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PURPOSE Secondary myeloid neoplasms (sMNs) remain the most serious long-term complications in patients with aplastic anemia (AA) and paroxysmal nocturnal hemoglobinuria (PNH). However, sMNs lack specific predictors, dedicated surveillance measures, and early therapeutic interventions. PATIENTS AND METHODS We studied a multicenter, retrospective cohort of 1,008 patients (median follow-up 8.6 years) with AA and PNH to assess clinical and molecular determinants of clonal evolution. RESULTS Although none of the patients transplanted upfront (n = 117) developed clonal complications (either sMN or secondary PNH), the 10-year cumulative incidence of sMN in nontransplanted cases was 11.6%. In severe AA, older age at presentation and lack of response to immunosuppressive therapy were independently associated with increased risk of sMN, whereas untreated patients had the highest risk among nonsevere cases. The elapsed time from AA to sMN was 4.5 years. sMN developed in 94 patients. The 5-year overall survival reached 40% and was independently associated with bone marrow blasts at sMN onset. Myelodysplastic syndrome with high-risk phenotypes, del7/7q, and ASXL1, SETBP1, RUNX1, and RAS pathway gene mutations were the most frequent characteristics. Cross-sectional studies of clonal dynamics from baseline to evolution revealed that PIGA/human leukocyte antigen lesions decreased over time, being replaced by clones with myeloid hits. PIGA and BCOR/L1 mutation carriers had a lower risk of sMN progression, whereas myeloid driver lesions marked the group with a higher risk. CONCLUSION The risk of sMN in AA is associated with disease severity, lack of response to treatment, and patients' age. sMNs display high-risk morphological, karyotypic, and molecular features. The landscape of acquired somatic mutations is complex and incompletely understood and should be considered with caution in medical management.

Differentially Expressed Proteins Deleted versus non-deleted clones

Supp Fig. S1-6, Supp Tables S1-S3, Supp Methods and References

Deletion of chromosome 6q is a well-recognized abnormality found in poor-prognosis T-cell acute lymphoblastic leukemia (T-ALL). Using integrated genomic approaches, we identified two candidate haploinsufficient genes contiguous at 6q14, SYNCRIP (encoding hnRNP-Q) and SNHG5 (that hosts snoRNAs), both involved in regulating RNA maturation and translation. Combined silencing of both genes, but not of either gene alone, accelerated leukemogeneis in a Tal1/Lmo1/Notch1-driven mouse model, demonstrating the tumor-suppressive nature of the two-gene region. Proteomic and translational profiling of cells in which we engineered a short 6q deletion by CRISPR/Cas9 genome editing indicated decreased ribosome and mitochondrial activities, suggesting that the resulting metabolic changes may regulate tumor progression. Indeed, xenograft experiments showed an increased leukemia-initiating cell activity of primary human leukemic cells upon coextinction of SYNCRIP and SNHG5. Our findings not only elucidate the nature of 6q deletion but also highlight the role of ribosomes and mitochondria in T-ALL tumor progression.Significance:The oncogenic role of 6q deletion in T-ALL has remained elusive since this chromosomal abnormality was first identified more than 40 years ago. We combined genomic analysis and functional models to show that the codeletion of two contiguous genes at 6q14 enhances malignancy through deregulation of a ribosome–mitochondria axis, suggesting the potential for therapeutic intervention.This article is highlighted in the In This Issue feature, p. 1494

Oncogenic alterations underlying B-cell acute lymphoblastic leukemia (B-ALL) in adults remain incompletely elucidated. To uncover novel oncogenic drivers, we performed RNA sequencing and whole-genome analyses in a large cohort of unresolved B-ALL. We identified a novel subtype characterized by a distinct gene expression signature and the unique association of 2 genomic microdeletions. The 17q21.31 microdeletion resulted in a UBTF::ATXN7L3 fusion transcript encoding a chimeric protein. The 13q12.2 deletion resulted in monoallelic ectopic expression of the homeobox transcription factor CDX2, located 138 kb in cis from the deletion. Using 4C-sequencing and CRISPR interference experiments, we elucidated the mechanism of CDX2 cis-deregulation, involving PAN3 enhancer hijacking. CDX2/UBTF ALL (n = 26) harbored a distinct pattern of additional alterations including 1q gain and CXCR4 activating mutations. Within adult patients with Ph− B-ALL enrolled in GRAALL trials, patients with CDX2/UBTF ALL (n = 17/723, 2.4%) were young (median age, 31 years) and dramatically enriched in females (male/female ratio, 0.2, P = .002). They commonly presented with a pro-B phenotype ALL and moderate blast cell infiltration. They had poor response to treatment including a higher risk of failure to first induction course (19% vs 3%, P = .017) and higher post-induction minimal residual disease (MRD) levels (MRD ≥ 10−4, 93% vs 46%, P < .001). This early resistance to treatment translated into a significantly higher cumulative incidence of relapse (75.0% vs 32.4%, P = .004) in univariate and multivariate analyses. In conclusion, we discovered a novel B-ALL entity defined by the unique combination of CDX2 cis-deregulation and UBTF::ATXN7L3 fusion, representing a high-risk disease in young adults.

International audience; Fanconi anemia (FA) patients experience chromosome instability, yielding hematopoietic stem/progenitor cell (HSPC) exhaustion and predisposition to poor-prognosis myeloid leukemia. Based on a longitudinal cohort of 335 patients, we performed clinical, genomic, and functional studies in 62 patients with clonal evolution. We found a unique pattern of somatic structural variants and mutations that shares features of BRCA-related cancers, the FA-hallmark being unbalanced, microhomology-mediated translocations driving copy-number alterations. Half the patients developed chromosome 1q gain, driving clonal hematopoiesis through MDM4 trisomy downmodulating p53 signaling later followed by secondary acute myeloid lukemia genomic alterations. Functionally, MDM4 triplication conferred greater fitness to murine and human primary FA HSPCs, rescued inflammation-mediated bone marrow failure, and drove clonal dominance in FA mouse models, while targeting MDM4 impaired leukemia cells in vitro and in vivo. Our results identify a linear route toward secondary leukemogenesis whereby early MDM4-driven downregulation of basal p53 activation plays a pivotal role, opening monitoring and therapeutic prospects.

Le processus de décrochage scolaire est un processus complexe, multidimensionnel, qui s’inscrit dans une temporalité de l’expérience scolaire et qui peut finir par déboucher sur la déscolarisation. Si de nombreuses études se sont orientées sur la diversité des manifestations du décrochage scolaire et des types de décrocheurs, l’objectif de notre étude est d’analyser, en amont, comment la socialisation entre pairs peut contribuer à susciter et/ou à accélérer ce processus de décrochage scolaire chez des adolescents de quatorze à seize ans. En effet, il nous paraît important d’inclure le rôle des pairs à propos de la vie scolaire dans la mesure où ils exercent une influence déterminante sur la socialisation et la construction identitaire de l’adolescent. L’étude a été réalisée par questionnaires auprès de 676 adolescents de quatorze à seize ans. L’outil appréhende d’une part la (dé)mobilisation scolaire et d’autre part, la position sociale via la qualité des relations aux pairs. Les résultats montrent que les élèves qui parviennent à ajuster leur mode de relations aux pairs (les « populaires ») sont les plus mobilisés scolairement. Leurs relations aux autres sont fondées sur la cohésion, le soutien, la solidarité et l’intimité. De même, les adolescents « en retrait », indépendants, autonomes et solitaires, sont également plutôt mobilisés dans leur scolarité. Enfin, les élèves ayant une représentation de l’amitié fondée sur la dépendance affective, les rapports de force, le conflit ou la conformité (les « négligés » et les « soumis ») sont démobilisés. Ils privilégient les apprentissages relationnels aux apprentissages intellectuels. The process of dropping out of school is a complex and multidimensional process, which is part of a temporality of academic experience and can eventually lead to dropping out. While many studies have focused on the diversity of manifestations of dropout and types of dropouts, the objective of our study is to analyze how socialization among peers can generate and / or accelerate dropout among adolescents. Indeed, it seems important to include the role of peers about school life because that they have a decisive influence on adolescent’s socialization and on his identity construction. Questionnaires were administered to 676 adolescents, aged 14 to 16 years. Academic engagement and social position (quality of peer’s relationships) were evaluated. Results reveal that students who are able to adjust their mode of relation to peers (“popular”) are more engaged academically. Similarly, adolescents “withdrawn”, independent, autonomous and solitary are also engaged in their schooling. Finally, students too dependent of group and with a representation of friendship based on power relations, conflict or conformity (“neglected” and “scapegoat”) are disengaged. They prefer relational learning to intellectual learning.

Despite therapeutic successes, AA patients (pts) exhibit a much higher risk of leukemic evolution than the general population. Secondary myeloid neoplasia (sMN) remains the most serious AA complication with major therapeutic and prognostic implications. Historically, multiple theories have been proposed as to the origin of leukemic evolution. For instance, sMN may be the consequence of a relentless autoimmune attack producing a maladaptive response to immunosuppression (IST). Alternatively, occurrence of leukemogenic drivers may be an event setting in motion initially successful and overshooting tumor surveillance reactions. Finally, sMN pathogenesis may be related to either CHIP evolution or acceleration of its progression. Each of these theories is supported by clinical-molecular features (e.g. HLA mutations, secondary PNH evolution, somatic mutations at presentation etc.). Here, we took advantage of a multicentric cohort of pts with AA (n=1010; to our knowledge the largest yet explored) and primary MN (n=3599) to define immunogenetic, iatrogenic and molecular determinants of MN progression. Among AA pts (M:F ratio 0.98; median age 34 years, IQR 20-54, median follow-up 89 months), the 5 and 10 years cumulative incidences of sMN were 6% and 11% respectively, with a median time to progression of 56 months (IQR 23-96). Analysis of available data showed that younger pts (3.5) were observed in 59% (vs 43% in pMDS, p=.02) due to the enrichment in poor/very poor cytogenetic risk groups. In particular, chr.7 abnormalities were the most frequent (54%, of which 88% were del7). By comparison, del7/7q was present in 8% of pMN cases (p At the time of AA onset only 18% of pts harbored somatic myeloid mutations with their presence/absence not influencing evolution, whereas mutations were found in 81% of sMN (1.7 mutations/patient, n=86/101). No difference in mutational burden was observed according to presence/absence of del7/7q, which constituted the founder lesion in 60% of cases, when the analysis was possible. ASXL1 (24% vs 14%, p=.01), RUNX1 (21% vs 11%, p=.008), SETBP1 (14% vs 3%, p AA malignant evolution is characterized by an orchestra of molecular events with an invariant genomic signature (e.g. CUX1, SETBP1, ASXL1). Immunogenetic and immune escape mechanisms may also play a role in shaping the fate of individual patients' trajectories towards PNH vs sMN progression, which may be considered a maladaptive escape event resulting from a bottlenecked hematopoiesis. Disclosures Sebert: BMS: Consultancy; Abbvie: Consultancy. Patel: Apellis: Consultancy, Other: educational talks, Speakers Bureau; Alexion: Consultancy, Other: educational talks, Speakers Bureau. Voso: Celgene: Consultancy, Research Funding, Speakers Bureau; Novartis: Speakers Bureau. Calado: Novartis Brasil: Honoraria; Alexion Brasil: Consultancy; AA&MDS International Foundation: Research Funding; Agios: Membership on an entity's Board of Directors or advisory committees; Instituto Butantan: Consultancy; Team Telomere, Inc.: Membership on an entity's Board of Directors or advisory committees. Peffault De Latour: Novartis: Consultancy, Honoraria, Research Funding; Pfizer: Consultancy, Honoraria, Research Funding; Amgen: Research Funding; Alexion Pharmaceuticals: Consultancy, Honoraria, Research Funding; Apellis Pharmaceuticals Inc: Consultancy, Honoraria; Swedish Orphan Biovitrum AB: Consultancy, Honoraria. Maciejewski: Regeneron: Consultancy; Alexion: Consultancy; Novartis: Consultancy; Bristol Myers Squibb/Celgene: Consultancy.

T-cell acute lymphoblastic leukemia (T-ALL) is a genetically heterogeneous disease that can be classified into different molecular genetic subtypes according to their mRNA gene expression profile. In this study, we applied RNA sequencing to investigate the full spectrum of miRNA expression in primary T-ALL patient samples, T-ALL leukemia cell lines and healthy donor thymocytes. Notably, this analysis revealed that genetic subtypes of human T-ALL also display unique miRNA expression signatures, which are largely conserved in human T-ALL cell lines with corresponding genetic background. Furthermore, small RNA-sequencing also unraveled the variety of isoforms that are expressed for each miRNA in T-ALL and showed that a significant number of miRNAs are actually represented by an alternative isomiR. Finally, comparison of CD34+ and CD4+CD8+ healthy donor thymocytes and T-ALL miRNA profiles allowed identifying several novel miRNAs with putative oncogenic or tumor suppressor functions in T-ALL. Altogether, this study provides a comprehensive overview of miRNA expression in normal and malignant T-cells and sets the stage for functional evaluation of novel miRNAs in T-ALL disease biology.

Deletion of chromosome 6q is a well-recognized abnormality found in poor-prognosis T-cell acute lymphoblastic leukemia (T-ALL). Using integrated genomic approaches, we identified two candidate haploinsufficient genes contiguous at 6q14, SYNCRIP (encoding hnRNP-Q) and SNHG5 (that hosts snoRNAs), both involved in regulating RNA maturation and translation. Combined silencing of both genes, but not of either gene alone, accelerated leukemogeneis in a Tal1/Lmo1/Notch1-driven mouse model, demonstrating the tumor-suppressive nature of the two-gene region. Proteomic and translational profiling of cells in which we engineered a short 6q deletion by CRISPR/Cas9 genome editing indicated decreased ribosome and mitochondrial activities, suggesting that the resulting metabolic changes may regulate tumor progression. Indeed, xenograft experiments showed an increased leukemia-initiating cell activity of primary human leukemic cells upon coextinction of SYNCRIP and SNHG5. Our findings not only elucidate the nature of 6q deletion but also highlight the role of ribosomes and mitochondria in T-ALL tumor progression. Significance: The oncogenic role of 6q deletion in T-ALL has remained elusive since this chromosomal abnormality was first identified more than 40 years ago. We combined genomic analysis and functional models to show that the codeletion of two contiguous genes at 6q14 enhances malignancy through deregulation of a ribosome–mitochondria axis, suggesting the potential for therapeutic intervention. This article is highlighted in the In This Issue feature, p. 1494

Touch Connection is a pair of digitally embroidered textile surfaces with vibrotactile capability. The surfaces are designed as a paired system, to enable people to connect, share touch responses and construct communal tactile experiences. It consists of two different fabric surfaces that use the textile techniques quilting and embroidery to create dramatic volume and texture.We explore the application of textural effects and vibrotactile feedback as materials for design that can be shaped and moulded to engage participants. The work explores the role these materials play in generating improvised action and emotional responses.

International audience; Bone marrow (BM) failure (BMF) in children and young adults is often suspected to be inherited, but in many cases diagnosis remains uncertain. We studied a cohort of 179 patients (from 173 families) with BMF of suspected inherited origin but unresolved diagnosis after medical evaluation and Fanconi anemia exclusion. All patients had cytopenias, and 12.0% presented ≥5% BM blast cells. Median age at genetic evaluation was 11 years; 20.7% of patients were aged ≤2 years and 36.9% were ≥18 years. We analyzed genomic DNA from skin fibroblasts using whole-exome sequencing, and were able to assign a causal or likely causal germ line mutation in 86 patients (48.0%), involving a total of 28 genes. These included genes in familial hematopoietic disorders (GATA2, RUNX1), telomeropathies (TERC, TERT, RTEL1), ribosome disorders (SBDS, DNAJC21, RPL5), and DNA repair deficiency (LIG4). Many patients had an atypical presentation, and the mutated gene was often not clinically suspected. We also found mutations in genes seldom reported in inherited BMF (IBMF), such as SAMD9 and SAMD9L (N = 16 of the 86 patients, 18.6%), MECOM/EVI1 (N = 6, 7.0%), and ERCC6L2 (N = 7, 8.1%), each of which was associated with a distinct natural history; SAMD9 and SAMD9L patients often experienced transient aplasia and monosomy 7, whereas MECOM patients presented early-onset severe aplastic anemia, and ERCC6L2 patients, mild pancytopenia with myelodysplasia. This study broadens the molecular and clinical portrait of IBMF syndromes and sheds light on newly recognized disease entities. Using a high-throughput sequencing screen to implement precision medicine at diagnosis can improve patient management and family counseling.

Perceived social support has been widely recognized as having beneficial effects on a person’s development, and adolescence is no exception. The objective of this article is to go beyond this “stereotypical” vision of friendship by showing that social support does not always have a positive and direct effect on adolescents’ academic achievement. We sought to understand whether this relationship is mediated by the educational goals and motives that student value and pursue. If these values do not coincide with the expectations and values that the academic institution seeks to transmit, the influence of social support can become harmful, especially with regard to the students’ academic achievement. Data were collected by means of questionnaires from 676 junior high school students in 9th grade from contrasting academic institutions. We chose to observe each participant’s subjective point of view of three dimensions: social support, academic achievement, and educational goals and motives. The results showed that the relationship between social support and attention and involvement in class was negative and mediated by students’ goals. The adolescents with a high level of social support mainly pursued social goals and neglected achievement and future goals. Consequently, these students were less attentive and involved in class. The results also showed that social support was predictive of students’ investment and interest in personal work and success, but only when the students pursued achievement and future goals.

Resume L’adolescent se trouve face a une double contrainte : le besoin de conformite extreme au groupe pour se sentir soutenu et reconnu, et la recherche d’independance face aux pressions du groupe, pour s’affirmer et s’autonomiser. Confrontes a ce paradoxe, comment les adolescents vont-ils se positionner dans leurs relations aux pairs et dans leur rapport a l’ecole ? Cette recherche a pour objectif l’etude du lien entre ces deux formes d’affirmation de soi au sein du groupe de pairs et la (de)mobilisation scolaire.676 collegiens de 14 a 16 ans ont ete interroges sur leurs relations aux pairs (recherche extreme de conformite ou recherche d’independance vis-a-vis du groupe), sur la valeur qu’ils accordent a l’ecole et sur leur mobilisation scolaire.Les analyses statistiques realisees montrent que plus la recherche d’independance ou de conformite est extreme, plus ces positionnements sont lies a la demobilisation scolaire. Ces derniers privilegient egalement les apprentissages sociaux et relationnels de l’ecole aux depens des apprentissages scolaires et intellectuels, ce qui les eloigne des logiques et attentes institutionnelles.

Bromodomain and Extra-Terminal inhibitors (BETi) such as OTX015 are active in Acute Myeloid Leukaemias (AML). Their activity on Leukemic Stem Cells (LSCs) is less documented. We interrogated the anti-LSC activity of OTX015 in a niche-like long-term culture in 26 primary AML samples and validated our findings in vivo. OTX015 impaired LSCs in AMLs harbouring Core Binding Factor or KMT2A gene fusions, NPM1 or chromatin/spliceosome genes mutations, but not in those with aneuploidy/TP53 mutations. In four patients, we dissected the transcriptomic footprint of Bet inhibition on LSCs versus blasts. Our results can instruct future clinical trials of BETi in AML.

MYC is a potent oncogene involved in ∼70% of human cancers, inducing tumorigenesis with high penetrance and short latency in experimental transgenic models. Accordingly, MYC is recognized as a major driver of T-cell acute lymphoblastic leukemia (T-ALL) in human and zebrafish/mouse models, and uncovering the context by which MYC-mediated malignant transformation initiates and develops remains a considerable challenge. Because MYC is a very complex oncogene, highly dependent on the microenvironment and cell-intrinsic context, we generated transgenic mice (tgMyc(spo)) in which ectopic Myc activation occurs sporadically (

Les problématiques actuelles liées à l’insertion professionnelle des jeunes ont amené chercheur.e.s et praticien.ne.s à s’intéresser à des compétences complémentaires aux compétences professionnelles : les compétences sociales. Dans cette perspective, des programmes destinés aux étudiant.e.s et favorisant le développement de ces compétences ont été créés. Cet article présente huit de ces programmes pour lesquels sont décrits, analysés et comparés objectifs, contenus (pédagogies, compétences ciblées, durée), évaluations et résultats. En soulignant les caractéristiques des programmes reconnues comme efficaces, mais également leurs limites au niveau théorique et méthodologique, l’objectif est de dégager des constats et propositions d’action utiles pour l’établissement de futurs programmes. The current problems relating to the professional integration of young people led researchers and practitioners to focus on complementary skills to professional skills: social skills. In this context, programs for students aimed at developing these skills have been created. This article presents eight of these programs and for each one, the specific objectives, contents (pedagogies, targeted social skills, and duration), assessments conducted and results are described, compared and analyzed. In highlighting the criteria of effectiveness of these programs and their limitations, the aim is to reveal findings and useful proposals for action for the implementation of future programs.

Touchplay is a compelling fabric surface that engages the tactile properties of physical materials and integrates them with abstract digital structures. It creates multi-sensorial encounters as a result of material affinities between sonic and tangible form. By inviting playful participation through familiar actions, participants learn to 'play the piece' and enjoy discovering and revealing sounds. The design and structure of the embroidered fabric surface encourages interaction with digitally embroidered discs that use colour, pattern and texture to distinguish them. The computational elements are approached as another material, crafted into the design of the piece during the making process. The evocative nature of sound results in a heightened emotional, resonant quality. Combining human voice with the tangible qualities of the fabric contributes to a compelling experience through touch, sound and textile.

Resume But de l’etude Cette recherche rend compte de la diversite des formes de relations aux pairs et de leur impact sur le developpement psychologique et socioaffectif de l’adolescent a travers son rapport a l’ecole. Plus precisement, cette etude examine l’influence du statut social sur le desinvestissement scolaire, chez des adolescents scolarises dans des colleges issus de l’« education prioritaire ». Population et methode Nous avons questionne 186 collegiens de 3e scolarises dans ces colleges. Une classification hierarchique descendante a ete realisee permettant de degager des profils types, selon leur statut social et le (des)investissement scolaire. Resultats Les adolescents ayant les relations les plus « extremes » (les populaires, les agressifs, les rejetes) sont ceux qui ont les plus grandes difficultes scolaires. Leurs relations, basees sur des rapports de force, la recherche de conformite ou la dependance affective leur octroient des comportements a-scolaires (dispersion, passivite scolaire…) et des valeurs trop eloignees des exigences scolaires. Les adolescents qui ont des relations equilibres avec leurs pairs reussissent mieux. Le soutien et la securite que leur offre le groupe leur permettent d’etre disponibles cognitivement pour les apprentissages. Les adolescents « solitaires » sont egalement de bons eleves : ces eleves fuient la pression sociale pour se concentrer sur les activites scolaires. Conclusion En definitive, ces resultats sous-tendent que le statut social de l’adolescent dans son groupe de pairs peut generer des problematiques individuelles affectant sa personnalite, sa confiance en soi, sa sociabilite, ses valeurs et, par consequent, son parcours scolaire.

Renal-cell carcinomas (RCC) are often resistant to conventional cytotoxic agents. Xenograft models are used for in vivo preclinical studies and drug development. The validity of these studies is highly dependent on the phenotypic and genotypic stability of the models. Here we assessed the stability of six aggressive human RCC xenografted in nude/NMRI mice. We compared the initial samples (P0), first (P1) and fifth (P5) passages for the following criteria: histopathology, immunohistochemistry for CK7, CD10, vimentin and p53, DNA allelic profiles using 10 microsatellites and CGH-array. Next we evaluated the response to sunitinib in primary RCC and corresponding xenografted RCC. We observed a good overall stability between primary RCC and corresponding xenografted RCC at P1 and P5 regarding histopathology and immunohistochemistry except for cytokeratin 7 (one case) and p53 (one case) expression. Out of 44 groups with fully available microsatellite data (at P0, P1 and P5), 66% (29 groups) showed no difference from P0 to P5 while 34% (15 groups) showed new or lost alleles. Using CGH-array, overall genomic alterations at P5 were not different from those of initial RCC. The xenografted RCC had identical response to sunitinib therapy compared to the initial human RCC from which they derive. These xenograft models of aggressive human RCC are clinically relevant, showing a good histological and molecular stability and are suitable for studies of basic biology and response to therapy.

Oncogenic subtypes in childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL) are used for risk-stratification. However, a significant number of BCPALLpatients are still genetically unassigned. Using array-CGH in a selected BCPALL cohort, we characterized a recurrent V(D)J-mediated intragenic deletion of the ERG gene (ERGdel). A breakpoint-specific PCR assay was designed and used to screen an independent non-selected cohort of 897 children aged 1-17 years treated for BCP-ALL in the EORTC-CLG 58951 trial. ERGdel was found in 29/897 patients (3.2%) and was mutually exclusive of known classifying genetic lesions, suggesting that it characterized a distinct leukemia entity. ERGdel was associated with higher age (median 7.0 years vs. 4.0 years, P=0.004), aberrant CD2 expression (43.5% vs. 3.7%, P

Oncogenic subtypes in childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL) are used for risk stratification. However, a significant number of BCP-ALL patients are still genetically unassigned. Using array-comparative genomic hybridization in a selected BCP-ALL cohort, we characterized a recurrent V(D)J-mediated intragenic deletion of the ERG gene (ERG(del)). A breakpoint-specific PCR assay was designed and used to screen an independent non-selected cohort of 897 children aged 1-17 years treated for BCP-ALL in the EORTC-CLG 58951 trial. ERG(del) was found in 29/897 patients (3.2%) and was mutually exclusive of known classifying genetic lesions, suggesting that it characterized a distinct leukemia entity. ERG(del) was associated with higher age (median 7.0 vs. 4.0 years, P=0.004), aberrant CD2 expression (43.5% vs. 3.7%, P

Metabolic syndrome (MS) is an emerging risk factor in hepatocellular carcinoma (HCC). HCC related to MS may occur either in advanced fibrosis or before the development of cirrhosis, suggesting involvement of different molecular pathways according to the features of background liver.To investigate genomic aberrations in HCC related to MS in order to identify new target genes involved in liver carcinogenesis.Chromosomal aberrations of HCC obtained from 20 patients with MS (HCC/MS) were studied by comparative genomic hybridisation and compared with HCC related to hepatitis C virus (HCV) infection (HCC/HCV, n=10) and, within the group of HCC with MS, according to the condition of the background liver (presence or absence of significant fibrosis).Among the most frequent chromosomal alterations observed in HCC, 6p21.1 amplification had a higher incidence in HCC/MS than in HCC/HCV (60% vs 20%, p0.01). Advanced fibrosis/cirrhosis in the peritumoral liver was the only clinicopathological factor associated with the 6p21.1 amplicon in HCC/MS. Increased expression of cullin7 (CUL7), a gene located at the 6p21.1 locus, was demonstrated in HCC with the 6p21.1 amplicon, in parallel with a decrease in cyclin D1 expression. CUL7 downregulation using siRNA transfection in hepatoma cell lines induced significant cyclin D1 expression (by promoting its degradation), decreased cell proliferation and increased apoptosis.This study demonstrates specific genomic alterations in HCC/MS and points to CUL7 as a novel gene potentially involved in liver carcinogenesis associated with MS, the amplification of which might influence cell proliferation.

Neurofibromatosis type 1 (NF1) is a tumor predisposition syndrome with a worldwide birth incidence of one in 2500. Genetic factors unrelated to the NF1 locus are thought to influence the number of plexiform neurofibromas (PNFs) in patients with NF1, but no factors have been identified to date.We used high-resolution array comparative genomic hybridization of tissue from 22 PNFs obtained from 18 NF1 patients to identify modifier genes involved in PNF development. We used a family-based association test for five previously identified cancer-susceptibility tag single-nucleotide polymorphisms (rs1063192, rs2151280, rs2218220, rs10757257, and rs7023329) located in chromosomal region 9p21.3 in 1105 subjects (740 NF1 patients and 365 non-affected relatives) from 306 families. To confirm the functional role of rs2151280, we used real-time quantitative reverse transcription-polymerase chain reaction to analyze the expression of cyclin-dependent kinase inhibitor 2A (CDKN2A), CDKN2B, alternate reading frame (ARF), and antisense noncoding RNA in the INK4 locus (ANRIL) in the peripheral blood of 124 NF1 patients. Relationships between CDKN2A, CDKN2B, ARF, and ANRIL expression and the rs2151280 genotype were tested by the Kruskal-Wallis test. All statistical tests were two-sided.In NF1-associated PNFs, 9p21.3 deletions (including the CDKN2A/B-ANRIL locus) were found as the only recurrent somatic alterations. Single-nucleotide polymorphism rs2151280 (located in ANRIL) was statistically significantly associated with the number of PNFs (P.001) in NF1 patients. In addition, allele T of rs2151280 was statistically significantly associated with reduced ANRIL transcript levels (P.001), suggesting that modulation of ANRIL expression mediates PNF susceptibility.Identification of ANRIL as a modifier gene in NF1 may offer clues to the molecular pathogenesis of PNFs, particularly neurofibroma formation, and emphasizes the unanticipated role of large noncoding RNA in activation of critical regulators of tumor development.

Compared with T-ALL diagnosis samples, samples obtained at relapse or after xenograft into immunodeficient mice exhibit additional genomic lesions in oncogenes and/or tumor suppressor genes; these lesions contribute to leukemia-initiating activity., Genomic studies in human acute lymphoblastic leukemia (ALL) have revealed clonal heterogeneity at diagnosis and clonal evolution at relapse. In this study, we used genome-wide profiling to compare human T cell ALL samples at the time of diagnosis and after engraftment (xenograft) into immunodeficient recipient mice. Compared with paired diagnosis samples, the xenograft leukemia often contained additional genomic lesions in established human oncogenes and/or tumor suppressor genes. Mimicking such genomic lesions by short hairpin RNA–mediated knockdown in diagnosis samples conferred a selective advantage in competitive engraftment experiments, demonstrating that additional lesions can be drivers of increased leukemia-initiating activity. In addition, the xenograft leukemias appeared to arise from minor subclones existing in the patient at diagnosis. Comparison of paired diagnosis and relapse samples showed that, with regard to genetic lesions, xenograft leukemias more frequently more closely resembled relapse samples than bulk diagnosis samples. Moreover, a cell cycle– and mitosis-associated gene expression signature was present in xenograft and relapse samples, and xenograft leukemia exhibited diminished sensitivity to drugs. Thus, the establishment of human leukemia in immunodeficient mice selects and expands a more aggressive malignancy, recapitulating the process of relapse in patients. These findings may contribute to the design of novel strategies to prevent or treat relapse.

The Ikaros (Ikzf1) gene, encoding a transcription regulator, is a major tumor suppressor in B-cell acute lymphoblastic leukemia (B-ALL). In the mouse, however, loss of Ikaros is primarily associated with T-ALL development. Whether Ikaros is also implicated in human T-ALL remains unclear. We studied Ikaros in 25 human T-ALL samples from diverse molecular subtypes at the mRNA, protein, sequence and genomic copy number level. We found that Ikaros was abnormal in only one sample: one allele was lost by genomic deletion, while proteins generated from the remaining allele were delocalized and concentrated at a single cytoplasmic structure. Thus, inactivation of Ikaros by deletion or mutation is rare in human T-ALL.

Abstract 121 The genetic landscape of B-cell precursor acute lymphoblastic leukemia (BCP-ALL) in children above 10 years and adolescents remains poorly defined. Specifically, more than half of these patients have none of the cytogenetic abnormalities that define oncogenic subtypes and underlie risk stratification. To uncover new genetic abnormalities in these unassigned cases, we studied 85 BCP-ALL from patients aged 10 to 17 diagnosed at St-Louis hospital (Paris, France), for which the main classifying genetic lesions were assessed (i.e. high hyperdiploidy, t(12;21)/ETV6-RUNX1, t(1;19)/TCF3-PBX1, t(9;22)/BCR-ABL1, iAMP21, MLL translocations, low hypodiploidy, and near haploidy). Fifty of these BCP-ALL presented no classifying genetic lesions. Paired leukemic and remission samples could be analysed by high density array-CGH (Agilent 1M arrays) in 17 of these unassigned cases. We focused on acquired, focal, and recurrent copy-number abnormalities. A mono-allelic intragenic deletion of the ETS-related Gene (ERG) was found in 3 cases. ERG belongs to the ETS family of transcription factors and is implicated in chromosomal translocations associated with several cancer types including acute myeloid leukemia. The possibility of a cryptic unbalanced translocation was ruled out in the 3 cases by FISH analysis. The deletions encompassed exons 3 to 7, or 3 to 9, and the breakpoints were tightly clustered. Based on the breakpoint sequences we designed a PCR assay that allowed us to screen ERG intragenic deletions in the whole cohort. ERG deletion was identified in 9 additional cases, none of them having any of the known classifying genetic lesions, bringing up to 25% (12 out of 50) the frequency of ERG deletion in unassigned BCP-ALL of children older than 10. These results suggested that ERG deletion characterized a novel oncogenic subtype of BCP-ALL. Of note, these results were consistent with independent data of Harvey et al. (2010) that reported ERG deletions in a distinct gene-expression cluster. To confirm and extend these findings in the whole population of paediatric BCP-ALL, we used our breakpoint-specific PCR assay to screen ERG deletions in an independent cohort of 822 unselected patients aged 1 to 17, enrolled in the EORTC 58951 trial. ERG deletion was identified in 31/822 (3.7%) patients. Again, none of them had another known classifying genetic lesion, confirming that ERG deletion characterizes a distinct oncogenic subtype. Patients with ERG deletion were significantly older compared to other patients (median 7.0 vs 4.0, p=0.002), but they had similar white blood counts at diagnosis. They had a favourable outcome, with a 8-year event free survival (EFS) of 82.4% and overall survival (OS) of 96.0%, which is similar to EFS of 83.4% and OS of 91.6% obtained for patients having no very high risk initial features (i.e. no t(9;22)/BCR-ABL1, MLL rearrangement or haploidy/low hypodiploidy). IKZF1 deletion is a cooperative genetic lesion that has been recently shown to be associated with a poor outcome in BCP-ALL. Remarkably, the incidence of IKZF1 deletions in patients with ERG deletion was significantly higher than in other BCR-ABL1-negative patients, especially when considering the IKZF1 intragenic deletion Δ4-7 (10/31, 32.3% vs 34/744, 4.6%, P Altogether, we have identified a novel oncogenic subtype of BCP-ALL characterized by ERG deletion. This subtype is frequently associated with IKZF1 deletions, suggesting a preferred oncogenic cooperation. Importantly, despite having older age and frequent IKZF1 deletions, which are factors usually predictive of a poor prognosis, patients with ERG deletion have a favourable outcome. Therefore, this genetic abnormality may be systematically assessed as part of the diagnostic work-up of BCP-ALL and taken into account when considering treatment stratification. Disclosures: No relevant conflicts of interest to declare.

Abstract 4188 Cumulative evidence indicates that MYC, one of the major downstream effectors of NOTCH1, is a critical component of T-cell acute lymphoblastic leukemia (T-ALL) oncogenesis and a potential candidate for targeted therapy. However, MYC is a complex oncogene, involving both fine protein dosage and cell-context dependency, and detailed understanding of MYC-mediated oncogenesis in T-ALL is still lacking. To better understand how MYC is interspersed in the complex T-ALL oncogenic networks, we performed the first thorough molecular and biochemical analysis of MYC activation in a comprehensive collection of primary adult and pediatric patient samples (n=163). RQ-PCR analysis first revealed that levels of MYC transcript are highly variable, and that high MYC expression can be generated in a large number of cases in absence of NOTCH1/FBXW7 mutations, suggesting the occurrence of multiple activation pathways in addition to NOTCH1. MYC protein levels were next assessed by western blot in a subpanel of 29 primary T-ALL samples. Although a large spectrum of protein expression was also observed, no direct correlation was apparent between transcript level and protein abundance. This discrepancy is in line with the frequent deregulation of factors controlling MYC protein turn-over in cancer cells. In Burkitt Lymphoma (BL) for example, mutations in the MYC-box (in and around the critical T58 phosphorylation site) impede efficient addressing to the proteasome, and results in oncogenic MYC protein stabilization. We sequenced the MYC-box region of 116 T-ALL samples and found no such mutations, suggesting that BL and T-ALL use different routes to achieve oncogenic MYC levels. An alternative candidate is the E3 ubiquitin ligase FBXW7 which has been shown to target both NOTCH1 and MYC for proteosomal degradation, and the loss-of-function mutations of which were previously shown to impair MYC protein stability. Although T-ALL harboring inactivating FBXW7 mutations were associated with high MYC protein level as expected, such mutants represented only 10% of the cases, and could thus not account for the numerous cases showing MYC protein accumulation in our series. This implied the frequent occurrence of other, alternative mechanisms of MYC stabilization in T-ALL. To identify these processes, we performed a detailed pan-genomic analysis of a T-ALL case showing high MYC protein levels at relapse but low MYC at diagnosis, despite identical transcript levels. Among a total of ten genomic aberrations found in this patient, only one difference was found between diagnosis and relapse, namely the progression of mono-allelic to bi-allelic deletion of a 10q23 region including the PTEN tumor suppressor gene. PTEN is considered the main negative regulator of the PI3K-AKT signaling. As AKT inactivates GSK3β, the serine-threonine kinase involved in MYC phosphorylation at T58, PTEN inactivation constitutes a likely candidate of MYC protein accumulation. To further test this hypothesis, we probed by western blot PTEN and MYC proteins in the previous subpanel of 29 T-ALL, and observed an inverse inter-relationship between PTEN and MYC protein levels. This anti-correlation was particularly clear in cases with abundant PTEN protein levels, where only faint MYC could be found despite relatively high transcription levels. Conversely, high MYC protein levels were in most cases associated with rather low PTEN protein levels. Finally, we functionally demonstrate that in T-ALL cell lines, the PI3K chemical inhibitor LY294002 which antagonizes AKT signaling and mimics PTEN function, is able to downmodulate MYC levels and activity. Altogether, our data reveals that post-transcriptional deregulation of MYC constitutes a major alternative pathway of MYC activation in T-ALL, operating partly via the PI3K/AKT axis through down-regulation of PTEN, and that NOTCH1 mutations might play a dual transcriptional and post-transcriptional role in this process. Our data lend further support to the significance of therapeutic targeting of MYC and/or the PTEN/AKT pathways, both in γ-secretase inhibitor-resistant and identified NOTCH1-independent/MYC-mediated T-ALL patients. Disclosures: No relevant conflicts of interest to declare.

This paper reports on results from the EU H2020 WEAR project, which between 2017-2019 has facilitated sustainable innovation processes in the field of smart textiles and wearable technology. 46 design-technology interdisciplinary projects and start-ups throughout Europe were selected through two Open Calls, and funded to develop creative solutions for a broad range of sustainability challenges within the relevant industries. The paper outlines collaboration between two WEAR teams (KOBA, Touch Craft) as makers and facilitators and their audiences through crafts and practical making of electronic textiles artefacts. In particular, it reports how engagement with communities (as customers and/or co-creators) was used to increase social cohesion and well-being as social sustainability potential. Findings highlight the importance of quality of interaction, in particular, ongoing, in-person exchange, either between crafter and customer (KOBA) or facilitator and crafts community groups (Touch Craft). It further emerged that social cohesion could contribute to finding solutions to environmental and economic challenges, through encouraging local production, made-to-order production and local business development.

Cumulative evidence indicates that MYC, one of the major downstream effectors of NOTCH1, is a critical component of T-cell acute lymphoblastic leukemia (T-ALL) oncogenesis and a potential candidate for targeted therapy. However, MYC is a complex oncogene, involving both fine protein dosage and cell-context dependency, and detailed understanding of MYC-mediated oncogenesis in T-ALL is still lacking. To better understand how MYC is interspersed in the complex T-ALL oncogenic networks, we performed a thorough molecular and biochemical analysis of MYC activation in a comprehensive collection of primary adult and pediatric patient samples. We find that MYC expression is highly variable, and that high MYC expression levels can be generated in a large number of cases in absence of NOTCH1/FBXW7 mutations, suggesting the occurrence of multiple activation pathways in addition to NOTCH1. Furthermore, we show that posttranscriptional deregulation of MYC constitutes a major alternative pathway of MYC activation in T-ALL, operating partly via the PI3K/AKT axis through down-regulation of PTEN, and that NOTCH1m might play a dual transcriptional and posttranscriptional role in this process. Altogether, our data lend further support to the significance of therapeutic targeting of MYC and/or the PTEN/AKT pathways, both in GSI-resistant and identified NOTCH1-independent/MYC-mediated T-ALL patients.

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