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  • 1
    In: Journal of Clinical Oncology, American Society of Clinical Oncology (ASCO), Vol. 37, No. 15_suppl ( 2019-05-20), p. TPS3159-TPS3159
    Abstract: TPS3159 Background: Molecular targeted agents (MTA) resulted in breakthroughs in selected niches. It is often assumed that tumor regression is consecutive to an oncogenic de-addiction effect. An emerging hypothesis suggests that genomic instability may be associated with poor response to MTA. Indeed, the accumulation of defects in multiple oncogenes or tumor suppressor genes may result in the activation of multiple oncogenic pathways. These multiple signaling would mechanically result in a limitation of the oncogenic de-addiction process. Another hypothesis, suggests that tumor heterogeneity could also be associated with poor outcome under MTA. Such heterogeneity could also result from the genomic instability, and be appraised by bioinformatic and functional approaches. In this study, we thought to investigate whether molecular profiles reflecting a low level of genomic alterations in genes causally implicated in oncogenesis could be associated with an exceptional response (ER) to MTA. Methods: This is an exploratory, multicenter, multicohort, prospective trial conducted in 264 adult patients, with advanced breast, lung, colorectal, ovarian, kidney cancers and melanoma, having presented an ER to an approved MTA. ER is defined using the definition chosen by the NCI which combines the three criteria: - complete or partial response, - lasting 〉 6 months, - and not expected in 〉 10% of the patients in this drug – organ situation. The primary objective is to assess whether ER can be associated with a low level of genomic instability in the tumor. Low genomic instability is defined by the presence of less than the 5th quantile of genomic alterations (mutations, amplifications, deletions) to be expected in the given tumor type as per TCGA database. For each tumor type, the null hypothesis H0: π = 0.05 will be tested, against the one-sided alternative hypothesis π 〉 0.05. For each of the 6 cohorts, a sample size of 44 patients is necessary to achieve 80% power at π = 15 with a one-sided level 5% test. Patients presenting an ER will be identified retrospectively, in a nationwide manner, then monthly reviewed and validated for inclusion by a panel of pathology experts. As of February 2019, 75 patients have been included. The identification of molecular traits associated with ER might serve the development of predictive classifiers for precision medicine. This study also represents a unique opportunity to better understand cancer biology. Clinical trial information: NCT02701907.
    Type of Medium: Online Resource
    ISSN: 0732-183X , 1527-7755
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    Language: English
    Publisher: American Society of Clinical Oncology (ASCO)
    Publication Date: 2019
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  • 2
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 83, No. 7_Supplement ( 2023-04-04), p. 1034-1034
    Abstract: Introduction Liquid biopsy (LB) is increasingly used in non-small cell lung cancer (NSCLC) for molecular diagnosis or resistance detection. Due to its non-invasive nature, it is often preferred over tissue biopsies, especially when sequential biopsies are warranted. Here, we report the clinical utility of liquid biopsy in patients with advanced NSCLC and ALK, ROS1 or RET fusions and its relevance to detect resistance after targeted therapy. Methods Between December 2020 and June 2022, 597 patients with advanced lung cancer had at least one liquid biopsy assessed by Foundation One CDx Liquid (panel of 324 genes) in a single institution. Plasma collection was performed in treatment-naïve patients and/or at time of progression. Clinical and molecular data were collected from patients with known fusions (ALK, ROS1, RET). LB were defined as “positive” if the fusion or resistance mutations were identified and “negative” in the absence of circulating-tumor DNA. The clinical utility of LB was evaluated as the proportion of positive results. The clinical relevance for resistance detection was defined as the proportion of LB that identified a putative resistance mechanism after targeted therapy. Results A total of 68 patients (29 ALK+, 22 RET+ and 17 ROS1+) with 83 LB were included. Patients were females in 50% of cases, had no smoking history in 58% of cases and had adenocarcinoma in 91% of cases. LB was positive in 55/83 (66%) cases overall, in 14/15 (93%) treatment-naïve patients and in 39/66 (59%) pre-treated patients. Factors significantly associated with a negative LB were limited disease progression (brain- or thoracic-only, p & lt;0.001) and ongoing treatment at time of LB collection (p & lt;0.05). Out of 47 LB performed at progression after targeted therapies, 7 (15%) found on-target resistance, 10 (21%) by-pass resistance, 10 (21%) no explainable resistance and 20 (43%) were negative. By-pass alterations included KRAS p.G12C/A mutations (N=2, ALK+), PIK3CA p.E545K/Q (N=3, ALK+), PTEN splice-site mutation (N=1, RET+), MYC amplifications (N=3, ALK+/RET+) and MET amplification (N=1, ROS1+). Conclusion LB was able to detect resistance mechanisms in one third of NSCLC patients with ALK, RET or ROS1 fusions. However, LB frequently failed to detect circulating-tumor DNA especially in patients with limited disease progression or with ongoing treatment at time of sample collection. Citation Format: Mihaela Aldea, Arianna Marinello, Marco Tagliamento, Filippo Dall'Olio, Damien Vasseur, Arnaud Bayle, Anas Gazzah, Miruna Grecea, Claudio Nicotra, Ludovic Lacroix, Santiago Ponce, Luc Friboulet, Fabrice Barlesi, Fabrice Andre, David Planchard, Etienne Rouleau, Antoine Italiano, Benjamin Besse. Clinical utility of liquid biopsy for molecular characterization and resistance detection in patients with advanced NSCLC and ALK, ROS1 or RET fusions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1034.
    Type of Medium: Online Resource
    ISSN: 1538-7445
    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2023
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  • 3
    In: Cell, Elsevier BV, Vol. 182, No. 6 ( 2020-09), p. 1401-1418.e18
    Type of Medium: Online Resource
    ISSN: 0092-8674
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    Language: English
    Publisher: Elsevier BV
    Publication Date: 2020
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  • 4
    In: JCO Precision Oncology, American Society of Clinical Oncology (ASCO), , No. 7 ( 2023-03)
    Abstract: High-risk clonal hematopoiesis (CH) is frequently incidentally found in patients with solid tumors undergoing plasma cell–free DNA sequencing. Here, we aimed to determine if the incidental detection of high-risk CH by liquid biopsy may reveal occult hematologic malignancies in patients with solid tumors. MATERIALS AND METHODS Adult patients with advanced solid cancers enrolled in the Gustave Roussy Cancer Profiling study (ClinicalTrials.gov identifier: NCT04932525 ) underwent at least one liquid biopsy (FoundationOne Liquid CDx). Molecular reports were discussed within the Gustave Roussy Molecular Tumor Board (MTB). Potential CH alterations were observed, and patients referred to hematology consultation in the case of pathogenic mutations in JAK2, MPL, or MYD88, irrespective of the variant allele frequency (VAF), or in DNMT3A, TET2, ASXL1, IDH1, IDH2, SF3B1, or U2AF1 with VAF ≥ 10%, while also considering patient cancer-related prognosis. TP53 mutations were discussed case-by-case. RESULTS Between March and October 2021, 1,416 patients were included. One hundred ten patients (7.7%) carried at least one high-risk CH mutation: DNMT3A (n = 32), JAK2 (n = 28), TET2 (n = 19), ASXL1 (n = 18), SF3B1 (n = 5), IDH1 (n = 4), IDH2 (n = 3), MPL (n = 3), and U2AF1 (n = 2). The MTB advised for hematologic consultation in 45 patients. Overall, 9 patients of 18 actually addressed had confirmed hematologic malignancies that were occult in six patients: two patients had myelodysplastic syndrome, two essential thrombocythemia, one a marginal lymphoma, and one a Waldenström macroglobulinemia. The other three patients were already followed up in hematology. CONCLUSION The incidental findings of high-risk CH through liquid biopsy may trigger diagnostic hematologic tests and reveal an occult hematologic malignancy. Patients should have a multidisciplinary case-by-case evaluation.
    Type of Medium: Online Resource
    ISSN: 2473-4284
    Language: English
    Publisher: American Society of Clinical Oncology (ASCO)
    Publication Date: 2023
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  • 5
    In: Cancer Discovery, American Association for Cancer Research (AACR), Vol. 12, No. 4 ( 2022-04-01), p. 958-983
    Abstract: Vaccination against coronavirus disease 2019 (COVID-19) relies on the in-depth understanding of protective immune responses to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). We characterized the polarity and specificity of memory T cells directed against SARS-CoV-2 viral lysates and peptides to determine correlates with spontaneous, virus-elicited, or vaccine-induced protection against COVID-19 in disease-free and cancer-bearing individuals. A disbalance between type 1 and 2 cytokine release was associated with high susceptibility to COVID-19. Individuals susceptible to infection exhibited a specific deficit in the T helper 1/T cytotoxic 1 (Th1/Tc1) peptide repertoire affecting the receptor binding domain of the spike protein (S1-RBD), a hotspot of viral mutations. Current vaccines triggered Th1/Tc1 responses in only a fraction of all subject categories, more effectively against the original sequence of S1-RBD than that from viral variants. We speculate that the next generation of vaccines should elicit Th1/Tc1 T-cell responses against the S1-RBD domain of emerging viral variants. Significance: This study prospectively analyzed virus-specific T-cell correlates of protection against COVID-19 in healthy and cancer-bearing individuals. A disbalance between Th1/Th2 recall responses conferred susceptibility to COVID-19 in both populations, coinciding with selective defects in Th1 recognition of the receptor binding domain of spike. See related commentary by McGary and Vardhana, p. 892. This article is highlighted in the In This Issue feature, p. 873
    Type of Medium: Online Resource
    ISSN: 2159-8274 , 2159-8290
    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2022
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  • 6
    In: Cell Death & Differentiation, Springer Science and Business Media LLC, Vol. 28, No. 12 ( 2021-12), p. 3297-3315
    Abstract: Patients with cancer are at higher risk of severe coronavirus infectious disease 2019 (COVID-19), but the mechanisms underlying virus–host interactions during cancer therapies remain elusive. When comparing nasopharyngeal swabs from cancer and noncancer patients for RT-qPCR cycle thresholds measuring acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in 1063 patients (58% with cancer), we found that malignant disease favors the magnitude and duration of viral RNA shedding concomitant with prolonged serum elevations of type 1 IFN that anticorrelated with anti-RBD IgG antibodies. Cancer patients with a prolonged SARS-CoV-2 RNA detection exhibited the typical immunopathology of severe COVID-19 at the early phase of infection including circulation of immature neutrophils, depletion of nonconventional monocytes, and a general lymphopenia that, however, was accompanied by a rise in plasmablasts, activated follicular T-helper cells, and non-naive Granzyme B + FasL + , Eomes high TCF-1 high , PD-1 + CD8 + Tc1 cells. Virus-induced lymphopenia worsened cancer-associated lymphocyte loss, and low lymphocyte counts correlated with chronic SARS-CoV-2 RNA shedding, COVID-19 severity, and a higher risk of cancer-related death in the first and second surge of the pandemic. Lymphocyte loss correlated with significant changes in metabolites from the polyamine and biliary salt pathways as well as increased blood DNA from Enterobacteriaceae and Micrococcaceae gut family members in long-term viral carriers. We surmise that cancer therapies may exacerbate the paradoxical association between lymphopenia and COVID-19-related immunopathology, and that the prevention of COVID-19-induced lymphocyte loss may reduce cancer-associated death.
    Type of Medium: Online Resource
    ISSN: 1350-9047 , 1476-5403
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2021
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  • 7
    In: Clinical Cancer Research, American Association for Cancer Research (AACR), ( 2023-07-19), p. OF1-OF14
    Abstract: The androgen receptor axis inhibitors (ARPI; e.g., enzalutamide, abiraterone acetate) are administered in daily practice for men with metastatic castration-resistant prostate cancer (mCRPC). However, not all patients respond, and mechanisms of both primary and acquired resistance remain largely unknown. Experimental Design: In the prospective trial MATCH-R (NCT02517892), 59 patients with mCRPC underwent whole-exome sequencing (WES) and/or RNA sequencing (RNA-seq) of samples collected before starting ARPI. Also, 18 patients with mCRPC underwent biopsy at time of resistance. The objectives were to identify genomic alterations associated with resistance to ARPIs as well as to describe clonal evolution. Associations of genomic and transcriptomic alterations with primary resistance were determined using Wilcoxon and Fisher exact tests. Results: WES analysis indicated that no single-gene genomic alterations were strongly associated with primary resistance. RNA-seq analysis showed that androgen receptor (AR) gene alterations and expression levels were similar between responders and nonresponders. RNA-based pathway analysis found that patients with primary resistance had a higher Hedgehog pathway score, a lower AR pathway score and a lower NOTCH pathway score than patients with a response. Subclonal evolution and acquisition of new alterations in AR-related genes or neuroendocrine differentiation are associated with acquired resistance. ARPIs do not induce significant changes in the tumor transcriptome of most patients; however, programs associated with cell proliferation are enriched in resistant samples. Conclusions: Low AR activity, activation of stemness programs, and Hedgehog pathway were associated with primary ARPIs’ resistance, whereas most acquired resistance was associated with subclonal evolution, AR-related events, and neuroendocrine differentiation.
    Type of Medium: Online Resource
    ISSN: 1078-0432 , 1557-3265
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2023
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  • 8
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 81, No. 13_Supplement ( 2021-07-01), p. 358-358
    Abstract: Background: The androgen receptor axis inhibitors (ARi) (e.g, enzalutamide, abiraterone acetate) are administered in daily practice for men with metastatic castration-resistant prostate cancer (mCRPC). However, not all patients respond, and mechanisms of both primary and acquired resistance remain largely unknown. Methods: In a prospective trial MATCH-R (NCT02517892), 55 mCRPC patients underwent whole exome sequencing (WES) (n=45) and RNA-sequencing (RNA-seq) (n=52) of metastatic biopsies before starting ARi. Also, 16 mCRPC patients underwent biopsy at time of resistance (WES=14, RNA-seq = 14). The objectives were to identify genomic alterations associated with resistance to ARi as well as to describe clonal evolution. Primary resistance was determined at 4 months of treatment using composite criteria for progression that included serum prostate specific antigen measurements, bone scan, CT imaging and symptom assessments. Acquired resistance was defined by occurrence of progressive disease after initial response or stable disease. Associations of genomic and transcriptomic alterations with primary resistance were determined using Wilcoxon and Fisher's exact tests. Results: At 4 months, 22/55 patients in the cohort had disease progression (primary resistance). No genomic alterations from WES analysis were significantly associated with primary resistance. Analysis of sequential biopsies suggests that mCRPC follows mainly a parallel evolution model and involve DNA-repair related mutational processes. At time of acquired resistance to ARi, most tumors acquired new drivers affecting AR pathway (e.g, AR, NCOR1/2) or lineage switching (e.g, RB1, PTEN, TP53). Using computational methods, we measured AR transcriptional function and performed gene set enrichment analysis to identify pathways whose activity state correlated with resistance. AR gene alterations and AR expression were similar between responding and non-responding patients. Transcriptional analysis demonstrated that multiple specific gene sets — including those linked to low AR transcriptional activity, stemness program, RB loss and homologous repair deficiency — were activated in both primary and acquired resistance. Conclusion: Resistance to AR axis inhibitors results from multiple transcriptional programs already activated in pre-treatment samples. Clonal evolution analysis along with RNA-seq data indicate the role of genomic instability and lineage switching in driving acquired resistance Citation Format: Naoual Menssouri, Loic Poiraudeau, Carole Helissey, Ludovic Bigot, Jonathan Sabio, Tony Ibrahim, Claudio Nicotra, Maud Ngocamus, Lambros Tselikas, Thierry De Baere, Etienne Rouleau, Ludovic Lacroix, Anne Chaucherau, Luc Friboulet, Ronan Flippot, Giulia Baciarello, Laurence Albiges, Emeline Colomba, Pernelle Lavaud, Stefan Michiels, Aline Maillard, Antoine Italiano, Fabrice Barlesi, Jean-Charles Soria, Jean-Yves Scoazec, christophe Massard, Benjamin Besse, Fabrice André, Karim Fizazi, Daniel Gautheret, Yohann Loriot. A prospective study of prostate cancer metastases identifies an androgen receptor activity-low, stemness program associated with resistance to androgen receptor axis inhibitors and unveils mechanisms of clonal evolution [abstract] . In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 358.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2021
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  • 9
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 82, No. 12_Supplement ( 2022-06-15), p. 1251-1251
    Abstract: Background: The discovery of immune checkpoint blockers (ICB) has revolutionized the systemic approach of the treatment of cancer. However, most patients receiving ICB do not derive benefit. Therefore, there is a crucial need to identify reliable predictive biomarkers of response to anti-PD-1/PD-L1 agents, both to develop precision medicine in cancer immunotherapy and to better understand mechanisms of sensitivity and resistance. One pathway that plays an important role in the regulation of immune cell reactivity is L-Arginine (Arg) metabolism, which is essential to T-cell activation. We therefore aimed at evaluating the association of baseline plasmatic level of Arg - serving as a surrogate of Arginase (Arg1) activity - and clinical benefit to ICB. Methods: Correlation with Arg levels and efficacy of ICB in the pre-clinical setting was assessed by using a syngeneic mouse model of colorectal cancer (MC38) known to be responsive to ICB. Correlation of Arg levels and clinical activity of ICB was assessed by analyzing the plasma samples obtained before treatment onset in two independent cohorts of patient with advanced cancer and included in two institutional molecular profiling programs (discovery cohort: BIP, NCT02534649, n=77; validation cohort: PREMIS, n=295, NCT03984318). In addition, using matched PBMCs-plasma samples, we analyzed the correlation between Arg level and features of PBMCs that were captured through multiplexed-flow cytometry analysis. Results: As expected, treatment of MC38-tumor bearing mice with anti-PD(L)1 antibodies demonstrated a strong anti-tumor effect with tumor rejection observed for app. 40% of mice (11 out of 28). The tumor rejection rate was significantly higher in mice with high baseline Arg level than in mice with low Arg level: 85.7% versus 23.8%, p=0.004. In both discovery and validation cohorts, low Arg level at baseline (42 & lt;µmol/L) was significantly associated with worse clinical benefit rate, progression-free survival (PFS) and overall survival (OS). Multivariate analysis showed that low baseline Arg level isd an independent prognostic factor for both PFS and OS. Finally, PBMCs immunophenotyping showed that low Arg level was significantly associated with increased PDL1 expression in several immune cell subsets from the myeloid lineage. Conclusions: Altogether, our results demonstrate that baseline Arg levels are highly predictive of ICB efficacy. Increase in PDL1 expression in myeloid cells upon Arg deprivation could partly underly its suppressive activity. Plasmatic Arg quantification can therefore represent an attractive biomarker to tailor novel therapeutic regimens targeting the Arginase pathway in combination with ICB. Citation Format: Jean-Philippe Guegan, Florent Peyraud, Aurelien Marabelle, Nathalie Chaput, Dominique Bodet, Laure Fontan, Anthony Gaultier, Imane NAFIA, Francois-Xavier Danlos, David Planchard, Caroline Robert, Caroline Even, Mohamed Khettab, Lambros Tselikas, Luc Friboulet, Jean-Charles Soria, Fabrice Andre, Fabrice Barlesi, Alban Bessede, Antoine Italiano. Low plasma Arginine level is associated with resistance to immune checkpoint blockers in patients with advanced cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1251.
    Type of Medium: Online Resource
    ISSN: 1538-7445
    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2022
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  • 10
    In: Cancer Research, American Association for Cancer Research (AACR), Vol. 80, No. 16_Supplement ( 2020-08-15), p. CT403-CT403
    Abstract: Background: The SARS-CoV-2 outbreak in Paris’ region significantly affected Gustave Roussy cancer center. Here, we report the Gustave Roussy experience during the SARS-CoV-2 outbreak. This outbreak has led to a rapid reorganization of cancer patients’ (pts) management, with two concurrent objectives. First, protect cancer pts, who may experience more severe form of the disease, from being infected by the SARS-CoV-2. Second, protect cancer pts from losing the chance to receive optimal, if not standard, cancer care. Methods: Cancer pts with suspected SARS-CoV-2 infection were admitted at Gustave Roussy starting March, 12th. Screening indications have been adapted over the time. All the COVID19 pts positively tested and managed at Gustave Roussy between March 14th (1st positive case) and April 15th have been included in a redcap database. Pts and underlying oncological and COVID19 diseases characteristics have been collected. Cancer and COVID-19 managements, and outcomes have been assessed. The primary endpoint of this analysis was the clinical deterioration, defined as the need for O2 supplementation of 6l/min or more, or death of any cause. Results: Overall, 7,251 cancer pts were managed at Gustave Roussy during this period of time, with 3616 being hospitalized. Based on our testing strategy, 1302 pts have been tested with 12% of them found positive for SARS-CoV-2. Among the first 137 cancer pts diagnosed with SARS-CoV-2, most cases were female (58%) with a median age of 61 years, including 36 pts (26%) ≥ 70 years. Most frequent underlying cancers were solid tumors (115) including breast (23), GI (18), head and neck (17), GU (17), GYN (17) malignancies or hemopathies (22). At time of COVID diagnosis, 79 pts (58%) had metastatic/active cancer and 56 pts (41%) were considered in remission/treated with curative intent. The diagnosis of SARS-CoV-2 infection was made by RT-PCR or thoracic CT scan alone in 93.4% and 6.6% of the cases, respectively. The majority of the pts was hospitalized (75%) and treated with HCQ/AZI (40; 30%) with inclusion in the ONCOVID trial (EudraCT: 2020-01250-21), IL-6 inhibitor (10), antiviral (6) or steroids (13). Fifteen pts were admitted in ICU (11%). Clinical deterioration occurred in 34 pts (24.8%) and was associated with hematological underlying disease, CRP at diagnosis of COVID19 & gt;50 and the use of cytotoxic chemotherapy within & lt;3mo. At data cut-off (April, 20th 2020), 95 (69.3%), 20 (14.6%), and 22 (16.1%) pts were discharged, had died, or were still hospitalized, respectively. All the deaths were considered related to the SARS-CoV-2 infection. Conclusions: Globally, the rate of the SARS-CoV-2 infection in our cancer patients’ population does not seem to be higher compared to the global population. We have not found evidence that COVID19 is more lethal or aggressive in cancer patients that underwent usual SARS-Cov-2 treatment. We believe that adequate testing and protective measures, along with the low rate of SARS-cov-2-treatment-related adverse events (5.5%), justify an optimal management of the cancer patients’ underlying tumor. Citation Format: Fabrice Barlesi, Stéphanie Foulon, Arnauld Bayle, Bertrand Gachot, Fanny Pommeret, Christophe Willekens, Annabelle Stoclin, Mansouria Merad, Franck GriscelliI, Jean-Baptise Micol, Roger Sun, Thomas Nihouarn, Corinne Balleygier, Fabrice André, Florian Scotte, Benjamin Besse, Jean-Charles Soria, Laurence Albiges. Outcome of cancer patients infected with COVID-19, including toxicity of cancer treatments [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr CT403.
    Type of Medium: Online Resource
    ISSN: 0008-5472 , 1538-7445
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    Language: English
    Publisher: American Association for Cancer Research (AACR)
    Publication Date: 2020
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