LAMC2 marks a tumor-initiating cell population with an aggressive signature in pancreatic cancer

[Background]: Tumor-initiating cells (TIC), also known as cancer stem cells, are considered a specific subpopulation of cells necessary for cancer initiation and metastasis; however, the mechanisms by which they acquire metastatic traits are not well understood.[Methods]: LAMC2 transcriptional levels were evaluated using publicly available transcriptome data sets, and LAMC2 immunohistochemistry was performed using a tissue microarray composed of PDAC and normal pancreas tissues. Silencing and tracing of LAMC2 was performed using lentiviral shRNA constructs and CRISPR/Cas9-mediated homologous recombination, respectively. The contribution of LAMC2 to PDAC tumorigenicity was explored in vitro by tumor cell invasion, migration, sphere-forming and organoids assays, and in vivo by tumor growth and metastatic assays. mRNA sequencing was performed to identify key cellular pathways upregulated in LAMC2 expressing cells. Metastatic spreading induced by LAMC2- expressing cells was blocked by pharmacological inhibition of transforming growth factor beta (TGF-β) signaling.[Results]: We report a LAMC2-expressing cell population, which is endowed with enhanced self-renewal capacity, and is sufficient for tumor initiation and differentiation, and drives metastasis. mRNA profiling of these cells indicates a prominent squamous signature, and differentially activated pathways critical for tumor growth and metastasis, including deregulation of the TGF-β signaling pathway. Treatment with Vactosertib, a new small molecule inhibitor of the TGF-β type I receptor (activin receptor-like kinase-5, ALK5), completely abrogated lung metastasis, primarily originating from LAMC2-expressing cells.[Conclusions]: We have identified a highly metastatic subpopulation of TICs marked by LAMC2. Strategies aimed at targeting the LAMC2 population may be effective in reducing tumor aggressiveness in PDAC patients. Our results prompt further study of this TIC population in pancreatic cancer and exploration as a potential therapeutic target and/or biomarker.This work was supported by: Marie Curie IF (H2020-MSCA-IF-2015, #703753), My First AIRC Grant (MFAG-2017, #20206), POR Campania FESR 2014/2020 (Project SATIN) to E.L.; AIRC IG grant 2018 n.21420 to A.D.L.; FIMP to D.D.C.; AECC (Proye18046BATL_002) to E.B.; My First AIRC Grant (MFAG grant #23029), WorldWide Cancer Research (Research grant #20–0188), EASI Genomics consortium (TNA project #15158) and the World Cancer Research Fund (Seed grant #2021–1769) to A.C

Alterations of the secretory pathway induced by a mutant p53/miR-30d axis

Cancer is, nowadays, among the most prevalent and deadly diseases worldwide. This term describes a group of pathologies characterized by an abnormal growth of a mass of cells harboring mutations in their DNA, resulting in uncontrolled growth, evasion from the cell control checkpoint mechanisms and spreading throughout the body. In recent years, the investigation of the interplay between cancer cells and the tumor micro-environment has gained a central spot in the comprehension of the neoplastic development and outgrowth. At a cellular level, the main hub regulating the communication with the surrounding tissues is the secretory pathway, which is deputed to the movement of proteins and lipids between the ER, the Golgi apparatus, and, through the secretory vesicles, to the extracellular space. Alterations in the functions of the secretory pathway could have an important role in helping the development and progression of the malignancies, fostering metastasis, invasion, altered secretion patterns and cytoskeletal remodeling.These phenotypes, among the others, have been linked by a vast amount of data to the mutated forms of p53, derived by missense point mutations in the TP53 gene which lose the oncosuppressive function of the wild-type form, and acquire, in many cases, novel pro-oncogenic features. mutp53s exert their pro-neoplastic functions through a plethora of interactors, both coding and non-coding. In our laboratory, we identified miR-30d as a new target gene of mutp53, which appears to be regulated through the interaction of this protein with the hypoxia-inducible factor HIF1\u3b1. Preliminary evidence show that miR-30d-regulated genes are enriched for factors involved in the unfolded protein response activation and protein secretion, suggesting an effect on the structure and functions of the secretory pathway. Our results show that miR-30d expression is able to blunt the activation of the UPR following drug-induced ER stress and concomitantly to induce major alterations in the secretory pathway organelles, mainly represented by a strong vesiculo-tubulation of the Golgi apparatus. Moreover, miR-30d, through the modulation of its direct and indirect targets, as its upstream regulators mutant p53 and HIF1\u3b1, is able to strongly promote the secretion of proteins by cancer and normal cells. Taken all together, the findings reported in this thesis suggest a role for this newly described mutant p53/HIF1\u3b1/miR-30d axis in the regulation of the structures and functions of the secretory pathway, and particularly on the Golgi apparatus

Macro Minerals and Trace Elements in Milk of Dairy Buffaloes and Cows Reared in Mediterranean Areas

Aim of this study was to evaluate the differences in Ca, P, K, Na, Mg, Zn, Fe, Cu, Mn, Se, Mo, Co, Li, B, Ti, Rb, and Sr concentrations in milk from buffaloes and cows reared in the same farm in Mediterranean areas and fed diets including the same ingredients. Individual milk samples were obtained from 32 Mediterranean buffaloes and 29 Italian Friesian cows and samples of milk, dietary ingredients and drinking water were analyzed for the investigated chemical elements by inductively coupled plasma-mass spectrometry. Data about milk element concentrations were processed by one-way analysis of variance. Buffalo milk contains higher concentrations of Ca, P, Mg, Zn, Fe, Cu, B, Ti, and Sr, and lower concentrations of K, Na, Mo, Li, and Rb compared to cow milk, whereas milk from both species contains similar concentrations of Mn, Se, and Co. The concentrations of the investigated elements in the diet were similar for both species and the differences observed between buffalo and cow milk were not dependent on environmental factors

7th European Symposium on South American Camelids and 3rd European Meeting on Fibre Animals - SYMPCAM 2017

Following the tradition of previous events, the Symposium will cover a broad range of topics related to the breeding and keeping of South American domestic and wild Camelids, wool sheep, Cashmere and Angora goats, Angora rabbit and all other fibre mammals in Europe and in the World. The invitation to participants interested in all fibre producing animals is expected to stimulate the exchange of knowledge and thus enabling their expertise to be implemented on a more general scale

Sterol regulatory element binding protein 1 couples mechanical cues and lipid metabolism

SREBP transcription factors activate lipid synthesis and generate raw materials to lipidate various proteins. Here, the authors show that a stiff cellular environment causes RhoA lipidation and acto-myosin contraction, which inhibits SREBP1 and connects the extracellular matrix to lipid metabolism

Organic Selenium induces ferroptosis in pancreatic cancer cells

Pancreatic ductal adenocarcinoma (PDA) cells reprogram both mitochondrial and lysosomal functions to support growth. At the same time, this causes significant dishomeostasis of free radicals. While this is compensated by the upregulation of detoxification mechanisms, it also represents a potential vulnerability.Here we demonstrate that PDA cells are sensitive to the inhibition of the mevalonate pathway (MVP), which supports the biosynthesis of critical antioxidant intermediates and protect from ferroptosis. We attacked the susceptibility of PDA cells to ferroptotic death with selenorganic compounds, including dibenzyl diselenide (DBDS) that exhibits potent pro-oxidant properties and inhibits tumor growth in vitro and in vivo. DBDS treatment induces the mobilization of iron from mitochondria enabling uncontrolled lipid peroxidation. Finally, we showed that DBDS and statins act synergistically to promote ferroptosis and provide evidence that combined treatment is a viable strategy to combat PDA

EMID2 is a novel biotherapeutic for aggressive cancers identified by in vivo screening

Background: New drugs to tackle the next pathway or mutation fueling cancer are constantly proposed, but 97% of them are doomed to fail in clinical trials, largely because they are identified by cellular or in silico screens that cannot predict their in vivo effect. Methods: We screened an Adeno-Associated Vector secretome library (> 1000 clones) directly in vivo in a mouse model of cancer and validated the therapeutic effect of the first hit, EMID2, in both orthotopic and genetic models of lung and pancreatic cancer. Results: EMID2 overexpression inhibited both tumor growth and metastatic dissemination, consistent with prolonged survival of patients with high levels of EMID2 expression in the most aggressive human cancers. Mechanistically, EMID2 inhibited TGFβ maturation and activation of cancer-associated fibroblasts, resulting in more elastic ECM and reduced levels of YAP in the nuclei of cancer cells. Conclusion: This is the first in vivo screening, precisely designed to identify proteins able to interfere with cancer cell invasiveness. EMID2 was selected as the most potent protein, in line with the emerging relevance of the tumor extracellular matrix in controlling cancer cell invasiveness and dissemination, which kills most of cancer patients

Additional file 1 of EMID2 is a novel biotherapeutic for aggressive cancers identified by in vivo screening

Supplementary Material 1: Table S1. Composition of the pilot AAV9 poo