Papers by Francesca Malentacchi
Frontiers in Immunology, Jan 26, 2022
Although accumulating data have investigated the effect of SARS-CoV-2 mutations on antibody neutr... more Although accumulating data have investigated the effect of SARS-CoV-2 mutations on antibody neutralizing activity, less is known about T cell immunity. In this work, we found that the ancestral (Wuhan strain) Spike protein can efficaciously reactivate CD4+ T cell memory in subjects with previous Alpha variant infection. This finding has practical implications, as in many countries only one vaccine dose is currently administered to individuals with previous COVID-19, independently of which SARS-CoV-2 variant was responsible of the infection. We also found that only a minority of Spike-specific CD4+ T cells targets regions mutated in Alpha, Beta and Delta variants, both after natural infection and vaccination. Finally, we found that the vast majority of Spike-specific CD4+ T cell memory response induced by natural infection or mRNA vaccination is conserved also against Omicron variant. This is of importance, as this newly emerged strain is responsible for a sudden rise in COVID-19 cases worldwide due to its increased transmissibility and ability to evade antibody neutralization. Collectively, these observations suggest that most of the memory CD4+ T cell response is conserved against SARS-CoV-2 variants of concern, providing an efficacious line of defense that can protect from the development of severe forms of COVID-19.
Fertility and Sterility, Feb 1, 2021
To study the molecular mechanisms involved in the appearance of the fibrotic trait in endometrios... more To study the molecular mechanisms involved in the appearance of the fibrotic trait in endometriosis by investigating whether the signaling pathway of the bioactive sphingolipid sphingosine 1-phosphate (S1P) was altered in endometriotic lesions. Design: Case-control laboratory study. Setting: University research institute and university hospital. Patient(s): A total of 75 women, with and without endometriosis, were included in the study. Interventions(s): Endometrial samples were obtained from women affected (n ¼ 15 endometrioma [OMA]; n ¼ 30 deep infiltrating endometriosis [DIE]) and not (n ¼ 30) by endometriosis by means of laparoscopic surgery, followed by clinical and imaging investigation and checking for the expression of fibrosis markers and genes implicated in S1P metabolism and signaling by means of real-time polymerase chain reaction. Main Outcome Measure(s): The role of the S1P signaling axis in endometriosis-associated fibrosis was studied in vitro, where RNA interference approaches were used to investigate if S1P synthesis by sphingosine kinases (SKs) and specific S1P receptors (S1PRs) are implicated in the profibrotic effect of the cytokine transforming growth factor (TGF) b1. Result(s): mRNA expression analysis of S1PR demonstrated a deep dysregulation of S1P signaling in endometriosis, characterized by increased expression of fibrosis markers: S1P 1 was transcriptionally more expressed in OMA, and S1P 3 and S1P 5 mRNA levels were significantly augmented in both OMA and DIE. SK1 and its activating protein calcium-and integrin-binding protein 1 (CIB1) were significantly up-regulated in OMA and DIE. A crucial role for the SK/S1PR axis in the profibrotic effect elicited by TGFb1 was highlighted in vitro. Conclusion(s): The S1P signaling axis may represent a useful biomarker or innovative pharmacologic target for endometriosis.
Anti-Cancer Drugs, Sep 1, 2020
Uterine carcinosarcomas are biphasic neoplasms consisting of mixed epithelial and mesenchymal ele... more Uterine carcinosarcomas are biphasic neoplasms consisting of mixed epithelial and mesenchymal elements, representing less than 5% of all uterine malignancies. Carcinosarcomas are rare, although the most common cause of uterine cancer-specific death. Few information is available on the pathogenesis, and molecular characterization is poorly investigated. Consequently, the treatment has not changed over the last years and is far too being tailored, consisting of surgery and traditional chemotherapy and radiotherapy. Molecular characterization of liquid biopsy by circulating tumor DNA (ctDNA)/circulating cell-free DNA (ccfDNA) evaluation in a patient with uterine carcinosarcoma. Here, we describe a case report of an 83-year-old woman with carcinosarcomas, stage T3aN0M0. Cancer cells did not express estrogen nor progesterone receptors, while p53 and p16 were positive. Molecular characterization of ccfDNA and of ctDNA was performed by quantitative PCR, amplification-refractory mutation system technology. The presence of phosphatidylInositol-4,5-bisphosphate 3-Kinase catalytic subunit alpha p.E545A mutation was detected in plasma. This approach may suggest the use of liquid biopsy and the development of specific targeted therapy for precision personalized medicine even in rare carcinosarcomas. Anti-Cancer Drugs 31: 880-883
Fertility and Sterility, Jun 1, 2021
OBJECTIVE To explore the link between sphingosine 1-phosphate (S1P) signaling and leiomyoma and t... more OBJECTIVE To explore the link between sphingosine 1-phosphate (S1P) signaling and leiomyoma and the possible S1P cross-talk with the fibrotic effect of activin A. DESIGN Case-control laboratory study. SETTING University institute and university hospital. PATIENT(S) Patients with uterine fibroids (n = 26). INTERVENTIONS(S) Tissue specimens of leiomyoma and normal myometrium were obtained from patients undergoing myomectomy or total hysterectomy. MAIN OUTCOME MEASURE(S) Expression of mRNA levels of the enzyme involved in S1P metabolism, S1P receptors, and S1P transporter Spns2 was evaluated in matched leiomyoma/myometrium specimens and cell populations. The effects of inhibition of S1P metabolism and signaling was evaluated on activin A-induced fibrotic action in leiomyoma cell lines. RESULT(S) The expression of the enzymes responsible for S1P formation, sphingosine kinase (SK) 1 and 2, and S1P2, S1P3, and S1P5 receptors was significantly augmented in leiomyomas compared with adjacent myometrium. In leiomyoma cells, but not in myometrial cells, activin A increased mRNA expression levels of SK1, SK2, and S1P2. The profibrotic action of activin A was abolished when SK1/2 were inhibited or S1P2/3 were blocked. Finally, S1P augmented by itself mRNA levels of fibrotic markers (fibronectin, collagen 1A1) and activin A in leiomyomas but not in myometrial cells. CONCLUSION(S) This study shows that S1P signaling is dysregulated in uterine fibroids and involved in activin A-induced fibrosis, opening new perspectives for uterine fibroid treatment.
Anti-Cancer Drugs, Jun 11, 2020
Endometrial cancer is the commonest gynecological cancer, the majority is endometrioid type, diag... more Endometrial cancer is the commonest gynecological cancer, the majority is endometrioid type, diagnosed at an early stage with 69-88% 5-year survival. Low-grade endometrial cancers have low recurrence rates and often do not receive adjuvant therapy; however, a subset of these patients will have poor outcomes and would benefit from adjuvant treatment has been challenging. We evaluate the circulating cell-free DNA (ccfDNA) in a patient with low-risk endometrial cancer in order to identify the presence of molecular markers associated with risk of recurrence. The evaluation of mutation profile was performed by next-generation sequencing (NGS) in primary tumor formalin-fixed paraffin-embedded (FFPE) tissue and in circulating tumor DNA (ctDNA). We identified a specific mutational profile in ctDNA, different from primary tumor tissue suggesting that the clone involved in the relapse may be different in comparison to the most represented in the primary tumor. These findings open new prospective and new wonderings. The molecular characterization of tissue may be useful for setting new target personalized therapy even in the treatment of endometrial cancer, moreover, endometrial cancer at low risk should be not underestimated for the incidence of relapse, and for this evaluation the molecular characterization may be useful. Moreover, these results suggest that the single analysis of primary tumors may be not sufficient for setting a specific personalized therapy targeted to avoid the relapse but may be necessary to join the molecular characterization of liquid biopsy to primary tissue. Anti-Cancer Drugs 31: 1091
Minerva ginecologica, 2021
In cervical cancer screening programs, women with abnormal cytology and confirmation by biopsy ar... more In cervical cancer screening programs, women with abnormal cytology and confirmation by biopsy are referred for colposcopy for histological evaluation. We characterized the presence and the genotype of HPV by Linear Array HPV genotyping assay in cytological samples collected from about 400 women undergoing conization, with reported high CIN grade after biopsy. The most prevalent genotype was HPV 16, with an increasing presence depending on the severity of the CIN and with the highest incidence in the 26-35 age range. In the group of younger women (<25) we found the highest percentage of CIN3 (39.3%) and the lowest of CIN1 (17.9%). An increase of CIN1 with increasing age was observed. A different distribution of HPV presence was observed depending on CIN grade (P<0.001): CIN1 HPV negative samples were 46.3%, CIN2: 5.8% and CIN3: 1.4%. Interesting, in the analyzed cohort, we observed the presence of 30% of CIN1. Moreover, within CIN1, 85% of them were associated to negative HPV detection, this observation suggested that the detection of HPV presence may be useful to identify low CIN grade that should be reconsidered for surgical treatment. These findings suggest implementing the protocol for the management of women with high risk precancer lesions, with a further HPV test before surgical treatment. The evaluation of HPV presence and genotype before conization might represent a useful tool in reducing or postpone the conization treatment.
BioMed Research International, 2018
The aim of this study was to develop a scoring system of the immunohistochemical (IHC) expression... more The aim of this study was to develop a scoring system of the immunohistochemical (IHC) expression of luteinizing hormone/human chorionic gonadotropin receptor (LHCG-R) in endometrial cancer (EC) patients. Nonconsecutive hysterectomy specimens containing EC collected from April 2013 to October 2015 were selected. Hematoxylin-eosin stained sections from each case were reviewed and representative sections from each tumor were selected. IHC staining was performed for the detection of LHCG-R. The percentage of stained cells and the staining intensity were assessed in order to develop an immunohistochemical score. Moreover, we examined the correlation of the score with grading and lymphovascular space invasion (LVSI). There was a statistically significant positive correlation between grading and IHC scoring (= 0.01) and a statistically significant positive correlation between LVSI and IHC score (< 0.01). In conclusion, we suggest that the immunohistochemical score presented here could be used as a marker of bad prognosis of EC patients. Nevertheless, further studies are needed in order to validate it. The study was registered in the Careggi Hospital public trials registry with the following number: 2013/0011391.
International Journal of Infectious Diseases, Dec 1, 2021
Background The emergence of SARS-CoV-2 variants of concern (VOCs) for increased transmissibility ... more Background The emergence of SARS-CoV-2 variants of concern (VOCs) for increased transmissibility and potentially capable of immune-escape, mandates for epidemiological surveillance. Genomic alterations present in VOCs can affect the results of RT-qPCR assays for routine diagnostic purposes, leading to peculiar profiles that can be used for rapid screening of variants. In this work, we report on a peculiar profile observed with the Allplex™ SARS-CoV-2/ FluA/ FluB/ RSV Assay and VOC Alpha (202012/01, lineage B.1.1.7, also named VOC-UK), which was the first identified SARS-CoV-2 VOC. Methods Samples were analyzed by two RT-qPCR assays: the Allplex™ SARS-CoV-2/ FluA/ FluB/ RSV Assay (ASFR, Seegene Technologies Inc; Seoul, South Korea), and the TaqPath COVID-19 RT-PCR (Thermo Fisher Scientifics, USA). Definition of SARS-CoV-2 variant was carried out by Sanger sequencing of relevant S-gene regions and, in some cases, by whole genome sequencing (WGS) using the ARTIC-nCoV workflow on a MiniION (Oxford Nanopore Technologies, Oxford, UK) or a MiSeq ILLUMINA platform (San Diego, California, US). Results Of 173 SARS-CoV-2-positive specimens, all those of lineage B.1.1.7 (N=71) showed an average Cq difference between the N and S gene of +11±2 (range, + 8/+15). None of the other specimens, including several different lineages (Wild-type for the analyzed regions, N=22; Gamma, N=63; Delta, N=9; B.1.258, N=3; B.1.160, N=3; B.1.177.7, N=1; B.1.1.420, N=1), exhibited a similar difference of Cq values. Conclusions The peculiar pattern of delayed N gene positivity could constitute a convenient method for screening of VOC Alpha, simultaneous to viral detection, when using the Allplex™ SARS-CoV-2/ FluA/ FluB/ RSV Assay.
Journal of Antimicrobial Chemotherapy, Dec 21, 2022
aligning with prior publications. 8,9 Conversely, Manesh et al. 10 found that crushed posaconazol... more aligning with prior publications. 8,9 Conversely, Manesh et al. 10 found that crushed posaconazole DR tablets given per feeding tube at a dose of 300 mg daily produced therapeutic levels after 2 weeks of therapy in 19 of 19 patients receiving treatment for rhino-orbito-cerebral mucormycosis. 10 Many factors can impact drug levels of crushed posaconazole DR tablets, spanning from the obvious patient factors (i.e. absorption) to terminal tube placement (e.g. J-tube versus G-tube) or the more nuanced factors (preparation and administration technique). The literature supporting this practice is evolving, but still limited. It is currently unknown how different manufacturers' DR matrices may impact serum concentrations if crushed, or how crushed DR tablets would compare in a head-to-head comparison against the oral suspension. Our experience adds to the growing body of evidence suggesting that crushing and administering posaconazole DR tablets via enteral feeding tubes is safe and often results in therapeutic levels. Importantly, doses may likely need to be up-or down-titrated when switching to or from the formulation of crushed posaconazole DR tablets, respectively. Clinicians should consider this route of administration, when necessary, in conjunction with careful dose titration and frequent TDM.
Oncology Reports, Dec 18, 2018
Endometrial cancer (EC) comprises a biological and clinical heterogeneous group of tumors. Severa... more Endometrial cancer (EC) comprises a biological and clinical heterogeneous group of tumors. Several genetic alterations are involved in the development and progression of EC, and may be used for targeted therapy, particularly in patients with advanced-stage EC. In the present study, a combined procedure was developed based on polymerase chain reaction (PCR)-high resolution melting analysis (HRMA) and Sanger sequencing for the evaluation of somatic mutations in selected phosphoinositide 3-kinase (PI3K) catalytic subunit α (PIK3CA; exons 1, 9 and 21) and phosphatase and tensin homolog (PTEN; exons 5, 6, 7 and 8) exons. This combined procedure has the specificity and sensitivity of the two techniques, and overcomes their limitations. A pilot study was performed on 18 selected homogenous EC samples, of grade 3 endometrioid subtype (G3 EEC). First, the feasibility of the combined procedure was investigated to properly identify the presence of somatic mutations on PIK3CA and PTEN, the variations identified were analyzed using Catalogue of Somatic Mutations in Cancer, PolyPhen-2 and Mutation Taster software, and the frequency of mutations/variations was determined in the selected samples. The evaluation of mutational load revealed that the majority of the G3 EEC samples exhibited PIK3CA mutations (39%) and PTEN mutations (67%), and the majority of the samples (83%) had mutations in at least one of the two genes, and 33% had mutations in the two genes. The results of the present pilot study suggested that the cost-effective combined PCR-HRMA and Sanger sequencing procedure may be suitable for identification of PTEN and PIK3CA mutations in G3 EEC and that their frequency was consistent in G3 EEC, indicating that the PI3K pathway serves a pivotal function that may have potential for defining targeted therapy for the treatment of G3 EEC.
Reproductive Sciences, Jan 6, 2020
The incidence of endometrial cancer (EC) is increasing in developed countries. The most frequent ... more The incidence of endometrial cancer (EC) is increasing in developed countries. The most frequent is the endometrioid subtype with usually good prognosis; nevertheless, some cases escape this paradigm and may have recurrence. A recent study from The Cancer Genome Atlas suggested to implement the EC analysis by molecular profile for improving diagnosis, prognosis, and therapeutic treatment. The present preliminary study was performed on 15 G3 endometrioid endometrial cancers (G3 EEC) for the identification of somatic mutations in a panel of specific exons in selected genes as ARID1A, CTNNB1, KRAS, PIK3CA, POLE, PTEN, and TP53. The combined procedure, based on the Sanger sequencing and PCR-high-resolution melting analysis, allowed the identification of variations of the selected gene panel in most of patients (93%) of our cohort. The overall evaluation of mutational load exhibited that the most frequent mutated genes were PTEN (93%), followed by PIK3CA (47%) suggesting a deep involvement of PI3K pathway alteration in G3 EEC. Mutations in TP53 (27%), ARID1A (27%), POLE (13%), and at the lower level in KRAS and CTNNB1 (7%) were also observed (exclusively in FIGO III stage patients). The evaluation of the mutations of our proposed panel (ARID1A, CTNNB1, KRAS, PIK3CA, POLE, PTEN, TP53) is suitable to improve the characterization of G3 EEC and could suggest targetable pathways for development of personalized treatments.
International Journal of Infectious Diseases
Journal of Antimicrobial Chemotherapy
aligning with prior publications. 8,9 Conversely, Manesh et al. 10 found that crushed posaconazol... more aligning with prior publications. 8,9 Conversely, Manesh et al. 10 found that crushed posaconazole DR tablets given per feeding tube at a dose of 300 mg daily produced therapeutic levels after 2 weeks of therapy in 19 of 19 patients receiving treatment for rhino-orbito-cerebral mucormycosis. 10 Many factors can impact drug levels of crushed posaconazole DR tablets, spanning from the obvious patient factors (i.e. absorption) to terminal tube placement (e.g. J-tube versus G-tube) or the more nuanced factors (preparation and administration technique). The literature supporting this practice is evolving, but still limited. It is currently unknown how different manufacturers' DR matrices may impact serum concentrations if crushed, or how crushed DR tablets would compare in a head-to-head comparison against the oral suspension. Our experience adds to the growing body of evidence suggesting that crushing and administering posaconazole DR tablets via enteral feeding tubes is safe and often results in therapeutic levels. Importantly, doses may likely need to be up-or down-titrated when switching to or from the formulation of crushed posaconazole DR tablets, respectively. Clinicians should consider this route of administration, when necessary, in conjunction with careful dose titration and frequent TDM.
Journal of lower genital tract disease, Jan 5, 2016
The aim of the study was to determine whether p16 positive/cervical intraepithelial neoplasia (CI... more The aim of the study was to determine whether p16 positive/cervical intraepithelial neoplasia (CIN) 2, 3, and cancer (p16 + CIN 2/3+) detected by colposcopy-directed or random biopsy differ by age, referral cytology, human papillomavirus (HPV) 16, and lesion size. Data from the Shenzhen Cervical Cancer Screening Trial II where, at colposcopy, women who had directed and random cervical biopsies were reviewed to find women with CIN 2, 3, or cancer; 227 such women identified had their paraffin-embedded tissue blocks recut, reviewed, and then immune stained for p16. Data were analyzed by χ, Fisher exact test, and linear regression. After histopathologic review and p16 staining of CIN 2, 175 women were diagnosed with p16 + CIN 2/3+. When compared with those diagnosed by colposcopy-directed biopsy (n = 138), those diagnosed by random biopsy (n = 37) were more likely to have Cytology-Lo (cytology of negative, atypical squamous cells of undetermined significance, or low-grade squamous intra...
Clinical Chemistry and Laboratory Medicine, 2015
Data in Brief, Mar 1, 2016
Within the EU-SPIDIA project (www.spidia.eu), the quality parameters of blood genomic DNA were de... more Within the EU-SPIDIA project (www.spidia.eu), the quality parameters of blood genomic DNA were defined [SPIDIA-DNA: an External Quality Assessment for the pre-analytical phase of blood samples used for DNA-based analyses-[1]; Influence of preanalytical procedures on genomic DNA integrity in blood samples: the SPIDIA experience-[2]; Combining qualitative and quantitative imaging evaluation for the assessment of genomic DNA integrity: the SPIDIA experience-[3]. DNA quality parameters were used to evaluate the laboratory performance within an External Quality Assessment (EQA) [Second SPIDIA-DNA External Quality Assessment (EQA): Influence of pre-analytical phase of blood samples on genomic DNA quality-[4]. These parameters included DNA purity and yield by UV spectrophotometric measurements, the presence of PCR interferences by Kineret software and genomic DNA integrity analysis by Pulsed Field Gel Electrophoresis.
Clinica Chimica Acta, Feb 1, 2016
Background: In order to develop evidence-based quality guidelines for the pre-analytical phase of... more Background: In order to develop evidence-based quality guidelines for the pre-analytical phase of blood samples used for DNA molecular testing, two pan-European External Quality Assessments (EQAs) were implemented within the European Commission funded project SPIDIA. Here we report the results of the 2nd SPIDIA EQA that has been implemented on the basis of the 1st DNA EQA with the inclusion of some stringent conditions related to blood storage temperature and time. Methods: SPIDIA facility sent to all the participants the same blood sample to be processed by their own procedure following SPIDIA suggestion for time and temperature storage. Evaluated genomic DNA (gDNA) quality parameters were: purity and yield by UV spectrophotometric analysis, PCR interferences by Kineret software and integrity by a dedicated algorithm. Results/conclusions: 188 applications have been collected from 26 European countries. A high variability of gDNA integrity was observed whereas purity, yield and PCR interferences had a narrow distribution within laboratories. A dedicated analysis on pre-analytical variables and the evaluated gDNA quality parameters showed that blood storage and DNA extraction procedures influence gDNA integrity. The performances of the participants were improved in comparison with the 1st SPIDIA-DNA EQA, probably due to adopted more stringent pre-analytical conditions.
8th European Congress of Endocrinology incorporating the British Endocrine Societies, Dec 4, 2006
Background: The aim of this study is to investigate the expressions of somatostatin receptor (SST... more Background: The aim of this study is to investigate the expressions of somatostatin receptor (SSTR), SSTR-2, SSTR-3, and SSTR-5, in pancreatic tissue and non-cancerous tissue and elucidate their clinical significance. Methods: The expression of somatostatin receptor subtypes SSTR-2, SSTR-3, and SSTR-5 messenger RNA (mRNA) in 108 cases of cancer tissue and adjacent tissue in patients with pancreatic cancer was detected by reverse transcriptase polymerase chain reaction (RT-PCR). Expression of SSTR-2, SSTR-3, and SSTR-5 mRNA was evaluated after specimens were taken from selected patients who underwent surgical resection by Whipple's operation. We speculated the clinical significance of the expression of somatostatin receptor (SSTR) subtype genes SSTR-2, SSTR-3, and SSTR-5 in pancreatic tissue and non-cancerous tissue and further elucidated their clinical significance. Results: The expression rates of SSTR-2 mRNA in cancer and adjacent tissue of 108 patients with pancreatic cancer were 81.5% (88/108) and 97.2% (105/108), respectively; SSTR-3 mRNA expression rates were 69.4% (75/108) and 55.6% (60/108). SSTR-5 mRNA expression rates were 13.0% (14/108) and 18.5% (20/108). Conclusion: We propose that SSTR-2 plays an important role in clinical implications for patients with pancreatic cancer undergoing somatostatin or its analog therapy.
Clinical Chemistry and Laboratory Medicine, 2015
Background: Circulating cell-free DNA (ccfDNA) has been confirmed as a useful biomarker in cancer... more Background: Circulating cell-free DNA (ccfDNA) has been confirmed as a useful biomarker in cancer and pre-natal clinical practice. One of the main critical points in using ccfDNA is a lack of standardisation for sample processing methods, storage conditions, procedures for extraction, and quantification that can affect ccfDNA quality and quantity. We report the results obtained from the SPIDIA-DNAplas, one of the EU SPIDIA (Standardisation and improvement of generic pre-analytical tools and procedures for in vitro diagnostics) subprojects based on the implementation of an External Quality Assessment scheme for the evaluation of the influence of the pre-analytical phase on ccfDNA. This is the first reported quality control scheme targeting ccfDNA for pre-analytical phase studies. Methods: Fifty-six laboratories throughout Europe were recruited. The participating laboratories received the same plasma sample and extracted ccfDNA by using their own procedures, at defined plasma storage conditions, and sent the isolated ccfDNA to the SPIDIA facility for analyses. Laboratory performance was evaluated by using specific quality parameters such as ccfDNA integrity (by multiplex PCR) and yield (by qPCR). Results: The analysis of the ccfDNA extracted by the laboratories showed that most of them (53 of 56) were able to recover ccfDNA but only 12.5% recovered non-fragmented ccfDNA. Extraction methods specifically designed for ccfDNA preserved the integrity profile. Conclusions: The evidence-based results of the SPIDIA-DNAplas EQA have been proposed as a basis for the development of a Technical Specification by the European Committee for standardisation (CEN).
European Urology Supplements, Apr 1, 2010
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Papers by Francesca Malentacchi