Relationship between telomere length, genetic traits and environmental/occupational exposures in bladder cancer risk by structural equation modelling

Background: Telomere length (TL) maintenance plays an important role in bladder cancer (BC) and prognosis. However the manifold influence of everyday life exposures and genetic traits on leucocyte TL (LTL), is not fully elucidated. Methods: Within the framework of a hospital-based case (n = 96)/control (n = 94) study (all Caucasian males), we investigated the extent to which LTL and BC risk were modulated by genetic polymorphisms and environmental and occupational exposures. Data on lifetime smoking, alcohol and coffee drinking, dietary habits and occupational exposures, pointing to aromatic amines (AAs) and polycyclic aromatic hydrocarbons (PAHs) were collected. Structural equation modelling (SEM) analysis appraised this complex relationships. Results: The SEM analysis indicates negative direct links (p < 0.05) between LTL with age, DNA adducts, alcohol and NAT2, and positive ones with coffee, MPO and XRCC3; and between BC risk (p < 0.01) with cigarettes, cumulative exposure to AAs and coffee, while are negative with LTL and age. There was evidence of indirect effects (p < 0.05) on BC risk, probably via LTL reduction, by age and NAT2 (positive link), MPO and XRCC3 (negative link). Conclusions: Our study supports evidence that LTL attrition is a critical event in BC. The new finding that LTL erosion depends on some preventable everyday life exposures genetically modulated, opens new perspectives in BC prevention

Dually Cross-Linked Core-Shell Structure Nanohydrogel with Redox–Responsive Degradability for Intracellular Delivery

A redox-responsive nanocarrier is a promising strategy for the intracellular drug release because it protects the payload, prevents its undesirable leakage during extracellular transport, and favors site-specific drug delivery. In this study, we developed a novel redox responsive core-shell structure nanohydrogel prepared by a water in oil nanoemulsion method using two biocompatible synthetic polymers: vinyl sulfonated poly(N-(2-hydroxypropyl) methacrylamide mono/dilactate)-polyethylene glycol-poly(N-(2-hydroxypropyl) methacrylamide mono/dilactate) triblock copolymer, and thiolated hyaluronic acid. The influence on the nanohydrogel particle size and distribution of formulation parameters was investigated by a three-level full factorial design to optimize the preparation conditions. The surface and core-shell morphology of the nanohydrogel were observed by scanning electron microscope, transmission electronmicroscopy, and further confirmed by Fourier transforminfrared spectroscopy and Raman spectroscopy from the standpoint of chemical composition. The redox-responsive biodegradability of the nanohydrogel in reducing environments was determined using glutathione as reducing agent. A nanohydrogel with particle size around 250 nm and polydispersity index around 0.1 is characterized by a thermosensitive shell which jellifies at body temperature and crosslinks at the interface of a redox-responsive hyaluronic acid core via theMichael addition reaction. The nanohydrogel showed good encapsulation efficiency for model macromolecules of different molecular weight (93% for cytochrome C, 47% for horseradish peroxidase, and 90% for bovine serum albumin), capacity to retain the peroxidase-like enzymatic activity (around 90%) of cytochrome C and horseradish peroxidase, and specific redox-responsive release behavior. Additionally, the nanohydrogel exhibited excellent cytocompatibility and internalization efficiency into macrophages. Therefore, the developed core-shell structure nanohydrogel can be considered a promising tool for the potential intracellular delivery of different pharmaceutical applications, including for cancer therapy

Mitigating cellular aging and enhancing cognitive functionality: visual arts-mediated Cognitive Activation Therapy in neurocognitive disorders

The growing phenomenon of population aging is redefining demographic dynamics, intensifying age-related conditions, especially dementia, projected to triple by 2050 with an enormous global economic burden. This study investigates visual arts-mediated Cognitive Activation Therapy (CAT) as a non-pharmacological CAT intervention targets both biological aging [leukocyte telomere length (LTL), DNA methylation age (DNAmAge)] and cognitive functionality. Aligning with a broader trend of integrating non-pharmacological approaches into dementia care. The longitudinal study involved 20 patients with mild to moderate neurocognitive disorders. Cognitive and functional assessments, and biological aging markers -i.e., LTL and DNAmAge- were analyzed before and after CAT intervention. Change in LTL was positively correlated with days of treatment (p =0.0518). LTL significantly elongated after intervention (p =0.0269), especially in men (p =0.0142), correlating with younger age (p =0.0357), and higher education (p =0.0008). DNAmAge remained instead stable post-treatment. Cognitive and functional improvements were observed for Copy of complex geometric figure, Progressive Silhouettes, Position Discrimination, Communication Activities of Daily Living—Second edition, Direct Functional Status (p < 0.0001) and Object decision (p =0.0594), but no correlations were found between LTL and cognitive gains. Visual arts-mediated CAT effectively mitigates cellular aging, especially in men, by elongating LTL. These findings underscore the potential of non-pharmacological interventions in enhancing cognitive and functional status and general well-being in dementia care. Further research with larger and longer-term studies is essential for validation

Exploring mechanisms of biological aging in susceptible subjects

The aim of the PhD project is to explore molecular mechanisms that characterize the process of biological aging with the main focus on the two most prominent early hallmarks of biological aging such as telomere length (TL) and epigenetic age, also defined as DNA methylation age (DNAmAge), in order to answer the following main questions: 1) If environmental and occupational exposures accelerate the biological aging in healthy subjects and in subjects affected by age-relate disease. 2) If, on the other hand, correct lifestyle including an intensive meditation/relaxing training in healthy subjects and patients with cardiovascular diseases, and heart transplantation in patients suffering from end-stage heart failure, slow down biological aging. To this aims we analyzed biological aging indicators in easily available human tissue (blood leucocytes) and in target organ (i.e., heart of donors and recipients and lung from induced sputum of Chronic Obstructive Pulmonary Disease (COPD) patients). Study populations consist of: 1) HEALTHY SUBJECTS EXPOSED TO GERONTOGENIC ENVIRONMENTAL AND OCCUPATIONAL EXPOSURES a) 71 night-shift workers and 84 day workers as control exposed to circadian rhythm disruption. b) 585 individuals from general population living in North-East Italy exposed to everyday exposure to carcinogenic polycyclic aromatic hydrocarbons (PAHs), which are fundamental constituents of air pollution. c) A prospective cohort study of women and men aged 70 years and older from the area of Treviso, which is characterized by the highest longevity in Italy. TRELONG longitudinal study population comprises 576 subjects at baseline, 300 and 200 subjects at T1 and T2 respectively, exposed to lifestyle and occupational exposures. 2) SUBJECTS AFFECTED BY AGE-RELATE DISEASES a) Bladder cancer (BC), a chronic disease characterized by the interaction between environmental/occupational and genetic risk factors. Study population includes newly diagnosed, histologically confirmed BC patients, admitted to the Urology Departments of two large hospitals from 1997 to 2000. Controls are 94 non-neoplastic urological patients matched to cases by age, period and hospital of admission. b) Idiopathic pulmonary fibrosis (IPF) and Chronic Obstructive Pulmonary Disease (COPD) as the most common manifestations of aging-mediated diseases. Study population consists of n=101 IPF patients (ATS/ERS/JRS/ALAT guidelines) and n=18 moderate COPD patients (GOLD 2019) all enrolled at the ambulatory of Pneumology and Respiratory Physiopathology Wards – Department of Cardiac, Thoracic and Vascular Sciences and Public Health, University of Padova. 3) HEALTHY SUBJECTS AND PATIENTS EXPOSED TO REJUVENATING FACTORS a) Patients after myocardial infarction (n=20) and healthy control individuals (n=10), age- and gender-matched, trained to meditation and relax for 60 days. b) 17 recipients receiving the heart from 17 donors in the period February 2018 - February 2019. The local Ethics Committee - University of Padova, approved the study protocols (3843/AO/16 and 3054/AO/14). Main finding stemming from our results revels that: 1) HEALTHY SUBJECTS EXPOSED TO GERONTOGENIC ENVIRONMENTAL AND OCCUPATIONAL EXPOSURES - Night-shift work is associated with increased systemic inflammation as proved by higher C-reactive protein (CRP) levels among night shift workers. LTL is reduced by CRP, while is positively associated with long pentraxin 3 (PTX3) that, by orchestrating an efficient governance of inflammatory processes, may protect telomere from attrition. This would make nocturnal workers more susceptible to premature LTL shortening and aging. - Certain preventable everyday life exposures to PAHs diminish LTL and even LmtDNAcn, in particular in males, acting through anti-B[a]PDE–DNA adduct formation. Our findings show that indoor activities and diet represent the primary determinants of PAHs exposure in increasing anti-B[a]PDE–DNA adduct levels that in turn decrease in presence of detoxifying GSTM1. - In TRELONG population LTL significantly declines over the years, from baseline to follow up, also considering only n=161 subjects whit all measurements at three different time points. 2) SUBJECTS AFFECTED BY AGE-RELATE DISEASES - LTL attrition is a critical event in BC. In particular, BC risk is increased directly by LTL attrition that depends on some preventable everyday life exposures genetically modulated. Furthermore, indirect effects on BC risk are evidenced via LTL reduction through age and genetic polymorphisms involved in modulating response to environmental exposure. - IPF patients in follow up present an increase in LTL positively related to the duration of antifibrotic treatment, with both pirfenidone and nintedanib, and with a decrease in lung function decline. These results would suggest that telomere shortening in IPF patients treated with antifibrotic drugs may be reversed leading to an increase in LTL. - In COPD patients: a) lung, i.e., pulmonary cells obtained from induced sputum, is biologically older than blood, as determined by TL and DNAmAge; b) blood age acceleration (AgeAcc) defined as the difference between DNAmAge and chronological age, but not TL, highly correlates with lung AgeAcc; c) blood AgeAcc significantly correlates with the main clinical features (CRP and FEV1) of COPD. 3) HEALTHY SUBJECTS AND PATIENTS EXPOSED TO REJUVENATING FACTORS - In healthy subjects but not in patients, DNAmAge is reduced after an intensive relaxing training. Differently, LTL is preserved in healthy subjects, while in patients it continues to decrease. However, the correlation between LTL and chronological age becomes positive after training in both groups. These findings would suggest that intensive relaxing practices influence different aging molecular mechanisms, i.e., DNAmAge and LTL, with a rejuvenating effect. - Biological donors’ heart age is found to be younger than chronological age, suggesting that donors’ cardiac tissues are biologically younger than chronologically measured. Furthermore, biological donors’ left atrium age is 5 years younger than recipients’ left atrium age. This would indicate that patients who underwent heart transplantation have received a younger heart. Further results and comments are described in each work reported in the specific Chapters. Our results contribute to reinforce the concept that biological aging may be modulated by a multiplicity of factors (environmental, occupational, lifestyle) making people more susceptible to premature aging or inducing an accelerated aging or, interestingly, eliciting a rejuvenation effect. In the era of the silver tsunami, identifying gerontogenic and rejuvenating factors is of paramount importance to develop anti-aging strategies for extending the number of healthy years of life

Exploring mechanisms of biological aging in susceptible subjects

The aim of the PhD project is to explore molecular mechanisms that characterize the process of biological aging with the main focus on the two most prominent early hallmarks of biological aging such as telomere length (TL) and epigenetic age, also defined as DNA methylation age (DNAmAge), in order to answer the following main questions: 1) If environmental and occupational exposures accelerate the biological aging in healthy subjects and in subjects affected by age-relate disease. 2) If, on the other hand, correct lifestyle including an intensive meditation/relaxing training in healthy subjects and patients with cardiovascular diseases, and heart transplantation in patients suffering from end-stage heart failure, slow down biological aging. To this aims we analyzed biological aging indicators in easily available human tissue (blood leucocytes) and in target organ (i.e., heart of donors and recipients and lung from induced sputum of Chronic Obstructive Pulmonary Disease (COPD) patients). Study populations consist of: 1) HEALTHY SUBJECTS EXPOSED TO GERONTOGENIC ENVIRONMENTAL AND OCCUPATIONAL EXPOSURES a) 71 night-shift workers and 84 day workers as control exposed to circadian rhythm disruption. b) 585 individuals from general population living in North-East Italy exposed to everyday exposure to carcinogenic polycyclic aromatic hydrocarbons (PAHs), which are fundamental constituents of air pollution. c) A prospective cohort study of women and men aged 70 years and older from the area of Treviso, which is characterized by the highest longevity in Italy. TRELONG longitudinal study population comprises 576 subjects at baseline, 300 and 200 subjects at T1 and T2 respectively, exposed to lifestyle and occupational exposures. 2) SUBJECTS AFFECTED BY AGE-RELATE DISEASES a) Bladder cancer (BC), a chronic disease characterized by the interaction between environmental/occupational and genetic risk factors. Study population includes newly diagnosed, histologically confirmed BC patients, admitted to the Urology Departments of two large hospitals from 1997 to 2000. Controls are 94 non-neoplastic urological patients matched to cases by age, period and hospital of admission. b) Idiopathic pulmonary fibrosis (IPF) and Chronic Obstructive Pulmonary Disease (COPD) as the most common manifestations of aging-mediated diseases. Study population consists of n=101 IPF patients (ATS/ERS/JRS/ALAT guidelines) and n=18 moderate COPD patients (GOLD 2019) all enrolled at the ambulatory of Pneumology and Respiratory Physiopathology Wards – Department of Cardiac, Thoracic and Vascular Sciences and Public Health, University of Padova. 3) HEALTHY SUBJECTS AND PATIENTS EXPOSED TO REJUVENATING FACTORS a) Patients after myocardial infarction (n=20) and healthy control individuals (n=10), age- and gender-matched, trained to meditation and relax for 60 days. b) 17 recipients receiving the heart from 17 donors in the period February 2018 - February 2019. The local Ethics Committee - University of Padova, approved the study protocols (3843/AO/16 and 3054/AO/14). Main finding stemming from our results revels that: 1) HEALTHY SUBJECTS EXPOSED TO GERONTOGENIC ENVIRONMENTAL AND OCCUPATIONAL EXPOSURES - Night-shift work is associated with increased systemic inflammation as proved by higher C-reactive protein (CRP) levels among night shift workers. LTL is reduced by CRP, while is positively associated with long pentraxin 3 (PTX3) that, by orchestrating an efficient governance of inflammatory processes, may protect telomere from attrition. This would make nocturnal workers more susceptible to premature LTL shortening and aging. - Certain preventable everyday life exposures to PAHs diminish LTL and even LmtDNAcn, in particular in males, acting through anti-B[a]PDE–DNA adduct formation. Our findings show that indoor activities and diet represent the primary determinants of PAHs exposure in increasing anti-B[a]PDE–DNA adduct levels that in turn decrease in presence of detoxifying GSTM1. - In TRELONG population LTL significantly declines over the years, from baseline to follow up, also considering only n=161 subjects whit all measurements at three different time points. 2) SUBJECTS AFFECTED BY AGE-RELATE DISEASES - LTL attrition is a critical event in BC. In particular, BC risk is increased directly by LTL attrition that depends on some preventable everyday life exposures genetically modulated. Furthermore, indirect effects on BC risk are evidenced via LTL reduction through age and genetic polymorphisms involved in modulating response to environmental exposure. - IPF patients in follow up present an increase in LTL positively related to the duration of antifibrotic treatment, with both pirfenidone and nintedanib, and with a decrease in lung function decline. These results would suggest that telomere shortening in IPF patients treated with antifibrotic drugs may be reversed leading to an increase in LTL. - In COPD patients: a) lung, i.e., pulmonary cells obtained from induced sputum, is biologically older than blood, as determined by TL and DNAmAge; b) blood age acceleration (AgeAcc) defined as the difference between DNAmAge and chronological age, but not TL, highly correlates with lung AgeAcc; c) blood AgeAcc significantly correlates with the main clinical features (CRP and FEV1) of COPD. 3) HEALTHY SUBJECTS AND PATIENTS EXPOSED TO REJUVENATING FACTORS - In healthy subjects but not in patients, DNAmAge is reduced after an intensive relaxing training. Differently, LTL is preserved in healthy subjects, while in patients it continues to decrease. However, the correlation between LTL and chronological age becomes positive after training in both groups. These findings would suggest that intensive relaxing practices influence different aging molecular mechanisms, i.e., DNAmAge and LTL, with a rejuvenating effect. - Biological donors’ heart age is found to be younger than chronological age, suggesting that donors’ cardiac tissues are biologically younger than chronologically measured. Furthermore, biological donors’ left atrium age is 5 years younger than recipients’ left atrium age. This would indicate that patients who underwent heart transplantation have received a younger heart. Further results and comments are described in each work reported in the specific Chapters. Our results contribute to reinforce the concept that biological aging may be modulated by a multiplicity of factors (environmental, occupational, lifestyle) making people more susceptible to premature aging or inducing an accelerated aging or, interestingly, eliciting a rejuvenation effect. In the era of the silver tsunami, identifying gerontogenic and rejuvenating factors is of paramount importance to develop anti-aging strategies for extending the number of healthy years of life

DNA methylation-based age prediction and telomere length reveal an accelerated aging in induced sputum cells compared to blood leukocytes: a pilot study in COPD patients.

none5noAging is the predominant risk factor for most degenerative diseases, including chronic obstructive pulmonary disease (COPD). This process is however very heterogeneous. Defining the biological aging of individual tissues may contribute to better assess this risky process. In this study, we examined the biological age of induced sputum (IS) cells, and peripheral blood leukocytes in the same subject, and compared these to assess whether biological aging of blood leukocytes mirrors that of IS cells. Biological aging was assessed in 18 COPD patients (72.4 ± 7.7 years; 50% males). We explored mitotic and non-mitotic aging pathways, using telomere length (TL) and DNA methylation-based age prediction (DNAmAge) and age acceleration (AgeAcc) (i.e., difference between DNAmAge and chronological age). Data on demographics, life style and occupational exposure, lung function, and clinical and blood parameters were collected. DNAmAge (67.4 ± 5.80 vs. 61.6 ± 5.40 years; p = 0.0003), AgeAcc (-4.5 ± 5.02 vs. -10.8 ± 3.50 years; p = 0.0003), and TL attrition (1.05 ± 0.35 vs. 1.48 ± 0.21 T/S; p = 0.0341) are higher in IS cells than in blood leukocytes in the same patients. Blood leukocytes DNAmAge (r = 0.927245; p = 0.0026) and AgeAcc (r = 0.916445; p = 0.0037), but not TL, highly correlate with that of IS cells. Multiple regression analysis shows that both blood leukocytes DNAmAge and AgeAcc decrease (i.e., younger) in patients with FEV1% enhancement (p = 0.0254 and p = 0.0296) and combined inhaled corticosteroid (ICS) therapy (p = 0.0494 and p = 0.0553). In conclusion, new findings from our work reveal a differential aging in the context of COPD, by a direct quantitative comparison of cell aging in the airway with that in the more accessible peripheral blood leukocytes, providing additional knowledge which could offer a potential translation into the disease management.noneManuela Campisi, Filippo Liviero, Piero Maestrelli, Gabriella Guarnieri, Sofia PavanelloCampisi, Manuela; Liviero, Filippo; Maestrelli, Piero; Guarnieri, Gabriella; Pavanello, Sofi

Exploring Epigenetic Age in Response to Intensive Relaxing Training: A Pilot Study to Slow Down Biological Age

DNA methylation (DNAm) is an emerging estimator of biological aging, i.e., the often-defined "epigenetic clock", with a unique accuracy for chronological age estimation (DNAmAge). In this pilot longitudinal study, we examine the hypothesis that intensive relaxing training of 60 days in patients after myocardial infarction and in healthy subjects may influence leucocyte DNAmAge by turning back the epigenetic clock. Moreover, we compare DNAmAge with another mechanism of biological age, leucocyte telomere length (LTL) and telomerase. DNAmAge is reduced after training in healthy subjects (p = 0.053), but not in patients. LTL is preserved after intervention in healthy subjects, while it continues to decrease in patients (p = 0.051). The conventional negative correlation between LTL and chronological age becomes positive after training in both patients (p < 0.01) and healthy subjects (p < 0.05). In our subjects, DNAmAge is not associated with LTL. Our findings would suggest that intensive relaxing practices influence different aging molecular mechanisms, i.e., DNAmAge and LTL, with a rejuvenating effect. Our study reveals that DNAmAge may represent an accurate tool to measure the effectiveness of lifestyle-based interventions in the prevention of age-related diseases

A rejuvenation effect of the antifibrotic therapy correlates with lung function improvement in IPF patients

Idiopathic pulmonary fibrosis (IPF) is an irreversible and fatal lung disease destined to increase as a sequel of the Severe Acute Respiratory Syndrome Coronavirus-2. Several studies implicate telomere shortening as hallmark in the pathomechanism of IPF. Our understanding of disease pathogenesis remains, however, incomplete, which hampers also the development of more effective drugs. Aims of this study are: 1) to analyze telomere length in blood leukocytes (LTL) in well phenotypically characterized IPF patients at diagnosis, before treatment with antifibrotic drugs, Pirfenidone and Nintedanib (T0); 2) to investigate in follow up (T1) a possible change in the rate of LTL by combining data on clinic, hematochemical tests, respiratory function and occupational exposure. We examined a group of 24 IPF patients that underwent to a therapy follow up (T1) (mean±SD 297±124 days). We observed an increase in LTL at T1 compared to LTL at T0 (mean±SD LTL (T/S), T1 vs T0, 1.29 ± 0.26 vs 1.19 ± 0.27; p=0.051). Multiple linear regression analysis reveals that the LTL increase, at T1, is significantly related to the time of treatment (p=0.002), LTL at T0 (p<0.0001) and to a slowing in lung function decline (FVC%pred) (p=0.054). The other variables considered including occupational exposure, pack-years, occupational risk factor, gender and body mass index are not significantly related. Our research would indicate a rejuvenation effect of the antifibrotic therapy by measuring the LTL that correlates with lung function improvement. This suggests that targeting fundamental mechanisms of cellular aging has the potential to interfere with the severity of the disease