The role of cancer predisposition syndrome in children and adolescents with very rare tumours

Germline predisposing pathogenic variants (GPVs) are present in approximately 8 to 10% of children with all cancer types. Very rare tumours (VRTs) represent many different diseases, defined with an annual incidence < 2 / 1,000,000, and correspond to 11% of all cancers in patients aged 0-14 years. Some of these VRTs, including cancer typical for adults, develop in children with a cancer predisposition syndrome (CPS). Classically, three situations lead to consider this association: Some patients develop a VRT for which histology itself strongly suggests a GPV related to a CPS; others are referred for germline genetic testing because of a family or personal history and finally, a systematic molecular genomic tumour analysis, reveals a PV typical to a CPS. Depending on the samples tested and type of analysis performed, information can be directly available about the germline status of such a PV. Depicting the association between CPS and VRT is clinically important as some of these tumour types require adapted therapy, sometimes in the frontline setting, and the proposal of a specific surveillance programme to detect other malignancies. The diagnosis of CPS necessitates a careful familial evaluation and genetic counselling regarding the risks faced by the child or other family members. The aim of this paper is to propose a literature review of solid VRTs occurring in paediatric and young adult patients associated with CPSs

Contrôle transcriptionnel de la spécification cellulaire dans le cerveau postérieur des Vertébrés

During embryonic development, the cell fates are specified by the expression of lineage factors whose expression must be finely regulated to ensure proper organ formation. To get insight into the control of lineage factors expression, we used the vertebrate hindbrain (rhombencephalon) as a system model. During development, the hindbrain is subdivided into seven segments, termed rhombomeres and noted from r1 to r7. Each segment corresponds to a coherent cell lineage. In particular, specification of r3 and r5 lineages is controlled by the zinc-finger transcription factor Krox20. When Krox20 function is abolished, r3 and r5 cells are not properly specified, hence modified neuronal fate. In the present PhD work, we aimed at deciphering the processes that control the number of Krox20-expressing cells, i.e. the size of r3 and r5. We focused in particular on the transcriptional control of Krox20 expression by studying the activity of Krox20 cis-regulatory elements. Two elements, termed B and C, are responsible for the initiation of Krox20 expression; a third one, noted A, amplifies and prolongs it through an autoregulatory activity. We showed (i) that cells commit to the r3/r5 fate only if they activate the element A, (ii) that the element A functions as a bistable switch, determined by the level of Krox20 initiation. These results were obtained by the analysis of a computational model, constrained by quantitative data derived from experiments on zebrafish embryos. Our model is built at the cell level and implement molecular events occuring at the level of element A. It thus establishes the relationship between enhancer activity and tissue-scale patterningAu cours du développement embryonnaire, le destin d'une cellule est spécifié par l'expression de gènes dits de lignage. Le contrôle de l'expression de ces gènes est essentiel à la cohérence du développement embryonnaire. Pour savoir comment s'effectue ce contrôle, nous avons choisi un modèle de spécification dans le cerveau postérieur des vertébrés, le rhombencéphale. Au cours de son développement, cet organe comprend sept groupes de cellules homogènes, appelés rhombomères (r) et notés de r1 à r7, qui subissent des processus de spécification distincts. La voie de spécification de r3 et r5 est la mieux connue car elle est contrôlée par un facteur unique, Krox20. En l'absence de ce facteur, les cellules de r3 et r5 ne sont pas correctement spécifiées et voient leur destin neuronal modifié. Dans ce travail de thèse, nous dévoilons les mécanismes qui permettent de contrôler le nombre de cellules exprimant le gène Krox20, donc la taille de r3 et r5. Ces mécanismes contrôlent la dynamique de transcription de Krox20, en régulant l'activité de ses éléments régulateurs. Deux éléments, B et C, sont responsables de l'initiation de l'expression de Krox20 ; un troisième, noté A, l'amplifie et la prolonge grâce à une activité autorégulatrice. Nous montrons (i) que les cellules n'acquièrent l'identité r3/r5 que si elles activent l'élément A, (ii) que l'activation de l'élément A suit un mode tout-ou-rien, selon le niveau d'initiation de Krox20. Ces conclusions ont été obtenues par l'analyse d'un modèle mathématique, contraint par des données expérimentales obtenues chez le poisson-zèbre, et suffisamment résolutif pour décrire l'activation de A à l'échelle moléculaire

Dissection of a Krox20 positive feedback loop driving cell fate choices in hindbrain patterning

Although feedback loops are essential in development, their molecular implementation and precise functions remain elusive. Using enhancer knockout in mice, we demonstrate that a direct, positive autoregulatory loop amplifies and maintains the expression of Krox20, a transcription factor governing vertebrate hindbrain segmentation. By combining quantitative data collected in the zebrafish with biophysical modelling that accounts for the intrinsic stochastic molecular dynamics, we dissect the loop at the molecular level. We find that it underpins a bistable switch that turns a transient input signal into cell fate commitment, as we observe in single cell analyses. The stochasticity of the activation process leads to a graded input–output response until saturation is reached. Consequently, the duration and strength of the input signal controls the size of the hindbrain segments by modulating the distribution between the two cell fates. Moreover, segment formation is buffered from severe variations in input level. Finally, the progressive extinction of Krox20 expression involves a destabilization of the loop by repressor molecules. These mechanisms are of general significance for cell type specification and tissue patterning

Pediatric spinal pilocytic astrocytomas form a distinct epigenetic subclass from pilocytic astrocytomas of other locations and diffuse leptomeningeal glioneuronal tumours

Abstract Pediatric spinal low-grade glioma (LGG) and glioneuronal tumours are rare, accounting for less 2.8–5.2% of pediatric LGG. New tumour types frequently found in spinal location such as diffuse leptomeningeal glioneuronal tumours (DLGNT) have been added to the World Health Organization (WHO) classification of tumours of the central nervous system since 2016, but their distinction from others gliomas and particularly from pilocytic astrocytoma (PA) are poorly defined. Most large studies on this subject were published before the era of the molecular diagnosis and did not address the differential diagnosis between PAs and DLGNTs in this peculiar location. Our study retrospectively examined a cohort of 28 children with LGGs and glioneuronal intramedullary tumours using detailed radiological, clinico-pathological and molecular analysis. 25% of spinal PAs were reclassified as DLGNTs. PA and DLGNT are nearly indistinguishable in histopathology or neuroradiology. 83% of spinal DLGNTs presented first without leptomeningeal contrast enhancement. Unsupervised t-distributed stochastic neighbor embedding (t-SNE) analysis of DNA methylation profiles showed that spinal PAs formed a unique methylation cluster distinct from reference midline and posterior fossa PAs, whereas spinal DLGNTs clustered with reference DLGNT cohort. FGFR1 alterations were found in 36% of spinal tumours and were restricted to PAs. Spinal PAs affected significantly younger patients (median age 2 years old) than DLGNTs (median age 8.2 years old). Progression-free survival was similar among the two groups. In this location, histopathology and radiology are of limited interest, but molecular data (methyloma, 1p and FGFR1 status) represent important tools differentiating these two mitogen-activated protein kinase (MAPK) altered tumour types, PA and DLGNT. Thus, these molecular alterations should systematically be explored in this type of tumour in a spinal location