المؤلفون: Beatrice Fontana, Giulia Gallerani, Irene Salamon, Ilaria Pace, Roberta Roncarati, Manuela Ferracin

المصدر: Frontiers in Oncology, Vol 13 (2023)

مصطلحات موضوعية: ARID1A, tumor suppressor gene, oncogene, solid tumors, SWI/SNF complex, synthetic lethality, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: ARID1A belongs to a class of chromatin regulatory proteins that function by maintaining accessibility at most promoters and enhancers, thereby regulating gene expression. The high frequency of ARID1A alterations in human cancers has highlighted its significance in tumorigenesis. The precise role of ARID1A in cancer is highly variable since ARID1A alterations can have a tumor suppressive or oncogenic role, depending on the tumor type and context. ARID1A is mutated in about 10% of all tumor types including endometrial, bladder, gastric, liver, biliopancreatic cancer, some ovarian cancer subtypes, and the extremely aggressive cancers of unknown primary. Its loss is generally associated with disease progression more often than onset. In some cancers, ARID1A loss is associated with worse prognostic features, thus supporting a major tumor suppressive role. However, some exceptions have been reported. Thus, the association of ARID1A genetic alterations with patient prognosis is controversial. However, ARID1A loss of function is considered conducive for the use of inhibitory drugs which are based on synthetic lethality mechanisms. In this review we summarize the current knowledge on the role of ARID1A as tumor suppressor or oncogene in different tumor types and discuss the strategies for treating ARID1A mutated cancers.

وصف الملف: electronic resource

العلاقة: https://www.frontiersin.org/articles/10.3389/fonc.2023.1136248/fullTest; https://doaj.org/toc/2234-943XTest

الوصول الحر: https://doaj.org/article/045e62a029d341bdbf028d9de0a72e38Test

المؤلفون: Tania Rossi, Davide Angeli, Giovanni Martinelli, Francesco Fabbri, Giulia Gallerani

المصدر: Frontiers in Genetics, Vol 13 (2022)

مصطلحات موضوعية: RNA-sequencing, NGS, single cell analysis, rare cells, deparray, gene expression, Genetics, QH426-470

الوصف: Combining phenotypical and molecular characterization of rare cells is challenging due to their scarcity and difficult handling. In oncology, circulating tumor cells (CTCs) are considered among the most important rare cell populations. Their phenotypic and molecular characterization is necessary to define the molecular mechanisms underlying their metastatic potential. Several approaches that require cell fixation make difficult downstream molecular investigations on RNA. Conversely, the DEPArray technology allows phenotypic analysis and handling of both fixed and unfixed cells, enabling a wider range of applications. Here, we describe an experimental workflow that allows the transcriptomic investigation of single and pooled OE33 cells undergone to DEPArray analysis and recovery. In addition, cells were tested at different conditions (unfixed, CellSearch fixative (CSF)- and ethanol (EtOH)-fixed cells). In a forward-looking perspective, this workflow will pave the way for novel strategies to characterize gene expression profiles of rare cells, both single-cell and low-resolution input.

وصف الملف: electronic resource

العلاقة: https://www.frontiersin.org/articles/10.3389/fgene.2022.1012191/fullTest; https://doaj.org/toc/1664-8021Test

الوصول الحر: https://doaj.org/article/f735916f23a7440e8cff7e6d89efc77dTest

المؤلفون: Ivan Vannini, Milena Urbini, Mattia Melloni, Tania Rossi, Giulia Gallerani, Michela Palleschi, Irene Azzali, Maurizio Puccetti, Giovanni Martinelli, Francesco Fabbri

المصدر: Frontiers in Medicine, Vol 9 (2022)

مصطلحات موضوعية: metaplastic breast cancer, extracellular vesicles, next generation sequencing (NGS), plasma, metastasis, Medicine (General), R5-920

الوصف: Metaplastic breast cancer (MpBC) is a rare tumor representing 1% of all breast malignancies. The prognosis of this histologic subtype is actually poor and there are no current clear-cut therapeutic guidelines. Hence, despite its uniqueness, its aggressive prognostic profile strongly encourages further studies to identify new markers and therapeutic targets. Herein, we report a case of 32-years-old patient affected with of triple negative spindle-shaped MpBC. The research of molecular targets on the primary tumor did not allow performing an effective therapeutic choice. Extracellular Vesicles (EVs) are under intense study as new potential pathophysiological markers and targets for therapeutic applications, in different tumors for their role in tumor onset, progression and aggressiveness. Here, we examined the involvement of EVs in this case, to look into the MpBC microenvironment willing to identify new potential molecular targets, pathways of aggressiveness, and markers of prognosis and therapeutic efficacy. Firstly, we characterized MpBC patient EV dimensions and surface proteins. Moreover, we analyzed the EV RNA cargo supposed to be delivered to nearby and distant recipient cells. Interestingly, we observed a dysregulation EV-contained miRNAs, which could determine an increased expression of oncogenes in the tumor microenvironment, probably enabling cancer progression. These data suggest that the characterization of miRNA cargo of EVs could be important for the identification of new markers and for the application of future new target therapies.

وصف الملف: electronic resource

العلاقة: https://www.frontiersin.org/articles/10.3389/fmed.2022.827206/fullTest; https://doaj.org/toc/2296-858XTest

الوصول الحر: https://doaj.org/article/740785c21b1b424f876aa70650354903Test

المؤلفون: Erika Bandini, Tania Rossi, Emanuela Scarpi, Giulia Gallerani, Ivan Vannini, Samanta Salvi, Irene Azzali, Mattia Melloni, Sara Salucci, Michela Battistelli, Patrizia Serra, Roberta Maltoni, William C. Cho, Francesco Fabbri

المصدر: Frontiers in Molecular Biosciences, Vol 8 (2021)

مصطلحات موضوعية: breast cancer, extracellular vesicles, plasma, liquid biopsy, biomarkers, Biology (General), QH301-705.5

الوصف: Breast cancer (BC) is the most commonly diagnosed malignant tumor in women worldwide, and the leading cause of cancer death in the female population. The percentage of patients experiencing poor prognosis along with the risk of developing metastasis remains high, also affecting the resistance to current main therapies. Cancer progression and metastatic development are no longer due entirely to their intrinsic characteristics, but also regulated by signals derived from cells of the tumor microenvironment. Extracellular vesicles (EVs) packed with DNA, RNA, and proteins, are the most attractive targets for both diagnostic and therapeutic applications, and represent a decisive challenge as liquid biopsy-based markers. Here we performed a study based on a multiplexed phenotyping flow cytometric approach to characterize BC-derived EVs from BC patients and cell lines, through the detection of multiple antigens. Our data reveal the expression of EVs-related biomarkers derived from BC patient plasma and cell line supernatants, suggesting that EVs could be exploited for characterizing and monitoring disease progression.

وصف الملف: electronic resource

العلاقة: https://www.frontiersin.org/articles/10.3389/fmolb.2021.732900/fullTest; https://doaj.org/toc/2296-889XTest

الوصول الحر: https://doaj.org/article/a2883136b93b4a32a4c6096fa762fe88Test

المؤلفون: Tania Rossi, Davide Angeli, Michela Tebaldi, Pietro Fici, Elisabetta Rossi, Andrea Rocca, Michela Palleschi, Roberta Maltoni, Giovanni Martinelli, Francesco Fabbri, Giulia Gallerani

المصدر: Cancers, Vol 14, Iss 16, p 3925 (2022)

مصطلحات موضوعية: metastatic breast cancer, circulating tumor cells, copy number aberrations, single nucleotide variants, single cell sequencing, next generation sequencing, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Circulating tumor cells’ (CTCs) heterogeneity contributes to counteract their introduction in clinical practice. Through single-cell sequencing we aim at exploring CTC heterogeneity in metastatic breast cancer (MBC) patients. Single CTCs were isolated using DEPArray NxT. After whole genome amplification, libraries were prepared for copy number aberration (CNA) and single nucleotide variant (SNV) analysis and sequenced using Ion GeneStudio S5 and Illumina MiSeq, respectively. CTCs demonstrate distinctive mutational signatures but retain molecular traces of their common origin. CNA profiling identifies frequent aberrations involving critical genes in pathogenesis: gains of 1q (CCND1) and 11q (WNT3A), loss of 22q (CHEK2). The longitudinal single-CTC analysis allows tracking of clonal selection and the emergence of resistance-associated aberrations, such as gain of a region in 12q (CDK4). A group composed of CTCs from different patients sharing common traits emerges. Further analyses identify losses of 15q and enrichment of terms associated with pseudopodium formation as frequent and exclusive events. CTCs from MBC patients are heterogeneous, especially concerning their mutational status. The single-cell analysis allows the identification of aberrations associated with resistance, and is a candidate tool to better address treatment strategy. The translational significance of the group populated by similar CTCs should be elucidated.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2072-6694/14/16/3925Test; https://doaj.org/toc/2072-6694Test

الوصول الحر: https://doaj.org/article/6b267d453eb0428997c3b1fb92b67a08Test

المؤلفون: Tania Rossi, Michela Palleschi, Davide Angeli, Michela Tebaldi, Giovanni Martinelli, Ivan Vannini, Maurizio Puccetti, Francesco Limarzi, Roberta Maltoni, Giulia Gallerani, Francesco Fabbri

المصدر: Frontiers in Medicine, Vol 8 (2021)

مصطلحات موضوعية: metaplastic breast cancer, circulating tumor cells, next generation sequencing, copy number aberration, metastasis, liquid biopsy, Medicine (General), R5-920

الوصف: Circulating tumor cells (CTCs) are a rare population of cells found in the bloodstream and represent key players in the metastatic cascade. Their analysis has proved to provide further core information concerning the tumor. Herein, we aim at investigating CTCs isolated from a 32-year-old patient diagnosed with triple negative spindle-shaped metaplastic breast cancer (MpBC), a rare tumor poorly responsive to therapies and with a dismal prognosis. The molecular analysis performed on the primary tumor failed to underline effective actionable targets to address the therapeutic strategy. Besides the presence of round-shaped CTCs, cells with a spindle shape were present as well, and through molecular analysis, we confirmed their malignant nature. This aspect was coherent with the primary tumor histology, proving that CTCs are released regardless of their morphology. Copy number aberration (CNA) profiling and variant analysis using next-generation sequencing (NGS) showed that these cells did not harbor the alterations exhibited by the primary tumor (PIK3CA G1049A mutation, MYC copy number gain). However, despite the great heterogeneity observed, the amplification of regions involved in metastasis emerged (8q24.22–8q24.23). Our findings support the investigation of CTCs to identify alterations that could have a role in the metastatic process. To the best of our knowledge, this is the first examination of CTCs in an MpBC patient.

وصف الملف: electronic resource

العلاقة: https://www.frontiersin.org/articles/10.3389/fmed.2021.689895/fullTest; https://doaj.org/toc/2296-858XTest

الوصول الحر: https://doaj.org/article/6f8eb4e4161c4b86b5ae70f408d550adTest

المؤلفون: Alberto Visioli, Fabrizio Giani, Nadia Trivieri, Riccardo Pracella, Elide Miccinilli, Maria Grazia Cariglia, Orazio Palumbo, Andrea Arleo, Fabio Dezi, Massimiliano Copetti, Laura Cajola, Silvia Restelli, Valerio Papa, Antonio Sciuto, Tiziana Pia Latiano, Massimo Carella, Dino Amadori, Giulia Gallerani, Riccardo Ricci, Sergio Alfieri, Graziano Pesole, Angelo L. Vescovi, Elena Binda

المصدر: EBioMedicine, Vol 44, Iss , Pp 346-360 (2019)

مصطلحات موضوعية: Medicine, Medicine (General), R5-920

الوصف: Background: Despite their lethality and ensuing clinical and therapeutic relevance, circulating tumor cells (CTCs) from colorectal carcinoma (CRC) remain elusive, poorly characterized biological entities. Methods and findings: We perfected a cell system of stable, primary lines from human CRC showing that they possess the full complement of ex- and in-vivo, in xenogeneic models, characteristics of CRC stem cells (CCSCs). Here we show how tumor-initiating, CCSCs cells can establish faithful orthotopic phenocopies of the original disease, which contain cells that spread into the circulatory system. While in the vascular bed, these cells retain stemness, thus qualifying as circulating CCSCs (cCCSCs). This is followed by the establishment of lesions in distant organs, which also contain resident metastatic CCSCs (mCCSCs). Interpretation: Our results support the concept that throughout all the stages of CRC, stemness is retained as a continuous property by some of their tumor cells. Importantly, we describe a useful standardized model that can enable isolation and stable perpetuation of human CRC's CCSCs, cCCSCs and mCCSCs, providing a useful platform for studies of CRC initiation and progression that is suitable for the discovery of reliable stage-specific biomarkers and the refinement of new patient-tailored therapies. Fund: This work was financially supported by grants from “Ministero della Salute Italiano”(GR-2011-02351534, RC1703IC36 and RC1803IC35) to Elena Binda and from “Associazione Italiana Cancro” (IG-14368) Angelo L. Vescovi. None of the above funders have any role in study design, data collection, data analysis, interpretation, writing the project. Keywords: Human colorectal carcinoma (CRC), Stemness, CRC stem cells, CRC circulating stem cells, CRC metastatic stem cells, CRC biology and biomarkers, Anti-CRC SCs strategies

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S2352396419302889Test; https://doaj.org/toc/2352-3964Test

الوصول الحر: https://doaj.org/article/1f869219eef84cd3b7742e7ad0525533Test

المؤلفون: Giulia Gallerani, Tania Rossi, Martina Valgiusti, Davide Angeli, Pietro Fici, Sara De Fanti, Erika Bandini, Claudia Cocchi, Giovanni Luca Frassineti, Massimiliano Bonafè, Francesco Fabbri

المصدر: Cancers, Vol 13, Iss 24, p 6369 (2021)

مصطلحات موضوعية: circulating tumor cells, single cell analysis, CNA profiling, esophageal cancer, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Background: Here, we monitored the evolution of CTCs spread in 11 patients affected by locally advanced EC who were undergoing therapy. Methods: In this perspective study, we designed multiple blood biopsies from individual patients: before and after neoadjuvant chemo-radio therapy and after surgery. We developed a multi-target array, named Grab-all assay, to estimate CTCs for their epithelial (EpCAM/E-Cadherin/Cytokeratins) and mesenchymal/stem (N-Cadherin/CD44v6/ABCG2) phenotypes. Identified CTCs were isolated as single cells by DEPArray, subjected to whole genome amplification, and copy number aberration (CNA) profiles were determined. Through bioinformatic analysis, we assessed the genomic imbalance of single CTCs, investigated specific focal copy number changes previously reported in EC and aberrant pathways using enrichment analysis. Results: Longitudinal monitoring allowed the identification of CTCs in at least one time-point per patient. Through single cell CNA analysis, we revealed that CTCs showed significantly dynamic genomic imbalance during treatment. Individual CTCs from relapsed patients displayed a higher degree of genomic imbalance relative to disease-free patients’ groups. Genomic aberrations previously reported in EC occurred mostly in post-neoadjuvant therapy CTCs. In-depth analysis showed that networks enrichment in all time-point CTCs were inherent to innate immune system. Transcription/gene regulation, post-transcriptional and epigenetic modifications were uniquely affected in CTCs of relapsed patients. Conclusions: Our data add clues to the comprehension of the role of CTCs in EC aggressiveness: chromosomal aberrations on genes related to innate immune system behave as relevant to the onset of CTC-status, whilst pathways of transcription/gene regulation, post-transcriptional and epigenetic modifications seem linked to patients’ outcome.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2072-6694/13/24/6369Test; https://doaj.org/toc/2072-6694Test

الوصول الحر: https://doaj.org/article/58677ef45a964cd98d70233308bcbd85Test

المؤلفون: Tania Rossi, Giulia Gallerani, Giovanni Martinelli, Roberta Maltoni, Francesco Fabbri

المصدر: Biomedicines, Vol 9, Iss 9, p 1242 (2021)

مصطلحات موضوعية: breast cancer, liquid biopsy, circulating tumor cells, heterogeneity, Biology (General), QH301-705.5

الوصف: Breast cancer (BC) is a disease characterized by high degrees of heterogeneity at morphologic, genomic, and genetic levels, even within the same tumor mass or among patients. As a consequence, different subpopulations coexist and less represented clones may have a selective advantage, significantly influencing the outcome of BC patients. Circulating tumor cells (CTCs) represent a rare population of cells with a crucial role in metastatic cascade, and in recent years have represented a fascinating alternative to overcome the heterogeneity issue as a “liquid biopsy”. However, besides the raw enumeration of these cells in advanced epithelial tumors, there are no CTC-based assays applied in the clinical practice to improve personalized medicine. In this review, we report the latest findings in the field of CTCs for intra-tumoral heterogeneity unmasking in BC, supporting the need to deepen their analysis to investigate their role in metastatic process and include the molecular characterization in the clinical practice. In the future, CTCs will be helpful in monitoring patients during treatment, as well as to better address therapeutic strategies.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2227-9059/9/9/1242Test; https://doaj.org/toc/2227-9059Test

الوصول الحر: https://doaj.org/article/7c8355c6a07b469cbcdd1bbabfb6ba81Test

المؤلفون: Tania Rossi, Giulia Gallerani, Davide Angeli, Claudia Cocchi, Erika Bandini, Pietro Fici, Michele Gaudio, Giovanni Martinelli, Andrea Rocca, Roberta Maltoni, Francesco Fabbri

المصدر: Cancers, Vol 12, Iss 9, p 2490 (2020)

مصطلحات موضوعية: CTCs, breast cancer, single-cell analysis, CNA, NGS, liquid biopsy, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Circulating tumor cells (CTCs) are a rare population of cells representing a key player in the metastatic cascade. They are recognized as a validated tool for the identification of patients with a higher risk of relapse, including those diagnosed with breast cancer (BC). However, CTCs are characterized by high levels of heterogeneity that also involve copy number alterations (CNAs), structural variations associated with gene dosage changes. In this study, single CTCs were isolated from the peripheral blood of 11 early-stage BC patients at different time points. A label-free enrichment of CTCs was performed using OncoQuick, and single CTCs were isolated using DEPArray. Libraries were prepared from single CTCs and DNA extracted from matched tumor tissues for a whole-genome low-coverage next-generation sequencing (NGS) analysis using the Ion Torrent S5 System. The analysis of the CNA burden highlighted that CTCs had different degrees of aberration based on the time point and subtype. CTCs were found even six months after surgery and shared CNAs with matched tumor tissue. Tumor-associated CNAs that were recurrent in CTCs were patient-specific, and some alterations involved regions associated with BC and survival (i.e., gains at 1q21-23 and 5p15.33). The enrichment analysis emphasized the involvement of aberrations of terms, associated in particular with interferon (IFN) signaling. Collectively, our findings reveal that these aberrations may contribute to understanding the molecular mechanisms involving CTC-related processes and their survival ability in occult niches, supporting the goal of exploiting their application in patients’ surveillance and follow-up.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2072-6694/12/9/2490Test; https://doaj.org/toc/2072-6694Test

الوصول الحر: https://doaj.org/article/95779f9640ee4c7cbddfabbdf3e99a06Test