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Article ; Online: Digital Droplet PCR in Hematologic Malignancies

Sara Galimberti / Serena Balducci / Francesca Guerrini / Marzia Del Re / Rossella Cacciola

Diagnostics, Vol 12, Iss 1305, p

A New Useful Molecular Tool

2022 Volume 1305

Abstract: Digital droplet PCR (ddPCR) is a recent version of quantitative PCR (QT-PCR), useful for measuring gene expression, doing clonality assays and detecting hot spot mutations. In respect of QT-PCR, ddPCR is more sensitive, does not need any reference curve ... ...

Abstract	Digital droplet PCR (ddPCR) is a recent version of quantitative PCR (QT-PCR), useful for measuring gene expression, doing clonality assays and detecting hot spot mutations. In respect of QT-PCR, ddPCR is more sensitive, does not need any reference curve and can quantify one quarter of samples already defined as “positive but not quantifiable”. In the IgH and TCR clonality assessment, ddPCR recapitulates the allele-specific oligonucleotide PCR (ASO-PCR), being not adapt for detecting clonal evolution, that, on the contrary, does not represent a pitfall for the next generation sequencing (NGS) technique. Differently from NGS, ddPCR is not able to sequence the whole gene, but it is useful, cheaper, and less time-consuming when hot spot mutations are the targets, such as occurs with IDH1 , IDH2 , NPM1 in acute leukemias or T315I mutation in Philadelphia-positive leukemias or JAK2 in chronic myeloproliferative neoplasms. Further versions of ddPCR, that combine different primers/probes fluorescences and concentrations, allow measuring up to four targets in the same PCR reaction, sparing material, time, and money. ddPCR is also useful for quantitating BCR-ABL1 fusion gene, WT1 expression, donor chimerism, and minimal residual disease, so helping physicians to realize that “patient-tailored therapy” that is the aim of the modern hematology.
Keywords	digital PCR ; quantitative PCR ; multiplexing PCR ; MRD ; clonality ; NGS ; Medicine (General) ; R5-920
Language	English
Publishing date	2022-05-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Relapsed/Refractory Chronic Lymphocytic Leukemia Patients Treated with Fixed Duration Venetoclax-Rituximab

Edoardo Benedetti / Claudia Baratè / Fabrizio Mavilia / Emilia Bramanti / Riccardo Morganti / Valentina Guerri / Giulia Cervetti / Enrico Capochiani / Ilaria Bertaggia / Salvatore Massimo Stella / Ginevra Traverso / Benedetto Bruno / Sara Galimberti

Journal of Clinical Medicine, Vol 12, Iss 1772, p

Assessment of Response with Ultrasound, and Relationship with Minimal Residual Disease

2023 Volume 1772

Abstract: A fixed duration of venetoclax-rituximab (VenR) resulted in a significant benefit of both PFS and in the attainment of an undetectable minimal residual disease (uMRD) compared with bendamustine-rituximab in relapsed/refractory (R/R) chronic lymphocytic ... ...

Abstract	A fixed duration of venetoclax-rituximab (VenR) resulted in a significant benefit of both PFS and in the attainment of an undetectable minimal residual disease (uMRD) compared with bendamustine-rituximab in relapsed/refractory (R/R) chronic lymphocytic leukemia (CLL) patients. The 2018 International Workshop on CLL guidelines, outside the context of clinical trials, suggested ultrasonography (US) as a possible imaging technique to evaluate visceral involvement, and palpation to evaluate superficial lymph nodes (SupLNs). In this real-life study we prospectively enrolled N = 22 patients. Patients were assessed by US, to determine nodal and splenic response in R/R CLL patients treated with a fixed duration VenR. We found an overall response rate, complete remission, partial remission, and stable disease, of 95.4%, 68%, 27.3%, and 4.5%, respectively. Responses were also correlated with risk categories. The time to response, and the time to clearance of the disease in the spleen, in abdominal LN (AbdLNs), and in SupLNs were discussed. Responses were independent from LN size. The correlation between response rate with MRD were also investigated. US allowed to detect a substantial CR rate correlated with uMRD.
Keywords	chronic lymphocytic leukemia ; ultrasound sonography ; venetoclax-rituximab ; Medicine ; R
Subject code	610
Language	English
Publishing date	2023-02-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Precision Medicine in Lymphoma by Innovative Instrumental Platforms

Antonello Di Paolo / Elena Arrigoni / Giacomo Luci / Federico Cucchiara / Romano Danesi / Sara Galimberti

Frontiers in Oncology, Vol

2019 Volume 9

Abstract: In recent years, many efforts have been addressed to the growing field of precision medicine in order to offer individual treatments to every patient on the basis of his/her genetic background. Formerly adopted to achieve new disease classifications as ... ...

Abstract	In recent years, many efforts have been addressed to the growing field of precision medicine in order to offer individual treatments to every patient on the basis of his/her genetic background. Formerly adopted to achieve new disease classifications as it is still done, innovative platforms, such as microarrays, genome-wide association studies (GWAS), and next generation sequencing (NGS), have made the progress in pharmacogenetics faster and cheaper than previously expected. Several studies in lymphoma patients have demonstrated that these platforms can be used to identify biomarkers predictive of drug efficacy and tolerability, discovering new possible druggable proteins. Indeed, GWAS and NGS allow the investigation of the human genome, finding interesting associations with putative or unexpected targets, which in turns may represent new therapeutic possibilities. Importantly, some objective difficulties have initially hampered the translation of findings in clinical routines, such as the poor quantity/quality of genetic material or the paucity of targets that could be investigated at the same time. At present, some of these technical issues have been partially solved. Furthermore, these analyses are growing in parallel with the development of bioinformatics and its capabilities to manage and analyze big data. Because of pharmacogenetic markers may become important during drug development, regulatory authorities (i.e., EMA, FDA) are preparing ad hoc guidelines and recommendations to include the evaluation of genetic markers in clinical trials. Concerns and difficulties for the adoption of genetic testing in routine are still present, as well as affordability, reliability and the poor confidence of some patients for these tests. However, genetic testing based on predictive markers may offers many advantages to caregivers and patients and their introduction in clinical routine is justified.
Keywords	precision medicine ; pharmacogenetics ; lymphoma ; innovative platforms ; microarray ; GWAS ; Neoplasms. Tumors. Oncology. Including cancer and carcinogens ; RC254-282
Subject code	300
Language	English
Publishing date	2019-12-01T00:00:00Z
Publisher	Frontiers Media S.A.
Document type	Article ; Online
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Article ; Online: Repeated SARS-CoV-2 vaccination in cancer patients treated with immune checkpoint inhibitors: induction of high-avidity anti-RBD neutralizing antibodies.

Caruso, Teresita / Salani, Francesca / Catanese, Silvia / Pratesi, Federico / Mercinelli, Chiara / Motta, Giuseppe / Genovesi, Virginia / Bonato, Adele / Sara, Galimberti / Masi, Gianluca / Migliorini, Paola

International journal of clinical oncology

2023 Volume 28, Issue 3, Page(s) 363–369

Abstract: Background: Cancer patients are more vulnerable to COVID-19 and are thus given high priority in vaccination campaigns. In solid cancer patients treated with checkpoint inhibitors, we evaluated the amount of anti-RBD and neutralizing antibodies and ... ...

Abstract	Background: Cancer patients are more vulnerable to COVID-19 and are thus given high priority in vaccination campaigns. In solid cancer patients treated with checkpoint inhibitors, we evaluated the amount of anti-RBD and neutralizing antibodies and antibody avidity after two or three doses of the vaccine. Methods: Thirty-eight solid cancer patients, 15 untreated hematological patients and 21 healthy subjects were enrolled in the study. Blood was collected before the first dose (T0), 21 days after the second (T2) and in 18 solid cancer patients also 15 days after the third dose of vaccine (T3). IgG, IgM and IgA anti-RBD antibodies were detected by ELISA. Neutralizing antibodies were measured testing the inhibition of RBD binding to ACE2. Antibody avidity was evaluated in 18 patients by a urea avidity ELISA. Results: IgG anti-RBD antibodies were produced in 65.8% of the cancer patients at T2, and in 60% of hematological patients at levels lower than healthy controls. IgM and IgA anti-RBD antibodies were also produced in 5.3% and 21% cancer patients, respectively. At T3, a significant increase in anti-RBD IgG levels was observed. Neutralizing antibodies were produced in 68.4% of cancer patients as compared with 93% of untreated hematological patients and 100% of controls, at titers lower than in healthy subjects. At T3, neutralizing antibodies and avidity of IgG anti-RBD increased; 6/18 patients negative at T2 developed neutralizing antibodies at T3. Conclusion: The data indicate that in cancer patients mRNA vaccine induces high avidity anti-RBD antibodies and neutralizing antibodies that increase after the third dose. The process of induction and selection of high-affinity antibodies is apparently unaffected by the treatment with anti-PD-1 or anti-PD-L1 antibodies.
MeSH term(s)	Humans ; Immune Checkpoint Inhibitors ; COVID-19 Vaccines/therapeutic use ; SARS-CoV-2 ; COVID-19/prevention & control ; Vaccination ; Antibodies, Neutralizing ; Immunoglobulin A ; Immunoglobulin G ; Immunoglobulin M ; Antibodies, Viral ; Neoplasms/drug therapy
Chemical Substances	Immune Checkpoint Inhibitors ; COVID-19 Vaccines ; Antibodies, Neutralizing ; Immunoglobulin A ; Immunoglobulin G ; Immunoglobulin M ; Antibodies, Viral
Language	English
Publishing date	2023-01-23
Publishing country	Japan
Document type	Journal Article
ZDB-ID	1400227-9
ISSN	1437-7772 ; 1341-9625
ISSN (online)	1437-7772
ISSN	1341-9625
DOI	10.1007/s10147-023-02295-0
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Article ; Online: Correction to: Repeated SARS-CoV-2 vaccination in cancer patients treated with immune checkpoint inhibitors: induction of high-avidity anti-RBD neutralizing antibodies.

International journal of clinical oncology

2023 Volume 28, Issue 3, Page(s) 370

Language	English
Publishing date	2023-02-18
Publishing country	Japan
Document type	Published Erratum
ZDB-ID	1400227-9
ISSN	1437-7772 ; 1341-9625
ISSN (online)	1437-7772
ISSN	1341-9625
DOI	10.1007/s10147-023-02310-4
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Article ; Online: Chronic Myeloid Leukemia and Pregnancy

Elisabetta Abruzzese / Stefano Aureli / Francesco Bondanini / Mariavita Ciccarone / Elisabetta Cortis / Antonello Di Paolo / Cristina Fabiani / Sara Galimberti / Michele Malagola / Alessandra Malato / Bruno Martino / Malgorzata Monika Trawinska / Domenico Russo / Paolo de Fabritiis

Journal of Clinical Medicine, Vol 11, Iss 1801, p

When Dreams Meet Reality. State of the Art, Management and Outcome of 41 Cases, Nilotinib Placental Transfer

2022 Volume 1801

Abstract: The overwhelming success of tyrosine kinase inhibitor (TKI) therapy in chronic myeloid leukemia (CML) patients has opened a discussion among medical practitioners and the lay public on the real possibility of pregnancy and conception in females and males ...

Abstract	The overwhelming success of tyrosine kinase inhibitor (TKI) therapy in chronic myeloid leukemia (CML) patients has opened a discussion among medical practitioners and the lay public on the real possibility of pregnancy and conception in females and males with CML. In the past 10 years this subject has acquired growing interest in the scientific community and specific knowledge has been obtained “from bench to bedside”. Embryological, pharmacological, and pathophysiological studies have merged with worldwide patient databases to provide a roadmap to a successful pregnancy and birth in CML patients. Male conception does not seem to be affected by TKI therapy, since this class of drugs is neither genotoxic nor mutagenic, however, caution should be used specially with newer drugs for which little or no data are available. In contrast, female patients should avoid TKI therapy specifically during the embryonic stage of organogenesis (5–12 weeks) because TKIs can be teratogenic. In the last 15 years, 41 pregnancies have been followed in our center. A total of 11 male conceptions and 30 female pregnancies are described. TKI treatment was generally terminated as soon as the pregnancy was discovered (3–5 weeks), to avoid exposure during embryonic period and to reduce the risk of needing treatment in the first trimester. Eleven pregnancies were treated with interferon, imatinib or nilotinib during gestation. Nilotinib plasma levels in cord blood and maternal blood at delivery were studied in 2 patients and reduced or absent placental crossing of nilotinib was observed. All of the patients were managed by a multidisciplinary team of physicians with obligatory hematological and obgyn consultations. This work provides an update on the state of the art and detailed description of pregnancy management and outcomes in CML patients.
Keywords	CML ; pregnancy ; placental transfer ; TKIs ; PEG-IFN ; conception ; Medicine ; R
Subject code	610
Language	English
Publishing date	2022-03-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: A Prospective Cross-Sectional Study on the Comparison of Ultrasound Assessment vs. Palpation in Chronic Lymphocytic Leukemia Patients in the Era of Targeted Therapy

Edoardo Benedetti / Fabrizio Mavilia / Claudia Baratè / Emilia Bramanti / Riccardo Morganti / Giulia Cervetti / Enrico Capochiani / Benedetto Bruno / Matteo Pelosini / Salvatore Massimo Stella / Sara Galimberti

Journal of Clinical Medicine, Vol 11, Iss 3206, p

2022 Volume 3206

Abstract: Background. In IWCLL guidelines, progressive splenomegaly and lymphadenopathy are signs of active disease. In this study, we have tested the hypotheses if US could be a reliable tool for both superficial lymphnodes (SupLNs) and splenic assessment in ... ...

Abstract	Background. In IWCLL guidelines, progressive splenomegaly and lymphadenopathy are signs of active disease. In this study, we have tested the hypotheses if US could be a reliable tool for both superficial lymphnodes (SupLNs) and splenic assessment in chronic lymphocytic leukemia (CLL) patients. Methods. We enrolled N = 75 patients. SupLN and the spleen were assessed by two independent physicians (M1 and M2) by palpation and by a third physician (M3) with ultrasound sonography (US) using two different sonographers (US1 and US2). The results of M1 vs. M2 assessment, US1 vs. US2, palpation vs. US were compared. The echostructure of N = 1037 SupLN and of the spleen was also investigated. Results. The dimensions of SupLNs assessed by MD1 vs. MD2 were statistically discordant. Splenic size was concordant. There was concordance between US1 and US2 SupLN and splenic assessment. US found a higher number of pathological SupLN (Cohen’s Kappa < 0.1) than palpation, which misses remarkable-sized SupLNs. LN echostructure and splenic involvement patterns were described. Conclusions. US is a reliable, radiation-free tool useful in clinical practice to assess SupLN and splenic involvement in CLL.
Keywords	lymph node ; ultrasound sonosgraphy ; spleen ; chronic lymphocytic leukemia ; Medicine ; R
Subject code	610
Language	English
Publishing date	2022-06-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article ; Online: Real-life Diagnostic and Therapeutic Approach to CLL

Monica Bocchia / Alberto Bosi / Enrico Capochiani / Stefania Ciolli / Romano Danesi / Sara Galimberti / Alessandro Gozzetti / Sabrina Moretti / Ubaldo Occhini / Mario Petrini

Farmeconomia: Health Economics and Therapeutic Pathways, Vol 21, Iss

A New Proposal from an Expert Panel in Tuscany Region

2020 Volume 1

Abstract: BACKGROUND: In the last years genomic and somatic alterations have shown to play a pivotal role in the pathogenesis of chronic lymphocytic leukemia (CLL) and new prognostic factors have been identified accordingly. AIM: To describe a real-life diagnostic ...

Abstract	BACKGROUND: In the last years genomic and somatic alterations have shown to play a pivotal role in the pathogenesis of chronic lymphocytic leukemia (CLL) and new prognostic factors have been identified accordingly. AIM: To describe a real-life diagnostic and therapeutic approach to CLL that takes into account the role of genomic and somatic prognostic factors in the risk stratification of developing progressive disease, and treatment decision. METHODS: This new proposal has been developed and validated by ten key opinion leaders from Tuscany Region during two Expert Meetings. The approach suggested comes from their experience in daily clinical practice and is supported by guidelines recommendations, clinical trials results, and drugs prescribing conditions in Italy. RESULTS: Beside TP53 deletion or mutated status, the Expert Panel highlighted the importance of the IGHV mutation status characterization, since the diagnosis, in order to identify patients who will have a more aggressive progression. Furthermore, just before starting treatment, to obtain useful prognostic information and indication in the selection of the therapy, they recommend cytogenetic analysis for the detection of del(11q), trisomy 12, del(13q), del(17p), conventional karyotyping of stimulated CLL cells, TP53 sequencing, and molecular genetic analysis to detect IGHV mutation status. CONCLUSIONS: The Expert Panel recognized the limitations associated with traditional staging systems in identifying patients who will have a more aggressive disease course and predicting response to treatment and suggested a real-life diagnostic and therapeutic approach to CLL to update the current patient management in light of recent advances that have improved understanding of CLL.
Keywords	chronic lymphocytic leukemia ; prognostic factors ; tp53 deletion ; ighv mutation status ; Medicine (General) ; R5-920
Subject code	610
Language	English
Publishing date	2020-02-01T00:00:00Z
Publisher	SEEd Medical Publishers
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Proteasome Inhibitors as a Possible Therapy for SARS-CoV-2

Lucia Longhitano / Daniele Tibullo / Cesarina Giallongo / Giacomo Lazzarino / Nicola Tartaglia / Sara Galimberti / Giovanni Li Volti / Giuseppe Alberto Palumbo / Arcangelo Liso

International Journal of Molecular Sciences, Vol 21, Iss 3622, p

2020 Volume 3622

Abstract: The COVID-19 global pandemic is caused by SARS-CoV-2, and represents an urgent medical and social issue. Unfortunately, there is still not a single proven effective drug available, and therefore, current therapeutic guidelines recommend supportive care ... ...

Abstract	The COVID-19 global pandemic is caused by SARS-CoV-2, and represents an urgent medical and social issue. Unfortunately, there is still not a single proven effective drug available, and therefore, current therapeutic guidelines recommend supportive care including oxygen administration and treatment with antibiotics. Recently, patients have been also treated with off-label therapies which comprise antiretrovirals, anti-inflammatory compounds, antiparasitic agents and plasma from convalescent patients, all with controversial results. The ubiquitin–proteasome system (UPS) is important for the maintenance of cellular homeostasis, and plays a pivotal role in viral replication processes. In this review, we discuss several aspects of the UPS and the effects of its inhibition with particular regard to the life cycle of the coronaviruses (CoVs). In fact, proteasome inhibition by various chemical compounds, such as MG132, epoxomycin and bortezomib, may reduce the virus entry into the eucariotic cell, the synthesis of RNA, and the subsequent protein expression necessary for CoVs. Importantly, since UPS inhibitors reduce the cytokine storm associated with various inflammatory conditions, it is reasonable to assume that they might be repurposed for SARS-CoV-2, thus providing an additional tool to counteract both virus replication as well as its most deleterious consequences triggered by abnormal immunological response.
Keywords	SARS-CoV-2 ; proteasome inhibitors ; endoplasmic stress ; UPR response ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999 ; covid19
Subject code	616
Language	English
Publishing date	2020-05-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article ; Online: Defining TCRγδ lymphoproliferative disorders by combined immunophenotypic and molecular evaluation

Antonella Teramo / Andrea Binatti / Elena Ciabatti / Gianluca Schiavoni / Giulia Tarrini / Gregorio Barilà / Giulia Calabretto / Cristina Vicenzetto / Vanessa Rebecca Gasparini / Monica Facco / Iacopo Petrini / Roberto Grossi / Nadia Pisanti / Stefania Bortoluzzi / Brunangelo Falini / Enrico Tiacci / Sara Galimberti / Gianpietro Semenzato / Renato Zambello

Nature Communications, Vol 13, Iss 1, Pp 1-

2022 Volume 11

Abstract: Tγδ large granular lymphocyte leukemia (Tγδ LGLL) is a rare lymphoproliferative neoplasm characterized by the expansion of T large granular lymphocytes expressing γδ TCR. Here, based on deep sequencing analysis of the clonotype repertoire, the authors ... ...

Abstract	Tγδ large granular lymphocyte leukemia (Tγδ LGLL) is a rare lymphoproliferative neoplasm characterized by the expansion of T large granular lymphocytes expressing γδ TCR. Here, based on deep sequencing analysis of the clonotype repertoire, the authors show that leukemic Tγδ cells are characterized by recurrent public clonotypes that are diversified between symptomatic and asymptomatic patients.
Keywords	Science ; Q
Language	English
Publishing date	2022-06-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
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