IRCCS San Raffaele Scientific Institute Showcase

Rationale: to evaluate Fibroblast Growth Factor 21 (FGF21) as a possible biomarkers for depressive symtopms in elderly. FGF21 is a stress-induced hepatokine involved in inflammation and neuroendocrine regulation, processes implicated in depression. Methods: We investigated the association between FGF21 and depressive symptoms in community-dwelling older adults. Data were obtained from 52 older individuals (median age 70; 61.5% women) within the FRASNET cohort who underwent longitudinal assessments (2017 for baseline –2024 for follow up). Depressive symptoms were evaluated using the 15-item Geriatric Depression Scale (GDS). Regression models adjusted for age, sex, and body mass index were used to assess associations between FGF21 and depressive symptoms. Results: Circulating FGF21 levels declined significantly over time (p = 0.03) but remained higher in individuals with depressive symptoms at both baseline and follow-up. Elevated baseline FGF21 predicted higher GDS scores at follow-up (adjusted B = 0.003, 95% CI 0.000–0.006, p = 0.049). Discriminatory performance for elevated depressive symptoms was modest (AUC = 0.66). Conclusions: Higher baseline FGF21 levels were associated with greater depressive symptom burden at follow-up. These findings should be considered preliminary and hypothesis-generating. Further studies using diagnostic outcomes and larger samples are warranted.

Rationale: To identify circulating biomarkers of fraily. Participants were eligible if they were ≥65 years old, able to walk >500 m without assistance, and had a life expectancy >6 months. Exclusion criteria included severe cognitive impairment (Mini-Mental State Examination [MMSE] <18), inability to provide informed consent, or severe health conditions such as uncontrolled hypertension, recent fractures of the upper or lower extremities, or myocardial infarction within the previous year. For this analysis, individuals recruited in nursing homes and those with missing data required to compute frailty or body composition were excluded. Data were collected at baseline (2017) and follow-up (2024). Frailty was assessed using three instruments: the Frailty Index (FI), the modified Frailty Phenotype (FP), and the Clinical Frailty Scale (CFS). Nutritional status was evaluated with the Mini Nutritional Assessment–Short Form (MNA-SF) and categorized as malnourished (≤7), at risk (8–11), or well-nourished (12–14). Sarcopenia was screened using the SARC-F questionnaire and confirmed according to EWGSOP2 criteria. Muscle strength was assessed using the SPPB chair-stand test (>15 s indicating reduced strength). Muscle mass was measured by bioelectrical impedance analysis (BIA), with cut-offs <32.9 % in men and <23.9 % in women. Sarcopenic obesity was defined according to ESPEN/EASO criteria as the coexistence of reduced muscle strength, low muscle mass, and excess fat mass (≥30 % in men and ≥42 % in women). Physical activity was assessed with the Physical Activity Scale for the Elderly (PASE; <76 indicating low activity). Cognitive function was measured using the MMSE (≤24 indicating impairment), and depressive symptoms using the Geriatric Depression Scale (GDS-15). Functional status was evaluated through Activities of Daily Living (ADL) and Instrumental ADL (IADL), and fall risk using the Tinetti scale. Biomarker analyses were performed in plasma samples from 52 of 228 participants due to assay costs and sample availability. Participants were randomly selected among those reassessed at follow-up and were representative of the overall cohort. Blood samples were collected in EDTA tubes from non-fasting subjects, processed within 24 h, aliquoted, and stored at −80 °C. Biomarker concentrations were measured using the ELLA™ automated immunoassay system (Bio-Techne, USA). Continuous variables were reported as mean ± SD or median (IQR), and categorical variables as frequencies (%). Baseline and follow-up comparisons used paired t-tests or Wilcoxon tests. Associations between biomarkers and clinical outcomes were examined using regression models adjusted for age and sex. GDF-15 and FGF-21 concentrations were log-transformed. ROC analyses evaluated the ability of baseline biomarkers to discriminate adverse outcomes at follow-up. Optimal thresholds were determined using the Youden index. Analyses were performed using IBM SPSS Statistics version 25.

Rationale: to identify circulating biomarkers for malnutrition in elderly. Background: Malnutrition is a prevalent geriatric syndrome, with multifactorial etiology and consequences for health and independence. Inflammation contributes to nutritional decline, yet conventional inflammatory markers often lack sensitivity for identifying malnutrition risk. The soluble receptor for advanced glycation end-products (sRAGE), a modulator of inflammatory responses, has emerged as a biomarker of disease risk and adverse outcomes in various conditions. Objectives: to evaluate the association between circulating sRAGE levels and nutritional status in community-dwelling older adults. Methods: This prospective observational study was conducted within the FRASNET cohort. Fifty-two community-dwelling older adults underwent multidimensional geriatric assessments during two time periods: 2017–2020 and 2023–2024. Serum sRAGE levels were measured at both timepoints. Nutritional status was assessed using the Mini Nutritional Assessment Short Form (MNA-SF). Associations between sRAGE and clinical parameters were evaluated through linear regression models adjusted for age and sex. The diagnostic performance of sRAGE in identifying malnutrition was assessed using ROC curve analysis. Results: Higher baseline sRAGE levels were significantly associated with lower BMI (β = –0.003, p = 0.036), reduced calf circumference (β= –0.002, p = 0.04), and poorer nutritional status as revealed by MNA-SF scores (β= –0.001, p = 0.03) at follow-up. Associations were more pronounced in women. ROC analysis indicated good diagnostic accuracy for identifying malnutrition risk, with an AUC of 0.85. The optimal sRAGE cut-off value for malnutrition risk was 1,362.5 pg/mL. Conclusions: Higher sRAGE levels were prospectively associated with poorer nutritional outcomes in older adults, particularly in women. sRAGE may aid early identification of inflammation-related malnutrition risk.

Machine Learning (ML) applied to healthcare Real World Data (RWD) may improve patient management. RWD, however, requires extensive preprocessing to make it ML-ready. The aim of this resource is to explore the impact of preprocessing on ML models applied to RWD from type 2 diabetes patients visits. A GitHub repo is available with the code related to the experiments, to show all the steps we prepared, the decisions we took and alternative pipelines we created. Work done in partial fulfillment of my PhD dissertation: "Leveraging real-world data with machine learning to disentangle the complexity of multimorbid internal medicine patients" Linked in the paper: Montagna M, Rabadzhiev AS, Traverso A, Setola E, Draetta E, Dimonte A, Barbieri S, Fabiani B, Piemonti L, Esposito A, Tacchetti C and Rovere Querini P (2026) From raw data to actionable insights: preprocessing real-world data for machine learning in diabetes care. Front. Digit. Health 8:1685842. doi: 10.3389/fdgth.2026.1685842

Preterm (PRET) neonates are characterized by reduced mature hematopoietic cell count and increased risk of infection, but the biological features of their Hematopoietic stem/progenitor cells (HSPC) compartment, responsible to produce immune cells, are largely unknown. Here, we exploited an optimized workflow to evaluate the immunophenotype, clonogenic potential and differentiation properties of peripheral blood (PB) HSPC from 35 PRET (< 32 weeks gestation) and 26 TERM neonates at birth. Compared to TERM, PRET displayed increased circulation of HSPC, Hematopoietic Stem Cells (HSC) and Multi-Potent Progenitors (MPP). Functionally, TERM and PRET HSPC showed a comparable clonogenic potential, although with a skew towards erythroid colonies of PRET precursors, and similar multi-lineage production. Strikingly, PRET-HSC+MPP and TERM-HSC+MPP displayed equivalent differentiation efficiency and were respectively enriched in uni-erythroid, in line with the erythroid-biased fetal hematopoiesis within the liver, and in uni-lymphoid clones, possibly reflecting the BM output. Finally, PRET with subsequent late-onset neonatal sepsis, who were even more premature, had lower counts of mature lymphoid and myeloid cells but not of HSPC subpopulations. Altogether, PRET- and TERM-HSPC showed comparable in vitro clonogenic and differentiation efficiency. In summary, the functional properties of neonatal HSPCs evolve with the gestational age, from more erythroid-biased fate at earlier stages towards a more myeloid/lymphoid-skewed composition due to distinct hematopoietic niche factors acting on HSPCs with similar differentiation properties. Moreover, our data support the hypothesis that a higher degree of immaturity of the hematopoietic system might increase the susceptibility to develop septic events in preterm newborns.

Mutations in UMOD, encoding uromodulin, lead to Autosomal Dominant Tubulointerstitial Kidney Disease (ADTKD), a genetic cause of kidney failure. UMOD mutations have a common gain-of-toxic-function effect, causing mutant uromodulin retention in the endoplasmic reticulum (ER). This leads to ER stress, alteration of protein homeostasis and mitochondrial dynamics, defective autophagy and increased cell death. Calorie restriction exerts a beneficial role in diseases characterized by accumulation of pathogenic protein and inflammation, by modulating several pathways, including autophagy induction and suppression of inflammation and fibrosis. Given the relevance of these features in ADTKD, we investigated the effect of calorie restriction on disease onset and progression. Transgenic mice expressing C147W uromodulin (TgUmodC147W) were subjected to a moderate (30%) calorie restriction regimen for 15 or 24 weeks, starting at different stages of disease progression. Calorie restriction restored autophagy, as shown by decreased P62 punctae and quenched mTOR activation specifically in mutant uromodulin expressing cells, and it recovered expression of key ER-phagy receptor genes, with a concomitant, striking reduction of mutant uromodulin ER retention. In pre-symptomatic TgUmodC147W mice, calorie restriction alleviated epithelial cell stress. This, likely along with a direct anti-inflammatory effect of calorie restriction, prevented inflammation and progressive decline of kidney function. At this early disease stage, calorie restriction ameliorated the already established kidney damage and reduced fibrosis, suggesting reversal of ADTKD phenotype. Calorie restriction was also effective in significantly delaying disease progression in TgUmodC147W mice with advanced disease and already compromised kidney function. Calorie restriction enhanced autophagy and uromodulin degradation, counteracting the primary effect of UMOD mutations, and significantly ameliorated kidney disease onset and progression. Complete data files relative to the manuscript "Cratere MG, Perrone B, Canciani B, Schaeffer C, Rampoldi L. Calorie Restriction Leads to Degradation of Mutant Uromodulin and Ameliorates Inflammation and Fibrosis in UMOD-Related Kidney Disease. J Am Soc Nephrol. 2026 Feb 3. doi: 10.1681/ASN.0000001032. Epub ahead of print. PMID: 41632531".

We defined an antidepressant potentiation treatment with add-on low-dose interleukin 2 (IL-2). IL-2 is a T-cell growth factor which has proven anti-inflammatory efficacy in autoimmune conditions, increasing thymic production of naïve CD4 + T cells, and possibly correcting the partial T cell defect observed in mood disorders. We performed a single-center, randomised, double-blind, placebo-controlled phase II trial evaluating the safety, clinical efficacy and biological responses of low-dose IL-2 in depressed patients with major depressive (MDD) or bipolar disorder (BD). 36 consecutively recruited inpatients at the Mood Disorder Unit were randomised in a 2:1 ratio to receive either aldesleukin (12 MDD and 12 BD) or placebo (6 MDD and 6 BD). Active treatment significantly potentiated antidepressant response to ongoing SSRI/SNRI treatment in both diagnostic groups, and expanded the population of T regulatory, T helper 2, and percentage of Naive CD4+/CD8 + immune cells. Changes in cell frequences were rapidly induced in the first five days of treatment, and predicted the later improvement of depression severity. No serious adverse effect was observed. This is the first randomised control trial (RCT) evidence supporting the hypothesis that treatment to strengthen the T cell system could be a successful way to correct the immuno-inflammatory abnormalities associated with mood disorders, and potentiate antidepressant response.

Hematopoietic stem/progenitor cell (HSPC) aging has long been associated with myeloid skewing, reduced clonal output, and impaired regenerative capacity, but quantitative immunophenotypic and functional analysis across the human lifespan has been lacking. Here, we provide a comprehensive phenotypic, transcriptional, and functional dissection of human hematopoiesis from youth to advanced age. Although primitive hematopoietic stem cell (HSC) numbers were stable during aging, overall cellularity declined, especially for erythroid and lymphoid lineages. HSPC from older individuals exhibited repopulating frequencies comparable to those from younger donors in both primary and secondary xenografts; however, aged HSC displayed impaired differentiation, chromatin and cell-cycle dysregulation, and poor tolerance to activation-induced proliferative stress, resulting in DNA damage and senescence-like features post-xenotransplantation. Importantly, imposing proliferative stress on young human HSPCs in vivo recapitulated key aging-associated phenotypic and functional declines. Together, our findings identify dysregulated activation responses as a defining feature of HSPC aging and establish proliferative stress–based xenotransplantation models as powerful platforms for investigating age-related hematopoietic dysfunctions. This work is published on Journal of Experimental Medicine (doi: 10.1084/jem.20251805) All the data generated in this study have been deposited in the San Raffaele Open Research Data Repository under accession code doi: 10.17632/72ty5v9djn.1 The RNA and ATAC sequencing data generated and discussed in this study have been deposited in NCBI's Gene Expression Omnibus (Edgar et al., 2002) and are accessible through GEO SuperSeries GSE311225, which contains the following data: GSE243327 (RNA-seq), GSE311221 (ATAC-seq). The code used to process and to generate the images of RNAseq data in this manuscript is available at: http://www.bioinfotiget.it/gitlab/custom/LetteraScala_Ageing/bulk_RNAseq. WES data generated and discussed in this study are available at the European Nucleotide Archive (ENA) under the following accession code: PRJEB93902

Using a multi-polygenic score approach, we characterized the relationship between genome-wide information and the history of PPD in patients with mood disorders, with the hypothesis that multiple polygenic risk scores (PRSs) could potentially influence the development of PPD. The PLS linear regression in the whole sample defined a model explaining 27.12% of the variance in the presence of PPD history, 56.73% of variance among MDD, and 42.96% of variance in BD. Our findings highlight that multiple genetic factors related to circadian rhythms, inflammation, and psychiatric diagnoses are top contributors to the prediction of PPD. Specifically, in MDD, the top contributing PRS was monocyte count, while in BD, it was chronotype, with PRSs for inflammation and psychiatric diagnoses significantly contributing to both groups.

The neurobiological differences between women who have experienced a peripartum episode and those who have only had episodes outside of this period are not well understood. Our findings demonstrate that women who have experienced a peripartum episode are neurobiologically distinct from women who have no history of PPD in a cluster within the basal ganglia, an area important for motivation, decision making, and emotional processing. Furthermore, we show that the genetic load for estradiol has a differing effect in this area based on PPD status, which supports the claim that PPD is associated with sensitivity to sex steroid hormones.