1,149 research outputs found
Recycling Parrondo games
We consider a deterministic realization of Parrondo games and use periodic
orbit theory to analyze their asymptotic behavior.Comment: 12 pages, 9 figure
Directed deterministic classical transport: symmetry breaking and beyond
We consider transport properties of a double delta-kicked system, in a regime
where all the symmetries (spatial and temporal) that could prevent directed
transport are removed. We analytically investigate the (non trivial) behavior
of the classical current and diffusion properties and show that the results are
in good agreement with numerical computations. The role of dissipation for a
meaningful classical ratchet behavior is also discussed.Comment: 10 pages, 20 figure
Hereditary pancreatic cancer: A retrospective single-center study of 5143 Italian families with history of BRCA-related malignancies
The identification of BRCA mutations plays a crucial role in the management of hereditary cancer prevention and treatment. Nonetheless, BRCA-testing in pancreatic cancer (PC) patients is not universally introduced in clinical practice. A retrospective analysis was conducted, firstly, to evaluate the rate of BRCA-positive families among those presenting a family history of PC besides breast and/or ovarian cancer. Secondly, the relationship between BRCA pathogenic variants and PC risk was evaluated. Finally, the characteristics of PC developed in BRCA families were described. Among 5143 family trees reporting breast and/or ovarian cancer cases, 392 showed a family history of PC. A total of 35 families (24.5% selected by the Modena Criteria and 21.3% by the NCCN Criteria) were positive to BRCA testing. Among the BRCA1 mutations, 36.8% were found within a region defined by c.3239\u2013c.3917, whilst 43.7% of BRCA2 mutations were located within c.7180\u2013c.8248. This study confirmed that an increase in the rate of positive tests in families with PC when associated to breast and/or ovarian tumors. Moreover, this analysis indicated two possible Pancreatic Cancer Cluster Regions that should be verified in future research. Finally, PC in families with breast and/or ovarian cancer history, particularly in BRCA families, were diagnosed at younger age and showed better one-year overall survival
STRATEGIES TO PREDICT TREATMENT RESPONSE AND SELECT THERAPIES IN METASTATIC BREAST CANCER PATIENTS USING A NEXT GENERATION SEQUENCING MULTI-GENE PANEL
The standard of care for many
patients with advanced breast
cancer (BC )is gradually
evolving from empirical
treatment based on clinicalpathological
characteristics to
the use of targeted approaches
based on the molecular profile
of the tumor.
In the last decade, an
increasing number of
molecularly targeted drugs
have been developed for the
treatment of metastatic BC.
These drugs target specific
molecular abnormalities that
confer to cancer cells a survival
advantage. Interestingly,
the ability to perform multigene
testing for a range of
molecular alterations may
provide an opportunity to
clarify the mechanisms of
treatment response, to find the
strategies to overcome
treatment resistance and thus,
to identify patients who are
more likely to develop relapse
and who may be candidates for
matched targeted therapies.
The main aim of this study is to
find prognostic and predictive
molecular biomarkers for the
management of metastatic BC
patients in clinical practice
STRATEGIES TO PREDICT TREATMENT RESPONSE AND SELECT THERAPIES IN METASTATIC BREAST CANCER PATIENTS USING A NEXT GENERATION SEQUENCING (NGS) MULTI-GENE PANEL
The standard of care for many
patients with advanced breast
cancer (BC )is gradually
evolving from empirical
treatment based on clinicalpathological
characteristics to
the use of targeted approaches
based on the molecular profile
of the tumor.
In the last decade, an
increasing number of
molecularly targeted drugs
have been developed for the
treatment of metastatic BC.
These drugs target specific
molecular abnormalities that
confer to cancer cells a survival
advantage [1]. Interestingly,
the ability to perform multigene
testing for a range of
molecular alterations may
provide an opportunity to
clarify the mechanisms of
treatment response, to find the
strategies to overcome
treatment resistance and thus,
to identify patients who are
more likely to develop relapse
and who may be candidates for
matched targeted therapies
[2-3].
The main aim of this study is to
find prognostic and predictive
molecular biomarkers for the
management of metastatic BC
patients in clinical practice. MATERIALS AND
METHODS
The amplicon-sequencing
analyses took advantage of the Ion
AmpliSeq™ technology (Thermo
Fisher, Waltham, MA, USA). A
custom panel was designed with
the help of the Designer online
tool (www.ampliseq.com),
which was employed to generate
optimized primers encompassing
the coding DNA sequences (with
100bp of exon padding and the
UTRs regions) of 25 genes in the
Human Reference Genome
(hg19); these genes were selected
searching and screening scientific
literature for treatments
resistance in BC and are reported
in Table 1. Primer pairs were
divided into two pools to
optimize multiplex PCR
conditions and the coverage, that
assessed to 89.02%. The
customized Ion AmpliSeq panel
was employed on samples from 7
primary BC samples and matched
metastatic sites (3 skin, 3 lymph
node and 1 lung metastases).
They were all processed using the
Ion AmpliSeq Library Kit 2.0,
starting from 15 nanograms of
FFPE extracted DNA/pool.
Samples were barcoded with the
Ion Express Kit to optimize
matched patients pooling on the
same 318 Chip v2 sequencing
chip. The template-positive Ion
Sphere Particles were sequenced
on a Personal Genome Machine
(Thermo Fisher, Waltham, MA,
USA). RESULTS The mutation profiles of paired primary and
secondary tumors of the seven patients enrolled in
this study are presented in Table 2. Ten different
genes (PTEN, PIK3CA, mTOR, ERBB2, ERBB3,
MET, INPP4B, MAP2K1, CDK6, KRAS) in 6
different patients showed possible damaging
variants as shown in Table 2.
• Four patients (number 1, 3, 5 and 6) showed no
additional or different mutations in secondary
tumors if compared to primary samples.
• In patient number 2, the metastatic site
presented new mutations if compared to the
primary tumor.
• Finally in patient number 4 and 7 we did not
detect in metastases some of the mutations
found in the primary tumor. DISCUSSION
In 5 patients (71,4%) the mutational status of primary tumor could explain treatment resistance and thus
predict relapse, in one patient the mutational status of the new subclones could be relevant for guiding
differently the subsequent treatment choices.
In 2 patients (28,5%) we were not able to detect in metastases some of the mutations found in the primary
tumor. This could be explained by considering the clonal evolution of metastases.
These preliminary data suggest that the multi-gene panel analysis of primary and secondary tumors may help
clinicians:
• in discriminating BC patients HR+ and/or HER2+ with mutations predicting an increased risk of adjuvant
treatment resistance and thus relapse
• in guiding treatment selection strategies in the metastatic setting.
The study is still open and we are currently recruiting other patients.The main aim of this study is to find prognostic and predictive molecular biomarkers for the management of metastatic BC patients in clinical practice. The preliminary data suggest that the multi-gene panel analysis of primary and secondary tumors may help clinicians: • in discriminating BC patients HR+ and/or HER2+ with mutations predicting an increased risk of adjuvant treatment resistance and thus relapse • in guiding treatment selection strategies in the metastatic setting. The study is still open and we are currently recruiting other patients
Targeted cancer exome sequencing reveals recurrent mutations in myeloproliferative neoplasms
With the intent of dissecting the molecular complexity of Philadelphia-negative myeloproliferative neoplasms (MPN), we designed a target enrichment panel to explore, using next-generation sequencing (NGS), the mutational status of an extensive list of 2,000 cancer-associated genes and microRNAs. The genomic DNA of granulocytes and in-vitro-expanded CD3+ T-lymphocytes, as a germline control, was target-enriched and sequenced in a learning cohort of 20 MPN patients using Roche 454 technology. We identified 141 genuine somatic mutations, most of which were not previously described. To test the frequency of the identified variants, a larger validation cohort of 189 MPN patients was additionally screened for these mutations using Ion Torrent AmpliSeq NGS. Excluding the genes already described in MPN, for 8 genes (SCRIB, MIR662, BARD1, TCF12, FAT4, DAP3, POLG, and NRAS), we demonstrated a mutation frequency between 3 and 8%.
We also found that mutations at codon 12 of NRAS (NRASG12V and NRASG12D) were significantly associated, for primary myelofibrosis (PMF), with highest DIPSS-plus score categories. This association was then confirmed in 66 additional PMF patients composing a final dataset of 168 PMF showing an NRAS mutation frequency of 4.7%, which was associated with a worse outcome, as defined by the DIPSS plus score
AMPLICON-BASED NGS: AN EFFECTIVE APPROACH FOR THE MOLECULAR DIAGNOSIS OF EPIDERMOLYSIS BULLOSA
Background: Epidermolysis Bullosa (EB) is caused by mutations in genes that encode proteins belonging to the epidermal-dermal
junction assembly. Due to the extreme clinical/genetic heterogeneity of the disease, the current methods available
for diagnosing EB involve immunohistochemistry of bioptic samples and transmission electron microscopy followed
by single candidate gene Sanger Sequencing (SS), which are labour intensive and expensive clinical pathways.
Objectives: According to the recently published recommendations for the EB diagnosis and treatment, the assessment of
the mutational landscape is now a fundamental step for developing a comprehensive diagnostic path. Next-Generation
Sequencing (NGS) via the parallel ultra-deep sequencing of many genes represents a proper method for reducing the
processing time and costs of EB diagnostics.
Methods: We developed an EB disease-comprehensive AmpliSeq panel to accomplish the NGS on the Ion Torrent PGM
platform. The panel was performed on ten patients with known genetic diagnoses and was then employed in eight family
trios with unknown molecular footprints.
Results: The panel was successful in finding the causative mutations in all ten of the patients with known mutations, fully
confirming the SS data and providing proof of concept of the sensitivity, specificity, and accuracy of this procedure. In
addition to being consistent with the clinical diagnosis, it was also effective in the trios, identifying all of the variants, including
ones that the SS missed or de novo mutations.
Conclusions: The NGS and AmpliSeq were shown to be an effective approach for the diagnosis of EB, resulting in a costand
time-effective 72-hour procedure
An explainable model of host genetic interactions linked to COVID-19 severity
We employed a multifaceted computational strategy to identify the genetic factors contributing to increased risk of severe COVID-19 infection from a Whole Exome Sequencing (WES) dataset of a cohort of 2000 Italian patients. We coupled a stratified k-fold screening, to rank variants more associated with severity, with the training of multiple supervised classifiers, to predict severity based on screened features. Feature importance analysis from tree-based models allowed us to identify 16 variants with the highest support which, together with age and gender covariates, were found to be most predictive of COVID-19 severity. When tested on a follow-up cohort, our ensemble of models predicted severity with high accuracy (ACC = 81.88%; AUCROC = 96%; MCC = 61.55%). Our model recapitulated a vast literature of emerging molecular mechanisms and genetic factors linked to COVID-19 response and extends previous landmark Genome-Wide Association Studies (GWAS). It revealed a network of interplaying genetic signatures converging on established immune system and inflammatory processes linked to viral infection response. It also identified additional processes cross-talking with immune pathways, such as GPCR signaling, which might offer additional opportunities for therapeutic intervention and patient stratification. Publicly available PheWAS datasets revealed that several variants were significantly associated with phenotypic traits such as "Respiratory or thoracic disease", supporting their link with COVID-19 severity outcome.A multifaceted computational strategy identifies 16 genetic variants contributing to increased risk of severe COVID-19 infection from a Whole Exome Sequencing dataset of a cohort of Italian patients
The polymorphism L412F in TLR3 inhibits autophagy and is a marker of severe COVID-19 in males
The polymorphism L412F in TLR3 has been associated with several infectious diseases. However, the mechanism underlying this association is still unexplored. Here, we show that the L412F polymorphism in TLR3 is a marker of severity in COVID-19. This association increases in the sub-cohort of males. Impaired macroautophagy/autophagy and reduced TNF/TNFα production was demonstrated in HEK293 cells transfected with TLR3L412F-encoding plasmid and stimulated with specific agonist poly(I:C). A statistically significant reduced survival at 28 days was shown in L412F COVID-19 patients treated with the autophagy-inhibitor hydroxychloroquine (p = 0.038). An increased frequency of autoimmune disorders such as co-morbidity was found in L412F COVID-19 males with specific class II HLA haplotypes prone to autoantigen presentation. Our analyses indicate that L412F polymorphism makes males at risk of severe COVID-19 and provides a rationale for reinterpreting clinical trials considering autophagy pathways. Abbreviations: AP: autophagosome; AUC: area under the curve; BafA1: bafilomycin A1; COVID-19: coronavirus disease-2019; HCQ: hydroxychloroquine; RAP: rapamycin; ROC: receiver operating characteristic; SARS-CoV-2: severe acute respiratory syndrome coronavirus 2; TLR: toll like receptor; TNF/TNF-α: tumor necrosis factor
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