The clinical management of metastatic colorectal cancer (mCRC) faces major challenges. Here, we show that nilotinib, a clinically approved drug for chronic myeloid leukaemia, strongly inhibits human CRC cell invasion in vitro and reduces their metastatic potential in intrasplenic tumour mouse models. Nilotinib acts by inhibiting the kinase activity of DDR1, a receptor tyrosine kinase for collagens, which we identified as a RAS‐independent inducer of CRC metastasis. Using quantitative phosphoproteomics, we identified BCR as a new DDR1 substrate and demonstrated that nilotinib prevents DDR1‐mediated BCR phosphorylation on Tyr177, which is important for maintaining β‐catenin transcriptional activity necessary for tumour cell invasion. DDR1 kinase inhibition also reduced the invasion of patient‐derived metastatic and circulating CRC cell lines. Collectively, our results indicate that the targeting DDR1 kinase activity with nilotinib may be beneficial for patients with mCRC.

Purpose: Use of circulating tumor DNA for diagnosis is limited regarding the low number of target molecules in early stage tumors. HPV-associated carcinomas represent a privileged model using circulating viral DNA (ctHPV DNA) as a tumor marker. However, the plurality of HPV genotypes represents a challenge. The NGSbased CaptHPV approach is able to characterize any HPV DNA sequence. To assess the ability of this method to establish the diagnosis of HPV-associated cancer via a blood sample, we analyzed ctHPV DNA in HPV-positive or HPV-negative carcinomas.Experimental Design: Patients ( 135) from France and Senegal with carcinoma developed in the uterine cervix (74), oropharynx (25), oral cavity (19), anus (12), and vulva (5) were prospectively registered. Matched tumor tissue and blood samples (10 ml) were taken before treatment and independently analyzed using the CaptHPV method.Results: HPV prevalence in tumors was 60.0% (81/135) (15 different genotypes).Viral analysis of plasmas compared to tumors was available for 134 patients. In the group of 80 patients with HPV-positive tumors, 77 were also positive in plasma (sensitivity 95.0%); in the group of 54 patients with HPV-negative tumors, one was positive in plasma (specificity 98.1%). In most cases, the complete HPV pattern observed in tumors could be established from the analysis of ctHPV DNA. Conclusions:In patients with carcinoma associated with any HPV genotype, a complete viral genome characterization can be obtained via the analysis of a standard blood sample. This should favor the development of non-invasive diagnostic tests providing the identification of personalized tumor markers.

BackgroundApproximately 900 children/adolescents are treated with radiotherapy (RT) every year in France. However, among the 80% of survivors, the cumulative incidence of long-term morbidity – including second malignancies - reach 73.4% thirty years after the cancer diagnosis. Identifying a priori the subjects at risk for RT sequelae is a major challenge of paediatric oncology. Individual radiosensitivity (IRS) of children/adolescents is unknown at this time, probably with large variability depending on the age when considering the changes in metabolic functions throughout growth. We previously retrospectively showed that unrepaired DNA double strand breaks (DSB) as well a delay in the nucleoshuttling of the pATM protein were common features to patients with RT toxicity. We aim to validate a high performance functional assay for IRS prospectively.Methods/designARPEGE is a prospective open-label, non-randomized multicentre cohort study. We will prospectively recruit 222 children/adolescents who require RT as part of their routine care and follow them during 15 years. Prior RT we will collect blood and skin samples to raise a primary dermal fibroblast line to carry out in blind the IRS assay. As a primary objective, we will determine its discriminating ability to predict the occurrence of unusual early skin, mucous or hematological toxicity. The primary endpoint is the measurement of residual double-strand breaks 24 h after ex vivo radiation assessed with indirect immunofluorescence (γH2AX marker). Secondary endpoints include the determination of pATM foci at 10 min and 1 h (pATM marker) and micronuclei at 24 h. In parallel toxicity including second malignancies will be reported according to NCI-CTCAE v4.0 reference scale three months of the completion of RT then periodically during 15 years. Confusion factors such as irradiated volume, skin phototype, previous chemotherapy regimen, smoking, comorbities (diabetes, immunodeficiency, chronic inflammatory disease...) will be reported.DiscussionARPEGE would be the first study to document the distribution of IRS in the pediatric subpopulation. Screening hypersensitive patients would be a major step forward in the management of cancers, opening a way to personalized pediatric oncology.Trial registrationID-RCB number: 2015-A00975–44, ClinicalTrials.gov Identifier: NCT02827552 Registered 7/6/2016.

Tyrosine kinases (TK) are frequently deregulated in human cancer and they play important roles in tumor progression. Since then, they have become valuable therapeutic targets and several inhibitors are currently used in the clinic. For example, the BCR-ABL inhibitor nilotinib is currently used for the treatment of patients with chronic myeloid leukemia (CML). Here we show that this antileukemic drug also inhibits invasive properties of colorectal cancer (CRC) cells. Inhibition was observed at the same dose-range for growth inhibition of CML cells. Nilotinib also strongly reduced liver metastasis induced by injection of CRC cells in the spleen of nude mice. Since we could not detect Abl deregulation in CRC cells, we speculated the involvement of an alternative target to be identified. Interestingly, a previous chemical-proteomic approach identified DDR1 as a novel target of nilotinib (Rix et al, Blood, 2007). DDR1 is a poorly-characterized TK and a receptor for collagen, a major component of the extracellular matrix. Accordingly, we found that DDR1 catalytic activity regulates cell invasion and metastasis of CRC cells. In addition, we demonstrate that nilotinib pharmacological activity is mediated by the inhibition of DDR1 in CRC cells: expression of the nilotinib resistant DDR1/T701I mutant renders CRC cells resistant to nilotinib treatment. Finally, we found that DDR1 catalytic activity is significantly increased in metastatic nodules compared to the primary tumor and the healthy tissue of the same patient. Altogether, these data suggest that the targeting of DDR1 by nilotinib may be of therapeutic value in metastatic CRC. Citation Format: Priscillia Tosti, Cédric Leroy, Valérie Simon, Bruno Robert, Josiane Pierre, Audrey Sirvent, Serge Roche. The antileukemic drug nilotinib inhibits the invasive activity and the metastatic potential of colorectal cancer cells by targeting the receptor tyrosine kinase DDR1. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2085. doi:10.1158/1538-7445.AM2013-2085 Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.