Archivio Istituzionale della Ricerca- Università del Piemonte Orientale
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Bipartite Genetically Encoded Biosensors to Sense Calcium Ion Dynamics at Membrane–Membrane Contact Sites
No full textSelf-complementing bipartite fluorescent proteins (FPs) are useful tools for the detection of protein–protein proximity and for localizing fluorophores to membrane–membrane contact sites. Here, we report versions of circularly permuted green FP (GFP), red FP (RFP), and mNeonGreen (NG), which are split into a large fragment composed of nine β-strands and a small fragment composed of two β-strands. In each case, the large and small fragments can associate in live cells to form the complete 11-stranded FP β-barrel. We further converted each of these three self-complementing FPs into bipartite calcium ion (Ca2+) biosensors. We demonstrate that appropriately targeted versions of these split FPs, and split FP-based biosensors, can be functionally assembled at membrane–membrane contact sites. We employ the bipartite NG-based Ca2+biosensor for visualization of pharmacologically induced Ca2+release at mitochondria-endoplasmic reticulum contact sites (MERCs)
Cancer-associated fibroblast driven paracrine IL-6/STAT3 signaling promotes migration and dissemination in invasive lobular carcinoma
Get PDFBackground: Invasive lobular carcinoma (ILC) is the second most common histological subtype of breast cancer after invasive ductal carcinoma of no special type (NST), accounting for 10–15% of diagnoses. Despite the myriad molecular, histological and clinical differences between ILC and NST tumors, patients are treated in the same way, and although prognosis initially is good, ILC patients have poorer long-term outcomes. Understanding the differences between these two subtypes and identifying ILC-enriched therapeutic targets is necessary to improve patient care. Methods: Human and mouse cancer-associated fibroblasts (CAFs), ILC cell lines and patient-derived organoids were used for in vitro and in vivo studies, including western blotting, migration, organotypic invasion assays and dissemination in zebrafish embryos. RNASeq was used to identify CAF and interleukin-6 (IL-6)-derived gene signatures. Bioinformatic analysis of public databases and immunohistochemical of human tumor microarrays was carried out. Results: We identified IL-6 as a paracrine CAF-derived factor that activates Signal-Transducer-and-Activator-of-Transcription-3 (STAT3) in human and mouse ILC models. Analysis of human breast tumors showed that the IL-6/JAK/STAT3 pathway is enriched in ER + ILC compared to ER + NST. A 42-gene CAF dependent IL-6 gene signature and 64-gene consensus IL-6 gene signature were generated and were significantly enriched in ER + ILC, with many of the genes overexpressed in ILC tumors. IL-6 treatment suppressed downstream estrogen signaling and also led to the acquisition of a more mesenchymal-like phenotype associated with increased migration and invasion. Finally, IL-6 treatment significantly increased ILC cell dissemination following injection into zebrafish embryos. Conclusions: CAF-derived IL-6 drives paracrine activation of the IL6/JAK/STAT3 signaling pathway which is enriched in ILC. This leads to the acquisition of pro-tumorigenic phenotypes, highlighting the pathway as a potential therapeutic target in ILC
Cancer-specific Mortality After Radical Nephrectomy and Thrombectomy for Renal Cell Carcinoma with Tumor Thrombus According to Histological Subtypes
No full textBackground and objective: We tested the effect of histological subtypes on cancer-specific mortality (CSM) after radical nephrectomy (RN) with thrombectomy for nonmetastatic renal cell carcinoma with tumor thrombus (RCC-TT) within the Surveillance, Epidemiology and End Results database (2010–2021). Methods: We used propensity score matching (PSM), multivariable competing-risk regression (CRR) analyses, and cumulative incidence functions to test differences in CSM between papillary RCC (pRCC) and clear-cell RCC (ccRCC), and between chromophobe RCC (chRCC) and ccRCC. Key findings and limitations: Of 5516 patients with nonmetastatic RCC-TT, 180 (3.3%), 186 (3.4%), and 5150 (93%) harbored pRCC, chRCC, and ccRCC, respectively. TT was limited to the renal vein in 135 (75%), 168 (91%), and 4457 (87%) patients with pRCC, chRCC, and ccRCC, respectively. After 1:3 PSM, 180 (100%) pRCC cases were compared with 540 (11%) ccRCC cases. The 5-yr CSM rate was 43% in pRCC versus 26% in ccRCC. Multivariable CRR revealed that pRCC independently predicted higher CSM relative to ccRCC (hazard ratio [HR] 1.81, 95% confidence interval [CI] 1.31–2.50; p < 0.001). After 1:3 PSM, 186 (100%) chRCC cases were compared with 558 (11%) ccRCC cases. The 5-yr CSM rate was 14% in chRCC versus 19% in ccRCC. Multivariable CRR revealed no significant difference in CSM between chRCC and ccRCC (HR 0.62, 95% CI 0.37–1.03; p = 0.07). Limitations include the retrospective nature of the study and a lack of data on disease progression and subsequent treatment. Conclusions and clinical implications: For patients with RCC-TT treated with RN with thrombectomy, pRCC independently predicted a 1.8-fold higher CSM rate in comparison to ccRCC, with no significant difference in CSM rates between chRCC and ccRCC. Adjuvant therapy should be considered for patients with pRCC or chRCC in the TT setting
Determination of the spin and parity of all-charm tetraquarks
No full textThe traditional quark model1,2 accounts for the existence of baryons, such as protons and neutrons, which consist of three quarks, as well as mesons, composed of a quark–antiquark pair. Only recently has substantial evidence started to accumulate for exotic states composed of four or five quarks and antiquarks3. The exact nature of their internal structure remains uncertain4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28–29. Here we report the first measurement of quantum numbers of the recently discovered family of three all-charm tetraquarks30, 31–32, using data collected by the CMS experiment at the Large Hadron Collider from 2016 to 2018 (refs. 33,34). The angular analysis techniques developed for the discovery and characterization of the Higgs boson35, 36–37 have been applied to the new exotic states. Here we show that the quantum numbers for parity P and charge conjugation C symmetries are found to be +1. The spin J of these exotic states is determined to be consistent with 2ħ, while 0ħ and 1ħ are excluded at 95% and 99% confidence levels, respectively. The JPC = 2++ assignment implies particular configurations of constituent spins and orbital angular momenta, which constrain the possible internal structure of these tetraquarks
Neuroglia in neurodegeneration: Alzheimer, Parkinson, and Huntington disease
No full textThe conspicuous rise of chronic neurodegenerative diseases, including Alzheimer (AD), Parkinson (PD), and Huntington (HD) diseases, is currently without disease-modifying therapies and accompanied by an excessive rate of unsuccessful clinical trials. This reflects a profound lack of understanding of the pathogenesis of these diseases, indicating that the current paradigms guiding disease modeling and drug development are in need of reconsideration. The role of neuroglia, namely astrocytes, microglial cells, and oligodendrocytes, in the pathogenesis of neurodegenerative diseases emerged during the last decades. This chapter provides the state-of-the-art update on the changes of astrocytes, microglial cells, and oligodendrocytes in AD, PD, and HD. A growing body of evidence suggests that homeostatic and defensive functions of glial cells are compromised at different disease stages, leading to increased susceptibility of neurons to noxious stimuli, eventually resulting in their malfunction and degeneration. Investments are needed in the generation of novel preclinical models suitable for studying glial pathology, in “humanizing” research, and in-depth investigation of glial cell alterations to slow down and, possibly, halt and prevent the rise of neurodegenerative disease. Targeting glial cells opens new therapeutic avenues to treat AD, PD, and HD
Multiomics approach identifies dysregulated lipidomic and proteomic networks in Parkinson’s disease patients mutated in TMEM175
No full textParkinson’s disease (PD) represents one of the most frequent neurodegenerative disorders for which clinically useful biomarkers remain to be identified and validated. Here, we adopted an untargeted omics approach to disclose lipidomic, metabolomic and proteomic alterations in plasma and in dermal fibroblasts of PD patients carrying mutations in TMEM175 gene. We revealed a wide dysregulation of lysosome, autophagy, and mitochondrial pathways in these patients, supporting a role of this channel in regulating these cellular processes. The most significant altered lipid classes were Fatty acyls, Glycerophospholipids and Phosphosphingolipids. The plasma level of Phosphatidylcholines (PC) and Phosphatidylinositol (PI) 34:1 significantly correlated with an earlier age at onset of the disease in TMEM175 patients (p = 0.008; p = 0.006). In plasma we also observed altered amino acids metabolic pathways in PD patients. We highlighted that increased level of L-glutamate strongly correlated (p < 0.001) with the severity of motor and non-motor symptoms in PD_TMEM175 patients. In dermal fibroblasts, we disclosed alterations of proteins involved in lipids biosynthesis (PAG15, PP4P1, GALC, FYV1, PIGO, PGPS1, PLPP1), in the insulin pathway (IGF2R), in mitochondrial metabolism (ACD10, ACD11, ACADS) and autophagy (RAB7L). Interestingly, we quantified 43 lysosomal or lysosomal-related proteins, which were differentially modulated between TMEM175 patients and controls. Integrative correlation analysis of proteome and lipidome of PD_TMEM175 cellular models identified a strong positive correlation of 13 proteins involved in biosynthetic processes with PC and Ceramides. Altogether, these data provide novel insights into the molecular and metabolic alterations underlying TMEM175 mutations and may be relevant for PD prediction, diagnosis and treatment
ASO Author Reflections: Rethinking Recurrence Patterns After Resection of Large Hepatocellular Carcinoma: Time to Overcome the Current Staging Paradigm?
No full textAmyotrophic Lateral Sclerosis Prevalence Projection in 2040: A Less Rare Disease
No full textObjective: To project ALS prevalence across multiple countries through 2040, accounting for both population aging and increased survival. Methods: Data from the Piemonte and Valle d'Aosta ALS register (PARALS) was used to estimate the trends in incidence and prevalence from 2005 to 2019. Survival trends over this period were also assessed. The observed annual increase was then projected into future years up to 2040. Concurrently, the incidence for each future year was calculated using population projections. Finally, the prevalence rate for each year was estimated as the product of the projected incidence and the projected survival. We also estimated survival for fifteen countries by dividing prevalence by incidence, based on available data, and applied the same increase observed in PARALS to project prevalence in these countries up to 2040. Results: Using data from 3294 patients, we determined that ALS survival increased by 0.06 years annually from 2005 to 2019 in Piemonte and Valle d'Aosta. Considering changes in incidence due to population aging, the prevalence is projected to reach 15.72 per 100,000 population by 2040 in this area, while rising by a median of 24.9% across multiple countries worldwide. If a new drug could provide a 6-month increase in survival starting in 2025, disease prevalence would rise by 37.8% by 2040. We provided a web interface so users can model different data and assumptions. Interpretation: ALS prevalence is projected to increase significantly over the next decades. This underscores the need for careful planning and allocation of public health resources
Non-Animal Hyaluronic Acid from Tremella fuciformis: A New Source with a Structure and Chemical Profile Comparable to Hyaluronic Acid
Get PDFTremella fuciformis is high in polysaccharides, which have a structure made up of a straight chain of (1→3) α-D-mannan and side chains of glucuronic acid, xylose, and fucose. This study aimed to evaluate whether the non-animal hyaluronic acid extracted from Tremella fuciformis can maintain the chemical and physical characteristics of hyaluronic acid that ensure its biological functionality. Chemical and physical analyses such as hyaluronic content, screening of metals, purity, pH, nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (ATR/FTIR), and MALDI-TOF were performed. Chemical characterisation revealed that the most abundant polysaccharide in the extract was hyaluronic acid, accounting for ca. 87.76%, with a molecular weight above 2000 kDa. In addition, ATR/FTIR and NMR spectroscopy and MALDI-TOF analysis confirmed that Tremella fuciformis extract is a source of non-animal hyaluronic acid. In summary, every molecular attribute examined played a significant role in determining the functional qualities of the extract, indicating that a thoughtful choice of extraction technique can enhance its advantages