Mario Negri Institute for Pharmacological Research

The combination of 5-fluorouracil and irinotecan (FOLFIRI) remains a standard-of-care treatment for metastatic colorectal cancer (mCRC) yet benefits only about half of patients. Using patient-derived xenografts, we investigated the biological underpinnings of this heterogeneous response. FOLFIRI-resistant models showed transcriptional upregulation of innate immunity and mitochondrial metabolism genes, together with reduced expression of the DNA polymerase POLD1. Sensitive counterparts exhibited a BRCAness-like phenotype with genomic scars of homologous recombination (HR) deficiency, not caused by genetic or epigenetic loss of HR genes but by low abundance of the RAD51 recombinase. In tumoroids, forced RAD51 overexpression attenuated HR deficiency-related scars and chemotherapy-induced damage, whereas HR inhibition through ATM blockade enhanced drug sensitivity. The predictive relevance of key response determinants was validated in clinical samples. This work illuminates functional, nongenetic facets of BRCAness in mCRC and introduces actionable biomarkers and targets, offering prospects to improve clinical decision-making and broaden therapeutic options for chemorefractory patients. SIGNIFICANCE:FOLFIRI response biomarkers in mCRC are lacking. Evidence in patient-derived xenografts, tumoroids, and patients shows that chemosensitivity arises from functional relaxation, rather than (epi)genetic inactivation, of the HR DNA repair pathway. Integrative analyses yield a chemopredictive algorithm centered on the expression of the RAD51 recombinase, with potential to refine patient stratification.

This review comprehensively examines the variability and uncertainty associated with test guideline (TG)-conform genotoxicity data and explores the respective implications for the integration of non-animal-methods (NAMs) into regulatory frameworks. Historical amendments to OECD TGs are mapped to reveal the method’s evolution that improves the scientific quality of the data but also explains data heterogeneity within available databases. An analysis of the major genotoxicity databases ECVAM, ISSMIC, and OASIS demonstrates substantial variability in genotoxicity calls. Using the EFSA genotoxicity database, which currently harbours the best-curated (meta-) data, we estimate that 22–77

Timely and accurate diagnosis of Alzheimer’s disease (AD) in clinical practice is a great challenge, especially during early disease stages with subtle or mild symptoms of cognitive decline. Moreover, robust and more accessible blood-based screening tests for early diagnosis are needed. In this study, we investigated the core AD blood biomarkers — amyloid beta 42 (Aβ42) and 40 (Aβ40) peptides, phosphorylated tau 181 (p-Tau181), neurofilament light chain (NfL), and total tau (t-Tau) — and extracellular vesicle (EVs) size and concentration in individuals characterized by different stages of cognitive decline to identify biochemical markers of dementia for early diagnosis. A total of n = 800 human plasma samples were analyzed. Plasma levels of NfL, t-Tau, p-Tau181, Aβ42, Aβ40 and plasma EVs were evaluated in n = 217 elderly healthy subjects (CTRL), in individuals with subjective cognitive complaints (SCC, n = 48), pre-mild cognitive impairment (pre-MCI, n = 58) and mild cognitive impairment (MCI, n = 426), and in n = 51 probable AD dementia patients (AD-dem), using ultrasensitive Single Molecule Array technology (Simoa®) and nanoparticle tracking analysis (NTA). Logistic regression and Receiver Operating Characteristic (ROC) analyses were employed. Plasma NfL displayed increased levels in AD-dem and MCI patients, while p-Tau181, Aβ42/Aβ40 ratio, Aβ42/p-Tau181 ratio, and EVs plasma levels were altered since the early stages of the pathology: in particular, p-Tau181 levels increased as cognitive symptoms worsened, already in the SCC and pre-MCI groups compared to CTRL, while the ratio of EVs concentration and size (EVs ratio) was decreased in all groups compared to CTRL. Plasma p-Tau181 best classified AD-dem patients from CTRL with an area under the curve (AUC) equal to 0.87, while EVs ratio best differentiated SCC from CTRL (AUC = 0.78). Combining p-Tau181 and EVs ratio with Aβ42/Aβ40 ratio and NfL, respectively, significantly improved the classification of pre-MCI and MCI from CTRL (AUCcomb = 0.79 and AUCcomb = 0.85). Combining biomarkers did not improve accuracy in discriminating MCI from SCC, pre-MCI and AD-dem. p-Tau181 and EVs ratio are promising biomarkers for the identification of individuals at risk of degenerative dementia. Combining the core AD plasma biomarkers with EVs ratio can aid in diagnosing the early stages of AD dementia.

Obesity and metabolic syndrome (MS) represent two of the most pressing global public health challenges, often coexisting and synergistically increasing the risk of cardiovascular and renal complications. Over the past decade, advances in our understanding of the pathophysiological mechanisms underlying chronic kidney disease (CKD) have reshaped our approach to patients with coexisting obesity and CKD. The identification of distinct phenotypes associated with an elevated risk of CKD progression and cardiovascular events, along with the development of novel therapeutic strategies, has contributed to a paradigm shift in clinical management. Currently, therapeutic options such as sodium-glucose co-transporter 2 inhibitors, glucagon-like peptide-1 receptor agonists and their combination with the non-steroidal mineralocorticoid receptor antagonist finerenone-when added to standard renin-angiotensin-aldosterone system blockade-are available and have shown potential for cardiorenal protection in patients with obesity and/or MS and CKD. In this review, we highlight the complex interplay between obesity and CKD, emphasizing the need for an integrated, multidisciplinary approach. The use of multitargeted therapeutic strategies holds promise in substantially altering the natural course of disease progression, ultimately improving patient outcomes.

This multicenter study focuses on the latest (2020-2024) trends in candidemia in 155 Italian ICUs, stressing its frequency and lethality, with a shift toward non-albicans, fluconazole-resistant species, namely C. parapsilosis. In this study, resistance to fluconazole prolonged ICU stays without increasing mortality, while non-catheter-related episodes had a more severe prognosis. Candidemia is the leading fungal bloodstream infection (BSI) in the ICU and carries high mortality. After the COVID-19 pandemic, non-albicans, fluconazole-resistant species increased in Europe, yet contemporary ICU-acquired data are scarce. The primary outcome of this study was to evaluate the incidence of ICU-acquired candidemia per 1,000 “at risk” ICU admissions (ICU length of stay [LOS] ≥ 48 h). Secondary analyses compared ICU-acquired candidemia with ICU-acquired bacterial BSI and assessed species distribution, fluconazole susceptibility, and outcomes by fluconazole resistance and by source of infection. Data were drawn from the PROSAFE network, a prospective multicentric observational project conducted across 192 Italian ICUs (2014–2023). Among 179,590 ICU admissions, 95,662 were at risk. ICU-acquired candidemia occurred in 373 patients (3.9‰ of at-risk admissions). Compared with ICU-acquired bacterial BSI, candidemia patients had longer intra-ICU and hospital LOS (32 vs 24 days and 44 vs 36 days, respectively; p < 0.001) and higher mortality (intra-ICU 41.3