Papers by Lucia Anna Muscarella
PLOS ONE, Dec 5, 2013
Hyperparathyroidism Jaw-Tumour Syndrome (HPT-JT) is characterized by primary hyperparathyroidism ... more Hyperparathyroidism Jaw-Tumour Syndrome (HPT-JT) is characterized by primary hyperparathyroidism (PHPT), maxillary/mandible ossifying fibromas and by parathyroid carcinoma in 15% of cases. Inactivating mutations of the tumour suppressor CDC73/HRPT2 gene have been found in HPT-JT patients and also as genetic determinants of sporadic parathyroid carcinoma/atypical adenomas and, rarely, typical adenomas, in familial PHPT. Here we report the genetic and molecular analysis of the CDC73/HRPT2 gene in three patients affected by PHPT due to atypical and typical parathyroid adenomas, in one case belonging to familial PHPT. Flag-tagged WT and mutant CDC73/ HRPT2 proteins were transiently transfected in HEK293 cells and functional assays were performed in order to investigate the effect of the variants on the whole protein expression, nuclear localization and cell overgrowth induction. We identified four CDC73/HRPT2 gene mutations, three germline (c.679_680delAG, p.Val85_Val86del and p.Glu81_Pro84del), one somatic (p.Arg77Pro). In three cases the mutation was located within the Nucleolar Localisation Signals (NoLS). The three NoLS variants led to instability either of the corresponding mutated protein or mRNA or both. When transfected in HEK293 cells, NoLS mutated proteins mislocalized with a predeliction for cytoplasmic or nucleo-cytoplasmic localization and, finally, they resulted in overgrowth, consistent with a dominant negative interfering effect in the presence of the endogenous protein.
Annals of Oncology, Apr 1, 2019
Advances in Anatomic Pathology, May 1, 2018
Less than 250 extraneuraxial hemangioblastomas occurring in paraneuraxial or peripheral sites hav... more Less than 250 extraneuraxial hemangioblastomas occurring in paraneuraxial or peripheral sites have been reported to date, sporadically or in the setting of von Hippel-Lindau disease. Seventeen such cases underwent molecular genetic analysis, using either the patient's peripheral blood in 9 cases or paraffin embedded tumor tissue in the rest. VHL gene mutations were documented in 3/9 cases in which DNA from peripheral blood lymphocytes was used, all with clinically manifest von Hippel-Lindau disease; instead, no VHL gene alterations were found in all of the 8 cases with sporadic extraneuraxial hemangioblastoma in which DNA from tumor tissue was analyzed. Our aim is to investigate the molecular genetic profile of the VHL gene in extraneuraxial hemangioblastoma using paraffin embedded tumor tissues. The clinical features, histopathology, and molecular investigations of 10 extraneuraxial hemangioblastomas (7 females, 3 males; median age: 47 years) are presented herein. The histopathologic diagnosis was supported by immunohistochemistry (10/10) and electron microscopy (4/10). Molecular genetic analysis was conducted (10/ 10) for VHL gene mutations, LOH, and gene promoter methylation. Two of the present cases were already published with only limited or no molecular investigations. Four tumors of the present series were paraneuraxial, and 6 peripheral (2 involved soft tissues, and 4 the kidney). One tumor was von Hippel-Lindau disease-associated, 1 was classified as "hemangioblastoma-only VHLD", 7 were sporadic, and one was unknown. All were histopathologically analogous to their counterpart located inside the central nervous system. Immunophenotypically, all tumors expressed vimentin, S-100, NSE, and alpha-inhibin (10/10). Ultrastructurally, unbound lipid droplets filled the cytoplasms of the stromal cells. Molecular analysis revealed 3 inactivating mutations (1 germline, two somatic) in the coding sequence of the VHL gene in 2 different extraneuraxial hemangioblastomas, and LOH in 4 (two as a double hit), all non-renal extraneuraxial hemangioblastomas. Methylation analysis failed to disclose promoter methylation in any case. In conclusion, we report eight new cases from the wide category of extraneuraxial hemangioblastomas (4 paraneuraxial, and 4 renal), one of which was von Hippel-Lindau disease-associated and 7 sporadic. VHL gene alterations were found not only in the von Hippel-Lindau disease-associated tumor, butfor the first timealso in 3 sporadic ones, two of which with novel mutations.
European Journal of Cancer, Oct 1, 2022
Oxidative Medicine and Cellular Longevity, 2018
Oxidative and electrophilic changes in cells are mainly coordinated by the KEAP1/NRF2 (Kelch-like... more Oxidative and electrophilic changes in cells are mainly coordinated by the KEAP1/NRF2 (Kelch-like erythroid-derived cap-n-collar homology-(ECH-) associated protein-1/nuclear factor (erythroid-derived 2)-like 2) axis. The physical interaction between these two proteins promotes the expression of several antioxidant defense genes in response to exogenous and endogenous insults. Recent studies demonstrated that KEAP1/NRF2 axis dysfunction is also strongly related to tumor progression and chemo-and radiotherapy resistance of cancer cells. In solid tumors, the KEAP1/NRF2 system is constitutively activated by the loss of KEAP1 or gain of NFE2L2 functions that leads to its nuclear accumulation and enhances the transcription of many cytoprotective genes. In addition to point mutations, epigenetic abnormalities, as aberrant promoter methylation, and microRNA (miRNA) and long noncoding RNA (lncRNA) deregulation were reported as emerging mechanisms of KEAP1/NRF2 axis modulation. This review will summarize the current knowledge about the epigenetic mechanisms that deregulate the KEAP1/NRF2 cascade in solid tumors and their potential usefulness as prognostic and predictive molecular markers.
Epigenetics, Mar 1, 2011
2-related factor 2), which plays a pivotal role in the cellular response to oxidative stress. Und... more 2-related factor 2), which plays a pivotal role in the cellular response to oxidative stress. Under basal conditions, Nrf2 is retained in the cytoplasm by the binding with Keap1 and it is maintained at a reduced level by the Keap1-dependent ubiquitination and proteasomal degradation systems. On exposure to oxidative stress, Keap1-dependent ubiquitin ligase activity is inhibited and Nrf2 can translocate to the nucleus, where it binds to consensus sequences on the target genes. Several evidences suggest that Keap1/Nrf2 system not only protects normal cells from carcinogenetic agents, but also promotes the survival of transformed cells under deleterious conditions. Initial reports uncovered mutations in the KEAP1 gene in non-small cell lung cancer (NSCLC) leading to a permanent Nrf2 activation. Mutations were often associated with loss of heterozygosity at the KEAP1 gene locus suggesting that biallelic inactivation is a common event in NSCLC. Somatic mutations ©2 0 1 1 L a n d e s B i o s c i e n c e . D o n o t d i s t r i b u t e .
International Journal of Immunopathology and Pharmacology, Jul 1, 2012
Alterations in hormone secretion and cytokine levels have been evidenced in many neoplastic disea... more Alterations in hormone secretion and cytokine levels have been evidenced in many neoplastic diseases. In this study we have evaluated the circadian profile of growth hormone (GH), insulin-like growth factor-l (IGF-l), interleukin-2 (IL2), melatonin (MEL) and cortisol (COR) serum levels in nonsmall cell lung cancer patients. Blood was sampled every 4 h for 24 h in 11 healthy (H) men (ages 35-53 years) and 9 men with stage 2, 3 or 4 non-small cell lung cancer (C) (ages 43-63 years). Serum GH, total IGF1, IL2, MEL and COR were measured and examined for group differences, trends, and rhythm characteristics. 24-h means were significantly higher in C234 vs H for GH, GH/IGF1, IL2 and COR, and lower for IGF1, but IL2 and COR were not different for C23 vs H. A linear regression across 4 groups (H, C2, C3, C4) found a positive trend for COR, GH, GH/IGFI and IL2, and a negative trend for IGFI. A linear regression run between the 24-h mean levels of GH, IGF1, COR, MEL and IL2 in healthy subjects evidenced a statistically significant positive trend between MEL and GH (R=0.281, p=0.022) and in cancer patients showed a statistically significant negative trend between GH and IGFI (R=0.332, p=O.Ol), COR and IGFI (R=0.430, p=O.OOl), and a statistically significant positive trend between the 24-h mean of COR and GH (R=0.304, p=0.02). Rhythms in MEL and COR (peaks near 01:OOh and 08:00h, respectively) indicated identical synchronization to the light-dark cycle for both groups. A circadian rhythm was detected in GH and GH/IGFI for C23 and H, with IGFI and IL2 non-rhythmic in any group. In conclusion, an increasing trend and progressive loss of circadian rhythmicity in GH and GH/IGF1, an increasing trend in cortisol and IL2, and a decreasing trend in IGFI in C, reflect a complex chain of events that could be involved in progression of neoplastic disease. A therapeutic strategy needs to take into account circadian patterns and complex interactions ofthe multiple functions that characterize the hormone and cytokine levels in the frame cancer progression.
We discovered a novel somatic gene fusion, CD74–NRG1, by transcriptome sequencing of 25 lung aden... more We discovered a novel somatic gene fusion, CD74–NRG1, by transcriptome sequencing of 25 lung adenocarcinomas of never smokers. By screening 102 lung adenocarcinomas negative for known oncogenic alterations, we found four additional fusion-positive tumors, all of which were of the invasive mucinous subtype. Mechanistically, CD74–NRG1 leads to extracellular expression of the EGF-like domain of NRG1 III-β3, thereby providing the ligand for ERBB2–ERBB3 receptor complexes. Accordingly, ERBB2 and ERBB3 expression was high in the index case, and expression of phospho-ERBB3 was specifically found in tumors bearing the fusion (P < 0.0001). Ectopic expression of CD74–NRG1 in lung cancer cell lines expressing ERBB2 and ERBB3 activated ERBB3 and the PI3K–AKT pathway, and led to increased colony formation in soft agar. Thus, CD74–NRG1 gene fusions are activating genomic alterations in invasive mucinous adenocarcinomas and may offer a therapeutic opportunity for a lung tumor subtype with, so f...
XLSX file 75K, This is an excel file containing 9 sheets, one for each of the 9 supplementary tab... more XLSX file 75K, This is an excel file containing 9 sheets, one for each of the 9 supplementary tables (S1 to S9) mentioned in the main text
Cancers
A range of different techniques are available for predictive biomarker testing for non-small-cell... more A range of different techniques are available for predictive biomarker testing for non-small-cell lung cancer (NSCLC) clinical management. International guidelines suggest next-generation sequencing (NGS) as the preferred procedure, but other reverse transcriptase-polymerase chain reaction (RT-PCR)-based methods are rapidly evolving. In this study, we evaluated the reliability and accuracy of the IdyllaTM GeneFusion assay, a rapid and fully automated platform able to simultaneously detect ALK, ROS1, RET and NTRK1/2/3 and MET ex14 skipping mutations and compared its performance with routine reference methods. The cohort included thirty-seven NSCLCs plus two parotid gland carcinomas, previously characterized for the above alterations through either IHC, FISH, RT-PCR or NGS. In 36 of 39 cases, the Idylla GeneFusion assay and the reference methods were concordant (overall agreement: 92.3%). Tumor sections stored at room temperature for up to 60 days and 17 cases older than 2 years were ...
Genes, Chromosomes and Cancer
Small cell lung cancer (SCLC) is treated as a homogeneous disease, although the expression of NEU... more Small cell lung cancer (SCLC) is treated as a homogeneous disease, although the expression of NEUROD1, ASCL1, POU2F3, and YAP1 identifies distinct molecular subtypes. The MYC oncogene, amplified in SCLC, was recently shown to act as a lineage‐specific factor to associate subtypes with histological classes. Indeed, MYC‐driven SCLCs show a distinct metabolic profile and drug sensitivity. To disentangle their molecular features, we focused on the co‐amplified PVT1, frequently overexpressed and originating circular (circRNA) and chimeric RNAs. We analyzed hsa_circ_0001821 (circPVT1) and PVT1/AKT3 (chimPVT1) as examples of such transcripts, respectively, to unveil their tumorigenic contribution to SCLC. In detail, circPVT1 activated a pro‐proliferative and anti‐apoptotic program when over‐expressed in lung cells, and knockdown of chimPVT1 induced a decrease in cell growth and an increase of apoptosis in SCLC in vitro. Moreover, the investigated PVT1 transcripts underlined a functional co...
Molecular Cytogenetics, 2017
regions of several autosomes associated with marked sequence variation. Sequence of a sorted 9;14... more regions of several autosomes associated with marked sequence variation. Sequence of a sorted 9;14 translocation chromosome in a human endometrial carcinoma not only characterized the exact breakpoint but also identified an expressed fusion gene. Our study of alpaca and other camelids has used sequence from sorted alpaca chromosomes to determine the identity of alpaca evolutionary rearrangements by comparison with the published human genome and the human:dromedary comparative map. These examples illustrate some of the possibilities of sorting for chromosome sequencing to be expected in the future.
Frontiers in Oncology
DNA methylation is the most recognized epigenetic mark that leads to a massive distortion in canc... more DNA methylation is the most recognized epigenetic mark that leads to a massive distortion in cancer cells. It has been observed that a large number of DNA aberrant methylation events occur simultaneously in a group of genes, thus providing a growth advantage to the cell in promoting cell differentiation and neoplastic transformation. Due to this reason, methylation profiles have been suggested as promising cancer biomarkers. Here, we designed and performed a first step of validation of a novel targeted next generation sequencing (NGS) panel for methylation analysis, which can simultaneously evaluate the methylation levels at CpG sites of multiple cancer-related genes. The OPERA_MET-A methylation panel was designed using the Ion AmpliSeq™ technology to amplify 155 regions with 125-175 bp mean length and covers a total of 1107 CpGs of 18 cancer-related genes. The performance of the panel was assessed by running commercially available fully methylated and unmethylated control human gen...
Journal of Thoracic Oncology, 2009
Endocrine-Related Cancer, 2021
Parathyroid carcinoma (PC) is one of the rarest and aggressive malignancies of the endocrine syst... more Parathyroid carcinoma (PC) is one of the rarest and aggressive malignancies of the endocrine system. In some instances, the histological diagnosis remains uncertain unless there is evidence of gross local invasion or secondary spread. The identification of molecular markers could improve the diagnostic accuracy of these lesions. The expression of 740 genes involved in the tumor progression processes was assessed in 8 parathyroid adenomas (PAs), 17 non-metastatic and 10 metastatic PCs using NanoString technology. Clustering analysis and Ingenuity Pathway Analysis (IPA) were interrogated to compare the gene expression profiles among the three analyzed groups and to evaluate the potential role of differentially expressed genes, respectively. The 103 differentially expressed genes between metastatic PCs and PAs are able to discriminate perfectly the two groups from a molecular point of view. The molecular signatures identified in non-metastatic PCs vs PAs and in metastatic PCs vs non-me...
Annals of Oncology, 2019
<jats:title>Abstract</jats:title> <jats:sec> <jats:title>Background</j... more <jats:title>Abstract</jats:title> <jats:sec> <jats:title>Background</jats:title> <jats:p>Neuregulin-1 gene (NRG1) fusions function as oncogenic drivers across various solid tumors, most notably in invasive mucinous adenocarcinoma (IMA) of the lung, and represent a rational potential target for treatment. NRG1 is a growth factor, which binds to ErbB3 or ErbB4 inducing the formation of ErbB3 or 4-containing homo- or heterodimers and activating downstream ErbB-family signaling pathways. Therefore, the ErbB-family blocker afatinib may be a potential treatment option for patients with solid tumors harboring NRG1 fusions.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>We report a case series of all known patients with NRG1 fusion-positive solid tumors who were treated with afatinib; afatinib therapy is ongoing for some patients.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>To date, 18 patients with NRG1 fusion-positive solid tumors have been treated with afatinib (Table). These include 12 cases of non-small cell lung cancer (NSCLC; 7 of which were reported as IMA), 5 cases of gastrointestinal (GI) cancer (primarily pancreatic ductal adenocarcinoma [PDAC]) and 1 case of ovarian cancer. Various NRG1 fusion partners were identified, most commonly CD74 in patients with NSCLC (n = 7; 58%) and ATP1B1 in patients with gastrointestinal cancer (n = 3; 60%). Best response with afatinib among patients with NSCLC was partial response (PR) lasting 24 months (10 months among those specifically with IMA of the lung). Patients with PDAC experienced PR of 3 and 5.5 months' duration, and one patient has an ongoing PR after 7 months. One patient with cholangiocarcinoma had a PR lasting 8 months; another patient with ovarian cancer had stable disease (SD) of unknown duration.</jats:p> </jats:sec> <jats:sec> <jats:title>Conclusions</jats:title> <jats:p>Afatinib is a potential treatment option for some patients with solid tumors harboring NRG1 fusions. The efficacy and safety of afatinib will be evaluated in ongoing/planned prospective non-randomized clinical trials of targeted drugs in patients with advanced cancer with potentially actionable genomic variants (NCT02925234 and NCT02693535).</jats:p> </jats:sec> <jats:sec> <jats:title>Editorial acknowledgement</jats:title> <jats:p>Greg Plosker of GeoMed, an Ashfield company, part of UDG Healthcare plc.</jats:p> </jats:sec> <jats:sec> <jats:title>Legal entity responsible for the study</jats:title> <jats:p>The authors.</jats:p> </jats:sec> <jats:sec> <jats:title>Funding</jats:title> <jats:p>Boehringer Ingelheim.</jats:p> </jats:sec> <jats:sec> <jats:title>Disclosure</jats:title> <jats:p>Y. Goto: Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (self): Eli Lilly; Advisory / Consultancy, Speaker Bureau / Expert testimony: Chugai; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (self): Taiho Pharmaceutical; Advisory / Consultancy, Speaker Bureau / Expert testimony: Boehringer Ingelheim; Speaker Bureau / Expert testimony, Research grant / Funding (self): Ono Pharmaceutical; Speaker Bureau / Expert testimony, Research grant / Funding (self): Bristol-Myers Squibb; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (self): Pfizer; Speaker Bureau / Expert testimony: MSD; Speaker Bureau / Expert testimony: Shionogi Pharma; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (self): Novartis; Advisory / Consultancy: AstraZeneca; Advisory / Consultancy: Glaxo Smith Kline; Research grant / Funding (self): AbbVie; Research grant / Funding (self): Daiichi Sankyo; Research grant / Funding (self): Kyorin. J. Cadranel: Advisory / Consultancy, Research grant / Funding (institution), Non-remunerated activity/ies: AZ; Advisory / Consultancy, Non-remunerated activity/ies: BI; Advisory / Consultancy: MSD; Advisory / Consultancy: BMS; Advisory / Consultancy: Takeda; Advisory / Consultancy, Research grant / Funding (institution): Pfizer; Advisory / Consultancy: Lilly; Advisory / Consultancy, Non-remunerated activity/ies: Roche; Advisory / Consultancy, Research grant / Funding (institution): Novartis. B.A. Weinberg: Speaker Bureau / Expert testimony: Lilly; Speaker Bureau / Expert testimony: Bayer; Research grant / Funding (institution): Novartis; Research grant / Funding (institution): Ipsen; Travel / Accommodation / Expenses: Caris Life Sciences; Travel / Accommodation / Expenses: Boehringer Ingelheim. M. Duruisseaux: Honoraria (self): Roche; Honoraria (self): Takeda; Honoraria (self): Pfizer; Honoraria (self): Novartis; Honoraria (self): AstraZeneca; Honoraria (self): BMS; Honoraria (self): MSD; Honoraria (self): AbbVie; Honoraria (self): Boerhinger ingelheim. S.V. Liu: Advisory / Consultancy: Apollomics; Advisory / Consultancy, Research grant /…
Journal of Thoracic Oncology, 2019
Background: The third generation TKI (osimertinib) according to AURA III trial results achieved l... more Background: The third generation TKI (osimertinib) according to AURA III trial results achieved longer PFS compared to standard platinum based chemotherapy in patient's resistant to 1-2 generation TKIs due to T790M mutation (8.5 vs 4.2 months). The evolution of resistance profile during thins therapy can be analyzed based on ctDNA. Method: In this study patients with metastatic EGFR mutated NSCLC, with a confirmed disease progression during treatment with 1/2 generation TKIs, T790M positive which included patients received osimertinib 80 mg daily. Before the treatment and then every 2 months, whole blood was taken, for qualitative assessment of ctDNA dynamics by RT-PCR. The aim of the study was to assess the relationship between the disappearance of T790M + ctDNA and the time to progression on osimertinib. Result: From August 2016 to December 2018 22 patients with T790M positive progression were identified. 18/22 (81.9%) were women, 4/22 (18.1%) -men. The mean age was 61.2 years (50-75). Only 1/22 had a smoking history >30 pack/years. Primary activating mutations in EGFR gene were ex19del, L858R and G719S + S768I in 16, 5 and 1 patients respectively. Median PFS on the first line TKI was 21.7 months (CI 95%, 10.8 e 53.3). In 59.1% (13/22) progressive disease was characterized by the appearance of new metastases and in 40.9% (9/22) by the growth of previously identified metastases. 22 patients were evaluable for response. PR and SD were achieved in 11/20 (50%) and 10/20 (45/5%) respectively. Median PFS was in a whole group 16.7 months (CI 95%, 11.4 -22.0). T790M in ctDNA was negative after 2 months of osimertinib treatment in 12/22 patients. Median PFS was 18.9 months (CI 95%, 14.8e19.7) in patients with undetectable T790M in ctDNA after 2 month of therapy compared to 8.0 months (CI 95%, 4.2 e 11.8) in patients remaining ctDNA T790M positive. No clinical factors were associated with the disappearance of ctDNA by statistical analysis. The disappearance of T790M + ctDNA after 2 months osimertinib therapy is predictive of greater PFS in patients with EGFR mutation positive NSCLC, receving of 2nd line.
Annals of Oncology, 2019
Background Small Cell Lung Cancer (SCLC) is a highly metastatic form of lung cancer and the thera... more Background Small Cell Lung Cancer (SCLC) is a highly metastatic form of lung cancer and the therapies remain unchanged since 40 years. Therefore, the discovery of novel transcriptomic alterations contributes to develop more effective targeted therapies. Among them, RLF/MYCL1 was described as a recurrent chimera implicated in tumor growth. The non-protein-coding RNA PVT1, linked to MYC expression, and its circular variant circPVT1 are also dysregulated in several malignancies and associated to tumor progression. Nevertheless, their functional and clinical role was only partially explored. We investigated these transcriptomic alterations in SCLC patients and cell lines, aiming at exploring a possible prognostic significance. Methods 12 SCLC cell lines harboring MYC, MYCN or MYCL1 amplifications were tested by RNA-sequencing and RT-PCR for the recurrence of RLF/MYCL1. Moreover, the relative expression of PVT1/circPVT1 was evaluated by RT-qPCR. To investigate whether these transcriptomi...
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Papers by Lucia Anna Muscarella