Immunosenescence, inflammation and immunotherapies

Résumé : Les immunothérapies antitumorales ciblant et bloquant les points de contrôle immunologiques (immune checkpoint blockers, ICB) sont devenues incontournables dans l'arsenal thérapeutique des patients atteints d'un cancer. Elles ont démontré des résultats cliniques très intéressants dans le cancer bronchique non à petites cellules avancé (CBNPC), mais le taux de réponse objective reste faible. Des biomarqueurs tumoraux (expression de PD-L1 par les cellules tumorales, charge mutationnelle tumorale) existent pour prédire la réponse à ces traitements mais il n'existe pas de biomarqueurs circulants non invasifs validés en clinique. Certaines études montrent une inneficacité des ICB chez les patients de plus de 75 ans, d'autres montrent un bénéfice supérieur à la chimiothérapie chez les patients agés de 65 à 75 ans.L'immunosénescence correspond au remodelage progressif du système immunitaire avec l'âge. La sénescence lymphocytaire est caractérisée par une faible activité proliférative, l'expression de CD57 et KLRG1 et la perte d'expression de CD28 par les lymphocytes T (LT). Les LT sénescents augmentent au cours du vieillissement, surtout chez les individus ayant une stimulation antigénique chronique telle que les infections virales chroniques ou un contexte inflammatoire chronique (cancer). Il a été montré que l'expression du CD28 était nécessaire pour restaurer les réponses T CD8 antitumorales pendant un traitement par ICB.Dans ce contexte, nous avons cherché à caractériser phénotypiquement et fonctionnellement les lymphocytes T sénescents chez des patients atteints d'un CBNPC avant traitement par ICB. Nous avons mis en évidence en cytométrie de flux une population de circulante de lymphocytes T CD8+ sénescents CD28-CD57+KLRG1+ dont la proportion, supérieure à 39.5% des CD8+ totaux, permettait d'attribuer au patient un statut SIP (senescent immune phenotype) positif (SIP+). Ces lymphocytes T8 sénescents avaient une faible capacité proliférative et produisaient des cytokines inflammatoires (IFN-γ, TNF-α) et peu d'IL-2. Les patients SIP+ avaient une survie globale et une survie sans progression sous ICB significativement plus courte que les patients SIP-.Nous nous sommes intéressés à plusieurs étiologies secondaires d'immunosénescence chez des patients atteints de CBNPC. Nous n'avons pas mis en évidence d'association entre le statut SIP et une inflammation ou un stress oxydatif systémique, ni une signature IFN de type I, connue pour entraîner une dysfonction lymphocytaire dans certaines infections virales chroniques. Cependant, nous avons mis en évidence un lien avec l'immunisation contre le CMV : la majorité des patients SIP+ sont CMV+ mais le CMV n'est pas suffisant puisque seulement 30% des patients CMV+ sont SIP+.En conclusion, le statut SIP est associé à la résistance aux ICB chez des patients atteints de CBNPC, n'est pas associé à une inflammation ou un stress oxydatif spécifique mais à une immunisation contre le CMV.Abstract : Immune checkpoint blockers (ICB) have brought a paradigm shift to non-small cell lung cancer (NSCLC) treatment. Efficacy data in the ederly population emerging from randomized clinical trials are often conclicting and suggest an absence of benefit in patients older than 75 years. Most data come from small subgroup analyses, lacking adequate statistical power to drive definitive conclusions.Aging is associated with several structural and functional changes in the immune system, which are called “immunosenescence”. T cell senescence refers to poor proliferative capacity, production of pro-inflammatory cytokines, loss of CD28 and increased expression of immune senescence markers such as CD57 and KLRG1. Persistent antigenic stimulation induced by cancer, infections, chronic inflammatory diseases and chemotherapy may also induce senescence in both innate and adaptive immune systems.In this context, we sought to phenotypically and functionnaly characterize senescent T cells in NSCLC patient upon ICB-treatment. We defined SIP as the proportion of CD28-CD57+KLRG1+CD8+ circulating T cells. We showed that a high pretreatment SIP (>39.5%, SIP+) was associated with resistance to ICB in patients with aNSCLC. In in vitro experiments, SIP cells were poorly proliferative and expressed pro-inflammatory cytokines but low levels of IL-2.We investigated several secondary etiologies of immunosenescence in NSCLC patients. We found no association between SIP status and systemic inflammation or oxidative stress, nor a type I IFN signature, which is known to cause lymphocyte dysfunction in some chronic viral infections. CMV+ patient's rate was significantly higher in SIP+ compared to SIP- patients (91.4% vs 50%, p<0.0001). Among CMV+ patients, SIP+ represents only 30%.In conclusion, SIP status is associated with ICB resistance in NSCLC patients, is not associated with inflammation or specific oxidative stress but with immunisation against CMV

The use of immunotherapy in older patients with advanced non-small cell lung cancer

Immune checkpoint blockers (ICBs) have a pivotal role in the management of non-small cell lung cancer (NSCLC), both as single agent and in combination strategies, providing a meaningful clinical and survival benefit.Older patients are underrepresented in clinical trials, including those involving immunotherapy, even though almost half of the patients with newly diagnosed NSCLC are aged 70 years or older. Moreover, due to selection biases, usually "fit" patients are preferably enrolled. This results in a lack of evidence regarding the use of ICBs in the older population, particularly when referring to chemo-immunotherapy regimens.Since ICBs are indeed of paramount importance in the treatment of patients with NSCLC, efforts are needed to optimize their use also in the older population. This entails furthermore taking into account additional features including the degree of fitness of the patient and the different health domains that can be affected by aging.This review aims to delve into the current evidences about the efficacy and toxicity of ICBs in monotherapy and in combination in older patients with advanced NSCLC, the role of the comprehensive geriatric assessment in supporting the selection of patients receiving immunotherapy, as well as the value of immunosenescence in modulating the activity of these drugs

Integrating Circulating Biomarkers in the Immune Checkpoint Inhibitor Treatment in Lung Cancer

Immune checkpoint inhibitors are now a cornerstone of treatment for non-small cell lung cancer (NSCLC). Tissue-based assays, such as Programmed cell death protein 1 (PD-L1) expression or mismatch repair deficiency/microsatellite instability (MMRD/MSI) status, are approved as treatment drivers in various settings, and represent the main field of research in biomarkers for immunotherapy. Nonetheless, responses have been observed in patients with negative PD-L1 or low tumor mutational burden. Some aspects of biomarker use remain poorly understood and sub-optimal, in particular tumoral heterogeneity, time-evolving sampling, and the ability to detect patients who are unlikely to respond. Moreover, tumor biopsies offer little insight into the host&rsquo;s immune status. Circulating biomarkers offer an alternative non-invasive solution to address these pitfalls. Here, we summarize current knowledge on circulating biomarkers while using liquid biopsies in patients with lung cancer who receive treatment with immune checkpoint inhibitors, in terms of their potential as being predictive of outcome as well as their role in monitoring ongoing treatment. We address host biomarkers, notably circulating immune cells and soluble systemic immune and inflammatory markers, and also review tumor markers, including blood-based tumor mutational burden, circulating tumor cells, and circulating tumor DNA. Technical requirements are discussed along with the current limitations that are associated with these promising biomarkers

Baseline circulating soluble factors as predictors of immune-related adverse events (irAEs) in patients (pts) with metastatic clear cell renal carcinoma (mRCC) treated with nivolumab: A translational NIVOREN GETUG-AFU 26 study.

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Baseline circulating unswitched memory B cells and B-cell related soluble factors are associated with overall survival in patients with clear cell renal cell carcinoma treated with nivolumab within the NIVOREN GETUG-AFU 26 study

International audienceBACKGROUND: The phase II NIVOREN GETUG-AFU 26 study reported safety and efficacy of nivolumab in patients with metastatic clear cell renal cell carcinoma (m-ccRCC) in a ’real-world setting’. We conducted a translational-research program to determine whether specific circulating immune-cell populations and/or soluble factors at baseline were predictive of clinical outcomes in patients with m-ccRCC treated with nivolumab within the NIVOREN study. METHODS: Absolute numbers of 106 circulating immune-cell populations were prospectively analyzed in patients treated at a single institution within the NIVOREN trial with available fresh-whole-blood, using dry formulation panels for multicolor flow cytometry. In addition, a panel of 14 predefined soluble factors was quantified for each baseline plasma sample using the Meso-Scale-Discovery immunoassay. The remaining patients with available plasma sample were used as a validation cohort for the soluble factor quantification analysis. Tumor immune microenvironment characterization of all patients included in the translational program of the study was available. The association of blood and tissue-based biomarkers, with overall survival (OS), progression-free survival (PFS) and response was analyzed. RESULTS: Among the 44 patients, baseline unswitched memory B cells (NSwM B cells) were enriched in responders (p=0.006) and associated with improved OS (HR=0.08, p=0.002) and PFS (HR=0.54, p=0.048). Responders were enriched in circulating T follicular helper (Tfh) (p=0.027) and tertiary lymphoid structures (TLS) (p=0.043). Circulating NSwM B cells positively correlated with Tfh (r=0.70, p&lt;0.001). Circulating NSwM B cells correlated positively with TLS and CD20 +B cells at the tumor center (r=0.59, p=0.044, and r=0.52, p=0.033) and inversely correlated with BCA-1/CXCL13 and BAFF (r=-0.55 and r=-0.42, p&lt;0.001). Tfh cells also inversely correlated with BCA-1/CXCL13 (r=-0.61, p&lt;0.001). IL-6, BCA-1/CXCL13 and BAFF significantly associated with worse OS in the discovery (n=40) and validation cohorts (n=313). CONCLUSION: We report the first fresh blood immune-monitoring of patients with m-ccRCC treated with nivolumab. Baseline blood concentration of NSwM B cells was associated to response, PFS and OS in patients with m-ccRCC treated with nivolumab. BCA-1/CXCL13 and BAFF, inversely correlated to NSwM B cells, were both associated with worse OS in discovery and validation cohorts. Our data confirms a role for B cell subsets in the response to immune checkpoint blockade therapy in patients with m-ccRCC. Further studies are needed to confirm these findings

Systemic short chain fatty acids limit antitumor effect of CTLA-4 blockade in hosts with cancer

International audienceGut microbiota composition influences the clinical benefit of immune checkpoints in patients with advanced cancer but mechanisms underlying this relationship remain unclear. Molecular mechanism whereby gut microbiota influences immune responses is mainly assigned to gut microbial metabolites. Short-chain fatty acids (SCFA) are produced in large amounts in the colon through bacterial fermentation of dietary fiber. We evaluate in mice and in patients treated with anti-CTLA-4 blocking mAbs whether SCFA levels is related to clinical outcome. High blood butyrate and propionate levels are associated with resistance to CTLA-4 blockade and higher proportion of Treg cells. In mice, butyrate restrains anti-CTLA-4-induced up-regulation of CD80/CD86 on dendritic cells and ICOS on T cells, accumulation of tumor-specific T cells and memory T cells. In patients, high blood butyrate levels moderate ipilimumab-induced accumulation of memory and ICOS + CD4 + T cells and IL-2 impregnation. Altogether, these results suggest that SCFA limits anti-CTLA-4 activity

Predicting immunotherapy outcomes under therapy in patients with advanced NSCLC using dNLR and its early dynamics

Background: dNLR at the baseline (B), defined by neutrophils/[leucocytes-neutrophils], correlates with immune-checkpoint inhibitor (ICI) outcomes in advanced non–small-cell lung cancer (aNSCLC). However, dNLR is dynamic under therapy and its longitudinal assessment may provide data predicting efficacy. We sought to examine the impact of dNLR dynamics on ICI efficacy and understand its biological significance. Patients and methods: aNSCLC patients receiving ICI at 17 EU/US centres were included [Feb/13-Jun/18]. As chemotherapy-only group was evaluated (NCT02105168). dNLR was determined at (B) and at cycle2 (C2) [dNLR≤3 = low]. B+C2 dNLR were combined in one score: good = low (B+C2), poor = high (B+C2), intermediate = other situations. In 57 patients, we prospectively explored the immunophenotype of circulating neutrophils, particularly the CD15+CD244-CD16low cells (immature) by flow cytometry. Results: About 1485 patients treatment with ICI were analysed. In ICI-treated patients, high dNLR (B) (~1/3rd) associated with worse progression-free (PFS)/overall survival (OS) (HR 1.56/HR 2.02, P < 0.0001) but not with chemotherapy alone (N = 173). High dNLR at C2 was associated with worse PFS/OS (HR 1.64/HR 2.15, P < 0.0001). When dNLR at both time points were considered together, those with persistently high dNLR (23%) had poor survival (mOS = 5 months (mo)), compared with high dNLR at one time point (22%; mOS = 9.2mo) and persistently low dNLR (55%; mOS = 18.6mo) (P < 0.0001). The dNLR impact remained significant after PD-L1 adjustment. By cytometry, high rate of immature neutrophils (B) (30/57) correlated with poor PFS/OS (P = 0.04; P = 0.0007), with a 12-week death rate of 49%. Conclusion: The dNLR (B) and its dynamics (C2) under ICI associate with ICI outcomes in aNSCLC. Persistently high dNLR (B+C2) correlated with early ICI failure. Immature neutrophils may be a key subpopulation on ICI resistance

Predicting immunotherapy outcomes under therapy in patients with advanced NSCLC using dNLR and its early dynamics

Background: dNLR at the baseline (B), defined by neutrophils/[leucocytes-neutrophils], correlates with immune-checkpoint inhibitor (ICI) outcomes in advanced non–small-cell lung cancer (aNSCLC). However, dNLR is dynamic under therapy and its longitudinal assessment may provide data predicting efficacy. We sought to examine the impact of dNLR dynamics on ICI efficacy and understand its biological significance. Patients and methods: aNSCLC patients receiving ICI at 17 EU/US centres were included [Feb/13-Jun/18]. As chemotherapy-only group was evaluated (NCT02105168). dNLR was determined at (B) and at cycle2 (C2) [dNLR≤3 = low]. B+C2 dNLR were combined in one score: good = low (B+C2), poor = high (B+C2), intermediate = other situations. In 57 patients, we prospectively explored the immunophenotype of circulating neutrophils, particularly the CD15+CD244-CD16low cells (immature) by flow cytometry. Results: About 1485 patients treatment with ICI were analysed. In ICI-treated patients, high dNLR (B) (~1/3rd) associated with worse progression-free (PFS)/overall survival (OS) (HR 1.56/HR 2.02, P < 0.0001) but not with chemotherapy alone (N = 173). High dNLR at C2 was associated with worse PFS/OS (HR 1.64/HR 2.15, P < 0.0001). When dNLR at both time points were considered together, those with persistently high dNLR (23%) had poor survival (mOS = 5 months (mo)), compared with high dNLR at one time point (22%; mOS = 9.2mo) and persistently low dNLR (55%; mOS = 18.6mo) (P < 0.0001). The dNLR impact remained significant after PD-L1 adjustment. By cytometry, high rate of immature neutrophils (B) (30/57) correlated with poor PFS/OS (P = 0.04; P = 0.0007), with a 12-week death rate of 49%. Conclusion: The dNLR (B) and its dynamics (C2) under ICI associate with ICI outcomes in aNSCLC. Persistently high dNLR (B+C2) correlated with early ICI failure. Immature neutrophils may be a key subpopulation on ICI resistance

The purinergic receptor P2X7 and the NLRP3 inflammasome are druggable host factors required for SARS-CoV-2 infection

International audienceAbstract Purinergic receptors and NOD-like receptor protein 3 (NLRP3) inflammasome regulate inflammation and viral infection, but their effects on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remain poorly understood. Here, we report that the purinergic receptor P2X7 and NLRP3 inflammasome are cellular host factors required for SARS-CoV-2 infection. Lung autopsies from patients with severe coronavirus disease 2019 (COVID-19) reveal that NLRP3 expression is increased in host cellular targets of SARS-CoV-2 including alveolar macrophages, type II pneumocytes and syncytia arising from the fusion of infected macrophages, thus suggesting a potential role of NLRP3 and associated signaling pathways to both inflammation and viral replication. In vitro studies demonstrate that NLRP3-dependent inflammasome activation is detected upon macrophage abortive infection. More importantly, a weak activation of NLRP3 inflammasome is also detected during the early steps of SARS-CoV-2 infection of epithelial cells and promotes the viral replication in these cells. Interestingly, the purinergic receptor P2X7, which is known to control NLRP3 inflammasome activation, also favors the replication of D614G and alpha SARS-CoV-2 variants. Altogether, our results reveal an unexpected relationship between the purinergic receptor P2X7, the NLRP3 inflammasome and the permissiveness to SARS-CoV-2 infection that offers novel opportunities for COVID-19 treatment