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    ABSTRACT: Multicellular tumor spheroids represent a 3D in vitro model that mimics solid tumor essential properties including assembly and development of extracellular matrix and nutrient, oxygen and proliferation gradients. In the present study, we analyze the impact of 3D spatial organization of HER2-overexpressing breast cancer cells on the response to Trastuzumab. We cultured human mammary adenocarcinoma cell lines as spheroids with the hanging drop method and we observed a gradient of proliferating, quiescent, hypoxic, apoptotic and autophagic cells towards the inner core. This 3D organization decreased Trastuzumab sensitivity of HER2 over-expressing cells compared to monolayer cell cultures. We did not observe apoptosis induced by Trastuzumab but found cell arrest in G0/G1 phase. Moreover, the treatment downregulated the basal apoptosis only found in tumor spheroids, by eliciting protective autophagy. We were able to increase sensitivity to Trastuzumab by autophagy inhibition, thus exposing the interaction between apoptosis and autophagy. We confirmed this result by developing a resistant cell line that was more sensitive to autophagy inhibition than the parental BT474 cells. In summary, the development of Trastuzumab resistance relies on the balance between death and survival mechanisms, characteristic of 3D cell organization. We propose the use of spheroids to further improve the understanding of Trastuzumab antitumor activity and overcome resistance.
    PLoS ONE 09/2015; 10(9):e0137920. DOI:10.1371/journal.pone.0137920
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    ABSTRACT: The suitability of human mesenchymal stem cells (hMSCs) in regenerative medicine relies on retention of their proliferative expansion potential in conjunction with the ability to differentiate toward multiple lineages. Successful utilisation of these cells in clinical applications linked to tissue regeneration requires consideration of biomarker expression, time in culture and donor age, as well as their ability to differentiate towards mesenchymal (bone, cartilage, fat) or non-mesenchymal (e.g., neural) lineages. To identify potential therapeutic suitability we examined hMSCs after extended expansion including morphological changes, potency (stemness) and multilineage potential. Commercially available hMSC populations were expanded in vitro for > 20 passages, equating to > 60 days and > 50 population doublings. Distinct growth phases (A-C) were observed during serial passaging and cells were characterised for stemness and lineage markers at representative stages (Phase A: P+5, approximately 13 days in culture; Phase B: P+7, approximately 20 days in culture; and Phase C: P+13, approximately 43 days in culture). Cell surface markers, stem cell markers and lineage-specific markers were characterised by FACS, ICC and Q-PCR revealing MSCs maintained their multilineage potential, including neural lineages throughout expansion. Co-expression of multiple lineage markers along with continued CD45 expression in MSCs did not affect completion of osteogenic and adipogenic specification or the formation of neurospheres. Improved standardised isolation and characterisation of MSCs may facilitate the identification of biomarkers to improve therapeutic efficacy to ensure increased reproducibility and routine production of MSCs for therapeutic applications including neural repair.
    PLoS ONE 09/2015; 10(9):e0137255. DOI:10.1371/journal.pone.0137255
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    ABSTRACT: We report the clinical, electroencephalo-graphic and neuroradiologic findings of a patient with supernumerary der(22) syndrome – partial 22 trisomy, derived from a (11;22) maternal translocation – and a wide spectrum of cerebral malformation. Magnetic resonance study evidenced a partial midline defect (hypoplasia of the corpus callosum, mild dilatation of the mid portion of the occipital horn of the lateral ventricles and a mild enlargement of the frontal horns) and a malrotation of the body and the tail of both hippocampi, which present a vertical position. This patient was severely mentally retarded and he was affected by drug-resistant focal epilepsy: the seizures were of fronto-temporal origin with possible secondary generalization. The electroencephalographic studies showed a pattern similar to that observed in other cortical malformations. About 30% of the patients with der(22) have cerebral malformation but none presents an hippocampal malrotation. Moreover, this defect seems to be linked to the epilepsy. (J Pediatr Neurol 2003; 1(1): 39-42).
    Journal of pediatric neurology: JPN 07/2015; 01(01). DOI:10.1055/s-0035-1557168


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    Via Verdi 8, 10100, Turin, Piedmont, Italy
  • Head of Institution
    Ajani Gianmaria
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Top publications last week by reads

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