MicroRNAs and Cancer Stem Cells in Medulloblastoma

In this chapter, we are describing the biology of medulloblastoma influenced by several genes/pathways which concur to its pathogenesis. Of note, several levels of regulation are mediated by miRNAs functions, which we dissect their “state of art” to underline their crucial roles on controlling cancer development. In brain tumours, literatures data, are supporting the values of Cancer Stem Cells (tumor propagating cells) and their functions for tumour recurrence for future therapeutic treatments. Thus, we link the potential use of miRNAs as “shuttle” to impair Cancer Stem Cells in medulloblastoma

Detection of erbB2 copy number variations in plasma of patients with esophageal carcinoma

<p>Abstract</p> <p>Background</p> <p>Mortality is high in patients with esophageal carcinoma as tumors are rarely detected before the disease has progressed to an advanced stage. Here, we sought to isolate cell-free DNA released into the plasma of patients with esophageal carcinoma, to analyze copy number variations of marker genes in the search for early detection of tumor progression.</p> <p>Methods</p> <p>Plasma of 41 patients with esophageal carcinoma was prospectively collected before tumor resection and chemotherapy. Our dataset resulted heterogeneous for clinical data, resembling the characteristics of the tumor. DNA from the plasma was extracted to analyze copy number variations of the <it>erbB2 </it>gene using real-time PCR assays.</p> <p>Results</p> <p>The real-time PCR assays for <it>erbB2 </it>gene showed significant (<it>P </it>= 0.001) copy number variations in the plasma of patients with esophageal carcinoma, as compared to healthy controls with high sensitivity (80%) and specificity (95%). These variations in <it>erbB2 </it>were negatively correlated to the progression free survival of these patients (<it>P </it>= 0.03), and revealed a further risk category stratification of patients with low VEGF expression levels.</p> <p>Conclusion</p> <p>The copy number variation of <it>erbB2 </it>gene from plasma can be used as prognostic marker for early detection of patients at risk of worse clinical outcome in esophageal cancer.</p

Alterations in ALK/ROS1/NTRK/MET drive a group of infantile hemispheric gliomas

© The Author(s) 2019. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.Infant gliomas have paradoxical clinical behavior compared to those in children and adults: low-grade tumors have a higher mortality rate, while high-grade tumors have a better outcome. However, we have little understanding of their biology and therefore cannot explain this behavior nor what constitutes optimal clinical management. Here we report a comprehensive genetic analysis of an international cohort of clinically annotated infant gliomas, revealing 3 clinical subgroups. Group 1 tumors arise in the cerebral hemispheres and harbor alterations in the receptor tyrosine kinases ALK, ROS1, NTRK and MET. These are typically single-events and confer an intermediate outcome. Groups 2 and 3 gliomas harbor RAS/MAPK pathway mutations and arise in the hemispheres and midline, respectively. Group 2 tumors have excellent long-term survival, while group 3 tumors progress rapidly and do not respond well to chemoradiation. We conclude that infant gliomas comprise 3 subgroups, justifying the need for specialized therapeutic strategies.info:eu-repo/semantics/publishedVersio

Cytogenetic Prognostication Within Medulloblastoma Subgroups

PURPOSE: Medulloblastoma comprises four distinct molecular subgroups: WNT, SHH, Group 3, and Group 4. Current medulloblastoma protocols stratify patients based on clinical features: patient age, metastatic stage, extent of resection, and histologic variant. Stark prognostic and genetic differences among the four subgroups suggest that subgroup-specific molecular biomarkers could improve patient prognostication. PATIENTS AND METHODS: Molecular biomarkers were identified from a discovery set of 673 medulloblastomas from 43 cities around the world. Combined risk stratification models were designed based on clinical and cytogenetic biomarkers identified by multivariable Cox proportional hazards analyses. Identified biomarkers were tested using fluorescent in situ hybridization (FISH) on a nonoverlapping medulloblastoma tissue microarray (n = 453), with subsequent validation of the risk stratification models. RESULTS: Subgroup information improves the predictive accuracy of a multivariable survival model compared with clinical biomarkers alone. Most previously published cytogenetic biomarkers are only prognostic within a single medulloblastoma subgroup. Profiling six FISH biomarkers (GLI2, MYC, chromosome 11 [chr11], chr14, 17p, and 17q) on formalin-fixed paraffin-embedded tissues, we can reliably and reproducibly identify very low-risk and very high-risk patients within SHH, Group 3, and Group 4 medulloblastomas. CONCLUSION: Combining subgroup and cytogenetic biomarkers with established clinical biomarkers substantially improves patient prognostication, even in the context of heterogeneous clinical therapies. The prognostic significance of most molecular biomarkers is restricted to a specific subgroup. We have identified a small panel of cytogenetic biomarkers that reliably identifies very high-risk and very low-risk groups of patients, making it an excellent tool for selecting patients for therapy intensification and therapy de-escalation in future clinical trials

Failure of human rhombic lip differentiation underlies medulloblastoma formation

Medulloblastoma (MB) comprises a group of heterogeneous paediatric embryonal neoplasms of the hindbrain with strong links to early development of the hindbrain 1–4. Mutations that activate Sonic hedgehog signalling lead to Sonic hedgehog MB in the upper rhombic lip (RL) granule cell lineage 5–8. By contrast, mutations that activate WNT signalling lead to WNT MB in the lower RL 9,10. However, little is known about the more commonly occurring group 4 (G4) MB, which is thought to arise in the unipolar brush cell lineage 3,4. Here we demonstrate that somatic mutations that cause G4 MB converge on the core binding factor alpha (CBFA) complex and mutually exclusive alterations that affect CBFA2T2, CBFA2T3, PRDM6, UTX and OTX2. CBFA2T2 is expressed early in the progenitor cells of the cerebellar RL subventricular zone in Homo sapiens, and G4 MB transcriptionally resembles these progenitors but are stalled in developmental time. Knockdown of OTX2 in model systems relieves this differentiation blockade, which allows MB cells to spontaneously proceed along normal developmental differentiation trajectories. The specific nature of the split human RL, which is destined to generate most of the neurons in the human brain, and its high level of susceptible EOMES +KI67 + unipolar brush cell progenitor cells probably predisposes our species to the development of G4 MB

Pyrimido[5,4-d]Pyrimidine or pyrimidine derivates compounds and uses there of in the treatment of cancer

In ottemperanza all’articolo 198 CPI si allega: Relazione in lingua italiana concernente l’invenzione della domanda di brevetto PCT dal titolo “PYRIMIDO[5,4-d]PYRIMIDINE OR PYRIMIDINE DERIVATIVES COMPOUNDS AND USES THEREOF IN THE TREATMENT OF CANCER”, a nome ZOLLO Massimo, GALEONE Aldo, VIRGILIO Antonella, SPANO Daniela, DE ANTONELLIS Pasquale L’invenzione concerne pirimido[5,4-d]pirimidino o pirimidino derivati e loro usi nel trattamento del cancro. In particolare, l’invenzione concerne pirimido[5,4-d]pirimidino o pirimidino derivati e loro usi nel trattamento del cancro iper-esprimente h-prune, ossia di cancro metastatico, come ad esempio i carcinoma metastatici alla mammella, alla prostata, al colon, al pancreas, ai polmoni, al fegato, medulloblastoma gastrico e esofageo, neuroblastoma, glioma, oligondendrioma, astrocitoma, sarcoma, linfoma, mieloma, leucemie, cancro alle ovaie. Costituiscono oggetto specifico della presente invenzione pirimido[5,4-d]pirimidino or pirimidino derivati aventi formula (I) e (II), o loro sali, ad esempio cloridrati: (I) (II) in cui X è H o 4-nitrobenzil etano-1,2-diilbis(metilcarbammato). I gruppi OX possono avere configurazione R o S nella formula I e configurazione R o S uguale o diversa tra loro nella formula II. Preferibilmente, i derivati della presente invenzione sono i seguenti: (R)-1-(4-((4-metossibenzil)ammino)pirimidin-2-il)pirrolidin-3-ol (composto denominato nel testo come AA7.1), (R)-1-(4-((4-metossibenzil)ammino)pirimidin-2-il)pirrolidin-3-il 4-nitrobenzil etan-1,2-diilbis(metilcarbammato) (composto denominato nel testo come AA7.1 hypoxia), (3S,3'S)-1,1'-(4,8-bis(4-metossibenzilammino)pirimido[5,4-d]pirimidin-2,6-diil)dipirrolidin-3-olo (composto denominato nel testo come composto 8 o AA2.9, (3R,3'R)-1,1'-(4,8-bis(4-metossibenzilammino)pirimido[5,4-d]pirimidino-2,6-diil)dipirrolidin-3-olo (composto denominato nel testo come composto 9 o AA2.8). Nella domanda di brevetto è riportata la descrizione dettagliata degli esperimenti condotti dagli inventori per testare l’efficacia dei composti sopra menzionati, ossia la loro efficacia nell’inibire l’attività PDE di h-prune, la migrazione e la proliferazione cellulare, utilizzando modelli animali di cancro metastatico. In particolare, gli inventori hanno messo a punto un modello animale per l’identificazione e lo studio di composti in grado di inibire l’attività di h-prune. In modo specifico, il modello animale è stato denominato Mouse Loricrin-prune/Vegfr2-luc ed è caratterizzato dal comprendere un transgene di h-prune (GenBank: NM_021222.1 dalla Base 137 alla 1495) con un epitopo flag tag la cui trascrizione è condotta da un promotore della loricrina (GenBank: S77319.1) e un transgene contenente un promotore murino VEGFR2 e un cDNA di luciferasi modificata di lucciola (Promega pGL-3). L’invenzione concerne anche composizioni farmaceutiche contenenti i composti sopra menzionati e loro usi, oltre al procedimento per la preparazione di detti composti e del modello animale

Novel pyrimidopyrimidine derivatives for inhibition of cellular proliferation and motility induced by h-prune in breast cancer

The human (h)-prune protein is a member of the DHH protein superfamily and it has a cAMP phosphodiesterase activity. Its overexpression in breast, colorectal and gastric cancers correlates with depth of invasion and a high degree of lymph-node metastasis. One mechanism by which h-prune stimulates cell motility and metastasis processes is through its phosphodiesterase activity, which can be suppressed by dipyridamole, a pyrimido[5,4-d]pyrimidine analogue. To obtain new and more potent agents that have high specificity towards inhibition of this h-prune activity, we followed structureeactivity-relationship methodologies starting from dipyridamole and synthesised eight new pyrimidoepyrimidine derivatives. We analysed these newly generated compounds for specificity towards h-prune activities in vitro in cellular models using scintillation proximity assay for cAMP-PDE activity, cell index in cell proliferation assays and transwell methodology for two-dimensional cell migration in a top-down strategy of selection. Our findings show that two pyrimido[5,4-d]pyrimidine compounds are more effective than dipyridamole in two highly metastatic cellular models of breast cancer in vitro. Future studies will assess their therapeutic effectiveness against breast and other cancers where there is over-expression of hprune, and in ad-hoc, proof of concept, animal models

The miR-17-92 MicroRNA Cluster regulates multiple components of the TGF-beta Pathway in neuroblastoma

The miR-17-92 microRNA cluster is often activated in cancer cells, but the identity of its targets remains elusive. Using SILAC and quantitative mass spectrometry, we examined the effects of activation of the miR-17-92 cluster on global protein expression in neuroblastoma (NB) cells. Our results reveal cooperation between individual miR-17-92 miRNAs and implicate miR-17-92 in multiple hallmarks of cancer, including proliferation and cell adhesion. Most importantly, we show that miR-17-92 is a potent inhibitor of TGF-beta signaling. By functioning both upstream and downstream of pSMAD2, miR-17-92 activation triggers downregulation of multiple key effectors along the TGF-beta signaling cascade as well as direct inhibition of TGF-beta-responsive genes