Antonella Bertucci - Academia.edu (original) (raw)

Papers by Antonella Bertucci

Retinal Cases & Brief Reports, 2007

To describe a patient with primary central nervous system lymphoma (PCNSL) who developed intraocu... more To describe a patient with primary central nervous system lymphoma (PCNSL) who developed intraocular recurrence after the onset of optic nerve disorder. A 46-year-old woman developed bilateral central vision loss 1 year after the onset of PCNSL. Ophthalmologic examination, including perimetry, magnetic resonance imaging, and determination of visual evoked potential, was performed. These examinations revealed a diagnosis of bilateral optic nerve disorder, and optic nerve disorder spontaneously resolved. Six months later, the patient had blurred vision in the left eye. Funduscopic examination revealed dense vitreous opacity and geographic subretinal exudates. Cytologic analysis of vitreous obtained by pars plana vitrectomy led to a diagnosis of recurrent intraocular lymphoma. The patient again developed brain recurrence 8 months after chemotherapy for intraocular recurrence. The clinical course in this case suggests that lymphoma cells infiltrated from the brain into the eye along the optic nerve.

Cancer Research, Apr 4, 2023

High-Grade Serous Ovarian Cancer (HGSOC) is the most fatal of gynecological cancers. The recurren... more High-Grade Serous Ovarian Cancer (HGSOC) is the most fatal of gynecological cancers. The recurrence rate of HGSOC after treatment is more than 80%. The available treatment options for HGSOC are limited; standard of care includes surgery combined with chemotherapy, with radiation therapy in some advanced cases. Both chemotherapy and radiation therapy are known to induce tumor cell aggressiveness. Many cancer studies have demonstrated that cancer stem cells (CSCs) and the epithelial-mesenchymal transition (EMT) promote cancer aggressiveness, invasion, metastasis, and drug resistance. Therefore, to better understand the cause of the poor prognosis in HGSOC, our objective is to scrutinize the role of CSCs and EMT in chemo- and radiation resistance. To lead our inquiry, we utilized the SORE6 reporter to identify Sox2/Oct4 expression, which indicates the stemness, and the Zeb1 3’ UTR reporter to detect EMT. Using flow cytometry, we quantified the reporter activity in 10 ovarian cancer cell types, including chemo-sensitive and chemo-resistant ones, considering both cell lines and patient-derived cells. Chemo-resistant cells are grown under cisplatin treatment. We exposed some of these ovarian cancer cell lines to 0, 1, 2, 4 and 8Gy of 250 MeV proton and 6 MeV photon beams, then analyzed the reporter activity 72 hours post-radiation. Our findings exhibit that chemo-resistant cells express higher stemness and EMT levels than chemo-sensitive cells and have higher resistance to radiation. These reporters can detect radiation-induced stemness and EMT. Furthermore, stemness and EMT levels positively correlate with the radiation dosage increment. We conclude that these reporters are an efficient surrogate for detection of stemness and EMT and can be used to test strategies for prevention of therapy-induced aggressive phenotypes. Citation Format: Yeonkyu Jung, Aaron Keniston, Ashley Antonissen, Ann Morcos, Antonella Bertucci, Marcelo Vazquez, Juli Unternaehrer. High grade serous ovarian cancer: Detecting stemness and EMT levels in response to chemo and radiation therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1089.

PLOS ONE, May 31, 2023

Human exposure to low-to-moderate dose ionizing radiation (LMD-IR) is increasing via environmenta... more Human exposure to low-to-moderate dose ionizing radiation (LMD-IR) is increasing via environmental, medical, occupational sources. Acute exposure to LMD-IR can cause subclinical damage to cells, resulting in altered gene expression and cellular function within the human brain. It has been difficult to identify diagnostic and predictive biomarkers of exposure using traditional research models due to factors including lack of 3D structure in monolayer cell cultures, limited ability of animal models to accurately predict human responses, and technical limitations of studying functional human brain tissue. To address this gap, we generated brain/cerebral organoids from human induced pluripotent stem cells to study the radiosensitivity of human brain cells, including neurons, astrocytes, and oligodendrocytes. While organoids have become popular models for studying brain physiology and pathology, there is little evidence to confirm that exposing brain organoids to LMD-IR will recapitulate previous in vitro and in vivo observations. We hypothesized that exposing brain organoids to proton radiation would (1) cause a time-and dose-dependent increase in DNA damage, (2) induce cell type-specific differences in radiosensitivity, and (3) increase expression of oxidative stress and DNA damage response genes. Organoids were exposed to 0.5 or 2 Gy of 250 MeV protons and samples were collected at 30 minute, 24 hour, and 48 hour timepoints. Using immunofluorescence and RNA sequencing, we found time-and dose-dependent increases in DNA damage in irradiated organoids; no changes in cell populations for neurons, oligodendrocytes, and astrocytes by 24 hours; decreased expression of genes related to oligodendrocyte lineage, astrocyte lineage, mitochondrial function, and cell cycle progression by 48 hours; increased expression of genes related to neuron lineage, oxidative stress, and DNA damage checkpoint regulation by 48 hours. Our findings demonstrate the possibility of using organoids to characterize cell-specific radiosensitivity and early radiationinduced gene expression changes within the human brain, providing new avenues for

Frontiers in Oncology, Sep 2, 2022

High-risk human papillomaviruses (HPVs) cause virtually all cervical cancer cases and are also as... more High-risk human papillomaviruses (HPVs) cause virtually all cervical cancer cases and are also associated with other types of anogenital and oropharyngeal cancers. Normally, HPV exists as a circular episomal DNA in the infected cell. However, in some instances, it integrates into the human genome in such a way as to enable increased expression of viral oncogenes, thereby leading to carcinogenesis. Since viral integration requires breaks in both viral and human genomes, DNA damage likely plays a key role in this critical process. One potentially significant source of DNA damage is exposure to elevated doses of ionizing radiation. Natural background radiation is ubiquitous; however, some populations, including radiological workers, radiotherapy patients, and astronauts, are exposed to significantly higher radiation doses, as well as to different types of radiation such as particle radiation. We hypothesize that ionizing radiation-induced DNA damage facilitates the integration of HPV into the human genome, increasing the risk of developing HPV-related cancers in the exposed population. To test this, we first determined the kinetics of DNA damage in keratinocytes exposed to ionizing radiation (protons) by assessing g-H2AX foci formation using immunofluorescence (direct damage), and also measured ROS and 8-oxoG levels via DCFDA and Avidin-FITC (indirect damage).As anticipated, direct DNA damage was observed promptly, within 30 min, whereas indirect DNA damage was delayed due to the time required for ROS to accumulate and cause oxidative damage. Although radiation was lethal at high doses, we were able to establish an experimental system where radiation exposure (protons and X-rays) induced DNA damage dose-dependently without causing major cytotoxic effects as assessed by several cytotoxicity assays. Most importantly, we explored the impact of radiation exposure on integration frequency using a clonogenic assay and demonstrated that as predicted, proton-induced DNA damage promotes the integration of HPV-like foreign DNA in oral keratinocytes. Overall, the insights gained from this work enable us to better understand the contribution of radiation exposure and DNA damage to Frontiers in Oncology frontiersin.org 01

Frontiers in Oncology, Feb 10, 2023

Introduction: Treatment-related toxicity following either chemo-or radiotherapy can create signif... more Introduction: Treatment-related toxicity following either chemo-or radiotherapy can create significant clinical challenges for HNSCC cancer patients, particularly those with HPV-associated oropharyngeal squamous cell carcinoma. Identifying and characterizing targeted therapy agents that enhance the efficacy of radiation is a reasonable approach for developing de-escalated radiation regimens that result in less radiation-induced sequelae. We evaluated the ability of our recently discovered, novel HPV E6 inhibitor (GA-OH) to radio-sensitize HPV+ and HPV-HNSCC cell lines to photon and proton radiation. Methods: Radiosensitivity to either photon or proton beams was assessed using various assays such as colony formation assay, DNA damage markers, cell cycle and apoptosis, western blotting, and primary cells. Calculations for radiosensitivity indices and relative biological effectiveness (RBE) were based on the linear quadratic model. Results: Our results showed that radiation derived from both X-ray photons and protons is effective in inhibiting colony formation in HNSCC cells, and that GA-OH potentiated radiosensitivity of the cells. This effect was stronger in HPV+ cells as compared to their HPV-counterparts. We also found that GA-OH was more effective than cetuximab but less effective than cisplatin (CDDP) in enhancing radiosensitivity of HSNCC cells. Further tests indicated that the effects of GA-OH on the response to radiation may be mediated through cell cycle arrest, particularly in HPV+ cell lines. Importantly, the results also showed that GA-OH increases the apoptotic induction of radiation as measured by several apoptotic markers, even though radiation alone had little effect on apoptosis. Conclusion: The enhanced combinatorial cytotoxicity found in this study indicates the strong potential of E6 inhibition as a strategy to sensitize cells to radiation. Future research is warranted to further characterize the interaction of GA-OH derivatives and other E6-specific inhibitors with radiation, as well as its potential to improve the safety and effectiveness of radiation treatment for patients with oropharyngeal cancer.

Cancers, Mar 23, 2021

This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

Capitolo 3: Biodosimetria e biomonitoraggio 3.1 Biomarcatori dell'esposizione a radiazione ionizz... more Capitolo 3: Biodosimetria e biomonitoraggio 3.1 Biomarcatori dell'esposizione a radiazione ionizzante 3.2 Fattore F come indice della qualità della radiazione 3.3 Effetti biologici della radiazione terrestre 3.4 Rana esculenta come biodosimetro animale Capitolo 4: Progetto d'esperimento 4.1 Descrizione dell'esperimento 4.1.1 Protoni 4.1.2 Ioni ferro 4.1.3 Biodosimetria in astronauti 4.1.4 Rana esculenta 4.2 Sistema biologico: linfociti del sangue periferico 4.3 Irraggiamento dei campioni presso il Brookhaven National Laboratory 4.4 Irraggiamento dei campioni con raggi X 4.5 Ibridizzazione In Situ in Fluorescenza 4.6 Colorazione in Giemsa di cromosomi di anfibi 4.7 Sistema di acquisizione di immagini: R.A.I.C. 4.8 Visual R.A.I.C. 101 Capitolo 5: Analisi e discussione dei risultati 105 5.1.1 Analisi dei campioni: protoni 105 5.1.2 Analisi dei campioni: protoni alto-basso-rateo di dose 116 5.2 Analisi dei campioni: ioni Fe e raggi X 119 5.3 Analisi dei campioni: biodosimetria in astronauti 123 5.4 Analisi dei campioni: Rana esculenta 129 5.4.1. Messa a punto di un protocollo per colture linfocitarie 129 5.4.2. Analisi cromosomica su esemplari di controllo 131 III 5.4.3. Risultati dell'analisi su esemplari esposti a differenti dosi di raggi X

Radiation and Environmental Biophysics, Jan 20, 2016

Developing new methods for radiation biodosimetry has been identified as a high priority need in ... more Developing new methods for radiation biodosimetry has been identified as a high priority need in case of a radiological accident or nuclear terrorist attacks. A large-scale radiological incident would result in an immediate critical need to assess the radiation doses received by thousands of individuals. Casualties will be exposed to different doses and dose-rates due to their geographical position and sheltering conditions, and dose-rate is one of the principal factors that determine the biological consequences of a given absorbed dose. In these scenarios high-throughput platforms are required to identify the biological dose in a large number of exposed individuals for clinical monitoring and medical treatment. The RABiT (Rapid Automated Biodosimetry Tool) is designed to be completely automated from the input of blood sample into the machine to the output of a dose estimate. The primary goal of this paper was to quantify the dose-rate effects for RABiTmeasured micronuclei in vitro in human lymphocytes. Blood samples from healthy volunteers were exposed in vitro to different doses of X-rays to acute and protracted doses over a period up to 24 hours. The acute dose (ADR) was delivered at ∼1.03Gy/min and the low dose rate (LDR) exposure at ∼0.31Gy/min. The results showed that the yield of micronuclei decreases with decreasing dose-rate starting at 2Gy, whereas response was indistinguishable from that of acute exposure in the low dose region, up to 0.5Gy. The results showed a linear-quadratic dose-response relationship for the occurrence of micronuclei for the acute exposure and a linear dose-response relationship for the low dose-rate exposure.

Cytogenetic and Genome Research, Aug 12, 2023

Following a mass-casualty nuclear/radiological event, there will be an important need for rapid a... more Following a mass-casualty nuclear/radiological event, there will be an important need for rapid and accurate estimation of absorbed dose for biological triage. The cytokinesis-block micronucleus (CBMN) assay is an established and validated cytogenetic biomarker used to assess DNA damage in irradiated peripheral blood lymphocytes. Here, we describe an intercomparison experiment between two biodosimetry laboratories, located at Columbia University (CU) and Health Canada (HC) that performed different variants of the human blood CBMN assay to reconstruct dose in human blood, with CU performing the assay on isolated lymphocytes and using semi-automated scoring whereas HC used the more conventional whole blood assay. Although the micronucleus yields varied significantly between the two assays, the predicted doses closely matched up to 4 Gy-the range from which the HC calibration curve was previously established. These results highlight the importance of a robust calibration curve(s) across a wide age range of donors that match the exposure scenario as closely as possible and that will account for differences in methodology between laboratories. We have seen that at low doses, variability in the results may be attributed to variation in the processing whilst at higher doses the variation is dominated by inter-individual variation in cell proliferation. This inter-laboratory collaboration further highlights the usefulness of the CBMN endpoint to accurately reconstruct absorbed dose in human blood after ionizing radiation exposure.

Cancer Research, Jun 15, 2022

Tumor recurrence and metastasis contribute to the poor prognosis of ovarian cancer (OC) and gliob... more Tumor recurrence and metastasis contribute to the poor prognosis of ovarian cancer (OC) and glioblastoma multiforme (GBM). The aggressiveness of GBM and OC during recurrence creates a need to identify potential therapeutic targets. The cancer stem cell (CSC)-like phenotype and epithelial mesenchymal transition (EMT) have been shown to contribute to cancer aggressiveness, which makes CSCs and EMT viable targets for therapeutic intervention. EMT is a normal process during development and wound healing, but its reactivation in cancer cells contributes to metastasis. To detect CSC phenotypes in live cells without the use of antibodies, we are analyzing core stem cell transcription factors in which a SOX2/OCT4 response element, designated as SORE6, regulates a GFP reporter system. Also, to detect EMT we are using a GFP based reporter regulated by Zeb1 3’ UTR, a common EMT marker. Our aim is to detect changes in stemness and EMT in response to proton and photon ionizing radiation (IR). We hypothesize that proton and photon IR have similar effects on aggressiveness in OC and GBM. To investigate this, 2 GBM cancer cell lines, 1 breast cancer cell line, 1 OC cell line, and two patient derived samples were exposed to 0, 1, 2, 4 and 8Gy of 250 MeV proton and 6 MeV photon beams. 72 hours post IR exposure, flow cytometry analysis quantified reporter activity. We found an increase in stemness as radiation dosage increased in most cell lines. Results suggest that proton radiation increases stemness more than photon exposure. Citation Format: Aaron Keniston, Alondra Enciso, Yeonkyu Jung, Ashley Antonissen, Ryan Fuller, Ann Morcos, Antonella Bertucci, Marcello Vazquez, Juli Unternaehrer. Detection of stemness and EMT in response to radiation in glioblastoma and ovarian cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6396.

Cytogenetic and Genome Research

Microgravity Science and Technology, 2007

Cosmic radiation represents the major health hazard to crews of interplanetary missions. Moreover... more Cosmic radiation represents the major health hazard to crews of interplanetary missions. Moreover, as the duration of space flights is increasing, as envisaged by the International Space Station (ISS) and Mars mission programs, the risk associated with prolonged exposure to ionizing radiation also increases. Physical doses are measured during flight using passive thermo-luminescent detectors and plastic nuclear track detectors. These measurements provide the accumulated absorbed skin dose. Physical dosimetry is routinely performed in manned space mission; however, biodosimetry is also necessary for a precise assessment of biologically relevant radiation risk [1, 2]. Chromosomal aberrations (CAs) in peripheral blood lymphocytes (PBLs) are particularly suitable to this purpose, as they can provide both equivalent radiation dose and risk. An increase in chromosome damage in human lymphocyte cells is linked to both prior exposure and risk and can be easily measured using the Fluorescence in Situ Hybridization (FISH) technique. CAs in astronauts have been measured after long-term space missions in Low Earth Orbit using both Giemsa staining [3, 4] 206

Radiation and Environmental Biophysics, 2007

Protons are the most abundant element in the galactic cosmic radiation, and the energy spectrum p... more Protons are the most abundant element in the galactic cosmic radiation, and the energy spectrum peaks around 1 GeV. Shielding of relativistic protons is therefore a key problem in the radiation protection strategy of crewmembers involved in long-term missions in deep space. Hydrogen ions were accelerated up to 1 GeV at the NASA Space Radiation Laboratory, Brookhaven National Laboratory, New York. The proton beam was also shielded with thick (about 20 g/cm2) blocks of lucite (PMMA) or aluminium (Al). We found that the dose rate was increased 40-60% by the shielding and decreased as a function of the distance along the axis. Simulations using the General-Purpose Particle and Heavy-Ion Transport code System (PHITS) show that the dose increase is mostly caused by secondary protons emitted by the target. The modified radiation field after the shield has been characterized for its biological effectiveness by measuring chromosomal aberrations in human peripheral blood lymphocytes exposed just behind the shield block, or to the direct beam, in the dose range 0.5-3 Gy. Notwithstanding the increased dose per incident proton, the fraction of aberrant cells at the same dose in the sample position was not significantly modified by the shield. The PHITS code simulations show that, albeit secondary protons are slower than incident nuclei, the LET spectrum is still contained in the low-LET range (&amp;lt;10 keV/microm), which explains the approximately unitary value measured for the relative biological effectiveness.

crr-cu.org

... RARAF Staff (l-r): Andrew Harken, Gerhard Randers-Pehrson, Antonella Bertucci, Yanping Xu, Da... more ... RARAF Staff (l-r): Andrew Harken, Gerhard Randers-Pehrson, Antonella Bertucci, Yanping Xu, David Brenner, Brian Ponnaiya, Charles Geard, Stephen Marino, Sasha Lyulko, Alan Bigelow, Guy Garty and Helen Turner. Not shown: Gary Johnson. David J. Brenner, Ph.D., D.Sc. ...

na.infn.it

La diversa sensibilità individuale del tessuto sano alle radiazioni ionizzanti determina la neces... more La diversa sensibilità individuale del tessuto sano alle radiazioni ionizzanti determina la necessità di effettuare test di monitoraggio in pazienti sottoposti a radioterapia, al fine di personalizzare i piani di trattamento radioterapeutici. L'analisi delle aneuploidie è stata ...

... Title : Schermatura dei protoni relativistici. Language : English. Author, co-author : Durant... more ... Title : Schermatura dei protoni relativistici. Language : English. Author, co-author : Durante, M [&gt; &gt; &gt; &gt;. Bertucci, A [&gt; &gt; &gt; &gt;. Gialanella, G [&gt; &gt; &gt; &gt;. Grossi, G [&gt; &gt; &gt; &gt;. Manti, L [&gt; &gt; &gt; &gt;. Pugliese, Marie [Centre Hospitalier Universitaire de Liège - CHU &gt; &gt; Dermatopathologie &gt;]. ...

Radiation research, 2015

The biological risks associated with low-dose-rate (LDR) radiation exposures are not yet well def... more The biological risks associated with low-dose-rate (LDR) radiation exposures are not yet well defined. To assess the risk related to DNA damage, we compared the yields of two established biodosimetry end points, γ-H2AX and micronuclei (MNi), in peripheral mouse blood lymphocytes after prolonged in vivo exposure to LDR X rays (0.31 cGy/min) vs. acute high-dose-rate (HDR) exposure (1.03 Gy/min). C57BL/6 mice were total-body irradiated with 320 kVP X rays with doses of 0, 1.1, 2.2 and 4.45 Gy. Residual levels of total γ-H2AX fluorescence in lymphocytes isolated 24 h after the start of irradiation were assessed using indirect immunofluorescence methods. The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to determine apoptotic cell frequency in lymphocytes sampled at 24 h. Curve fitting analysis suggested that the dose response for γ-H2AX yields after acute exposures could be described by a linear dependence. In contrast, a linear-quadratic dose-respo...