Leader: Fabiola Olivieri (INRCA); Other collaborator(s): Miriam Capri (UNIBO); Nicola Baldini (UNIBO); Stefano Salvioli (UNIBO); Licia Iacoviello (NEUROMED), RTDA PNRR TBA, UNICATT
To analyze the functional biomarkers already available related to inflammaging and anti-inflammaging as well as biological age (IL-6, CXC-L9, Humanin, FGF21, GDF15, inflamma-miRs) and/or identified in spoke #2 in association with routine laboratory parameters and clinical information to characterize ageing phenotypes and to track aging trajectories in relation to multimorbidity, frailty, functional decline and death of geriatric patients. A specific focus on biomarkers of endothelial dysfunction, cellular senescence will be performed. To apply multivariate analyses and machine learning approaches. To test customized composite biomarkers signatures to estimate diagnostic/prognostic sensitivity/specificity in specific settings of geriatric patients and in samples of real-world population.
Brief description of the activities and of the intermediate results: In collaboration with researchers of Spoke 2, Partner INRCA reviewed data relative to biological aging clocks and omics-based data, suggesting different organ-specific aging rates. The results of this analysis have been published in Aging Research Reviews (Prattichizzo et al., 2024). Also, Partner INRCA reviewed the literature regarding circulating biomarkers of AD in combination with biomarkers of inflammaging/neuro-inflammaging. The results of this analysis have been published in Aging Research Reviews (Abbatecola et al., 2024).
As far as Partner UNIBO, Prof. Capri obtained the ethical committee approval for including the project CAROMIRNA-19 in Age-It. This project aims at the identification of a molecular signature for the carotid artery occlusion. Different cytokines have been tested i.e., FGF21, IL-6, sgp130, sIL1-Ralpha, Fibronectin, NfL, CXCL9, GDF8, Follistatin and Activin A. The most significant cytokines are GDF8, CXCL9 and IL-6. These data have been reported in a manuscript submitted to Frontiers in Endocrinology (Capri et al., submitted). Also, a review article has been published on the role of microRNAs in age-related vascular diseases (Ciavarella et al., 2024).
Furthermore, Partner UNIBO (Prof. Salvioli) has completed and published a study on the subject of GDF15 where they have demonstrated that GDF15 is associated to Perilipin 2 (PLIN2) knock down (KD) and is needed for the induction of cell senescence (Chiariello, Rossetti et al., 2024). Another study by the same group dealing with the diagnostic power of both circulating and intracellular forms of GDF15 in sarcopenic elderly patients has been submitted to Frontiers in Endocrinology (Chiariello et al., submitted). As far as the analyses of 4 core biomarkers (GDF15, FGF21, sRAGE and NfL) in the 100+ cohort, Partner UNIBO has analyzed so far the samples for FGF21 and GDF15.
Partner UNIBO (Prof. Baldini) has analyzed potential circulating biomarkers associated with osteosarcopenia and bone fragility. Specifically, serum levels of the inflammatory bone remodeling-related cytokines IL-6 and IL-8 have been quantified in two groups of patients undergoing hip replacement surgery. The serum levels of TNF-α as a further potential marker associated with osteosarcopenia and bone fragility.
Partner NEUROMED has identified within the CASSIOPEA population, a subgroup of 1,000 subjects aged over 65 at baseline (T0), who had repeated clinical evaluations, questionnaires, and blood samples at follow-up (T1). Additionally, in the Moli-sani population aged over 65 (n=5,831), a case-control study has been designed on subjects who developed Parkinson's disease or Alzheimer's disease at follow-up, compared with a control group. Moreover, hypoalbuminemia has been tested in the Moli-sani cohort as a easily measurable markers of frailty and predictor of mortality in individuals aged 65 years and older.
Main policy, industrial and scientific implications: drafting and projecting of a national event on HL, foreseen at the end of September/first part of October 2024
Partner INRCA has studied risk factors associated with survival in old type 2 diabetes (T2D) patients, 500 samples. In this study, all-cause 16-year mortality has been investigated in a real-world sample of older diabetic patients (age > 70 years) afferent to the outpatient facilities of INRCA hospital (Ancona, Italy). Routine laboratory parameters were included in the analysis. Machine learning approach was applied to identify the most relevant variable on risk mortality. Analysis of serum biomarkers NfL, GDF15, FGF21 and SRAGE in 1000 plasma samples from REPORTAGE INRCA study was performed. A review paper on Autonomic nervous system imbalance has been submitted and is now under review.
Partner UNIBO Salvioli’s group: The analysis of soluble biomarkers (GDF15, FGF21, NfL and sRAGE) is completed on the 100+ cohort. The association study with the Frailty Index calculated as deficit accumulation over 45 functional, clinical and hematochemical parameters is ongoing. The role of GDF15 is being investigated on K562 erythroblastoid cancer cell line and in dermal fibroblasts induced to adipocytic differentiation. In the latter, upon differentiation, a strong decrement of GDF15 is observed. Studies are in progress.
Partner UNIBO, Capri’s group: Current analysis of correlations between all the tested biomarkers in CAROMIRNA (IL-6, gp130, IL-6Ralpha, NfL, RAGEs, GDF8, Follistatin, Activin, Fibronectin, CXCL9, FGF21, GDF15) with frailty index (FI, using 33 hemato-biochemical parameters and others related to disease presence) suggest that FI correlates with GDF15 (ELLA Technique) in the population 1 of CAROMIRNA: r = 0.3595, p = 0.0209. However, further analysis needs to be performed in order to confirm the goodness of FI adopted. Analysis on Population 2 (patients with different level of carotid artery occlusion) is ongoing.
Partner UNIBO Baldini's group: investigation of the gut microbiota profile in a patient cohort undergoing hip replacement surgery:
- cases: patients with fracture from femoral neck fragility (total n = 10);
- controls: patients with severe osteoarthritis of the coxofemoral joint (total n = 10).
This analysis focused on the taxonomic composition of the microbiota at the phylum and family levels. While alpha and beta diversity did not differ between the two groups, we observed compositional variations, warranting further investigation. To explore potential associations between dietary patterns and gut microbiota composition in these patients, nutritional questionnaires completed by the case and control groups of patients have been analysed. A muscle questionnaire (SARC-F) to diagnose sarcopenia has also been analysed.
Partner NEUROMED in the two identified populations, is measuring the biomarkers selected in the first phase of the project, using the ELLA tool (Ella Automated Immunoassay Systems), according to the standardization procedures developed within the WP. Moreover, in the whole Moli-sani population it has analysed the association between blood markers of inflammation and sarcopenia with aging related disease.
In the framework of task 3.1, Partner INRCA performed the quantification of selected serum biomarkers NfL, GDF15, FGF21 and SRAGE through Ella™ Automated ELISA technique in 1000 plasma samples from REPORTAGE study. Multivariate statistical analysis and machine learning approach are currently applied to identify the most relevant combination of biomarkers/routinary parameters associated with frailty and/or short and long-term mortality risk. In this period, we also investigated the relevance of triglyceride glucose (TyG) index in predicting long-term mortality and major adverse cardiovascular events in patients affected by type 2 diabetes (T2D).A retrospective analysis was conducted on a cohort of 568 patients with T2D and 376 presumably healthy controls (CTR). Routine biomarkers were measured on serum samples using commercially available methods. Patients with T2D exhibited higher TyG Index values compared to CTR, with significant correlations between the TyG Index and markers of obesity, glucose metabolism, inflammation, and liver function. Overall, the TyG index was identified as a valuable prognostic marker for long-term risk of all-cause mortality and MACE in patients with T2D, supporting its use in clinical risk stratification. We are preparing the final draft.
Within the activity of Partner UNIBO, the analysis of the same four biomarkers has been performed in 463 community-dwelling people of different age, from 50 to 113 years of age and compared with the Frailty Index generated using 45 clinical/functional parameters. An algorithm has been generated that can identify frail people (FI<0.25) with an accuracy of 82%. Other activities of UNIBO included the evaluation of the following biomarkers: blood circulating miRNA 126-5p and 1271-5p on 10 patients with different level of carotid artery occlusion through real time qPCR; gp130, activin A, GDF-8, IL-6 Ralpha on 11 patients through kit ELISA technique and FGF21 with NfL through ELLA technique on 10 patients. Further biomarkers/cytokines, including miR-126-5p and miR-1271, will be assessed in the 2nd population as soon as reagents will arrive and recruitment will be completed. The 2nd population will permit to assess the prediction value of the identified molecular signature in Capri et al., 2024.
Partner UNISR took advantage of the data collected in the FRASNET study, a multicenter observational cohort study conducted in Italy from April 1, 2017, to July 31, 2019. Recruitment took place at recreational and cultural centers and retirement homes in the Milan and Monza Brianza regions of Italy, enrolling a total of 1,250 participants. Between 2023 and 2024, a telephone follow-up was carried out to assess participants' availability for the study's next phase, revealing that 62 participants had passed away and 235 had attended follow-up visits to date. The study included comprehensive assessments of clinical, physical, and biochemical parameters. The analysis of biobanked blood samples collected, as part of the Age-it project, has highlighted the potential of FGF-21, GDF-15, NfL, and sRAGE in understanding aging and frailty. Specific findings include positive correlations between NFL in 2017 age and frailty and negative correlation between NFL in 2017 and physical activity level in 2024 and ability in performing the instrumental activities of daily living in 2024. Moreover, we found a negative correlation between sRAGE in 2017 and nutritional status in 2024 and a negative correlation between GDF-15 in 2017 and physical activity, cognitive performance and balance in 2024. Finally, we found a negative correlation between FGF-21 in 2017 and physical activity and balance in 2024. Moreover, in a cohort of neurologic patients follow up at the Neurologic Clinic of the San Raffaele Hospital and affected by mild cognitive impairment, and dementia of various aetiology (vascular, Alzheimer, Fronto Temporal, Parkinson) we found a cross sectional correlation between GDF15 and MNA score, and a negative correlation between NFL and gait speed and a negative correlation between sRAGE and frailty index.
Partner NEUROMED has completed the analysis of FGF-21, GDF-15 e NFL in serum 661 samples of subjects over 65 years of age from the CASSIOPEA study at baseline and 598 at T1 follow-up. Moreover, the same biomarkers have been measured in 334 sample from subjects of the Moli-sani study who developed Alzheimer’s disease and 359 who developed Parkinson’s disease at follow-up.
Research products of the reported period related to this Task: