Publications by authors named "Francesco Fiorentino"

81 Publications

Mass spectrometry informs the structure and dynamics of membrane proteins involved in lipid and drug transport.

Curr Opin Struct Biol 2021 May 5;70:53-60. Epub 2021 May 5.

Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, Oxford, OX1 3QZ, UK; The Kavli Institute for Nanoscience Discovery, 3 South Parks Road, Oxford, OX1 3QU, UK. Electronic address:

Membrane proteins are important macromolecules that play crucial roles in many cellular and physiological processes. Over the past two decades, the use of mass spectrometry as a biophysical tool to characterise membrane proteins has grown steadily. By capturing these dynamic complexes in the gas phase, many unknown small molecule interactions have been revealed. One particular application of this research has been the focus on antibiotic resistance with considerable efforts being made to understand underlying mechanisms. Here we review recent advances in the application of mass spectrometry that have yielded both structural and dynamic information on the interactions of antibiotics with proteins involved in bacterial cell envelope biogenesis and drug efflux.
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http://dx.doi.org/10.1016/j.sbi.2021.03.014DOI Listing
May 2021

Cross-Validation of Next-Generation Sequencing Technologies for Diagnosis of Chromosomal Mosaicism and Segmental Aneuploidies in Preimplantation Embryos Model.

Life (Basel) 2021 Apr 12;11(4). Epub 2021 Apr 12.

Villa Mafalda, Reproductive Medicine, 00199 Rome, Italy.

Detection of mosaic embryos is crucial to offer more possibilities of success to women undergoing in vitro fertilization (IVF) treatment. Next Generation Sequencing (NGS)-based preimplantation genetic testing are increasingly used for this purpose since their higher capability to detect chromosomal mosaicism in human embryos. In the recent years, new NGS systems were released, however their performance for chromosomal mosaicism are variable. We performed a cross-validation analysis of two different NGS platforms in order to assess the feasibility of these techniques and provide standard parameters for the detection of such aneuploidies. The study evaluated the performance of Miseq Veriseq (Illumina, San Diego, CA, USA) and Ion Torrent Personal Genome Machine PGM ReproSeq (Thermo Fisher, Waltham, MA, USA) for the detection of whole and segmental mosaic aneuploidies. Reconstructed samples with known percentage of mosaicism were analyzed with both platforms and sensitivity and specificity were determined. Both platforms had high level of specificity and sensitivity with a Limit Of Detection (LOD) at ≥30% of mosaicism and a showed a ≥5.0 Mb resolution for segmental abnormalities. Our findings demonstrated that NGS methodologies are capable of accurately detecting chromosomal mosaicism and segmental aneuploidies. The knowledge of LOD for each NGS platform has the potential to reduce false-negative and false-positive diagnoses when applied to detect chromosomal mosaicism in a clinical setting.
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http://dx.doi.org/10.3390/life11040340DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8069536PMC
April 2021

What people really change after genetic testing (GT) performed in private labs: results from an Italian study.

Eur J Hum Genet 2021 Apr 12. Epub 2021 Apr 12.

Applied Research Division for Cognitive and Psychological Science, European Institute of Oncology (IEO), IRCCS, Milan, Italy.

Despite the widespread diffusion of direct-to-consumer genetic testing (GT), it is still unclear whether people who learn about their genetic susceptibility to a clinical condition change their behaviors, and the psychological factors involved. The aim of the present study is to investigate long-term changes in health-related choices, individual tendencies and risk attitudes in an Italian sample of GT users. In the context of the Mind the Risk study, which investigated a sample of Italian adults who underwent GT in a private laboratory, 99 clients participated in the follow up assessment. They completed a self-administered questionnaire investigating: (a) clinical history and motivation for testing, (b) lifestyle and risk behaviors, (c) individual tendencies toward health, and (d) risk-taking attitude and risk tolerance. Such variables were measured at three different time-points: T0-before GT, T1-at 6 months after genetic results, and T2-at 1 year from results. Results showed that, at baseline, participants who stated they intended to modify their behavior after GT results, effectively did so over time. This result held both for participants who received a positive or negative test result. In general, a healthier diet was the most frequently observed long-term behavioral change. As regards psychological variables, a risk-taking attitude and risk tolerance did not seem to affect the decision to change the lifestyle. Finally, we found an overall reduction in anxiety and worry over health over time, but also a reduction in the motivation for health promotion and prevention, health esteem, and positive expectations for their health in the future.
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http://dx.doi.org/10.1038/s41431-021-00879-wDOI Listing
April 2021

The Two-Faced Role of SIRT6 in Cancer.

Cancers (Basel) 2021 Mar 8;13(5). Epub 2021 Mar 8.

Department of Drug Chemistry & Technologies, Sapienza University of Rome, P. le A Moro 5, 00185 Rome, Italy.

Sirtuin 6 (SIRT6) is a NAD-dependent nuclear deacylase and mono-ADP-ribosylase with a wide spectrum of substrates. Through its pleiotropic activities, SIRT6 modulates either directly or indirectly key processes linked to cell fate determination and oncogenesis such as DNA damage repair, metabolic homeostasis, and apoptosis. SIRT6 regulates the expression and activity of both pro-apoptotic (e.g., Bax) and anti-apoptotic factors (e.g., Bcl-2, survivin) in a context-depending manner. Mounting evidence points towards a double-faced involvement of SIRT6 in tumor onset and progression since the block or induction of apoptosis lead to opposite outcomes in cancer. Here, we discuss the features and roles of SIRT6 in the regulation of cell death and cancer, also focusing on recently discovered small molecule modulators that can be used as chemical probes to shed further light on SIRT6 cancer biology and proposed as potential new generation anticancer therapeutics.
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http://dx.doi.org/10.3390/cancers13051156DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7962659PMC
March 2021

Discrepancy of p16 immunohistochemical expression and HPV RNA in penile cancer. A multiplex in situ hybridization/immunohistochemistry approach study.

Infect Agent Cancer 2021 Mar 31;16(1):22. Epub 2021 Mar 31.

Pathology Unit, Department of Mental and Physical Health and Preventive Medicine, University of Campania "L. Vanvitelli", Complesso di Santa Patrizia, Via Luciano Armanni, 5, 80138, Naples, Italy.

Background: The high-risk human papillomavirus (HPV) infection represents one of the main etiologic pathways of penile carcinogenesis in approximately 30-50 % of cases. Several techniques for the detection of HPV are currently available including Polymerase chain reaction-based techniques, DNA and RNA in situ hybridization (ISH), p16 immunohistochemistry (IHC). The multiplex HPV RNA ISH/p16 IHC is a novel technique for the simultaneous detection of HPV E6/E7 transcripts and p16INK4a overexpression on the same slide in a single assay. The main aim of this study was to evaluate the discrepancy of p16 IHC expression relatively to HPV RNA ISH in penile cancer tissue.

Methods: We collected a series of 60 PCs. HPV has been analysed through the RNA ISH, p16 IHC and the multiplex HPV RNA ISH/p16 IHC.

Results: The multiplex HPV RNA ISH /p16 IHC results in the series were in complete agreement with the previous results obtained through the classic p16 IHC and HPV RNA scope carried out on two different slides. The multiplex HPV RNA ISH /p16 IHC showed that HPV positivity in our series is more frequently in usual squamous cell carcinoma than in special histotypes (19 out of 60 - 15 %- versus 6 out of 60 - 10 %-), in high-grade than in moderate/low grade carcinomas (6 out of 60 - 10 %- versus 4 out of 60 - 6.7 %-). In addition, our data revealed that in 5 out of 20 cases with p16 high intensity expression is not associated with HPV RNA ISH positivity.

Conclusions: Our findings emphasize that the use of p16 as a surrogate of HPV positivity was unsuccessful in approximatively 8 % of cases analysed in our series. Indeed, p16 IHC showed a sensitivity of 100 % and a specificity of 71 %, with a positive predictive value (PPV) of 54 % and a negative predictive value of 100 %; when considering high intensity, p16 IHC showed a sensitivity of 100 %, a specificity of 89 %, with a PPV of 75 % and NPV of 100 %. Since HPV positivity could represent a relevant prognostic and predictive value, the correct characterization offered by this approach appears to be of paramount importance.
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http://dx.doi.org/10.1186/s13027-021-00361-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8011184PMC
March 2021

Using outcome data from one thousand mosaic embryo transfers to formulate an embryo ranking system for clinical use.

Fertil Steril 2021 May 6;115(5):1212-1224. Epub 2021 Mar 6.

Cooper Genomics, Livingston, New Jersey.

Objective: To study how the attributes of mosaicism identified during preimplantation genetic testing for aneuploidy relate to clinical outcomes, in order to formulate a ranking system of mosaic embryos for intrauterine transfer.

Design: Compiled analysis.

Setting: Multi-center.

Patient(s): A total of 5,561 euploid blastocysts and 1,000 mosaic blastocysts used in clinical transfers in patients undergoing fertility treatment.

Intervention(s): None.

Main Outcome Measure(s): Implantation (gestational sac), ongoing pregnancy, birth, and spontaneous abortion (miscarriage before 20 weeks of gestation).

Result(s): The euploid group had significantly more favorable rates of implantation and ongoing pregnancy/birth (OP/B) compared with the combined mosaic group or the mosaic group affecting only whole chromosomes (implantation: 57.2% vs. 46.5% vs. 41.8%; OP/B: 52.3% vs. 37.0% vs. 31.3%), as well as lower likelihood of spontaneous abortion (8.6% vs. 20.4% vs. 25%). Whole-chromosome mosaic embryos with level (percent aneuploid cells) <50% had significantly more favorable outcomes than the ≥50% group (implantation: 44.5% vs. 30.4%; OP/B: 36.1% vs. 19.3%). Mosaic type (nature of the aneuploidy implicated in mosaicism) affected outcomes, with a significant correlation between number of affected chromosomes and unfavorable outcomes. This ranged from mosaicism involving segmental abnormalities to complex aneuploidies affecting three or more chromosomes (implantation: 51.6% vs. 30.4%; OP/B: 43.1% vs. 20.8%). Combining mosaic level, type, and embryo morphology revealed the order of subcategories regarding likelihood of positive outcome.

Conclusion(s): This compiled analysis revealed traits of mosaicism identified with preimplantation genetic testing for aneuploidy that affected outcomes in a statistically significant manner, enabling the formulation of an evidence-based prioritization scheme for mosaic embryos in the clinic.
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http://dx.doi.org/10.1016/j.fertnstert.2020.11.041DOI Listing
May 2021

BET-Inhibitor I-BET762 and PARP-Inhibitor Talazoparib Synergy in Small Cell Lung Cancer Cells.

Int J Mol Sci 2020 Dec 16;21(24). Epub 2020 Dec 16.

Department of Biomedical Science, and National Institute of Biostructures and Biosystems, University of Sassari, 07100 Sassari, Italy.

Small cell lung cancer (SCLC) is an aggressive type of lung cancer with high mortality that is caused by frequent relapses and acquired resistance. Despite that several target-based approaches with potential therapeutic impact on SCLC have been identified, numerous targeted drugs have not been successful in providing improvements in cancer patients when used as single agents. A combination of targeted therapies could be a strategy to induce maximum lethal effects on cancer cells. As a starting point in the development of new drug combination strategies for the treatment of SCLC, we performed a mid-throughput screening assay by treating a panel of SCLC cell lines with BETi or AKi in combination with PARPi or EZH2i. We observed drug synergy between I-BET762 and Talazoparib, BETi and PARPi, respectively, in SCLC cells. Combinatorial efficacy was observed in -amplified and -wt SCLC cells over SCLC cells with impaired MYC signaling pathway or non-tumor cells. We indicate that drug synergy between I-BET762 and Talazoparib is associated with the attenuation HR-DSBR process and the downregulation of various players of DNA damage response by BET inhibition, such as CHEK2, PTEN, NBN, and FANCC. Our results provide a rationale for the development of new combinatorial strategies for the treatment of SCLC.
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http://dx.doi.org/10.3390/ijms21249595DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7766292PMC
December 2020

The complex relationship between female age and embryo euploidy.

Minerva Obstet Gynecol 2021 Feb 11;73(1):103-110. Epub 2020 Dec 11.

Center for Reproductive Medicine, Villa Mafalda, Rome, Italy.

Background: Female age is the strongest predictor of embryo chromosomal abnormalities and has a nonlinear relationship with the blastocyst euploidy rate: with advancing age there is an acceleration in the reduction of blastocyst euploidy. Aneuploidy was found to significantly increase with maternal age from 30% in embryos from young women to 70% in women older than 40 years old. The association seems mainly due to chromosomal abnormalities occurring in the oocyte. We aimed to elaborate a model for the blastocyst euploid rate for patients undergoing in-vitro fertilization/intra cytoplasmic sperm injection (IVF/ICSI) cycles using advanced machine learning techniques.

Methods: This was a retrospective analysis of IVF/ICSI cycles performed from 2014 to 2016. In total, data of 3879 blastocysts were collected for the analysis. Patients underwent PGT-Aneuploidy analysis (PGT-A) at the Center for Reproductive Medicine of European Hospital (Rome, Italy) have been included in the analysis. The method involved whole-genome amplification followed by array comparative genome hybridization. To model the rate of euploid blastocysts, the data were split into a train set (used to fit and calibrate the models) and a test set (used to assess models' predictive performance). Three different models were calibrated: a classical linear regression; a gradient boosted tree (GBT) machine learning model; a model belonging to the generalized additive models (GAM).

Results: The present study confirms that female age, which is the strongest predictor of embryo chromosomal abnormalities, and blastocyst euploidy rate have a nonlinear relationship, well depicted by the GBT and the GAM models. According to this model, the rate of reduction in the percentage of euploid blastocysts increases with age: the yearly relative variation is -10% at the age of 37 and -30% at the age of 45. Other factors including male age, female and male Body Mass Index, fertilization rate and ovarian reserve may only marginally impact on embryo euploidy rate.

Conclusions: Female age is the strongest predictor of embryo chromosomal abnormalities and has a non-linear relationship with the blastocyst euploidy rate. Other factors related to both the male and female subjects may only minimally affect this outcome.
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http://dx.doi.org/10.23736/S0026-4784.20.04740-1DOI Listing
February 2021

Two distinct TP53 mutations in HNSCC primary tumor: Only one circulates in the blood.

Oral Oncol 2021 04 21;115:105096. Epub 2020 Nov 21.

Oncogenomic and Epigenetic Unit, IRCSS Regina Elena National Cancer Institute, Via Elio Chianesi, 53, Rome 00144, Italy. Electronic address:

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http://dx.doi.org/10.1016/j.oraloncology.2020.105096DOI Listing
April 2021

Dynamics of an LPS translocon induced by substrate and an antimicrobial peptide.

Nat Chem Biol 2021 02 16;17(2):187-195. Epub 2020 Nov 16.

Department of Chemistry, University of Oxford, Oxford, UK.

Lipopolysaccharide (LPS) transport to the outer membrane (OM) is a crucial step in the biogenesis of microbial surface defenses. Although many features of the translocation mechanism have been elucidated, molecular details of LPS insertion via the LPS transport (Lpt) OM protein LptDE remain elusive. Here, we integrate native MS with hydrogen-deuterium exchange MS and molecular dynamics simulations to investigate the influence of substrate and peptide binding on the conformational dynamics of LptDE. Our data reveal that LPS induces opening of the LptD β-taco domain, coupled with conformational changes on β-strands adjacent to the putative lateral exit gate. Conversely, an antimicrobial peptide, thanatin, stabilizes the β-taco, thereby preventing LPS transport. Our results illustrate that LPS insertion into the OM relies on concerted opening movements of both the β-barrel and β-taco domains of LptD, and suggest a means for developing antimicrobial therapeutics targeting this essential process in Gram-negative ESKAPE pathogens.
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http://dx.doi.org/10.1038/s41589-020-00694-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7116625PMC
February 2021

Massive squamous differentiation and anaplastic features in eccrine porocarcinoma as rare and confounding elements.

Australas J Dermatol 2021 May 27;62(2):e320-e321. Epub 2020 Sep 27.

Division of Pathology, Department of Mental and Physical Health and Preventive Medicine, University of Campania "L., Naples, Italy.

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http://dx.doi.org/10.1111/ajd.13472DOI Listing
May 2021

Structural Basis of Tail-Anchored Membrane Protein Biogenesis by the GET Insertase Complex.

Mol Cell 2020 10 9;80(1):72-86.e7. Epub 2020 Sep 9.

Heidelberg University Biochemistry Center (BZH), Im Neuenheimer Feld 328, 69120 Heidelberg, Germany. Electronic address:

Membrane protein biogenesis faces the challenge of chaperoning hydrophobic transmembrane helices for faithful membrane insertion. The guided entry of tail-anchored proteins (GET) pathway targets and inserts tail-anchored (TA) proteins into the endoplasmic reticulum (ER) membrane with an insertase (yeast Get1/Get2 or mammalian WRB/CAML) that captures the TA from a cytoplasmic chaperone (Get3 or TRC40, respectively). Here, we present cryo-electron microscopy reconstructions, native mass spectrometry, and structure-based mutagenesis of human WRB/CAML/TRC40 and yeast Get1/Get2/Get3 complexes. Get3 binding to the membrane insertase supports heterotetramer formation, and phosphatidylinositol binding at the heterotetramer interface stabilizes the insertase for efficient TA insertion in vivo. We identify a Get2/CAML cytoplasmic helix that forms a "gating" interaction with Get3/TRC40 important for TA insertion. Structural homology with YidC and the ER membrane protein complex (EMC) implicates an evolutionarily conserved insertion mechanism for divergent substrates utilizing a hydrophilic groove. Thus, we provide a detailed structural and mechanistic framework to understand TA membrane insertion.
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http://dx.doi.org/10.1016/j.molcel.2020.08.012DOI Listing
October 2020

Lysine Acetyltransferase Inhibitors From Natural Sources.

Front Pharmacol 2020 12;11:1243. Epub 2020 Aug 12.

Department of Chemistry and Technology of Drugs, Sapienza University of Rome, Rome, Italy.

Acetylation of histone and non-histone protein lysine residues has been widely described as a critical modulator of several cell functions in humans. Lysine acetyltransferases (KATs) catalyse the transfer of acetyl groups on substrate proteins and are involved in multiple physiological processes such as cell signalling, metabolism, gene regulation, and apoptosis. Given the pivotal role of acetylation, the alteration of KATs enzymatic activity has been clearly linked to various cellular dysfunctions leading to several inflammatory, metabolic, neurological, and cancer diseases. Hence, the use KAT inhibitors (KATi) has been suggested as a potentially successful strategy to reverse or prevent these conditions. To date, only a few KATi have proven to be potential drug candidates, and there is still a keen interest in designing molecules showing drug-like properties from both pharmacodynamics and pharmacokinetics point of view. Increasing literature evidence has been highlighting natural compounds as a wide source of molecular scaffolds for developing therapeutic agents, including KATi. In fact, several polyphenols, catechins, quinones, and peptides obtained from natural sources (including nuts, oils, root extracts, and fungi metabolites) have been described as promising KATi. Here we summarize the features of this class of compounds, describing their modes of action, structure-activity relationships and (semi)-synthetic derivatives, with the aim of assisting the development of novel more potent, isoform selective and drug-like KATi.
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http://dx.doi.org/10.3389/fphar.2020.01243DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7434864PMC
August 2020

Assembly and regulation of the chlorhexidine-specific efflux pump AceI.

Proc Natl Acad Sci U S A 2020 07 7;117(29):17011-17018. Epub 2020 Jul 7.

Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, Oxford OX1 3QZ, United Kingdom

Few antibiotics are effective against , one of the most successful pathogens responsible for hospital-acquired infections. Resistance to chlorhexidine, an antiseptic widely used to combat , is effected through the proteobacterial antimicrobial compound efflux (PACE) family. The prototype membrane protein of this family, AceI ( chlorhexidine efflux protein I), is encoded for by the gene and is under the transcriptional control of AceR ( chlorhexidine efflux protein regulator), a LysR-type transcriptional regulator (LTTR) protein. Here we use native mass spectrometry to probe the response of AceI and AceR to chlorhexidine assault. Specifically, we show that AceI forms dimers at high pH, and that binding to chlorhexidine facilitates the functional form of the protein. Changes in the oligomerization of AceR to enable interaction between RNA polymerase and promoter DNA were also observed following chlorhexidine assault. Taken together, these results provide insight into the assembly of PACE family transporters and their regulation via LTTR proteins on drug recognition and suggest potential routes for intervention.
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http://dx.doi.org/10.1073/pnas.2003271117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7382281PMC
July 2020

TRF2 and VEGF-A: an unknown relationship with prognostic impact on survival of colorectal cancer patients.

J Exp Clin Cancer Res 2020 Jun 15;39(1):111. Epub 2020 Jun 15.

Oncogenomic and Epigenetic Unit, IRCCS - Regina Elena National Cancer Institute, Via Elio Chianesi 53, 00144, Rome, Italy.

Background: Colorectal cancer is one of most common tumors in developed countries and, despite improvements in treatment and diagnosis, mortality rate of patients remains high, evidencing the urgent need of novel biomarkers to properly identify colorectal cancer high-risk patients that would benefit of specific treatments. Recent works have demonstrated that the telomeric protein TRF2 is over-expressed in colorectal cancer and it promotes tumor formation and progression through extra-telomeric functions. Moreover, we and other groups evidenced, both in vitro on established cell lines and in vivo on tumor bearing mice, that TRF2 regulates the vascularization mediated by VEGF-A. In the present paper, our data evidence a tight correlation between TRF2 and VEGF-A with prognostic relevance in colorectal cancer patients.

Methods: For this study we sampled 185 colorectal cancer patients surgically treated and diagnosed at the Regina Elena National Cancer Institute of Rome and investigated the association between the survival outcome and the levels of VEGF-A and TRF2.

Results: Tissue microarray immunohistochemical analyses revealed that TRF2 positively correlates with VEGF-A expression in our cohort of patients. Moreover, analysis of patients' survival, confirmed in a larger dataset of patients from TCGA, demonstrated that co-expression of TRF2 and VEGF-A correlate with a poor clinical outcome in stage I-III colorectal cancer patients, regardless the mutational state of driver oncogenes.

Conclusions: Our results permitted to identify the positive correlation between high levels of TRF2 and VEGF-A as a novel prognostic biomarker for identifying the subset of high-risk colorectal cancer patients that could benefit of specific therapeutic regimens.
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http://dx.doi.org/10.1186/s13046-020-01612-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7294609PMC
June 2020

Comparison of euploidy rates of blastocysts in women treated with progestins or GnRH antagonist to prevent the luteinizing hormone surge during ovarian stimulation.

Hum Reprod 2020 06;35(6):1325-1331

Centre For Reproductive Medicine, European Hospital, Via Portuense, 700, 00149 Rome, Italy.

Study Question: Does the prevalence of euploid blastocysts differ between patients treated with progestin primed ovarian stimulation (PPOS) and those treated with conventional ovarian stimulation?

Summary Answer: The numbers of blastocysts and euploid blastocysts per patient and the number of euploid embryos per injected oocyte are similar for patients undergoing progestin-primed ovarian stimulation and for those undergoing conventional ovarian stimulation with GnRH antagonist.

What Is Known Already: New approaches to ovarian stimulation have been developed based on the use of drugs administrable by mouth instead of via injections. Attention has been dedicated to progestins to block the LH surge. Previous data regarding the number of oocytes retrieved and the number of good-quality embryos generated in PPOS have demonstrated similar outcomes when compared to conventional ovarian stimulation, even if some concerns regarding the quality of embryos have been advanced.

Study Design, Size, Duration: This is a prospective non-inferiority age-matched case-control study. In a period of 6 months, a total of 785 blastocysts from 1867 injected oocytes obtained from 192 patients were available for analysis.

Participants/materials, Setting, Methods: Infertile women undergoing IVF and preimplanation genetic testing (PGT) cycles were included. Forty-eight patients were treated with PPOS, and for each of them three age-matched historical controls (n = 144) treated with a GnRH antagonist protocol were selected. PGT was performed according to next-generation sequencing technology.

Main Results And The Role Of Chance: Basal characteristics were similar in the two groups; a substantial similarity of the main outcome measures in the two treatment groups has also been found. The rate of formation of euploid blastocysts per oocyte was 21% in both the two treatment groups. The percentage of patients with euploid embryos and the total number of euploid blastocysts per patient (median and interquartile range, IQR) in the PPOS group were 38.7 (25.5-52.9) and 2 (1.3-3.1), respectively. These figures were not significantly different in women treated with the GnRH antagonist protocol i.e. 42 (28-53.8) and 2.1 (1.3-2.9), respectively.

Limitations, Reasons For Caution: This was a case-control study which may limit the reliability of the main findings.

Wider Implications Of The Findings: Our results encourage the use of PPOS, especially for oocyte donation, for fertility preservation and for patients in which total freezing of embryos is foreseen, for those expected to be high responders or candidates for preimplantation genetic testing. However, studies aiming to investigate the effect of PPOS on the live birth rate are warranted.

Study Funding/competing Interest(s): None.
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http://dx.doi.org/10.1093/humrep/deaa068DOI Listing
June 2020

Cross-sectional analysis of circulating tumor DNA in primary colorectal cancer at surgery and during post-surgery follow-up by liquid biopsy.

J Exp Clin Cancer Res 2020 Apr 20;39(1):69. Epub 2020 Apr 20.

Oncogenomics Division, Eurofins Genoma Group, Via Castel Giubileo, 11, 00138, Rome, Italy.

Background: Liquid biopsy (LB) in early-stage, non-metastatic colorectal cancer (CRC) must be sensitive enough to detect extremely low circulating tumor DNA (ctDNA) levels. This challenge has been seldom and non-systematically investigated.

Methods: Next generation sequencing (NGS) and digital PCR (dPCR) were combined to test tumor DNAs (tDNAs) and paired ctDNAs collected at surgery from 39 patients, 12 of whom were also monitored during the immediate post-surgery follow up. Patients treated for metastatic disease (n = 14) were included as controls.

Results: NGS and dPCR concordantly (100% agreement) called at least one single nucleotide variant (SNV) in 34 tDNAs, estimated differences in allelic frequencies being negligible (±1.4%). However, despite dPCR testing, SNVs were only detectable in 15/34 (44.1%) ctDNAs from patients at surgery, as opposed to 14/14 (100%) metastatic patients. This was likely due to striking differences (average 10 times, up to 500) in ctDNA levels between groups. NGS revealed blood-only SNVs, suggesting spatial heterogeneity since pre-surgery disease stages, and raising the combined NGS/dPCR sensitivity to 58.8%. ctDNA levels at surgery correlated with neither tumor size, stage, grade, or nodal status, nor with variant abundance in paired tDNA. LB sensitivity reached 63.6% when ctDNA was combined with CEA. Finally, persistence and absence of ctDNA on the first conventional (month 3) post-surgery follow-up were associated with fast relapse and a disease-free status in 3 and 7 patients, respectively.

Conclusions: A simple clinical NGS/dPCR/CEA combination effectively addresses the LB challenge in a fraction of non-metastatic CRC patients.
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http://dx.doi.org/10.1186/s13046-020-01569-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7168847PMC
April 2020

Antiproliferative and proapoptotic effects of extract on Burkitt lymphoma cell line.

Tumour Biol 2020 Feb;42(2):1010428319901061

Department of Medical, Surgical and Experimental Sciences, University of Sassari, Sassari, Italy.

Burkitt lymphoma is a very aggressive B-cell non-Hodgkin lymphoma. Although remarkable progress has been made in the therapeutic scenario for patients with Burkitt lymphoma, search and development of new effective anticancer agents to improve patient outcome and minimize toxicity has become an urgent issue. In this study, the antitumoral activity of , a traditional herb obtained from plants collected on the Asinara Island, Italy, was evaluated in order to explore potential antineoplastic effects of its metabolites on Burkitt lymphoma. Raji human cell line was treated with increasing extract concentration for cytotoxicity screening and subsequent establishment of cell cycle arrest and apoptosis. Moreover, gene expression profiles were performed to identify molecular mechanisms involved in the anticancer activities of this medical plant. The extract exhibited powerful antiproliferative and cytotoxic activities on Raji cell line, showing a dose- and time-dependent decrease in cell viability, obtained by cell cycle arrest in the G2/M phase and an increase in cell apoptosis. The treatment with caused downregulation of genes involved in cell cycle and proliferation (c-MYC, CCND1) and inhibition of cell apoptosis (BCL2, BCL2L1, BCL11A). The extract causes strong anticancer effects on Burkitt lymphoma cell line. The molecular mechanisms underlying such antineoplastic activity are based on targeting and downregulation of genes involved in cell cycle and apoptosis. Our data suggest that natural metabolites should be further exploited as potential antineoplastic agents against Burkitt lymphoma.
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http://dx.doi.org/10.1177/1010428319901061DOI Listing
February 2020

Occult breast cancer in a female with benign lesions.

J Cancer Res Ther 2019 Jul-Sep;15(5):1170-1172

Department of Radiology, University of Foggia, Foggia; Department of Radiology, Scientific Institute Hospital "Casa Sollievo della Sofferenza," San Giovanni Rotondo, FG, Italy.

Occult breast cancer is a carcinoma discovered by the presence of axillary lymph node metastases without the detection of the primary breast tumor. The incidence of this very rare pathology is 0.3%-1.0%. The limited number of these cases does not allow for the precise management of this rare pathology and often, the breast cancer manifestation can take many years to become obvious. We report the case of a 35-year-old woman who presented to our department for annual breast screening examination, without any symptoms. At the time of visit, there were two right and one left tumefactions; unfixed and palpable. Ultrasonography examination confirmed the lesions to be benign. One year later, a palpable hypoechoic axillary left lesion appeared: a lymph node with doubtful morphology. On cytological examination, a biopsy was performed for the axillary left mass which showed irregular masses of large malignant cells with pleomorphism and mitotic figures that suggested a carcinoma. The management of this case is suggestive for cancer of unknown primary syndrome.
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http://dx.doi.org/10.4103/jcrt.JCRT_329_17DOI Listing
July 2020

A challenging diagnosis of mesenchymal neoplasm of the colon: colonic dedifferentiated liposarcoma with lymph node metastases-a case report and review of the literature.

Int J Colorectal Dis 2019 Oct 9;34(10):1809-1814. Epub 2019 Sep 9.

Department of Advanced Biomedical Sciences, Pathology Section, University of Naples Federico II, Via S. Pansini 5, 80131, Naples, Italy.

Purpose: We report a case of primitive colonic dedifferentiated liposarcoma along with lymph node metastases.

Methods: The patient's clinical, radiologic, surgical, and histologic data were reviewed, as well as the literature on colonic dedifferentiated liposarcoma with a focus on the incidence of lymph node metastasis in gastrointestinal sarcomas and on the differential diagnosis with other spindle cell tumors in the gastrointestinal tract.

Results: A 53-year-old man was referred to our hospital with a 3 year-history of pain on the right back that was refractory to drugs. He performed an abdominal computed tomography scan which revealed a colonic wall thickening in the hepatic flexure and a few serosal nodularities. With these findings, the patient underwent an extended right hemicolectomy. On histopathologic examination, it turned out to be a colonic dedifferentiated liposarcoma with lymph node metastases.

Conclusions: The present case was a challenging diagnosis both at presurgical and histopathological level because it strongly mimicked a colonic adenocarcinoma. This was due to non-specific clinical and radiological presentation, to the non-characteristic histologic morphology and to the misleading presence of lymph node metastases. Malignant stromal tumors of the gastrointestinal tract beyond gist are fairly rare entities. Colonic dedifferentiated liposarcoma must be kept in mind and must be considered in the differential diagnosis of gastrointestinal tumors.
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http://dx.doi.org/10.1007/s00384-019-03394-zDOI Listing
October 2019

Clinical outcomes after the transfer of blastocysts characterized as mosaic by high resolution Next Generation Sequencing- further insights.

Eur J Med Genet 2020 Feb 21;63(2):103741. Epub 2019 Aug 21.

Genoma, Roma, Italy.

Objective: To determine the pregnancy outcome potential of euploid, mosaic and aneuploid embryos.

Design: Retrospective study.

Setting: Reference genetics laboratories.

Patient(s): 2654 PGT-A cycles with euploid characterized embryo transfers, 253 PGT-A cycles with transfer of embryos characterized as mosaic, and 10 PGT-A cycles with fully abnormal embryo transfers.

Intervention(s): Blastocysts were assessed by trophectoderm (TE) biopsy followed by PGT-A via array CGH or NGS.

Main Outcome Measure(s): Implantation, miscarriage, ongoing implantation rates (OIR), and karyotype if available, were compared between different embryo groups, and between the two PGT-A techniques.

Results: The Ongoing Pregnancy Rate (OPR)/transfer was significantly higher for NGS-classified euploid embryos (85%) than for aCGH ones (71%) (p < 0.001), but the OPR/cycle was similar (63% vs 59%). NGS-classified mosaic embryos resulted in 37% OPR/cycle (p < 0.001 compared to euploid). Mosaic aneuploid embryos with <40% abnormal cells in the TE sample had an OIR of 50% compared to 27% for mosaics with 40-80% abnormal cells in the TE, and 9% for complex mosaic embryos. All the karyotyped ongoing pregnancies (n = 29) were euploid. Transfers of embryos classified as aneuploid via aCGH (n = 10) led to one chromosomally abnormal pregnancy.

Conclusion(s): NGS-classified euploid embryos yielded higher OIRs but similar OPRs/cycle compared to aCGH. NGS-classified mosaic embryos had reduced potential to reach term, compared to euploid embryos. If they did reach term, those with karyotype results available were euploid. Embryos carrying uniform aneuploidies affecting entire chromosomes were mostly unable to implant after transfer, and the one that implanted ended up in a chromosomally abnormal live birth.
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http://dx.doi.org/10.1016/j.ejmg.2019.103741DOI Listing
February 2020

The Different Effects of Substrates and Nucleotides on the Complex Formation of ABC Transporters.

Structure 2019 04 21;27(4):651-659.e3. Epub 2019 Feb 21.

Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3QZ, UK. Electronic address:

The molybdate importer (ModBC-A of Archaeoglobus fulgidus) and the vitamin B importer (BtuCD-F of Escherichia coli) are members of the type I and type II ABC importer families. Here we study the influence of substrate and nucleotide binding on complex formation and stability. Using native mass spectrometry we show that the interaction between the periplasmic substrate-binding protein (SBP) ModA and the transporter ModBC is dependent upon binding of molybdate. By contrast, vitamin B disrupts interactions between the transporter BtuCD and the SBP BtuF. Moreover, while ATP binds cooperatively to BtuCD-F, and acts synergistically with vitamin B to destabilize the BtuCD-F complex, no effect is observed for ATP binding on the stability of ModBC-A. These observations not only highlight the ability of mass spectrometry to capture these importer-SBP complexes but allow us to add molecular detail to proposed transport mechanisms.
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http://dx.doi.org/10.1016/j.str.2019.01.010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6453779PMC
April 2019

Polyethylene Glycol Epirubicin-Loaded Transcatheter Arterial Chemoembolization Procedures Utilizing a Combined Approach with 100 and 200 μm Microspheres: A Promising Alternative to Current Standards.

J Vasc Interv Radiol 2019 03 31;30(3):305-313. Epub 2019 Jan 31.

Vascular and Interventional Radiology Unit, Department of Diagnostic Service, Sapienza University of Rome, Rome, Italy.

Purpose: To report clinical effectiveness, toxicity profile, and prognostic factors of combined 100 μm ± 25 and 200 μm ± 50 epirubicin-loaded polyethylene glycol (PEG) microsphere drug-eluting embolic transcatheter arterial chemoembolization protocol in patients with hepatocellular carcinoma.

Materials And Methods: In this prospective, single-center, single-arm study with 18 months of follow-up, 36 consecutive patients (mean age 69.9 y ± 10.8; 26 men, 10 women; 54 naïve lesions) were treated. Embolization was initiated with 100 μm ± 25 microspheres, and if stasis (10 heart beats) was not achieved, 200 μm ± 50 microspheres were administered. Each syringe (2 mL) of PEG microsphere was loaded with 50 mg of epirubicin. Results were evaluated using Modified Response Evaluation Criteria In Solid Tumors with multidetector computed tomography/magnetic resonance imaging at 1, 3-6, 9-12, and 15-18 months. Toxicity profile was assessed by laboratory testing before and after the procedure. Complications were recorded. Postembolization syndrome (PES) was defined as onset of fever/nausea/pain after the procedure. Patient/lesion characteristics and treatment results were correlated with predicted outcome using regression analysis. Child-Pugh score was A in 86.1% of patients (31/36) and B in 13.9% (5/36).

Results: In 10 of 21 lesions, < 2 cm in diameter (47.5%) stasis was achieved with 100 μm ± 25 microspheres only, whereas all other lesions required adjunctive treatment with 200 μm ± 50 microspheres. Reported adverse events were grade 1 acute liver bile duct injury (3/39 cases, 7.7%) and PES (grade 2; 3/39 cases, 7.7%). Complete response (CR) at 1, 3-6, 9-12, and 15-18 months was 61.1%, 65.5%, 63.63%, and 62.5%. Objective response (CR + partial response) at 1, 3-6, 9-12, and 15-18 months was 83.3%, 65.85%, 63.63%, and 62.5%. No single factor (laboratory testing, etiology, patient status, hepatic status, tumor characteristics, administration protocol) predicted outcomes except for albumin level at baseline for CR (P < .05, odds ratio = 1.09).

Conclusions: The combined microsphere sizing strategy was technically feasible and yielded promising results in terms of effectiveness and toxicity.
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http://dx.doi.org/10.1016/j.jvir.2018.10.026DOI Listing
March 2019

The pros and cons of preimplantation genetic testing for aneuploidy: clinical and laboratory perspectives.

Fertil Steril 2018 08;110(3):353-361

Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine, Weill Cornell Medical College, New York, New York.

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http://dx.doi.org/10.1016/j.fertnstert.2018.06.002DOI Listing
August 2018

BET inhibition is an effective approach against KRAS-driven PDAC and NSCLC.

Oncotarget 2018 Apr 10;9(27):18734-18746. Epub 2018 Apr 10.

Vall d'Hebron Institute of Oncology (VHIO), Edifici Cellex, Hospital Vall d'Hebron, Barcelona, Spain.

Effectively treating KRAS-driven tumors remains an unsolved challenge. The inhibition of downstream signaling effectors is a way of overcoming the issue of direct targeting of mutant KRAS, which has shown limited efficacy so far. Bromodomain and Extra-Terminal (BET) protein inhibition has displayed anti-tumor activity in a wide range of cancers, including KRAS-driven malignancies. Here, we preclinically evaluate the effect of BET inhibition making use of a new BET inhibitor, BAY 1238097, against Pancreatic Ductal Adenocarcinoma (PDAC) and Non-Small Cell Lung Cancer (NSCLC) models harboring RAS mutations both and . Our results demonstrate that BET inhibition displays significant therapeutic impact in genetic mouse models of KRAS-driven PDAC and NSCLC, reducing both tumor area and tumor grade. The same approach also causes a significant reduction in cell number of a panel of RAS-mutated human cancer cell lines (8 PDAC and 6 NSCLC). In this context, we demonstrate that while BET inhibition by BAY 1238097 decreases MYC expression in some cell lines, at least in PDAC cells its anti-tumorigenic effect is independent of MYC regulation. Together, these studies reinforce the use of BET inhibition and prompt the optimization of more efficient and less toxic BET inhibitors for the treatment of KRAS-driven malignancies, which are in urgent therapeutic need.
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http://dx.doi.org/10.18632/oncotarget.24648DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5922351PMC
April 2018

Lysine acetyltransferase inhibitors: structure-activity relationships and potential therapeutic implications.

Future Med Chem 2018 05 20;10(9):1067-1091. Epub 2018 Apr 20.

Department of Drug Chemistry & Technologies, Sapienza University of Rome, P.le A Moro 5, 00185 Rome, Italy.

Lysine acetylation is a post-translational modification of both histone and nonhistone proteins that is catalyzed by lysine acetyltransferases and plays a key role in numerous biological contexts. The dysregulation of this enzyme activity is implicated in many human pathologies such as cancer, neurological and inflammatory disorders. Many lysine acetyltransferase inhibitors (KATi) have been developed so far, but there is still the need for new, more potent, metabolically stable and selective KATi as chemical tools for studying KAT biology and/or as potential therapeutic agents. This review will examine the features of KAT enzymes and related diseases, with particular emphasis on KATi (bisubstrate analogs, natural compounds and synthetic derivatives), analyzing their mechanism of action, structure-activity relationships, pharmacokinetic/pharmacodynamic properties and potential future applications.
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http://dx.doi.org/10.4155/fmc-2017-0244DOI Listing
May 2018

Adult-Type Rhabdomyoma of the Larynx: Clinicopathologic Study of an Uncommon Tumor in a Rare Location.

Case Rep Otolaryngol 2017 26;2017:7186768. Epub 2017 Nov 26.

Department of Sense Organs, Sapienza University of Rome, Rome, Italy.

Rhabdomyoma is an uncommon benign mesenchymal tumor with skeletal muscle differentiation that may occur either in the heart or in extracardiac sites. Even though the head and neck region is the most common area of extracardiac rhabdomyoma, the larynx is rarely involved. We present the case of an 85-year-old woman who reported a 10-day history of breathing difficulties, dysphagia, and dysphonia. A computed tomography scan of the head and neck showed a contrast-enhanced, solid hypopharyngeal-laryngeal neoplasm with well-defined margins causing subtotal obliteration of the right pyriform sinus and a reduction in air lumen of the laryngeal vestibule. The patient underwent complete endoscopic removal of the lesion; histologic examination revealed an adult-type rhabdomyoma based on the histologic features and the immunoreactivity of the neoplastic cells for desmin, myoglobin, and muscle-specific actin but not for cytokeratin, S-100, CD68R, chromogranin-A, and synaptophysin. Since clinical and imaging features are not specific for rhabdomyoma, histologic examination and immunohistochemical analyses play a central role in the differential diagnosis of the adult-type rhabdomyoma from other laryngeal neoplasms. A correct diagnosis is mandatory to avoid inappropriate treatment.
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http://dx.doi.org/10.1155/2017/7186768DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5727691PMC
November 2017

Extent of chromosomal mosaicism influences the clinical outcome of in vitro fertilization treatments.

Fertil Steril 2018 01 28;109(1):77-83. Epub 2017 Nov 28.

Reproductive Medicine, European Hospital, Rome, Italy.

Objective: To assess whether the extent of chromosomal mosaicism can influence the success rate of IVF treatments.

Design: Prospective study.

Setting: Private genetic and assisted reproduction centers.

Patient(s): The transfer of mosaic embryos was offered to 77 women for which IVF resulted in no euploid embryos available for transfer.

Intervention(s): All embryos were cultured to blastocyst stage; trophectoderm biopsy was performed on day 5/6 of development. Comprehensive chromosome screening was performed using either next-generation sequencing or array-comparative genomic hybridization methodologies.

Main Outcome Measure(s): The clinical outcome obtained after transfer of mosaic embryos with low (<50%) and high (≥50%) aneuploidy percentage was compared with that resulting from a control group of 251 euploid blastocysts.

Result(s): A significantly higher implantation rate (48.9% vs. 24.2%), clinical pregnancy rate/ET (40.9% vs. 15.2%), and live-birth rate (42.2% vs. 15.2%) were observed comparing embryos with mosaicism <50% and ≥50%. Mosaic embryos with high aneuploidy percentage (≥50%) showed a significantly lower clinical pregnancy rate/ET (15.2% vs. 46.4%), implantation rate (24.4% vs. 54.6%), and live-birth rate (15.2% vs. 46.6%) than euploid blastocysts. In contrast, embryos with lower aneuploidy percentage (<50%) have a clinical outcome similar to euploid embryos.

Conclusion(s): The results of this study further confirm that mosaic embryos can develop into healthy euploid newborns. We demonstrated that the extent of mosaicism influences the IVF success rate. Mosaic embryos with low aneuploidy percentage have higher chances of resulting in the birth of healthy babies compared with embryos with higher mosaicism levels.
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http://dx.doi.org/10.1016/j.fertnstert.2017.09.025DOI Listing
January 2018

Author's reply to Grati and Benn.

Prenat Diagn 2017 10;37(10):1053-1054

GENOMA - Molecular Genetics Laboratories, Rome, Italy.

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http://dx.doi.org/10.1002/pd.5136DOI Listing
October 2017