28 results on '"Salvianti F"'
Search Results
2. miR-20a-5p expression as a potential non-invasive diagnostic biomarker in patients with non-obstructive azoospermia
- Author
-
Cito, G., primary, Pinzani, P., additional, Fucci, R., additional, Picone, R., additional, Salvianti, F., additional, Giachini, C., additional, Falcone, P., additional, Micelli, E., additional, Cocci, A., additional, Verrienti, P., additional, Minervini, A., additional, Carini, M., additional, Coccia, M.E., additional, and Natali, A., additional
- Published
- 2020
- Full Text
- View/download PDF
3. 671P Prospective assessment of AR splice variant and PSMA detection on circulating tumour cells of mCRPC patients: Preliminary results of PRIMERA trial (NCT04188275)
- Author
-
Francolini, G., primary, Salvestrini, V., additional, Loi, M., additional, Mangoni, M., additional, Detti, B., additional, Pinzani, P., additional, Salvianti, F., additional, Desideri, I., additional, Aquilano, M., additional, Mariotti, M., additional, Garlatti, P., additional, Stocchi, G., additional, Ciccone, L.P., additional, Lucidi, S., additional, Salvatore, G., additional, Sottili, M., additional, Meattini, I., additional, and Livi, L., additional
- Published
- 2020
- Full Text
- View/download PDF
4. SC10 - miR-20a-5p expression as a potential non-invasive diagnostic biomarker in patients with non-obstructive azoospermia
- Author
-
Cito, G., Pinzani, P., Fucci, R., Picone, R., Salvianti, F., Giachini, C., Falcone, P., Micelli, E., Cocci, A., Verrienti, P., Minervini, A., Carini, M., Coccia, M.E., and Natali, A.
- Published
- 2020
- Full Text
- View/download PDF
5. CirculatingBRAFV600Ein the Diagnosis and Follow-Up of Differentiated Papillary Thyroid Carcinoma
- Author
-
Pupilli, C., primary, Pinzani, P., additional, Salvianti, F., additional, Fibbi, B., additional, Rossi, M., additional, Petrone, L., additional, Perigli, G., additional, De Feo, M. L., additional, Vezzosi, V., additional, Pazzagli, M., additional, Orlando, C., additional, and Forti, G., additional
- Published
- 2013
- Full Text
- View/download PDF
6. Molecular Analysis of Single Circulating Tumor Cells (CTCS) Isolated from Metastatic Breast Cancer (MBC) Patients (PTS)
- Author
-
Pestrin, M., primary, Galardi, F., additional, Salvianti, F., additional, De Luca, F., additional, Bessi, S., additional, Capaccioli, G., additional, Di Leo, A., additional, Giannini, A., additional, Pinzani, P., additional, and Pazzagli, M., additional
- Published
- 2013
- Full Text
- View/download PDF
7. Transplantation: basic science
- Author
-
Cantaluppi, V., primary, De Lena, M., additional, Beltramo, S., additional, Ferrario, S., additional, Dellepiane, S., additional, Figliolini, F., additional, Bruno, S., additional, Biancone, L., additional, Segoloni, G. P., additional, Tetta, C., additional, Camussi, G., additional, Prasad, N., additional, Jaisawal, A., additional, Yadav, B., additional, Agarwal, V., additional, Tripathi, D., additional, Nunez-Lozano, R., additional, Quiros, Y., additional, Sanchez-Gonzalez, P., additional, Perez de Obanos, M. P., additional, Ruiz, J., additional, Lopez-Hernandez, F. J., additional, Lopez-Novoa, J. M., additional, Yang, J. W., additional, Kim, J. S., additional, Lee, J. Y., additional, Park, H. C., additional, Han, B. G., additional, Choi, S. O., additional, Matsuyama, M., additional, Yoshimura, R., additional, Hayama, T., additional, Chargui, J., additional, Touraine, J.-L., additional, Yoshimura, N., additional, Zanazzi, M., additional, Carta, P., additional, Caroti, L., additional, Antognoli, G., additional, Pinzani, P., additional, Salvianti, F., additional, Villari, D., additional, Minetti, E., additional, Genina, A., additional, Ismail, W., additional, Soliman, A., additional, Ucar, H., additional, Akbas, H. S., additional, Yilmaz, V. T., additional, Aktas, A., additional, Suleymanlar, G., additional, Yucel, G., additional, Cappuccilli, M. L., additional, La Manna, G., additional, Capelli, I., additional, Baraldi, O., additional, Cuna, V., additional, Battaglino, G., additional, Todeschini, P., additional, Feliciangeli, G., additional, Scolari, M. P., additional, Stefoni, S., additional, Loiacono, E., additional, Votta, B., additional, Amore, A., additional, Ranghino, A., additional, Camilla, R., additional, Peruzzi, L., additional, Donadio, M. E., additional, Serriello, I., additional, Gallo, R., additional, Puccinelli, M. P., additional, Coppo, R., additional, Sahin, G., additional, Meltem Akay, O., additional, Uslu, S., additional, Bal, C., additional, Ugur Yalcin, A., additional, Gulbas, Z., additional, and George, J., additional
- Published
- 2013
- Full Text
- View/download PDF
8. Prospective evaluation of RASSF1A cell-free DNA as a biomarker of pre-eclampsia
- Author
-
Laura Cremonesi, Luca Valsecchi, Francesca Salvianti, Mario Pazzagli, Silvia Galbiati, Maddalena Smid, Annalisa Inversetti, Massimo Candiani, Maurizio Ferrari, Pamela Pinzani, Salvianti, F, Inversetti, A, Smid, M, Valsecchi, L, Candiani, Massimo, Pazzagli, M, Cremonesi, L, Ferrari, Maurizio, Pinzani, P, and Galbiati, S.
- Subjects
Adult ,medicine.medical_specialty ,RASSF1A cell-free DNA ,Preeclampsia ,Pre-Eclampsia ,Predictive Value of Tests ,Pregnancy ,medicine ,Humans ,Promoter Regions, Genetic ,Fetus ,Eclampsia ,Predictive marker ,business.industry ,Obstetrics ,Tumor Suppressor Proteins ,Obstetrics and Gynecology ,Gestational age ,Biomarker, Maternal plasma ,DNA ,DNA Methylation ,medicine.disease ,Epidemiologic Studies ,Reproductive Medicine ,Cell-free fetal DNA ,Immunology ,Gestation ,Biomarker (medicine) ,Female ,business ,Biomarkers ,Developmental Biology - Abstract
Introduction This study aims to quantify total and fetal cell-free DNA (cfDNA) in maternal plasma at different gestational ages and to assess whether this could represent a reliable predictive marker of pre-eclampsia (PE) before clinical onset. Methods We performed a qPCR assay to compare the cfDNA concentration of hypermethylated and unmethylated RASSF1A promoter gene sequences in maternal plasma among 3 groups of pregnant women. These included 17 women with overt PE, 33 women at risk for the disease subsequently differentiated into 9 who developed PE and 24 who did not, and 73 controls. All women at risk were consecutively sampled throughout the whole gestation. Results Both total and fetal cfDNA had a good diagnostic performance in distinguishing patients with overt PE from healthy controls. When comparing women at risk who developed PE to women at risk who did not, the predictive capability was satisfactory at a gestational age ranging from 17 to 30 weeks. This allowed establishing within this time interval a cut-off value of 735 GE/ml for total cfDNA (87.5% sensitivity and 70.0% specificity), and a cut-off value of 7.49 GE/ml for fetal cfDNA (100% sensitivity and 50% specificity). cfDNA levels turned positive several weeks before the onset of the disease: from 2 to 18 weeks for total cfDNA and from 8 to 17 weeks for fetal cfDNA. Discussion The simultaneous use of total and fetal cfDNA would allow an accurate monitoring and prevention of PE development thus suggesting that RASSF1A could represent a potential biomarker of PE.
- Published
- 2015
9. Atypical Spitz tumors in patients younger than 18 years
- Author
-
Angela Rita Sementa, Claudio Gambini, Paola Collini, Maria Elena Errico, Milena Paglierani, Francesca Salvianti, Rebecca Senetta, Gabrina Tragni, Carlo Tomasini, Franco Rongioletti, Vittoria Donofrio, Maria Cristina Montesco, Andrea Ferrari, Daniela Massi, Renata Boldrini, Massi, D, Tomasini, C, Senetta, R, Paglierani, M, Salvianti, F, Errico, Me, Donofrio, V, Collini, P, Tragni, G, Sementa, Ar, Rongioletti, F, Boldrini, R, Ferrari, A, Gambini, C, and Montesco, Mc
- Subjects
Male ,medicine.medical_specialty ,Pathology ,Skin Neoplasms ,Adolescent ,medicine.medical_treatment ,skin neoplasms ,Sentinel lymph node ,Dermatology ,Epithelioid and Spindle Cell ,preschool ,male ,nevus epithelioid and spindle cell ,Nevus, Epithelioid and Spindle Cell ,Medicine ,Humans ,In patient ,humans ,Child ,Preschool ,fluorescence in situ hybridization ,Nevus ,Retrospective Studies ,child ,medicine.diagnostic_test ,business.industry ,Melanoma ,atypical Spitz tumor ,pediatric age ,sentinel lymph nodevSpitz nevus ,Child, Preschool ,Female ,Infant ,Retrospective cohort study ,Pediatric age ,medicine.disease ,Spitz nevus ,infant ,retrospective studies ,female ,adolescent ,Lymphadenectomy ,business ,Fluorescence in situ hybridization - Abstract
Background Diagnosis and proper management of atypical Spitz tumors in pediatric age are still controversial. Objective We sought to investigate the clinicopathological and molecular features of atypical Spitz tumors in patients aged 18 years or younger. Methods We performed a retrospective clinicopathological and fluorescence in situ hybridization study on 50 pediatric atypical Spitz tumors. Results Parameters that were significantly correlated with a diagnosis of atypical Spitz tumors over Spitz nevus included asymmetry, level IV/V, lack of maturation, solid growth, nuclear pleomorphism, high nuclear-cytoplasmic ratio, atypical and deep mitoses, and more than 6 mitoses/mm 2 . In the atypical Spitz tumors group, a significantly higher mitotic rate was observed in prepuberal age ( P = .04). The 4-probe fluorescence in situ hybridization melanoma assay did not discriminate atypical Spitz tumors from Spitz nevi. Heterozygous 9p21 loss was found in 3 of 37 cases and homozygous 9p21 loss in 2 of 37 cases. Only 1 child experienced a fatal outcome, showing genetic abnormalities by melanoma fluorescence in situ hybridization probe and a heterozygous 9p21 deletion. Limitations The limited number of adverse outcomes did not allow the prognostic analysis of single morphologic features. Conclusion Pediatric atypical Spitz tumors are associated with minimal lethal potential. Atypical Spitz tumors require complete excision and careful follow-up while our data do not support any clinical benefit for the sentinel lymph node biopsy procedure and completion lymphadenectomy.
- Published
- 2015
10. Atypical Spitzoid melanocytic tumors:A morphological, mutational, and FISH analysis
- Author
-
Antonio Maiorana, Milena Paglierani, Silvana Lukic, Marco Santucci, Vincenzo Canzonieri, Daniela Massi, Francesca Salvianti, Luigino Dal Maso, Carlo Tomasini, Anna Maria Cesinaro, Vincenzo De Giorgi, Claudio Orlando, Lisa Simi, Pamela Pinzani, Stefania Bettelli, Massi, D, Cesinaro, Am, Tomasini, C, Paglierani, M, Bettelli, S, Dal Maso, L, Simi, L, Salvianti, F, Pinzani, P, Orlando, C, De Giorgi, V, Lukic, S, Maiorana, A, Santucci, M, and Canzonieri, V
- Subjects
Adult ,Male ,Proto-Oncogene Proteins B-raf ,Pathology ,medicine.medical_specialty ,Skin Neoplasms ,atypical Spitz nevus, atypical Spitzoid tumor, BRAFV600E, fluorescence in situ hybridization ,Adolescent ,Sentinel lymph node ,Lymph node biopsy ,Dermoscopy ,Dermatology ,Gene mutation ,Ether ,Young Adult ,BRAFV600E ,Formaldehyde ,Nevus, Epithelioid and Spindle Cell ,Biopsy ,medicine ,Humans ,Child ,Lymph node ,fluorescence in situ hybridization ,In Situ Hybridization, Fluorescence ,Acetic Acid ,Chromatography ,Ethanol ,medicine.diagnostic_test ,business.industry ,Micrometastasis ,Infant ,atypical Spitzoid tumor ,Middle Aged ,atypical Spitz nevus ,Prognosis ,Immunohistochemistry ,Genes, ras ,medicine.anatomical_structure ,Child, Preschool ,Female ,business ,Fluorescence in situ hybridization ,Comparative genomic hybridization - Abstract
Background: Identification of the clinical behavior of atypical Spitzoid tumors with conflicting histopathologic features remains controversial. Objective: We sought to assess whether molecular findings may be helpful in the diagnostic and prognostic assessment of atypical Spitzoid tumors. Methods: A total of 38 controversial, atypical Spitzoid lesions (1 mm in thickness) were analyzed for clinicopathological features, chromosomal alterations by fluorescence in situ hybridization (FISH) analysis (RREB1/MYB/CCND1/CEP6), BRAF(V600E) mutation by allele-specific real-time polymerase chain reaction confirmed by sequencing, and H-RAS gene mutation by direct sequencing. Results: Atypical Spitzoid lesions developed in 21 female and 17 male patients (mean age 22 years). Nine patients underwent sentinel lymph node biopsy and a sentinel lymph node micrometastasis was detected in 4 of these 9 cases. Four additional patients, who did not receive a sentinel lymph node biopsy, experienced bulky lymph node metastases and one experienced visceral metastases and death. Lesions from patients with lymph node involvement showed more deep mitoses (P < .01), less inflammation = .05), and more plasma cells (P = .04). FISH analysis demonstrated the presence of chromosomal alterations in 6 of 25 cases. Correlation with follow-up data showed that the only case with fatal outcome showed multiple chromosomal alterations by FISH analysis. BRAF(V600E) mutation was detected in 12 of 16 cases (75%) and H-RAS mutation on exon 3 was found in 3 of 11 cases (27%). Limitations: Our results require validation in a larger series with longer follow-up information. Conclusions: FISH assay may be of help in the prognostic evaluation of atypical Spitzoid tumors. Diagnostic significance of BRAF(V600E) and H-RAS mutations in this setting remains unclear. (J Am Acad Dermatol 2011;64:919-35)
- Published
- 2011
11. Early changes in circulating tumor DNA (ctDNA) predict treatment response in metastatic KRAS-mutated colorectal cancer (mCRC) patients.
- Author
-
Lavacchi D, Gelmini S, Calabri A, Rossi G, Simi L, Caliman E, Mancini I, Salvianti F, Petroni G, Guidolin A, Scolari F, Messerini L, Pillozzi S, Pinzani P, and Antonuzzo L
- Abstract
The detection of RAS mutations and co-mutations in liquid biopsy offers a novel paradigm for the dynamic management of metastatic colorectal cancer (mCRC) patients. Expanding the results of the prospective OMITERC (OMIcs application from solid to liquid biopsy for a personalized ThERapy of Cancer) project, we collected blood samples at specific time points from patients who received a first-line chemotherapy (CT) for KRAS-mutated mCRC. CTC quantification was performed by CellSearch® system. Libraries from cfDNA were prepared using the Oncomine™ Colon cfDNA Assay to detect tumour-derived DNA in cfDNA. The analysis involved >240 hotspots in 14 genes. Twenty patients with KRAS-mutated mCRC treated at the Medical Oncology Unit of Careggi University Hospital were prospectively enrolled. Nine patients had available data for longitudinal monitoring of cfDNA. After 6 weeks of first-line CT an increase of KRAS-mutated clone was reported in the only patient who did not obtain disease control, while all patients with decrease of KRAS clones obtained disease control. Overall, in patients with a short (<9 months) progression-free survival (PFS) we registered, at 6 weeks, an increase in cfDNA levels and in KRAS mutations or other co-mutations, i.e. PIK3CA, FBXW7, GNAS, and TP53. In selected cases, co-mutations were able to better anticipate radiological progressive disease (PD) than the increase of KRAS-mutated clones. In conclusion, our study confirms plasma ctDNA as a crucial tool for anticipating PD at an early time point and highlights the value of a comprehensive assessment of clonal dynamics to improve the management of patients with mCRC., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Lorenzo Antonuzzo reports financial support was provided by 10.13039/501100009888Tuscany Region. Lorenzo Anronuzzo reports a relationship with 10.13039/501100009888Tuscany Region that includes: funding grants. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Published by Elsevier Ltd.)
- Published
- 2023
- Full Text
- View/download PDF
12. Safe and Successful Surgical Outcome in Persons with Hemophilia A with and without Inhibitors Treated with Emicizumab: A Large, Single Center, Real-World Experience.
- Author
-
Castaman G, Linari S, Pieri L, Carulli C, Prosperi P, Tonelli P, Demartis F, Fjerza R, Attanasio M, Coppo M, and Salvianti F
- Abstract
Emicizumab is a humanized recombinant bispecific antibody, bridging together activated factor IX (FIXa) and factor X (FX), thus mimicking the activity of FVIII in vivo. Emicizumab is designed for long-term prophylaxis in patients with severe hemophilia A with and without inhibitors. This approach provides constant protection, with significant reduction in bleeding rate and improved quality of life. However, protection provided by emicizumab is not absolute, and clotting factor concentrates (FVIII, rFVIIa, aPCC) may be necessary for post-traumatic bleeding or surgery, with a potential thrombotic risk or difficulty in preventing bleeding. Real world evidence is still scanty, especially for managing major surgery. In this study, 75 surgeries were managed in 28 patients (27 major procedures in 15 patients and 48 minor procedures in 20 patients. In 17 patients without inhibitors, 30 minor surgeries were carried out by using FVIII in 5, with only a bleeding event, which was successfully treated with FVIII concentrate. Six major surgeries were uneventfully performed with FVIII concentrate. Eleven PWHA and high-titer inhibitors underwent 39 surgical procedures (18 minor and 21 major surgeries). Minor surgeries were mostly performed without prophylaxis with rFVIIa, with only a single bleeding complication. All 21 major surgeries were covered with a homogeneous protocol using rFVIIa. In four instances, bleeding complications occurred, treated with rFVIIa. Of them, a single patient only failed to respond and died because of an uncontrollable bleeding from a large ruptured retroperitoneal pseudotumor. Surgery in patients with emicizumab can be safely carried out with the use of appropriate replacement therapy protocols.
- Published
- 2023
- Full Text
- View/download PDF
13. Circulating tumour cells and cell-free DNA as a prognostic factor in metastatic colorectal cancer: the OMITERC prospective study.
- Author
-
Salvianti F, Gelmini S, Mancini I, Pazzagli M, Pillozzi S, Giommoni E, Brugia M, Di Costanzo F, Galardi F, De Luca F, Castiglione F, Messerini L, Pinzani P, and Antonuzzo L
- Subjects
- Aged, Aged, 80 and over, Colorectal Neoplasms genetics, Disease Progression, Female, High-Throughput Nucleotide Sequencing, Humans, Longitudinal Studies, Male, Middle Aged, Mutation, Neoplasm Metastasis, Prognosis, Prospective Studies, Biomarkers, Tumor genetics, Cell-Free Nucleic Acids genetics, Colorectal Neoplasms pathology, Neoplastic Cells, Circulating pathology, Proto-Oncogene Proteins p21(ras) genetics, Sequence Analysis, DNA methods
- Abstract
Background: Within the OMITERC prospective study (OMIcs application from solid to liquid biopsy for a personalised ThERapy of Cancer), we explored the prognostic role of liquid biopsy encompassing cell-free DNA (cfDNA) and circulating tumour cells (CTCs) in KRAS mutated metastatic colorectal cancer (mCRC)., Methods: We defined a workflow including pre-analytical and analytical procedures collecting blood before therapy and every 3 months until disease progression (PD). CTCs were counted by CellSearch® and isolated by DEPArray™. NGS sequencing of CTCs and cfDNA was performed using a panel of cancer/CRC related genes respectively., Results: KRAS mutational status was mostly concordant between tumour tissues and liquid biopsy. The percentage of cfDNA samples with mutations in CRC driver genes was in line with literature. In longitudinal monitoring circulating biomarkers anticipated or overlapped conventional diagnostic tools in predicting PD. The presence of CTCs at baseline was confirmed a negative prognostic marker., Conclusions: Cell-free DNA and CTCs are readily available candidates for clinical application in mCRC. While CTCs demonstrated a prognostic significance at baseline, cfDNA was confirmed an easily accessible material for monitoring the mutational status of the tumour over time. Moreover, in the longitudinal study, the two markers emerged as complementary in assessing disease progression.
- Published
- 2021
- Full Text
- View/download PDF
14. Prognostic and Monitoring Value of Circulating Tumor Cells in Adrenocortical Carcinoma: A Preliminary Monocentric Study.
- Author
-
Cantini G, Canu L, Armignacco R, Salvianti F, De Filpo G, Ercolino T, Nesi G, Maggi M, Mannelli M, Pinzani P, and Luconi M
- Abstract
Adrenocortical carcinoma (ACC), a rare and aggressive neoplasia, presents poor prognosis when metastatic at diagnosis and limited therapies are available. Specific and sensitive markers for early diagnosis and a monitoring system of therapy and tumor evolution are urgently needed. The liquid biopsy represents a source of tumor material within a minimally invasive blood draw that allows the recovery of circulating tumor cells (CTCs). CTCs have been recently shown to be detectable in ACC. In the present paper, we evaluated the prognostic value of CTCs obtained by size-filtration in a small pilot cohort of 19 ACC patients. We found CTCs in 68% of pre-surgery and in 38% of post-surgery blood samples. In addition, CTC clusters (CTMs) and cancer associated macrophages (CAMLs) were detectable in some ACC patients. The median number of CTCs significantly decreased after the mass removal. Finally, stratifying patients in high and low pre-surgery CTC number groups, assuming the 75th percentile CTC value as cut-off, CTCs significantly predicted patients' overall survival (log rank = 0.005), also in a multivariate analysis adjusted for age and tumor stage. In conclusion, though preliminary and performed in a small cohort of patients, our study suggests that CTC number may represent a promising marker for prognosis and disease monitoring in ACC.
- Published
- 2020
- Full Text
- View/download PDF
15. Analytical Evaluation of an NGS Testing Method for Routine Molecular Diagnostics on Melanoma Formalin-Fixed, Paraffin-Embedded Tumor-Derived DNA.
- Author
-
Mancini I, Simi L, Salvianti F, Castiglione F, Sonnati G, and Pinzani P
- Abstract
Next Generation Sequencing (NGS) is a promising tool for the improvement of tumor molecular profiling in view of the identification of a personalized treatment in oncologic patients. To verify the potentiality of a targeted NGS (Ion AmpliSeq™ Cancer Hotspot Panel v2), selected melanoma samples ( n = 21) were retrospectively analyzed on S5 platform in order to compare NGS performance with the conventional techniques adopted in our routine clinical setting (Sequenom MassARRAY system, Sanger sequencing, allele-specific real-time PCR). The capability in the identification of rare and low-frequency mutations in the main genes involved in melanoma ( BRAF and NRAS genes) was verified and integrated with the results deriving from other oncogenes and tumor suppressor genes. The analytical evaluation was carried out by the analysis of DNA derived from control cell lines and FFPE (Formalin-Fixed, Paraffin-Embedded) samples to verify that the achieved resolution of uncommon mutations and low-frequency variants was suitable to meet the technical and clinical requests. Our results demonstrate that the amplicon-based NGS approach can reach the sensitivity proper of the allele-specific assays together with the high specificity of a sequencing method. An overall concordance among the tested methods was observed in the identification of classical and uncommon mutations. The assessment of the quality parameters and the comparison with the orthogonal methods suggest that the NGS method could be implemented in the clinical setting for melanoma molecular characterization.
- Published
- 2019
- Full Text
- View/download PDF
16. The diagnostic potential of mutation detection from single circulating tumor cells in cancer patients.
- Author
-
Salvianti F and Pinzani P
- Subjects
- DNA Mutational Analysis standards, Genomics methods, Genomics standards, Humans, Liquid Biopsy, Sensitivity and Specificity, Single-Cell Analysis standards, Whole Genome Sequencing, Biomarkers, Tumor, DNA Mutational Analysis methods, Mutation, Neoplasms diagnosis, Neoplasms genetics, Neoplastic Cells, Circulating pathology, Single-Cell Analysis methods
- Abstract
Introduction: Circulating tumor cells (CTCs) have gained importance in the oncology field as biomarkers of tumor development. The most relevant observation that emerged from the recent studies on CTCs is their heterogeneity, which can be investigated by new technologies for single cell analysis. Areas covered: This review considers the most recent advances (limited to the last two years) in the mutational analysis of single CTCs with a critical point of view on the technical challenges still to be faced and the steps needed to reach a standardization of the procedures able to translate these new approaches into clinical practice. Expert commentary: CTCs represent a surrogate tumor sample obtained by a minimally invasive procedure allowing the serial monitoring of the patient during the follow-up period or after treatment. Notwithstanding that, the analysis of CTCs is not so widespread; in fact, a limited number of centers can be equipped and possess the expertise for the development of workflows able to identify, enrich and isolate CTCs from blood. Moreover, the lack of standardized procedures and guidelines limits the study of CTCs to 'research use only' approaches.
- Published
- 2017
- Full Text
- View/download PDF
17. Circulating tumor cells and microemboli can differentiate malignant and benign pulmonary lesions.
- Author
-
Mascalchi M, Maddau C, Sali L, Bertelli E, Salvianti F, Zuccherelli S, Matucci M, Borgheresi A, Raspanti C, Lanzetta M, Falchini M, Mazza E, Vella A, Luconi M, Pinzani P, and Pazzagli M
- Abstract
The presence of circulating tumor cells (CTC) or microemboli (CTM) in the peripheral blood can theoretically anticipate malignancy of solid lesions in a variety of organs. We aimed to preliminarily assess this capability in patients with pulmonary lesions of suspected malignant nature. We used a cell-size filtration method (ScreenCell) and cytomorphometric criteria to detect CTC/CTM in a 3 mL sample of peripheral blood that was taken just before diagnostic percutaneous CT-guided fine needle aspiration (FNA) or core biopsy of the suspicious lung lesion. At least one CTC/CTM was found in 47 of 67 (70%) patients with final diagnoses of lung malignancy and in none of 8 patients with benign pulmonary nodules. In particular they were detected in 38 (69%) of 55 primary lung cancers and in 9 (75%) of 12 lung metastases from extra-pulmonary cancers. Sensitivity of CTC/CTM presence for malignancy was 70.1% (95%CI: 56.9-83.1%), specificity 100%, positive predictive value 100% and negative predictive value 28.6% (95%CI: 11.9-45.3%). Remarkably, the presence of CTC/CTM anticipated the diagnosis of primary lung cancer in 3 of 5 patients with non-diagnostic or inconclusive results of FNA or core biopsy, whereas CTC/CTM were not observed in 1 patient with sarcoidosis and 1 with amarthocondroma. These results suggest that presently, due to the low sensitivity, the search of CTC/CTM cannot replace CT guided percutaneous FNA or core biopsy in the diagnostic work-up of patients with suspicious malignant lung lesions. However, the high specificity may as yet indicate a role in cases with non-diagnostic or inconclusive FNA or core biopsy results that warrants to be further investigated., Competing Interests: Competing Interests: The authors have declared that no competing interest exists.
- Published
- 2017
- Full Text
- View/download PDF
18. New insights in the clinical and translational relevance of miR483-5p in adrenocortical cancer.
- Author
-
Salvianti F, Canu L, Poli G, Armignacco R, Scatena C, Cantini G, Di Franco A, Gelmini S, Ercolino T, Pazzagli M, Nesi G, Mannelli M, Pinzani P, and Luconi M
- Abstract
Adrenocortical cancer (ACC) is a rare aggressive malignancy. Recent ACC integrated genomics analysis contributed to redefine the risk groups on molecular basis, including tumor microRNAs (miRs), detectable also in the bloodstream. We developed a quantitative real-time (RT) assay for the measurement of miR483 and miR483-5p absolute levels in plasma samples. miR483/miR483-5p levels were evaluated in plasma samples of 27 patients with ACC before surgery and at follow-up. Statistically significant differences in miR483-5p and miR483 levels were found between stage 1/2 and stage 3/4 ACCs in pre-surgery and post-surgery samples. ROC curve analysis of miR483-5p levels gave a prediction of the clinical stage (accuracy 0.917±0.084), with the best cut-off value of 0.221 ng/ml, prognosticating overall and recurrence-free survival. In a multivariate Cox analysis (HR 16.2, 95%CI[1.39-188.6, P<0.026]), miR483-5p was the only variable that significantly predicted recurrence, but not overall survival. In addition, miR483 and miR483-5p levels correlated with the number of circulating tumor cells (CTCs) detected in the same blood samples, independently of the timing of sampling. In conclusion, we demonstrated that miR483-5p absolute plasma levels in ACC patients are powerful molecular markers that may help in the follow-up of patients after surgery and chemotherapy, and contribute to more accurately classify and predict tumor progression., Competing Interests: CONFLICTS OF INTEREST The authors declare no conflicts of interest
- Published
- 2017
- Full Text
- View/download PDF
19. Integrity and Quantity of Total Cell-Free DNA in the Diagnosis of Thyroid Cancer: Correlation with Cytological Classification.
- Author
-
Salvianti F, Giuliani C, Petrone L, Mancini I, Vezzosi V, Pupilli C, and Pinzani P
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Case-Control Studies, DNA Fragmentation, Female, Humans, Liquid Biopsy, Male, Middle Aged, ROC Curve, Reproducibility of Results, Sensitivity and Specificity, Thyroid Neoplasms pathology, Thyroid Neoplasms surgery, Young Adult, Biomarkers, Tumor, Cell-Free Nucleic Acids, DNA, Neoplasm, Thyroid Neoplasms diagnosis, Thyroid Neoplasms genetics
- Abstract
Cell-free DNA (cfDNA) quantity and quality in plasma has been investigated as a non-invasive biomarker in cancer. Previous studies have demonstrated increased cfDNA amount and length in different types of cancer with respect to healthy controls. The present study aims to test the hypothesis that the presence of longer DNA strands circulating in plasma can be considered a biomarker for tumor presence in thyroid cancer. We adopted a quantitative real-time PCR (qPCR) approach based on the quantification of two amplicons of different length (67 and 180 bp respectively) to evaluate the integrity index 180/67. Cell-free DNA quantity and integrity were higher in patients affected by nodular thyroid diseases than in healthy controls. Importantly, cfDNA integrity index was higher in patients with cytological diagnosis of thyroid carcinoma (Thy4/Thy5) than in subjects with benign nodules (Thy2). Therefore, cfDNA integrity index 180/67 is a suitable parameter for monitoring cfDNA fragmentation in thyroid cancer patients and a promising circulating biomarker in the diagnosis of thyroid nodules., Competing Interests: The authors declare no conflict of interest.
- Published
- 2017
- Full Text
- View/download PDF
20. Mutational analysis of single circulating tumor cells by next generation sequencing in metastatic breast cancer.
- Author
-
De Luca F, Rotunno G, Salvianti F, Galardi F, Pestrin M, Gabellini S, Simi L, Mancini I, Vannucchi AM, Pazzagli M, Di Leo A, and Pinzani P
- Subjects
- Breast Neoplasms secondary, Female, Humans, Prognosis, Single-Cell Analysis, Survival Rate, Tumor Suppressor Protein p53 genetics, Biomarkers, Tumor genetics, Breast Neoplasms genetics, DNA Mutational Analysis methods, High-Throughput Nucleotide Sequencing methods, Mutation, Neoplastic Cells, Circulating pathology
- Abstract
Circulating Tumor Cells (CTCs) represent a "liquid biopsy" of the tumor potentially allowing real-time monitoring of cancer biology and therapies in individual patients.The purpose of the study was to explore the applicability of a protocol for the molecular characterization of single CTCs by Next Generation Sequencing (NGS) in order to investigate cell heterogeneity and provide a tool for a personalized medicine approach.CTCs were enriched and enumerated by CellSearch in blood from four metastatic breast cancer patients and singularly isolated by DEPArray. Upon whole genome amplification 3-5 single CTCs per patient were analyzed by NGS for 50 cancer-related genes.We found 51 sequence variants in 25 genes. We observed inter- and intra-patient heterogeneity in the mutational status of CTCs.The highest number of somatic deleterious mutations was found in the gene TP53, whose mutation is associated with adverse prognosis in breast cancer.The discordance between the mutational status of the primary tumor and CTCs observed in 3 patients suggests that, in advanced stages of cancer, CTC characteristics are more closely linked to the dynamic modifications of the disease status.In one patient the mutational profiles of CTCs before and during treatment shared only few sequence variants.This study supports the applicability of a non-invasive approach based on the liquid biopsy in metastatic breast cancer patients which, in perspective, should allow investigating the clonal evolution of the tumor for the development of new therapeutic strategies in precision medicine., Competing Interests: The authors declare no potential conflicts of interest.
- Published
- 2016
- Full Text
- View/download PDF
21. Tumor-Related Methylated Cell-Free DNA and Circulating Tumor Cells in Melanoma.
- Author
-
Salvianti F, Orlando C, Massi D, De Giorgi V, Grazzini M, Pazzagli M, and Pinzani P
- Abstract
Solid tumor release into the circulation cell-free DNA (cfDNA) and circulating tumor cells (CTCs) which represent promising biomarkers for cancer diagnosis. Circulating tumor DNA may be studied in plasma from cancer patients by detecting tumor specific alterations, such as genetic or epigenetic modifications. Ras association domain family 1 isoform A (RASSF1A) is a tumor suppressor gene silenced by promoter hypermethylation in a variety of human cancers including melanoma. The aim of the present study was to assess the diagnostic performance of a tumor-related methylated cfDNA marker in melanoma patients and to compare this parameter with the presence of CTCs. RASSF1A promoter methylation was quantified in cfDNA by qPCR in a consecutive series of 84 melanoma patients and 68 healthy controls. In a subset of 68 cases, the presence of CTCs was assessed by a filtration method (Isolation by Size of Epithelial Tumor Cells, ISET) as well as by an indirect method based on the detection of tyrosinase mRNA by RT-qPCR. The distribution of RASSF1A methylated cfDNA was investigated in cases and controls and the predictive capability of this parameter was assessed by means of the area under the ROC curve (AUC). The percentage of cases with methylated RASSF1A promoter in cfDNA was significantly higher in each class of melanoma patients (in situ, invasive and metastatic) than in healthy subjects (Pearson chi-squared test, p < 0.001). The concentration of RASSF1A methylated cfDNA in the subjects with a detectable quantity of methylated alleles was significantly higher in melanoma patients than in controls. The biomarker showed a good predictive capability (in terms of AUC) in discriminating between melanoma patients and healthy controls. This epigenetic marker associated to cfDNA did not show a significant correlation with the presence of CTCs, but, when the two parameters are jointly considered, we obtain a higher sensitivity of the detection of positive cases in invasive and metastatic melanomas. Our data suggest that cell-free tumor DNA and CTCs represent two complementary aspects of the liquid biopsy which may improve the diagnosis and the clinical management of melanoma patients.
- Published
- 2016
- Full Text
- View/download PDF
22. Single circulating tumor cell sequencing as an advanced tool in cancer management.
- Author
-
Salvianti F, Pazzagli M, and Pinzani P
- Subjects
- Biopsy, Humans, Mutation, Neoplasm Metastasis, Neoplasms genetics, Neoplasms pathology, Neoplastic Cells, Circulating metabolism, Neoplasms blood, Neoplastic Cells, Circulating pathology, Sequence Analysis, DNA, Single-Cell Analysis
- Abstract
Circulating tumor cells (CTCs) shed by the primary tumor and metastases are considered a real-time 'liquid biopsy', reflecting the disease complexity that evolves during progression, showing in its late stages different genetic, epigenetic and expression features. Consequently, heterogeneity and development of characteristic features upon disease progression are the two main goals that emerging technologies should account for in view of a clinical application. Single-cell analysis, now possible due to technological advances, may help elucidate tumor heterogeneity at the CTC level. This review focuses on the necessary steps for the analysis of CTCs at the single-cell level. A concise overview is given on the alternative methods referring in particular to studies on the mutational status of single CTCs from cancer patients.
- Published
- 2016
- Full Text
- View/download PDF
23. Feasibility of a workflow for the molecular characterization of single cells by next generation sequencing.
- Author
-
Salvianti F, Rotunno G, Galardi F, De Luca F, Pestrin M, Vannucchi AM, Di Leo A, Pazzagli M, and Pinzani P
- Abstract
The purpose of the study was to explore the feasibility of a protocol for the isolation and molecular characterization of single circulating tumor cells (CTCs) from cancer patients using a single-cell next generation sequencing (NGS) approach. To reach this goal we used as a model an artificial sample obtained by spiking a breast cancer cell line (MDA-MB-231) into the blood of a healthy donor. Tumor cells were enriched and enumerated by CellSearch(®) and subsequently isolated by DEPArray™ to obtain single or pooled pure samples to be submitted to the analysis of the mutational status of multiple genes involved in cancer. Upon whole genome amplification, samples were analysed by NGS on the Ion Torrent PGM™ system (Life Technologies) using the Ion AmpliSeq™ Cancer Hotspot Panel v2 (Life Technologies), designed to investigate genomic "hot spot" regions of 50 oncogenes and tumor suppressor genes. We successfully sequenced five single cells, a pool of 5 cells and DNA from a cellular pellet of the same cell line with a mean depth of the sequencing reaction ranging from 1581 to 3479 reads. We found 27 sequence variants in 18 genes, 15 of which already reported in the COSMIC or dbSNP databases. We confirmed the presence of two somatic mutations, in the BRAF and TP53 gene, which had been already reported for this cells line, but also found new mutations and single nucleotide polymorphisms. Three variants were common to all the analysed samples, while 18 were present only in a single cell suggesting a high heterogeneity within the same cell line. This paper presents an optimized workflow for the molecular characterization of multiple genes in single cells by NGS. The described pipeline can be easily transferred to the study of single CTCs from oncologic patients.
- Published
- 2015
- Full Text
- View/download PDF
24. Heterogeneity of PIK3CA mutational status at the single cell level in circulating tumor cells from metastatic breast cancer patients.
- Author
-
Pestrin M, Salvianti F, Galardi F, De Luca F, Turner N, Malorni L, Pazzagli M, Di Leo A, and Pinzani P
- Subjects
- Adult, Aged, Base Sequence, Breast Neoplasms pathology, Cell Line, Tumor, Class I Phosphatidylinositol 3-Kinases, DNA Mutational Analysis, Female, Humans, Middle Aged, Molecular Sequence Data, Neoplasm Metastasis, Reproducibility of Results, Breast Neoplasms enzymology, Breast Neoplasms genetics, Genetic Heterogeneity, Neoplastic Cells, Circulating pathology, Phosphatidylinositol 3-Kinases genetics, Single-Cell Analysis
- Abstract
Circulating Tumor Cells (CTCs) represent a "liquid biopsy of the tumor" which might allow real-time monitoring of cancer biology and therapies in individual patients. CTCs are extremely rare in the blood stream and their analysis is technically challenging. The CellSearch(®) system provides the enumeration of CTCs with prognostic significance in patients with metastatic breast cancer (mBC), but it does not allow their molecular characterization, which might be useful to identify therapeutically relevant targets for individualized treatment. Combining the CellSearch(®) and DEPArray™ technologies allows the recovery of single CTCs as a pure sample for molecular analysis. The purpose of the study was to investigate the heterogeneity of PIK3CA mutational status within single CTCs isolated from individual mBC patients. CTCs were enriched and enumerated by CellSearch(®) in blood samples collected from 39 mBC patients. In 20 out of 39 patients enriched samples with ≥5 CTCs were sorted using DEParray™ to isolate single CTCs or pools of CTCs to be submitted to Whole Genome Amplification (WGA) before sequencing analysis. In 18 out of 20 patients, it was possible to perform PIK3CA sequencing on exons 9 and 20. Twelve subjects were wild type (wt) for the PIK3CA gene. PIK3CA status could also be assessed in pools of CTCs in seven of these patients, with consistent wt status found. Six patients (33%) had a PIK3CA mutation identified. In 2 of the six patients, molecular heterogeneity was detected when mutational analysis was performed on more than one single CTC, including one patient with loss of heterozygosity on both single and pooled CTCs, and one patient with three different PIK3CA variants on single CTCs but PIK3CA wt status on pooled CTC samples. In six out of the 18 cases PIK3CA status was also evaluable on a primary tumor sample. In one of the six cases a discordance in PIK3CA status between the primary (wild-type) and the matched CTC (exon 20 mutation) was observed. This study demonstrates the feasibility of a non-invasive approach based on the liquid biopsy in mBC patients. Moreover, our data suggest the importance of characterizing CTCs at the single cell level in order to investigate the molecular heterogeneity within cells from the same patient., (Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
25. Circulating tumor cells detection and counting in uveal melanomas by a filtration-based method.
- Author
-
Mazzini C, Pinzani P, Salvianti F, Scatena C, Paglierani M, Ucci F, Pazzagli M, and Massi D
- Abstract
Uveal melanoma is one of the most deadly diseases in ophthalmology for which markers able to predict the appearance of metastasis are needed. The study investigates the role of circulating tumor cells (CTC) as a prognostic factor in this disease. We report the detection of circulating tumor cells by Isolation by Size of Epithelial Tumor cells (ISET) in a cohort of 31 uveal melanoma patients: we identified single CTCs or clusters of cells in 17 patients, while the control population, subjects with choroidal nevi, showed no CTC in peripheral blood. The presence of CTCs did not correlate with any clinical and pathological parameter, such as tumor larger basal diameter (LBD), tumor height and TNM. By stratifying patients in groups on the basis of the number of CTC (lower or higher than 10 CTC per 10 mL blood) and the presence of CTC clusters we found a significant difference in LBD (p = 0.019), Tumor height (p = 0.048), disease-free and overall survival (p < 0.05). In conclusion, we confirm the role of CTC as a negative prognostic marker in uveal melanoma patients after a long follow-up period. Further characterization of CTC will help understanding uveal melanoma metastasization and improve patient management.
- Published
- 2014
- Full Text
- View/download PDF
26. Detection of circulating tumor cells in patients with adrenocortical carcinoma: a monocentric preliminary study.
- Author
-
Pinzani P, Scatena C, Salvianti F, Corsini E, Canu L, Poli G, Paglierani M, Piccini V, Pazzagli M, Nesi G, Mannelli M, and Luconi M
- Subjects
- Adult, Disease Progression, Female, Humans, Male, Middle Aged, Prognosis, Adrenal Cortex Neoplasms pathology, Adrenocortical Carcinoma pathology, Neoplastic Cells, Circulating pathology
- Abstract
Context: Adrenocortical carcinoma (ACC) is a rare malignancy, the prognosis of which is mainly dependent on stage at diagnosis. The identification of disease-associated markers for early diagnosis and drug monitoring is mandatory. Circulating tumor cells (CTCs) are released into the bloodstream from primary tumor/metastasis. CTC detection in blood samples may have enormous potential for assisting in the diagnosis of malignancy, estimating prognosis, and monitoring the disease., Objective: The aim of the study was to investigate the presence of CTCs in blood samples of patients with ACC or benign adrenocortical adenoma (ACA)., Setting: We conducted the study at a university hospital., Intervention: CTC analysis was performed in blood samples from 14 ACC patients and 10 ACA patients. CTCs were isolated on the basis of cell size by filtration through ScreenCell devices, followed by identification according to validated morphometric criteria and immunocytochemistry., Main Outcome Measure: We measured the difference in CTC detection between ACC and ACA., Results: CTCs were detected in all ACC samples, but not in ACA samples. Immunocytochemistry confirmed the adrenocortical origin. When ACC patients were stratified according to the median value of tumor diameter and metastatic condition, a statistically significant difference was found in the number of CTCs detected after surgery. A significant correlation between the number of CTCs in postsurgical samples and clinical parameters was found for tumor diameter alone., Conclusions: Our findings provide the first evidence for adrenocortical tumors that CTCs may represent a useful marker to support differential diagnosis between ACC and ACA. The correlation with some clinical parameters suggests a possible relevance of CTC analysis for prognosis and noninvasive monitoring of disease progression and drug response.
- Published
- 2013
- Full Text
- View/download PDF
27. Multiparametric analysis of cell-free DNA in melanoma patients.
- Author
-
Salvianti F, Pinzani P, Verderio P, Ciniselli CM, Massi D, De Giorgi V, Grazzini M, Pazzagli M, and Orlando C
- Subjects
- Adult, Aged, Aged, 80 and over, Biomarkers, Tumor blood, Biomarkers, Tumor genetics, DNA Methylation, DNA, Neoplasm blood, Female, Humans, Male, Melanoma diagnosis, Melanoma genetics, Middle Aged, Neoplasm Staging, ROC Curve, Skin Neoplasms diagnosis, Skin Neoplasms genetics, Young Adult, DNA blood, Melanoma blood, Skin Neoplasms blood
- Abstract
Cell-free DNA in blood (cfDNA) represents a promising biomarker for cancer diagnosis. Total cfDNA concentration showed a scarce discriminatory power between patients and controls. A higher specificity in cancer diagnosis can be achieved by detecting tumor specific alterations in cfDNA, such as DNA integrity, genetic and epigenetic modifications.The aim of the present study was to identify a sequential multi-marker panel in cfDNA able to increase the predictive capability in the diagnosis of cutaneous melanoma in comparison with each single marker alone. To this purpose, we tested total cfDNA concentration, cfDNA integrity, BRAF(V600E) mutation and RASSF1A promoter methylation associated to cfDNA in a series of 76 melanoma patients and 63 healthy controls. The chosen biomarkers were assayed in cfDNA samples by qPCR. Comparison of biomarkers distribution in cases and controls was performed by a logistic regression model in both univariate and multivariate analysis. The predictive capability of each logistic model was investigated by means of the area under the ROC curve (AUC). To aid the reader to interpret the value of the AUC, values between 0.6 and 0.7, between 0.71 and 0.8 and greater than 0.8 were considered as indicating a weak predictive, satisfactory and good predictive capacity, respectively. The AUC value for each biomarker (univariate logistic model) was weak/satisfactory ranging between 0.64 (BRAF(V600E)) to 0.85 (total cfDNA). A good overall predictive capability for the final logistic model was found with an AUC of 0.95. The highest predictive capability was given by total cfDNA (AUC:0.86) followed by integrity index 180/67 (AUC:0.90) and methylated RASSF1A (AUC:0.89).An approach based on the simultaneous determination of three biomarkers (total cfDNA, integrity index 180/67 and methylated RASSF1A) could improve the diagnostic performance in melanoma.
- Published
- 2012
- Full Text
- View/download PDF
28. Application of a filtration- and isolation-by-size technique for the detection of circulating tumor cells in cutaneous melanoma.
- Author
-
De Giorgi V, Pinzani P, Salvianti F, Panelos J, Paglierani M, Janowska A, Grazzini M, Wechsler J, Orlando C, Santucci M, Lotti T, Pazzagli M, and Massi D
- Subjects
- Adult, Aged, Aged, 80 and over, Cell Line, Tumor, Cell Size, Female, Filtration methods, Humans, Immunohistochemistry, Male, Middle Aged, Monophenol Monooxygenase genetics, Neoplasm Metastasis pathology, Nevus, Pigmented pathology, Prognosis, RNA, Messenger blood, Reverse Transcriptase Polymerase Chain Reaction, Sensitivity and Specificity, Cell Separation methods, Melanoma secondary, Neoplastic Cells, Circulating pathology, Skin Neoplasms pathology
- Abstract
Analysis of circulating tumor cells (CTC) in the peripheral blood of cutaneous melanoma patients provides information on the metastatic process and potentially improves patient management. The isolation by size of epithelial tumor cells (ISET) is a direct method for CTC identification in which tumor cells are collected by filtration as a result of their large size. So far, ISET has been applied only to CTC detection from epithelial cancer patients, and the technique has never been applied to cutaneous melanoma patients. We herein investigated the presence of CTC by ISET in the peripheral blood of 140 subjects (87 with cutaneous melanomas, 10 subjects undergoing surgery for melanocytic nevi, 5 patients with non-melanoma skin tumors, and 38 healthy volunteers). The identification of the cells trapped in filters as CTC was supported by positivity for immunohistochemical markers and for tyrosinase mRNA by real-time RT-PCR. CTC were neither detected in the controls nor in the in situ melanoma group. In contrast, CTC were shown in 29% of patients with primary invasive melanoma and in 62.5% of metastatic melanoma patients (P<0.01). CTC detection correlated with the presence of mRNA tyrosinase in blood samples, assayed by real-time RT-PCR (P=0.001). CTC detection corroborated by suitable molecular characterization may assist in the identification and monitoring of more appropriate therapies in melanoma patients.
- Published
- 2010
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.