Appropriate cancer care requires a thorough understanding of the natural history of the disease, including the cell of origin, the pattern of clonal evolution, and the functional consequences of the mutations. Using deep sequencing of flow-sorted cell populations from patients with chronic lymphocytic leukemia (CLL), we established the presence of acquired mutations in multipotent hematopoietic progenitors. Mutations affected known lymphoid oncogenes, including BRAF, NOTCH1, and SF3B1. NFKBIE and EGR2 mutations were observed at unexpectedly high frequencies, 10.7% and 8.3% of 168 advanced-stage patients, respectively. EGR2 mutations were associated with a shorter time to treatment and poor overall survival. Analyses of BRAF and EGR2 mutations suggest that they result in deregulation of B-cell receptor (BCR) intracellular signaling. Our data propose disruption of hematopoietic and early B-cell differentiation through the deregulation of pre-BCR signaling as a phenotypic outcome of CLL mutations and show that CLL develops from a pre-leukemic phase. SIGNIFICANCE:The origin and pathogenic mechanisms of CLL are not fully understood. The current work indicates that CLL develops from pre-leukemic multipotent hematopoietic progenitors carrying somatic mutations. It advocates for abnormalities in early B-cell differentiation as a phenotypic convergence of the diverse acquired mutations observed in CLL. Cancer Discov; 4(9); 1088-1101

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Deletions of the long arm of chromosome 14 [del(14q)] are rare but recurrently observed in mature B-cell neoplasms, particularly in chronic lymphocytic leukemia (CLL). To further characterize this aberration, we studied 81 cases with del(14q): 54 of CLL and 27 of small lymphocytic lymphoma (SLL), the largest reported series to date. Using karyotype and fluorescence in situ hybridization (FISH), the most frequent additional abnormality was trisomy 12 (tri12), observed in 28/79 (35%) cases, followed by del13q14 (12/79, 15%), delTP53 (11/80, 14%) delATM (5/79, 6%), and del6q21 (3/76, 4%). IGHV genes were unmutated in 41/53 (77%) patients, with a high frequency of IGHV1-69 (21/52, 40%). NOTCH1 gene was mutated in 14/45 (31%) patients. There was no significant difference in cytogenetic and molecular abnormalities between CLL and SLL. Investigations using FISH and SNP-array demonstrated the heterogeneous size of the 14q deletions. However, a group with the same del(14)(q24.1q32.33) was identified in 48% of cases. In this group, tri12 (P = 0.004) and NOTCH1 mutations (P = 0.02) were significantly more frequent than in the other patients. In CLL patients with del(14q), median treatment-free survival (TFS) was 27 months. In conclusion, del(14q) is associated with tri12 and with pejorative prognostic factors: unmutated IGHV genes (with over-representation of the IGHV1-69 repertoire), NOTCH1 mutations, and a short TFS.

Homeoproteins of the Engrailed family are involved in the patterning of mesencephalic boundaries through a mechanism classically ascribed to their transcriptional functions. In light of recent reports on the paracrine activity of homeoproteins, including Engrailed, we asked whether Engrailed intercellular transfer was also involved in brain patterning and boundary formation. Using time-controlled activation of Engrailed combined with tools that block its transfer, we show that the positioning of the diencephalic-mesencephalic boundary (DMB) requires Engrailed paracrine activity. Both zebrafish Eng2a and Eng2b are competent for intercellular transfer in vivo, but only extracellular endogenous Eng2b, and not Eng2a, participates in DMB positioning. In addition, disruption of the Pbx-interacting motif in Engrailed, known to strongly reduce the gain-of-function phenotype, also downregulates Engrailed transfer, thus revealing an unsuspected participation of the Pbx interaction domain in this pathway.

Trisomy 12 (tri12) is the second most frequent chromosomal aberration (15%-20%) in chronic lymphocytic leukemia (CLL). Tri12 confers an intermediate prognosis but is a heterogeneous entity. We examined whether additional mutational or chromosomal alterations might impact tri12 patient outcomes. This retrospective study, carried out by the French Innovative Leukemia Organization, included 188 tri12 patients with comprehensive information on immunoglobulin heavy chain (IGHV) gene status, karyotypic/FISH abnormalities, and NOTCH1, TP53, SF3B1, and MYD88 mutations. The main cytogenetic abnormalities associated with tri12 were del(13q) (25%), additional trisomies (14%) (including tri19 (10%) and tri18 (4%)), 14q32 translocations (10%), del(17p) (6.5%), del(14q) (4%), and del(11q) (4%). Unmutated (UM) IGHV, NOTCH1, and TP53, mutations were identified in respectively 66%, 25%, and 8.5% of cases. Multivariate analyses showed that additional trisomies (HR = 0.43, 95% CI = 0.23-0.78, P = .01) were associated with a significantly longer time to first treatment in Binet stage A patients and with a lower risk of relapse (HR = 0.37, 95% CI = 0.15-0.9, P = .03) in the overall tri12 population. Binet stage B/C, TP53 disruption, and UM IGHV status were associated with a shorter time to next treatment, while Binet stage B/C (HR = 4, 95% CI = 1.6-4.9, P = .002) and TP53 disruption (HR = 5, 95% CI = 1.94-12.66, P = .001) conferred shorter overall survival in multivariate comparisons. These data indicate that additional cytogenetic and mutational abnormalities, and particularly additional trisomies, IGHV status, and TP53 disruption, influence tri12 patient outcomes and could improve risk stratification in this population.

The problem of the arterial vascularization of the human thalamus has been debated at length. Anatomical references concerning the thalamic arterial groups are contradictory and complex, preventing any solid application in practice. It is, therefore, difficult to produce reliable anatomical radio-clinical correlation. In this work, 12 adult human cerebellums (24 hemispheres) were dissected after intra-vascular injection. With care for clarification and standardization, the extra-parenchymal thalamic arteries were classified in six groups: pre-mamillary artery, perforating thalamic arteries, thalamo-geniculate arteries, perforating branches of the postero-medial, postero-lateral and anterior choroidal arteries. Variations in the pre-mamillary artery were rare. The origin of the perforating thalamic artery was unilateral in two of three cases. The origin of the thalamo-geniculate arteries arose between the posterior cerebral artery (53%) and the posterior choroidal arteries (43%). The postero-median choroidal artery was most often single and usually gave the perforating branches for the medial aspect of the thalamus. The postero-lateral choroidal artery was frequently multiple and essentially gave the perforating branches for the superior aspect of the thalamus. The pulvinarian branches most often rose from the postero-lateral choroidal arteries (two thirds of cases) and more rarely from the postero-median choroidal arteries (one third of cases). The anterior choroidal artery is a source of thalamic vascularization by its cisternal branches running towards the lateral thalamus. It can also participate in the vascularization of the pulvinar by the plexiform branches crossing the temporal horn of the lateral ventricle. This study has allowed definition of the intra-parenchymatous arterial map of the thalamus. This mapping is essential for producing anatomical radio-clinical correlations which are pertinent for therapeutic decisions.

Background Cytogenetic abnormalities are of key importance for predicting clinical course and response to therapy in patients with chronic lymphocytic leukemia (CLL). Trisomy 12 (tri12), the third most frequent chromosomal aberration in CLL patients (10-20%), is associated with an intermediate prognostic risk but represents a clinical heterogeneous entity. Recently, next generation sequencing have revealed recurrent mutations in genes that were unknown to be involved in CLL pathogenesis, including NOTCH1, MYD88, SF3B1, XPO1 and BIRC3. In patients harboring tri12, NOTCH1 mutations have been shown to be present in up to 25% of cases and to confer unfavorable outcome explaining in part the clinical heterogeneity of tri12 patients. To better understand the genetic basis and prognosis of tri12 patients, we performed a multicenter retrospective study combining extensive mutational and cytogenetic analysis. Methods Patients carrying tri12 were identified using fluorescence in situ hybridization (FISH) and/or chromosome banding (CB). Main clinical and biological characteristics were collected and included in univariate analysis of prognostic factors, comprising age, Binet stage (A vs. B-C), splenomegaly, lymphocyte doubling time (LDT), LDH, beta2microglobulin (B2M), CD38 expression, IGHV mutational status, percentage of interphase nuclei positive (INP) for tri12, additional FISH (del13q, del11q, del17p) or chromosomal aberrations and presence of complex karyotype (> 2 CB abnormalities). Search for mutations was performed by Sanger direct sequencing for TP53 (exons 5-10), NOTCH1 (exon 34), MYD88 (exons 14-16), SF3B1 (exons 14-16) and XPO1 (exons 14-15). Primary and secondary endpoints were time to first treatment (TFT), response to therapy, time to next treatment (TNT) and overall survival (OS). Results The study population comprised a total of 177 untreated patients including 112 and 75 patients with stage A and B-C CLL, respectively. The median age at diagnosis was 62 years old (range, 31-87) and 33% of patients were female. B2M was superior to 4 mg/L in 30/92 (32%) patients and LDH elevated in 65%. CD38 expression was positive (>30%) in 58% and IGHV status was unmutated in 60%. Among the whole study population, all patients were positive for tri12 by FISH and 158/165 by CB. Tri12 was associated by CB with tri19 in 21 patients (13.2%), tri18 in 12 patients (7.5%), tri3 in 1 patient (<1%), t(14;18) in 9 patients (5.7%), t(14;19) in 3 patients (2%) and del14q in 7 patients (4.5%). Complex karyotype was present in 42 patients (26%) and tri12 was the sole abnormality observed by CB in 71 patients (45%). Out of 170 patients analysed with the four probes by FISH, tri12 was the sole abnormality in 114 patients (67%) and was associated with del13q, del11q and del17p in, respectively, 44 (26%), 8 (4%) and 10 (6%) patients. The median percentage of INP for tri12 was 58% (range, 5-100). TP53, NOTCH1, MYD88 and SF3B1 mutations were tested in 113 patients and identified in, respectively, 9 (8%), 19 (17%), 1 (<1%) and 1 (<1%) patients. No mutation of XPO1 was observed. Among the stage A population of 112 patients, 64 (57%) needed treatment with a median TFT of 45 months (range, 1-170). A shorter TFT was significantly associated with a LDT inferior to one year (P=0.0001), and the presence of TP53 (P=0.04) mutation while the presence of another trisomy (P=0.03) was associated with a significantly longer TFT. By multivariate analysis, only TP53 mutation retain prognostic significance for TFT (HR=3.9, 95%CI=1.02-15.04). Among the 75 stage B-C patients, 62 were treated, 51 evaluable for response to therapy and 28 received a second line of treatment. The only prognostic variable associated with poor response to therapy was the presence of NOTCH1 mutation (P=0.01). Finally, regarding the whole tri12 population, Binet stage (A vs. B-C), splenomegaly, lymphocytosis, LDH, B2M, the percentage of INP and the IGHV unmutated status were associated with worst OS in univariate analysis. Conclusion The most frequent molecular abnormalities observed in our tri12 cohort were TP53 and NOTCH1 mutations that occurred in, respectively, 8 and 17% of cases. NOTCH1 mutations were associated with poor response to therapy and TP53 mutations with a shorter time to first treatment. Disclosures: Leblond: Mundipharma: Honoraria, Membership on an entity’s Board of Directors or advisory committees, Speakers Bureau; Janssen: Honoraria, Membership on an entity’s Board of Directors or advisory committees; Roche : Consultancy, Honoraria, Membership on an entity’s Board of Directors or advisory committees, Speakers Bureau.

The concept of Internet of Things (IoT) has changed the way we live by integrating commodity devices with cyberspace to automate our everyday tasks. Nowadays, IoT devices in the home environment are becoming ubiquitous with seamless connectivity and diverse application domains. Modern IoT devices have adopted a manyto-many connectivity model to enhance user experience and device functionalities compared to early IoT devices with standalone device setup and limited functionalities. However, the continuous connection between devices and cyberspace has introduced new cyber attacks targeting IoT devices and networks. Due to the resource-constrained nature of IoT devices as well as the opacity of the IoT framework, traditional intrusion detection systems cannot be applied here. In this paper, we introduce Sentinel, a novel intrusion detection system that uses kernel-level information to detect malicious attacks. Specifically, Sentinel collects low-level system information (CPU usage, RAM usage, total load, available swap, etc.) of each IoT device in a network and learns the pattern of device behavior to differentiate between benign and malicious events. We evaluated the efficacy and performance of Sentinel in different IoT platforms with multiple devices and settings. We also measured the performance of Sentinel against five types of real-life attacks. Our evaluation shows that Sentinel can detect different attacks to IoT devices and networks with high accuracy (over 95%) and secure the devices in different IoT platforms and configurations. Also, Sentinel achieves minimum overhead in power consumption, ensuring high compatibility in resource-constraint IoT devices. CCS CONCEPTS• Security and privacy → Intrusion detection systems.