BACKGROUND AND PURPOSE:Metastasis-directed stereotactic body radiotherapy (SBRT), in combination with systemic therapy, is increasingly used in patients with oligometastatic disease (OMD), supported by international guidelines. However, real-world data on health-related quality of life (HRQoL) following SBRT remain limited. This study aimed to describe baseline HRQoL, evaluate changes over time, and identify clinical factors associated with HRQoL and its evolution. MATERIALS AND METHODS:OligoCare is a pragmatic, prospective cohort study within the E2-RADIatE registry, including patients with oligometastatic breast, colorectal, non-small cell lung cancer (NSCLC), and prostate cancer treated with SBRT. An optional HRQoL sub-study assesses patient-reported outcomes at baseline and within 6 months post-SBRT. Analyses included descriptive assessment of baseline HRQoL, multivariable regression to identify factors associated with baseline global health status (GHS), and longitudinal evaluation of HRQoL changes. Clinically meaningful changes were defined using a minimal clinically important difference (MCID) of ≥ 10 points. RESULTS:Of 2,041 patients, 1,033 (50.6%) completed baseline HRQoL questionnaires and 450 (22%) had follow-up data. The median time to follow-up HRQoL assessment was 2.7 months (range 0.5-6.1). Median baseline GHS was 75. Higher baseline GHS was associated with prostate cancer and good performance status (p < 0.0001). At follow-up, mean GHS remained stable (71, SD 19). Among patients with follow-up data, 23% reported worsened HRQoL, 15% improved. No significant associations were found between HRQoL changes and clinical variables. CONCLUSION:Early findings support that SBRT for OMD maintains overall HRQoL in most patients, reinforcing its role in multimodal treatment without compromising patient-reported outcomes.

Background and purpose: Particle therapy (PT), including proton (PRT) and carbon ion radiotherapy (CIRT), offers physical and biological advantages over photon radiotherapy (XRT), particularly for radioresistant tumours such as glioblastoma and osteosarcoma. However, systematic preclinical comparisons using physiologically relevant models remain limited. Material and methods: T98G (glioblastoma), Saos-2 and U2-OS (osteosarcoma) cells were cultured as two-dimensional (2D) monolayers and three-dimensional (3D) spheroids and irradiated with XRT, PRT or CIRT at 2, 4 or 6 Gy. Clonogenic survival, metabolic activity (PrestoBlue), invasion and spheroid growth kinetics were quantified. Relative biological effectiveness (RBE) was derived from survival data, and spheroid sections were analysed histologically (H&E). Results: CIRT induced the strongest cytotoxic and anti-invasive effects across all models. In 2D cultures, the surviving fraction at 2 Gy decreased from 0.62 to 0.69 after XRT to 0.16–0.28 following CIRT (RBE = 2.1–2.4 vs. 1.1 for PRT; p < 0.0001). 3D spheroids exhibited overall higher radioresistance, yet CIRT markedly reduced growth and invasion, lowering normalised indices to 0.52 ± 0.12 (T98G) and 0.58 ± 0.09 (Saos-2) at 6 Gy, while photons often promoted invasion (> 1.2; p < 0.001). RBE values in 3D reached 3.6–4.0. H&E staining confirmed dose-dependent architectural disruption, with carbon ions inducing extensive necrosis and cellular degeneration. Interpretation: This study introduces a robust 3D preclinical platform for radiobiological assessment of particle therapy. CIRT consistently overcame intrinsic and microenvironment-mediated resistance, outperforming photons and protons in suppressing viability, invasion and spheroid integrity, thus reinforcing the translational relevance of 3D models and the therapeutic promise of carbon ion therapy for resistant malignancies.

The phosphorylation of H2AX at serine 139 (γH2AX) is a marker of DNA damage response, well characterized after ionizing radiation (IR) exposure. In this work we compared γH2AX activation in different human cell lines exposed to X-rays, up to 5 Gy, or UV-C light (below ionization threshold), up to 40 J/m2. We selected HeLa, Caco-2 and HaCaT cells as models with different origins and radiosensitivity, conventionally screened with the clonogenic assay. We integrated data from fluorescence microscopy and flow cytometry. Results highlight that γH2AX spatial pattern is determined by the genotoxic agent: X-rays induce discrete, countable foci (associated with DNA double-strand breaks), while UV-C leads to pan-nuclear signal distributions. Instead, dose-response curves are strongly dependent on the cell-line, showing linear or saturating trends, not attributable to the readout technique. After 5 Gy X-rays, γH2AX signal peaks within 1 hour. The residual intensity at 24 hours is compatible with the control for all cell lines; however, cells show different clonogenic survival fractions at 7 days. The kinetics of γH2AX positive cells following 20 J/m2 UV-C exposure is completely different, with a slower accumulation and persistent signaling from viable cells (not restricted to the S-phase) up to 24 hours, but with a loss of clonogenic potential in the long-term. These findings emphasize that γH2AX marker may be considered as a functional sensor of DNA damage response rather than a marker of specific DNA lesions, and that both radiation quality and cellular context must be considered for its correct interpretation.

OBJECTIVE:The results of radiotherapy (RT) in oligometastatic ovarian cancers (OCs) lead to the query whether it is possible to stratify patients based on tumor hallmarks to ensure the best-personalized RT treatment. To address this question, we designed a preclinical study to evaluate the effects of high and low linear energy transfer (LET) radiation while considering molecular features and alpha/beta ratios of different OC cell lines. METHODS:Exponentially growing human OVSAHO, OVCAR8, COV362, and OVCAR3 cells cultured in T-25 and T-75 flasks were exposed to different single physical doses of photons, protons, and carbon ion (CIRT) irradiation. We assessed ovarian cells' in vitro response using clonogenic survival (fitted using LQ model), migration by Boyden chamber assay, and invasion through BioCoat Matrigel invasion assay. RESULTS:Following photon irradiation, OVCAR3 was the most radioresistant and OVCAR8 the most radiosensitive cell line. OC cell migration decreased in a dose-dependent manner after irradiation, with CIRT showing the strongest effect, evident by the α/β ratio. The number of invading cells decreased following irradiation with all types. However, the greatest reduction was seen in CIRT, particularly at higher α/β ratios. Proton irradiation demonstrated similar potential to photons but did not match the effects of carbon ions in terms of survival, migration, and invasion. Conclusion: CIRT markedly reduced survival, migration, and invasion of OC cells, particularly in BRCA wild-type OVCAR3 emphasizing its potential to improve local control and lower metastasis risk. This preliminary study serves as a foundation for developing personalized clinical radiation strategies to treat oligometastatic OCs.

Introduction: Pancreatic cancer (PC) is one of the most aggressive and lethal malignancies, calling for enhanced research. Pancreatic ductal adenocarcinoma (PDAC) represents 70–80% of all cases and is known for its resistance to conventional therapies. Carbon-ion radiotherapy (CIRT) has emerged as a promising approach due to its ability to deliver highly localized doses and unique radiobiological properties compared to X-rays. In vitro radiobiology has relied on two-dimensional (2D) cell culture models so far; however, these are not sufficient to replicate the complexity of the in vivo tumor architecture. Three-dimensional (3D) models become a paradigm shift, surpassing the constraints of traditional models by accurately re-creating morphological, histological, and genetic characteristics as well as the interaction of tumour cells with the microenvironment. Materials and Methods: This study investigates the survival of pancreatic cancer cells in both 2D and spheroids, a 3D model, following photon, proton, and carbon-ion irradiation by means of clonogenic, MTT, spheroid growth, and vitality assays. Results: Our results demonstrate that carbon ions are more efficient in reducing cancer cell survival compared to photons and protons. In 2D cultures, carbon-ion irradiation reduced cell survival to approximately 15%, compared to 45% with photons and 30% with protons. In the 3D culture model, spheroid growth was similarly inhibited by carbon-ion irradiation; however, the overall survival rates were higher across all irradiation modalities compared to the 2D cultures. Carbon ions consistently showed the highest efficacy in reducing cell viability in both models. Conclusions: Our research highlights the pivotal role of 3D models in unraveling the complexities of pancreatic cancer radiobiology, offering new avenues for designing more effective and precise treatment protocols.

BACKGROUND: Breast cancer is the most frequently diagnosed malignancy amongst females. Physical activity acts as an excellent preventative therapy and plays a role in maintaining a good quality of life by reducing the incidences of comorbidity and tumor reoccurrences, as well as alleviating the side effects of pharmacological and radiation therapies. This study aims to evaluate the safety and efficacy of physical activity in improving quality of life and aerobic capacity in patients treated for breast cancer. METHODS: From May 2012 to May 2017, 140 patients treated for breast cancer were recruited on a voluntary basis. After the first consultation, 74 patients were evaluated at three months and 41 at nine months. A non-monitored individualized exercise protocol was prescribed according to the American College of Sport Medicine's guidelines (ACSM). Anthropometric analysis, VO2 peak test with ECG monitoring and the presence of medically unexplained symptoms (MUS) were assessed. RESULTS: In the 41 subjects followed for 9 months, no statistical differences were found in BMI and body composition. The VO2 peak value was noted to increase significantly in 24 subjects between the ages of 36 and 58, while in 17 patients over 58 years old the increase was not significant. MUS showed a downward trend over 9 months. CONCLUSIONS: Our study suggests that even a non-monitored physical exercise has positive effects on cardiopulmonary system and psychophysical well-being. A regular physical exercise maintained for at least 3 months is able to improve cardiovascular function. ( Cite this article as: Bidoglio F, Canepari M, De Simone A, Longa E, Poerio CS, Ceccaroli M, et al . Exercise therapy in breast cancer patients: effects on cardiorespiratory fitness and quality of life. Med Sport 2024;77:390-402. DOI: 10.23736/S0025-7826.24.04464-8)

Drepanocytosis is a genetic disease relevant for its epidemiological, clinical and socio-economic aspects. In our country the prevalence is highly uneven with peaks in former malaria areas, but migration flows in recent years have led to significant changes. In this document we review the screening programs currently existing in Italy with particular emphasis on newborn screening, which in other countries around the world, including within Europe, is at most universal and mandatory. The essential laboratory issues are reviewed, from sampling aspects (cord blood or peripheral), to the analytical (analytical methods dedicated to neonatal screening and adult carrier detection) and post analytical (reporting, informative) ones. An economic analysis based on data collected in the province of Modena is also proposed, clearly showing that neonatal screening is also beneficial from an economic point of view.

Purpose:Primary gynecological melanomas are uncommon with lower survival rates compared to cutaneous melanomas. Although melanocytes have been identified in a variety of mucosal membranes, little is known about their interactions or roles inside the mucosa layer. Melanin is a common pigment in nature and is endowed with several peculiar chemical, paramagnetic, and semiconductive characteristics. One of its latest explored functions is its interaction with ionizing radiation as a protective mechanism as well as its implication in the metastatic cascade of tumor cells. Materials and Methods:In this work, we analyzed in vitro the effects of different doses of photon and carbon ion irradiation on dendrite formation, pigmentation, migration, and invasion abilities of human mucosal melanoma cells of the vagina. We evaluated the morphology and melanin production of HMV-II cells exposed to photon and carbon ion beams with single doses between 0.5 and 10 Gy. Results:Our results showed that irradiation induces dendrite formation or elongation and pigmentation in HMV-II cells in a dose-type-dependent and radiation-type-dependent way but also a decrease in cell motility. Conclusion:The present study describes for the first time an induction of dendritic formation, melanin production, and alterations in migration and invasion abilities by low-linear energy transfer and high-linear energy transfer radiation in human mucosal melanoma cells, suggesting a radioprotective response to further possible exposures increasing the radioresistance of these cells.

PURPOSE:Intraoperative radiation therapy with electrons (IOERT) may represent a viable choice for partial breast reirradiation after repeat quadrantectomy for local recurrence (LR) for primary breast cancer (BC) in lieu of mastectomy. METHODS AND MATERIALS:A database collecting data on partial breast reirradiation with IOERT from 8 Italian centers was set up in 2016 to 2018, providing data on cumulative incidence (CumI) of second LR and survival with a long follow-up. RESULTS:From 2002 to 2015, 109 patients underwent the conservative retreatment. The median primary BC first LR interval was 11.1 years (range, 2.4-27.7). The median first LR size was 0.9 cm (range, 0.3-3.0), and 43.6% cases were luminal A. Median IOERT dose was 18 Gy (range, 12-21), and median collimator diameter was 4 cm (range, 3-6). Median follow-up duration was 11.7 years (IQR, 7.7-14.6). The second LR CumI was 12.2% (95% CI, 6.8%-19.2%) at 5 years and 32.3% at 10 years (95% CI, 22.8%-42.2%), occurring in the same site as the first LR in about half of the cases. Human epidermal growth factor receptor 2 status and collimator size were independent LR predictors. The 5- and 10-year overall survival rates were 95.2% and 88.3%, respectively, whereas 5- and 10-year BC-specific survival rates were 98% and 94.5%, respectively. The development of a second LR significantly reduced BC-specific survival (hazard ratio, 9.40; P < .001). Grade ≥3 fibrosis rate was 18.9%. Patient-reported cosmesis was good/excellent in 59.7% of the cases. CONCLUSIONS:Second LR CumI was within the range of the literature but higher than expected, opening questions on radiation field extension and fractionation schedule. Because a second LR worsened the outcome, salvage modality must be carefully planned.

Introduction: Decellularized extracellular matrix (ECM) bioscaffolds have emerged as a promising three-dimensional (3D) model, but so far there are no data concerning their use in radiobiological studies. Material and Methods: We seeded two well-known radioresistant cell lines (HMV-II and PANC-1) in decellularized porcine liver-derived scaffolds and irradiated them with both high- (Carbon Ions) and low- (Photons) Linear Energy Transfer (LET) radiation in order to test whether a natural 3D-bioscaffold might be a useful tool for radiobiological research and to achieve an evaluation that could be as near as possible to what happens in vivo. Results: Biological scaffolds provided a favorable 3D environment for cell proliferation and expansion. Cells did not show signs of dedifferentiation and retained their distinct phenotype coherently with their anatomopathological and clinical behaviors. The radiobiological response to high LET was higher for HMV-II and PANC-1 compared to the low LET. In particular, Carbon Ions reduced the melanogenesis in HMV-II and induced more cytopathic effects and the substantial cell deterioration of both cell lines compared to photons. Conclusions: In addition to offering a suitable 3D model for radiobiological research and an appropriate setting for preclinical oncological analysis, we can attest that bioscaffolds seemed cost-effective due to their ease of use, low maintenance requirements, and lack of complex technology

Abstract Background: The research into senescent cells, particularly their accumulation and contribution to aging and age-related diseases, has gained significant momentum in recent years. This interest is primarily driven by findings in mouse models, where the excessive buildup of senescent cells is linked to various age-related pathologies. Consequently, there has been a concerted effort in the scientific community to identify and develop senolytics. A notable development in this field is the identification of senolytics within the class of HSP90 inhibitors. These inhibitors, initially investigated for their potential in cancer therapy, have been repurposed due to their high selectivity for senescent cells. Our research has led to new compounds with mild inhibitory activity against HSP90α in response to these challenges. These compounds stand out for their lack of toxic side effects and their demonstrated senolytic activity across multiple cellular models. Objective: The primary objective of this research is to identify and characterize innovative HSP90 inhibitors that exhibit senolytic properties, specifically in the context of hormone-driven cancers. Methods and Results: Utilizing a structure-based virtual screening technique, we identified several compounds displaying strong affinity and predicted inhibitory activity against the target protein, HSP90α. Among these, two inhibitors designated K4 and K5, demonstrated notable senolytic activity in vitro without inducing cytotoxic effects in non-senescent cells. These compounds were tested for their efficacy in senolytic activity and displayed half-maximal inhibitory concentrations (IC50) of 155 nM and 111 nM, respectively. The experimental model for this study was the MCF7 cell line, which was treated with 4-Hydroxytamoxifen (Tam, 10uM for 96 hours) to induce a state of senescence. Subsequent growth curve analyses and beta-galactosidase assays established that Tam treatment effectively halted cell proliferation and induced a senescence state in about 40% of the cell population compared to the ethanol control. When treated with K4 and K5 (10 uM for an additional 96 hours), a significant reduction in the senescent cell population was observed, with K5 showing a particularly pronounced increase in the mortality rate of these cells (up to 2.5-fold compared to the DMSO control). Western blot analysis supported these results, suggesting the effectiveness of K5 combined with Tam in inhibiting proliferation and inducing cell death in MCF7 cells. Conclusion: Our findings propose a novel therapeutic approach for hormone-dependent breast cancer, combining the dual roles of tamoxifen and HSP90 inhibitors to target senescent cancer cells. This strategy could reduce tumor recurrence and contribute to the advancement of cancer treatment methods. Citation Format: Luca Cis, Michela Gottardi Zamperla, Veronica Barbi, Pietro Cigala Fulgosi, Sara De Martino, Aurora Aiello, Simona Nanni, Chiara Cencioni, Davide Pirolli, Paola Tabarelli De Fatis, Giovanni Battista Ivaldi, Marco Malavolta, Christian Steinkühler, Maria Cristina De Rosa, Antonella Farsetti, Sandra Atlante, Carlo Gaetano. Novel HSP90 inhibitors with senolytic activity for a targeted therapy in a hormone-induced breast cancer senescence model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2977.

Background and introduction: Optimal dose and fractionation in stereotactic body radiotherapy (SBRT) for oligometastatic cancer patients remain unknown. In this interim analysis of OligoCare, we analyzed factors associated with SBRT dose and fractionation. Materials and methods: Analysis was based on the first 1,099 registered patients. SBRT doses were converted to biological effective doses (BED) using alpha/beta of 10 Gy for all primaries, and cancer-specific alpha/beta of 10 Gy for nonsmall cell lung and colorectal cancer (NSCLC, CRC), 2.5 Gy for breast cancer (BC), or 1.5 Gy for prostate cancer (PC). Results: Of the interim analysis population of 1,099 patients, 999 (99.5 %) fulfilled inclusion criteria and received metastasis-directed SBRT for NSCLC (n = 195; 19.5 %), BC (n = 163; 16.3 %), CRC (n = 184; 18.4 %), or PC (n = 457; 47.5 %). Two thirds of patients were treated for single metastasis. Median number of fractions was 5 (IQR, 3-5) and median dose per fraction was 9.7 (IQR, 7.7-12.4) Gy. The most frequently treated sites were nonvertebral bone (22.8 %), lung (21.0 %), and distant lymph node metastases (19.0 %). On multivariate anal