1,720,977 research outputs found
ALK gene alterations in cancer: Biological aspects and therapeutic implications
No full textALK was first reported in 1994 as a translocation in anaplastic large cell lymphoma and then described with different abnormalities in a number of tumors. Recently, a shortly accumulated biomedical research clarified the numerous biological processes underlying its ability to support cancer development, growth and progression. Advent of precision medicine has finally provided unexpected advances, leading to the development of ALK-targeting inhibitors with superior efficacy as compared with standard chemotherapy regimens, as well as the identification of resistance mechanisms and the creation of 'next-generation' treatments. This review summarizes the current understanding of ALK-driven cancers from the oncogenesis and mutation frequency by The Cancer Genome Atlas database through the diagnostic approach, to an updated portrait of available tyrosine kinase inhibitors, considering their effectiveness in cancer treatment, the molecular reasons of therapeutic failure, and the actual and future ways to overcome resistances
Abstract 2012: In vitro validation of tumor-derived large extracellular vesicles isolation and characterization as suitable tool for liquid biopsy
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Rare dihydropyrimidine dehydrogenase variants and toxicity by floropyrimidines: A case report
No full textVariations in the activity, up to absolute deficiency, of the enzyme dihydropyrimidine dehydrogenase (DPD), result in the occurrence of adverse reactions to chemotherapy, and have been included among the pharmacogenetic factors underlying inter-individual variability in response to fluoropyrimidines. The study of single-nucleotide polymorphisms of the DPYD gene, which encodes the DPD enzyme, is one of the main parameters capable of predicting reduced enzymatic activity and the consequent influence on fluoropyrimidine treatment, in terms of reduction of both adverse reactions and therapeutic efficacy in disease control. In this paper, we describe a patient with metastatic breast cancer showing signs of increased toxicity following capecitabine therapy. The DPD enzyme activity analysis revealed a partial deficiency. The study of the most frequent polymorphisms of the DPYD gene suggested a wild-type genotype but indicated a novel variant c.1903A>G (p.Asn635Asp), not previously described, proximal to the splice donor site of exon 14. After excluding the potential pathogenic feature of the newly-identified variant, we performed cDNA sequencing of the entire DPYD coding sequence. This analysis identified the variants c.85T>C and c.496A>G, which were previously described as pivotal components of the haplotype associated with decreased enzyme activity and suggested that both variant alleles are related to DPD deficiency. The clinical case findings described in this study emphasize the importance of performing complete genetic analysis of the DPYD gene in order to identify rare and low frequency variants potentially responsible for toxic reactions to fluoropyrimidine treatment
Application of “omics” sciences to the prediction of bone metastases from breast cancer: State of the art
Full text linkBreast cancer (BC) is the most frequent malignancy and the first cause of cancer-related death in women.
The majority of patients with advanced BC develop skeletal metastases which may ultimately lead to serious complications, termed skeletal-related events, that often dramatically impact on quality of life and survival.
Therefore, the identification of biomarkers able to stratify BC patient risk to develop bone metastases (BM) is fundamental to define personalized diagnostic and therapeutic strategies, possibly at the earliest stages of the disease.
In this regard, the advent of “omics” sciences boosted the investigation of several putative biomarkers of BC osteotropism, including deregulated genes, proteins and microRNAs.
The present review revisits the current knowledge on BM development in BC and the most recent studies exploring potential BM-predicting biomarkers, based on the application of omics sciences to the study of primary breast malignancies
Characterization of a Rare Nonpathogenic Methylenetetrahydrofolatereductase (MTHFR) Gene Mutation p.Lys215del in a Southern Italian family
No full textCharacterization of the metastatic behaviour and gene expression profile (RNAseq) of different melanoma cell lines: a comprehensive in vivo model.
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Abstract 2808: Characterization of the metastatic behavior and gene expression profile (RNAseq) of different melanoma cell lines: a comprehensive in vivo model
No full textResults
Conclusions
Metastasis is the major cause of death in malignant
melanoma. Several factors, including clinicalpathological
and tumor biological features may restrain
prognosis. Molecular mechanisms regulating melanoma
progression and metastasis have been partially
discovered, and thus we developed an in vivo model of
metastatic melanoma to investigate potential genes
implicated in these events.
To evaluate the in vivo metastatic activity of five
different melanoma cell lines (LCP, LCM, WM266, SKMel28
and A375), we completed intra-cardiac (ic)
injection of 1x106 luminescent cells or PBS as control in
three NOD-SCID mice per cell line. After 3 weeks, mice
were studies by in vivo bioluminescence imaging (IVIS
Lumina LT) to measure the mean radiance
(p/sec/cm2/sr), that we arbitrarily considered as a
surrogate of the total metastatic tumor burden (t-MTB).
Animals were euthanized at the humane endpoints
achievement and underwent X-ray evaluation for bone
metastasis detection. The Kaplan-Meyer curves
described the time to sacrifice from ic injection. Mouse
necropsy identified metastatic organs that were
explanted and processed for both histology and gene
expression analysis. Melanoma cell lines were profiled
for gene expression (RNAseq) of 118 genes notably
involved in cancer progression and metastasis.
Quantitative real time PCR (qRT-PCR) explored the
expression of a restrict number of genes on formalinfixed
paraffin-embedded (FFPE) metastatic samples
from euthanized animals.
All melanoma cell lines demonstrated a metastatic
behaviour following ic injection with a variable attitude
to produce bone and visceral metastasis (Figure 1).
Mice injected with LCP, LCM and WM266 showed a
lower t-MTB and increased survival (Figure 2) as
compared to A375 and SK-Mel28. Thus, we arbitrarily
defined LCP, LCM and WM-266 cells ‘poorly metastatic’
(group A), while A375 and SK-Mel28 ‘highly metastatic’
(group B).
The principal component analysis (Figure 3A) and the
unsupervised hierarchical clustering of 118 gene
transcriptome analysis (not shown) revealed similar
gene expression profiling among cell lines grouped for
the metastatic attitude. The gene expression analysis
performed on FFPE samples (Figure 3B) identified five
deregulated genes (WNT5A, COL6A3, PTHLH, SOX9 and
SERPINE1) between A and B.
We describe the metastatic capacity of five melanoma
cell lines. Gene expression profiling revealed the
activation of five genes as putatively responsible for the
high aggressiveness of A375 and SK-Mel28 cells. These
results suggest to investigate these genes in a clinical
setting and their possible application as druggable
target for future therapeutic strategies
Characterization of the metastatic behavior and gene expression profile (RNAseq) of different melanoma cell lines: a comprehensive in vivo model.
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Next-generation Sequencing (NGS) Analysis on Single Circulating Tumor Cells (CTCs) with No Need of Whole-genome Amplification (WGA)
Full text linkBackground: Isolation and genotyping of circulating tumor cells (CTCs) is gaining an increasing interest by clinical researchers in oncology not only for investigative purposes, but also for concrete application in clinical practice in terms of diagnosis, prognosis and decision treatment with targeted therapies. For the mutational analysis of single CTCs, the most advanced biotechnology methodology currently available includes the combination of whole genome amplification (WGA) followed by next-generation sequencing (NGS). However, the sequence of these molecular techniques is time-consuming and may also favor operator-dependent errors, related to the procedures themselves that, as in the case of the WGA technique, might affect downstream molecular analyses. Materials and Methods: A preliminary approach of molecular analysis by NGS on a model of CTCs without previous WGA procedural step was performed. We set-up an artificial sample obtained by spiking the SK-MEL-28 melanoma cell line in normal donor peripheral whole blood. Melanoma cells were first enriched using an AutoMACS® (Miltenyi) cell separator and then isolated as single and pooled CTCs by DEPArrayTM System (Silicon Biosystems). NGS analysis, using the Ion AmpliSeqTM Cancer Hotspot Panel v2 (Life Technologies) with the Ion Torrent PGMTM system (Life Technologies), was performed on the SK-MEL-28 cell pellet, a single CTC previously processed with WGA and on 1, 2, 4 and 8 recovered CTCs without WGA pre-amplification. Results: NGS directly carried out on CTCs without WGA showed the same mutations identified in SK-MEL-28 cell line pellet, with a considerable efficiency and avoiding the errors induced by the WGA procedure. Conclusion: We identified a cost-effective, time-saving and reliable methodological approach that could improve the analytical accuracy of the liquid biopsy and appears promising in studying CTCs from cancer patients for both research and clinical purposes
Characterization of a rare nonpathogenic sequence variant (c.1905C>T) of the dihydropyrimidine dehydrogenase gene (DPYD)
Full text linkBackground
In the era of precision medicine, the suitability of fluoropyrimidine therapies in clinical oncology can be checked by pharmacogenetic investigations of single patients, thus optimizing resources and indicating the appropriate drugs to personalize their chemotherapy. For example, the presence of dihydropyrimidine dehydrogenase gene (DPYD) polymorphisms in cancer patients may lead to adverse effects when adopting fluoropyrimidine-based therapies.
Methods
We detected in a cancer patient a rare germline synonymous heterozygous variant of DPYD (c.1905C>T) in proximity to the exon 14 splice donor site. Because in silico analyses hypothesized potential deleterious effects of the splice site, we performed both quantitative and qualitative mRNA analyses to investigate the possible pathogenic nature of the variant.
Results
We did not detect any alterations in mRNA expression or in the cDNA sequence of DPYD gene transcript.
Conclusions
Our observations suggest that the c.1905C>T variant of DPYD does not have a pathogenic effect. Therefore, assessment of the clinical significance of rare sequence variants could emphasize the predictive value of DPYD gene alterations in identifying patients at potential risk for fluoropyrimidine-related toxicity
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