 78% HA-BSI were ICU-acquired. Median Sequential Organ Failure Assessment (SOFA) score was 8 [IQR 5; 11] at HA-BSI diagnosis. Most frequent sources of infection included pneumonia (26.7%) and intravascular catheters (26.4%). Most frequent pathogens were Gram-negative bacteria (59.0%), predominantly Klebsiella spp. (27.9%), Acinetobacter spp. (20.3%), Escherichia coli (15.8%), and Pseudomonas spp. (14.3%). Carbapenem resistance was present in 37.8%, 84.6%, 7.4%, and 33.2%, respectively. Difficult-to-treat resistance (DTR) was present in 23.5% and pan-drug resistance in 1.5%. Antimicrobial therapy was deemed adequate within 24 h for 51.5%. Antimicrobial resistance was associated with longer delays to adequate antimicrobial therapy. Source control was needed in 52.5% but not achieved in 18.2%. Mortality was 37.1%, and only 16.1% had been discharged alive from hospital by day-28. Conclusions HA-BSI was frequently caused by Gram-negative, carbapenem-resistant and DTR pathogens. Antimicrobial resistance led to delays in adequate antimicrobial therapy. Mortality was high, and at day-28 only a minority of the patients were discharged alive from the hospital. Prevention of antimicrobial resistance and focusing on adequate antimicrobial therapy and source control are important to optimize patient management and outcomes

The role of centre and country factors on process and outcome indicators in critically ill patients with hospital-acquired bloodstream infections

Purpose: The primary objective of this study was to evaluate the associations between centre/country-based factors and two important process and outcome indicators in patients with hospital-acquired bloodstream infections (HABSI). Methods: We used data on HABSI from the prospective EUROBACT-2 study to evaluate the associations between centre/country factors on a process or an outcome indicator: adequacy of antimicrobial therapy within the first 24 h or 28-day mortality, respectively. Mixed logistical models with clustering by centre identified factors associated with both indicators. Results: Two thousand two hundred nine patients from two hundred one intensive care units (ICUs) were included in forty-seven countries. Overall, 51% (n = 1128) of patients received an adequate antimicrobial therapy and the 28-day mortality was 38% (n = 839). The availability of therapeutic drug monitoring (TDM) for aminoglycosides everyday [odds ratio (OR) 1.48, 95% confidence interval (CI) 1.03-2.14] or within a few hours (OR 1.79, 95% CI 1.34-2.38), surveillance cultures for multidrug-resistant organism carriage performed weekly (OR 1.45, 95% CI 1.09-1.93), and increasing Human Development Index (HDI) values were associated with adequate antimicrobial therapy. The presence of intermediate care beds (OR 0.63, 95% CI 0.47-0.84), TDM for aminoglycoside available everyday (OR 0.66, 95% CI 0.44-1.00) or within a few hours (OR 0.51, 95% CI 0.37-0.70), 24/7 consultation of clinical pharmacists (OR 0.67, 95% CI 0.47-0.95), percentage of vancomycin-resistant enterococci (VRE) between 10% and 25% in the ICU (OR 1.67, 95% CI 1.00-2.80), and decreasing HDI values were associated with 28-day mortality. Conclusion: Centre/country factors should be targeted for future interventions to improve management strategies and outcome of HABSI in ICU patients

Presentation, management, and outcomes of older compared to younger adults with hospital-acquired bloodstream infections in the intensive care unit: a multicenter cohort study

Purpose: Older adults admitted to the intensive care unit (ICU) usually have fair baseline functional capacity, yet their age and frailty may compromise their management. We compared the characteristics and management of older (≥ 75 years) versus younger adults hospitalized in ICU with hospital-acquired bloodstream infection (HA-BSI). Methods: Nested cohort study within the EUROBACT-2 database, a multinational prospective cohort study including adults (≥ 18 years) hospitalized in the ICU during 2019-2021. We compared older versus younger adults in terms of infection characteristics (clinical signs and symptoms, source, and microbiological data), management (imaging, source control, antimicrobial therapy), and outcomes (28-day mortality and hospital discharge). Results: Among 2111 individuals hospitalized in 219 ICUs with HA-BSI, 563 (27%) were ≥ 75 years old. Compared to younger patients, these individuals had higher comorbidity score and lower functional capacity; presented more often with a pulmonary, urinary, or unknown HA-BSI source; and had lower heart rate, blood pressure and temperature at presentation. Pathogens and resistance rates were similar in both groups. Differences in management included mainly lower rates of effective source control achievement among aged individuals. Older adults also had significantly higher day-28 mortality (50% versus 34%, p < 0.001), and lower rates of discharge from hospital (12% versus 20%, p < 0.001) by this time. Conclusions: Older adults with HA-BSI hospitalized in ICU have different baseline characteristics and source of infection compared to younger patients. Management of older adults differs mainly by lower probability to achieve source control. This should be targeted to improve outcomes among older ICU patients