DLK1/DIO3 locus upregulation by a ß-catenin-dependent enhancer drives cell proliferation and liver tumorigenesis.

The CTNNB1 gene, encoding ß-catenin, is frequently mutated in hepatocellular carcinoma (HCC, ∼30%) and in hepatoblastoma (HB, >80%), in which DLK1/DIO3 locus induction is correlated with CTNNB1 mutations. Here, we aim to decipher how sustained ß-catenin activation regulates DLK1/DIO3 locus expression and the role this locus plays in HB and HCC development in mouse models deleted for Apc (ApcΔhep) or Ctnnb1-exon 3 (ß-cateninΔExon3) and in human CTNNB1-mutated hepatic cancer cells. We identified an enhancer site bound by TCF-4/ß-catenin complexes in an open conformation upon sustained ß-catenin activation (DLK1-Wnt responsive element [WRE]) and increasing DLK1/DIO3 locus transcription in ß-catenin-mutated human HB and mouse models. DLK1-WRE editing by CRISPR-Cas9 approach impaired DLK1/DIO3 locus expression and slowed tumor growth in subcutaneous CTNNB1-mutated tumor cell grafts, ApcΔhep HB and ß-cateninΔExon3 HCC. Tumor growth inhibition resulted either from increased FADD expression and subsequent caspase-3 cleavage in the first case or from decreased expression of cell cycle actors regulated by FoxM1 in the others. Therefore, the DLK1/DIO3 locus is an essential determinant of FoxM1-dependent cell proliferation during ß-catenin-driven liver tumorigenesis. Targeting the DLK1-WRE enhancer to silence the DLK1/DIO3 locus might thus represent an interesting therapeutic strategy to restrict tumor growth in primary liver cancers with CTNNB1 mutations.

Wnt, glucocorticoid and cellular prion protein cooperate to drive a mesenchymal phenotype with poor prognosis in colon cancer.

BACKGROUND: The mesenchymal subtype of colorectal cancer (CRC), associated with poor prognosis, is characterized by abundant expression of the cellular prion protein PrPC, which represents a candidate therapeutic target. How PrPC is induced in CRC remains elusive. This study aims to elucidate the signaling pathways governing PrPC expression and to shed light on the gene regulatory networks linked to PrPC. METHODS: We performed in silico analyses on diverse datasets of in vitro, ex vivo and in vivo models of mouse CRC and patient cohorts. We mined ChIPseq studies and performed promoter analysis. CRC cell lines were manipulated through genetic and pharmacological approaches. We created mice combining conditional inactivation of Apc in intestinal epithelial cells and overexpression of the human prion protein gene PRNP. Bio-informatic analyses were carried out in two randomized control trials totalizing over 3000 CRC patients. RESULTS: In silico analyses combined with cell-based assays identified the Wnt-ß-catenin and glucocorticoid pathways as upstream regulators of PRNP expression, with subtle differences between mouse and human. We uncover multiple feedback loops between PrPC and these two pathways, which translate into an aggravation of CRC pathogenesis in mouse. In stage III CRC patients, the signature defined by PRNP-CTNNB1-NR3C1, encoding PrPC, ß-catenin and the glucocorticoid receptor respectively, is overrepresented in the poor-prognosis, mesenchymal subtype and associates with reduced time to recurrence. CONCLUSIONS: An unleashed PrPC-dependent vicious circle is pathognomonic of poor prognosis, mesenchymal CRC. Patients from this aggressive subtype of CRC may benefit from therapies targeting the PRNP-CTNNB1-NR3C1 axis.

Downregulation of mitochondrial complex I induces ROS production in colorectal cancer subtypes that differently controls migration.

BACKGROUND: Colorectal cancer (CRC) can be classified into four molecular subtypes (CMS) among which CMS1 is associated with the best prognosis, while CMS4, the mesenchymal subtype, has the worst outcome. Although mitochondria are considered to be hubs of numerous signaling pathways, the study of mitochondrial metabolism has been neglected for many years. Mitochondrial Complex I (CI) plays a dual role, both in energy and reactive oxygen species (ROS) production. However, the possible contribution of CI to tumorigenesis in cancer remains unclear. The purpose of this study was to investigate the CI under the prism of the CMS classification of CRC in ex vivo models. METHODS: Biochemical dosages, bioenergetics analysis and western-blot were used to characterize CI expression, function and redox balance in LoVo and MDST8 cell lines, belonging to CMS1 and CMS4 subgroups, respectively. Cell proliferation and migration were assessed by xCELLigence technology. Overproduction or scavenging of mitochondrial ROS (mtROS) were performed to analyze the effect of mtROS on proliferation, migration, and mesenchymal markers. Focal adhesion kinase (FAK) and its activation were analyzed by immunofluorescence. We assessed the distribution of two CI scores in CRC cohorts according to CMS classification and their relevance for patient survival. RESULTS: We found that CI is downregulated in CMS4 cells and is associated with elevated mtROS. We establish for the first time that in these migrating cells, mtROS production is maintained at optimal levels not only through changes in CI activity but also by inactivation/acetylation of superoxide dismutase 2 (SOD2), a major mitochondrial antioxidant enzyme. We show that promoting or scavenging mtROS both mitigate CMS4 cells' migration. Our results also point to a mtROS-mediated focal adhesion kinase (FAK) activation, which likely sustains their migratory phenotype. Using cohorts of CRC patients, we document that the expression of CI is downregulated in the CMS4 subgroup, and that low CI expression is associated with poor prognosis. Patients' datasets reveal an inverse correlation between CI and the epithelial-mesenchymal transition (EMT) pathway. CONCLUSION: We showed that inhibition of CI contributes to heighten mtROS, which likely foster MDST8 migration and might account for the specific EMT signature of CMS4 tumors. These data reveal a novel role of mitochondrial CI in CRC, with biological consequences that may be targeted with anti- or pro-oxidant drugs in clinical practice.

A novel Chr1-miR-200 driven whole transcriptome signature shapes tumor immune microenvironment and predicts relapse in early-stage lung adenocarcinoma.

BACKGROUND: In Lung adenocarcinoma (LUAD), targeted therapies and immunotherapies have moved from metastatic to early stage and stratification of the relapse risk becomes mandatory. Here we identified a miR-200 based RNA signature that delineates Epithelial-to-mesenchymal transition (EMT) heterogeneity and predicts survival beyond current classification systems. METHODS: A miR-200 signature was identified using RNA sequencing. We scored the miR-200 signature by WISP (Weighted In Silico Pathology), used GSEA to identify pathway enrichments and MCP-counter to characterize immune cell infiltrates. We evaluate the clinical value of this signature in our series of LUAD and using TCGA and 7 published datasets. RESULTS: We identified 3 clusters based on supervised classification: I is miR-200-sign-down and enriched in TP53 mutations IIA and IIB are miR-200-sign-up: IIA is enriched in EGFR (p < 0.001), IIB is enriched in KRAS mutation (p < 0.001). WISP stratified patients into miR-200-sign-down (n = 65) and miR-200-sign-up (n = 42). Several biological processes were enriched in MiR-200-sign-down tumors, focal adhesion, actin cytoskeleton, cytokine/receptor interaction, TP53 signaling and cell cycle pathways. Fibroblast, immune cell infiltration and PDL1 expression were also significantly higher suggesting immune exhaustion. This signature stratified patients into high-vs low-risk groups, miR-200-sign-up had higher DFS, median not reached at 60 vs 41 months and within subpopulations with stage I, IA, IB, or II. Results were validated on TCGA data on 7 public datasets. CONCLUSION: This EMT and miR-200-related prognostic signature refines prognosis evaluation independently of tumor stage and paves the way towards assessing the predictive value of this LUAD clustering to optimize perioperative treatment.

Poloxamers Have Vaccine-Adjuvant Properties by Increasing Dissemination of Particulate Antigen at Distant Lymph Nodes.

Vaccine technology is still facing challenges regarding some infectious diseases, which can be addressed by innovative drug delivery systems. In particular, nanoparticle-based vaccines combined with new types of adjuvants are actively explored as a platform for improving the efficacy and durability of immune protection. Here, biodegradable nanoparticles carrying an antigenic model of HIV were formulated with two combinations of poloxamers, 188/407, presenting or not presenting gelling properties, respectively. The study aimed to determine the influence of poloxamers (as a thermosensitive hydrogel or a liquid solution) on the adaptive immune response in mice. The results showed that poloxamer-based formulations were physically stable and did not induce any toxicity using a mouse dendritic cell line. Then, whole-body biodistribution studies using a fluorescent formulation highlighted that the presence of poloxamers influenced positively the dissemination profile by dragging nanoparticles through the lymphatic system until the draining and distant lymph nodes. The strong induction of specific IgG and germinal centers in distant lymph nodes in presence of poloxamers suggested that such adjuvants are promising components in vaccine development.

Family Resilience as an Emerging Concept in Neonatology: Evidence From a Metasummary Review.

BACKGROUND: Family resilience can be observed through specific resilience-promoting processes, namely, shared belief systems, communication, and organizational processes, but the concept remains mostly unstudied in neonatology. This metasummary aims to evaluate the frequency of family resilience processes in qualitative scientific literature to illustrate how family resilience is exhibited in the neonatal intensive care unit (NICU) setting. METHODS: A search among 4 databases yielded 7029 results, which were reviewed for inclusion. Following Sandelowski and Barroso's qualitative metasummary method, findings from each study were independently coded and frequency effect size was calculated. RESULTS: Forty-six primary qualitative studies published between 2016 and 2022 conducted with parents of preterm infants who discussed their NICU hospitalization experience were included in this metasummary. All 9 of Walsh's family resilience processes were identified in the literature, and their frequency effect size ranged from 4% to 91%. Four additional themes emerged pertaining to specific family resilience behaviors exhibited by NICU families. CONCLUSION: This analysis sheds new light on the most recent qualitative evidence of parents' experiences in the NICU by analyzing it through the lens of family resilience and posits family resilience as a promising concept in relation to the predominance of the family-centered care philosophy in neonatal units.

For a better understanding of the relationship between caregivers and young adults living with a concomitant substance-related mental health and substance use disorder: an integrative review / Mieux comprendre la relation entre les intervenants et les jeunes adultes vivant avec un trouble concomitant de santé mentale et lié à l'usage de substances : une revue intégrative des écrits

Introduction: The importance of a quality relationship between young adults living with dual diagnosis and their health care providers is well documented. Context: Although this complex phenomenon was mostly studied from an individual perspective, the results indicated the systemic nature of this relationship. Objective: This study aims to better understand the relationship between young adults living with dual diagnosis and their health care providers, with a systemic perspective. Method: Six data bases were consulted; manual research in gray literature and references screening enhanced the process. Results: Of a total of 532 studies and reports identified, 44 were included in the review. Thematic data analysis was carried out, and two themes were identified: the health care system as a constraining environment; and the relationship at the heart of care. Discussion: This study confirms the joint role played by the young adult in question and their health care provider in developing and maintaining the relationship, by acknowledging the importance of the care, of mutual confidence, and of a hierarchic relationship. Conclusion: This integrative review provides a basis for future nursing interventions that foreground the relationship and take a systemic approach.

Introduction: Plusieurs écrits soulignent l'importance de la qualité de la relation entre de jeunes adultes présentant un trouble concomitant de santé mentale et lié aux substances, et leurs intervenants. Contexte: Ce phénomène complexe a toutefois été étudié surtout dans une perspective individuelle, alors que plusieurs résultats soutiennent le caractère systémique de cette relation. Objectif: Mieux comprendre, selon une perspective systémique, le phénomène de la relation entre ces jeunes adultes et leurs intervenants. Méthode: Une revue intégrative, encadrée par une approche systémique, a été effectuée à partir de six bases de données, d'une recherche manuelle de la littérature grise et d'une vérification des références. Résultats: 532 écrits ont été recensés, et 44 d'entre eux ont servi pour l'analyse thématique qui a fait ressortir deux thèmes : l'environnement contraignant du système de santé et la relation au cœur des soins. Discussion: Cette étude corrobore le rôle conjoint joué par le jeune et l'intervenant dans le développement et le maintien de leur relation, en reconnaissant l'importance des soins, de la confiance réciproque et de la relation hiérarchique. Conclusion: Les résultats peuvent constituer l'assise pour le développement d'interventions infirmières mettant à l'avant-plan la relation selon une perspective systémique.

For a better understanding of the relationship between caregivers and young adults living with a concomitant substance-related mental health and substance use disorder: An integrative review / Mieux comprendre la relation entre les intervenants et les jeunes adultes vivant avec un trouble concomitant de santé mentale et lié à l'usage de substances : une revue intégrative des écrits

INTRODUCTION: The importance of a quality relationship between young adults living with dual diagnosis and their health care providers is well documented. CONTEXT: Although this complex phenomenon was mostly studied from an individual perspective, the results indicated the systemic nature of this relationship. OBJECTIVE: This study aims to better understand the relationship between young adults living with dual diagnosis and their health care providers, with a systemic perspective. METHOD: Six data bases were consulted; manual research in gray literature and references screening enhanced the process. RESULTS: Of a total of 532 studies and reports identified, 44 were included in the review. Thematic data analysis was carried out, and two themes were identified: the health care system as a constraining environment; and the relationship at the heart of care. DISCUSSION: This study confirms the joint role played by the young adult in question and their health care provider in developing and maintaining the relationship, by acknowledging the importance of the care, of mutual confidence, and of a hierarchic relationship. CONCLUSION: This integrative review provides a basis for future nursing interventions that foreground the relationship and take a systemic approach.

Introduction: Plusieurs écrits soulignent l'importance de la qualité de la relation entre de jeunes adultes présentant un trouble concomitant de santé mentale et lié aux substances, et leurs intervenants. Contexte : ce phénomène complexe a toutefois été étudié surtout dans une perspective individuelle, alors que plusieurs résultats soutiennent le caractère systémique de cette relation. Objectif: Mieux comprendre, selon une perspective systémique, le phénomène de la relation entre ces jeunes adultes et leurs intervenants. Méthode: Une revue intégrative, encadrée par une approche systémique, a été effectuée à partir de six bases de données, d'une recherche manuelle de la littérature grise et d'une vérification des références. Résultats: 532 écrits ont été recensés, et 44 d'entre eux ont servi pour l'analyse thématique qui a fait ressortir deux thèmes : l'environnement contraignant du système de santé et la relation au cœur des soins. Discussion: Cette étude corrobore le rôle conjoint joué par le jeune et l'intervenant dans le développement et le maintien de leur relation, en reconnaissant l'importance des soins, de la confiance réciproque et de la relation hiérarchique. Conclusion: Les résultats peuvent constituer l'assise pour le développement d'interventions infirmières mettant à l'avant-plan la relation selon une perspective systémique.

Cellular prion protein dysfunction in a prototypical inherited metabolic myopathy.

Inherited fatty acid oxidation diseases in their mild forms often present as metabolic myopathies. Carnitine Palmitoyl Transferase 2 (CPT2) deficiency, one such prototypical disorder is associated with compromised myotube differentiation. Here, we show that CPT2-deficient myotubes exhibit defects in focal adhesions and redox balance, exemplified by increased SOD2 expression. We document unprecedented alterations in the cellular prion protein PrPC, which directly arise from the failure in CPT2 enzymatic activity. We also demonstrate that the loss of PrPC function in normal myotubes recapitulates the defects in focal adhesion, redox balance and differentiation hallmarks monitored in CPT2-deficient cells. These results are further corroborated by studies performed in muscles from Prnp-/- mice. Altogether, our results unveil a molecular scenario, whereby PrPC dysfunction governed by faulty CPT2 activity may drive aberrant focal adhesion turnover and hinder proper myotube differentiation. Our study adds a novel facet to the involvement of PrPC in diverse physiopathological situations.

Expression Pattern of the Pneumocystis jirovecii Major Surface Glycoprotein Superfamily in Patients with Pneumonia.

BACKGROUND: The human pathogen Pneumocystis jirovecii harbors 6 families of major surface glycoproteins (MSGs) encoded by a single gene superfamily. MSGs are presumably responsible for antigenic variation and adhesion to host cells. The genomic organization suggests that a single member of family I is expressed at a given time per cell, whereas members of the other families are simultaneously expressed. METHODS: We analyzed RNA sequences expressed in several clinical samples, using specific weighted profiles for sorting of reads and calling of single-nucleotide variants to estimate the diversity of the expressed genes. RESULTS: A number of different isoforms of at least 4 MSG families were expressed simultaneously, including isoforms of family I, for which confirmation was obtained in the wet laboratory. CONCLUSION: These observations suggest that every single P. jirovecii population is made of individual cells with distinct surface properties. Our results enhance our understanding of the unique antigenic variation system and cell surface structure of P. jirovecii.

Co-invalidation of Prnp and Sprn in FVB/N mice affects reproductive performances and highlight complex biological relationship between PrP and Shadoo.

Shadoo and PrP belongs to the same protein family, whose biological function remains poorly understood. Previous experiments reported potential functional redundancies or antagonisms between these two proteins, depending on the tissue analysed. While knockdown experiments suggested the requirement of Shadoo in the absence of PrP during early mouse embryogenesis, knockout ones, on the contrary, highlighted little impact, if any, of the double-knockout of these two loci. In the present study, we reinvestigated the phenotype associated with the concomitant knockout of these two genes using newly produced FVB/N Sprn knockout mice. In this genetic background, the combined two genes' knockout induces intra-uterine growth retardations, likely resulting from placental failures highlighted by transcriptomic analyses that revealed potential redundant or antagonist roles of these two proteins in different developmental-related pathways. It also induced an increased perinatal-lethality and ascertained the role of these two loci in the lactation process.

A new AMPK activator, GSK773, corrects fatty acid oxidation and differentiation defect in CPT2-deficient myotubes.

Carnitine palmitoyl transferase 2 (CPT2) deficiency is one of the most common inherited fatty acid oxidation (FAO) defects and represents a prototypical mitochondrial metabolic myopathy. Recent studies have suggested a pivotal role of adenosine monophosphate-activated protein kinase (AMPK) in skeletal muscle plasticity and mitochondrial homeostasis. Thus, we tested the potential of GSK773, a novel direct AMPK activator, to improve or correct FAO capacities in muscle cells from patients harboring various mutations. We used controls' and patients' myotubes and studied the parameters of FAO metabolism, of mitochondrial quantity and quality and of differentiation. We found that AMPK is constitutively activated in patients' myotubes, which exhibit both reduced FAO and impaired differentiation. GSK773 improves or corrects several metabolic hallmarks of CPT2 deficiency (deficient FAO flux and C16-acylcarnitine accumulation) by upregulating the expression of CPT2 protein. Beneficial effects of GSK773 are also likely due to stimulation of mitochondrial biogenesis and induction of mitochondrial fusion, by decreasing dynamin-related protein 1 and increasing mitofusin 2. GSK773 also induces a shift in myosin heavy chain isoforms toward the slow oxidative type and, therefore, fully corrects the differentiation process. We establish, through small interfering RNA knockdowns and pharmacological approaches, that these GSK773 effects are mediated through peroxisome proliferator-activated receptor gamma co-activator 1-alpha, reactive oxygen species and p38 mitogen-activated protein kinase, all key players of skeletal muscle plasticity. GSK773 recapitulates several important features of skeletal muscle adaptation to exercise. The results show that AMPK activation by GSK773 evokes the slow, oxidative myogenic program and triggers beneficial phenotypic adaptations in FAO-deficient myotubes. Thus, GSK773 might have therapeutic potential for correction of CPT2 deficiency.

The Cellular Prion Protein Controls Notch Signaling in Neural Stem/Progenitor Cells.

The prion protein is infamous for its involvement in a group of neurodegenerative diseases known as Transmissible Spongiform Encephalopathies. In the longstanding quest to decipher the physiological function of its cellular isoform, PrPC , the discovery of its participation to the self-renewal of hematopoietic and neural stem cells has cast a new spotlight on its potential role in stem cell biology. However, still little is known on the cellular and molecular mechanisms at play. Here, by combining in vitro and in vivo murine models of PrPC depletion, we establish that PrPC deficiency severely affects the Notch pathway, which plays a major role in neural stem cell maintenance. We document that the absence of PrPC in a neuroepithelial cell line or in primary neurospheres is associated with drastically reduced expression of Notch ligands and receptors, resulting in decreased levels of Notch target genes. Similar alterations of the Notch pathway are recovered in the neuroepithelium of Prnp-/- embryos during a developmental window encompassing neural tube closure. In addition, in line with Notch defects, our data show that the absence of PrPC results in altered expression of Nestin and Olig2 as well as N-cadherin distribution. We further provide evidence that PrPC controls the expression of the epidermal growth factor receptor (EGFR) downstream from Notch. Finally, we unveil a negative feedback action of EGFR on both Notch and PrPC . As a whole, our study delineates a molecular scenario through which PrPC takes part to the self-renewal of neural stem and progenitor cells. Stem Cells 2017;35:754-765.

Iron oxide nanoparticle surface decorated with cRGD peptides for magnetic resonance imaging of brain tumors.

In this article, a specific targeting Magnetic Resonance Imaging (MRI) nanoplatform, composed by iron oxide nanoparticle (NP) with cRGD peptides as targeting agent onto NP surface, is explored for the diagnosis of brain tumors by MRI using intracranial U87MG mice xenograft tumor. This article is part of a Special Issue entitled "Recent Advances in Bionanomaterials" Guest Editor: Dr. Marie-Louise Saboungi and Dr. Samuel D. Bader.

Non-aqueous sol-gel synthesis of ultra small persistent luminescence nanoparticles for near-infrared in vivo imaging.

Ultra-small ZnGa2 O4 :Cr(3+) nanoparticles (6 nm) that exhibit near-infrared (NIR) persistent luminescence properties are synthesized by using a non-aqueous sol-gel method assisted by microwave irradiation. The nanoparticles are pegylated, leading to highly stable dispersions under physiological conditions. Preliminary in vivo studies show the high potential for these ultra-small ZnGa2 O4 :Cr(3+) nanoparticles to be used as in vivo optical nanotools as they emit without the need for in situ excitation and, thus, avoid the autofluorescence of tissues.

Clinical Challenges of Consensus Molecular Subtype CMS4 Colon Cancer in the Era of Precision Medicine.

Over the past decade, our understanding of the diversity of colorectal cancer has expanded significantly, raising hopes of tailoring treatments more precisely for individual patients. A key achievement in this direction was the establishment of the consensus molecular classification, particularly identifying the challenging consensus molecular subtype (CMS) CMS4 associated with poor prognosis. Because of its aggressive nature, extensive research is dedicated to the CMS4 subgroup. Recent years have unveiled molecular and microenvironmental features at the tissue level specific to CMS4 colorectal cancer. This has paved the way for mechanistic studies and the development of preclinical models. Simultaneously, efforts have been made to easily identify patients with CMS4 colorectal cancer. Reassessing clinical trial results through the CMS classification lens has improved our understanding of the therapeutic challenges linked to this subtype. Exploration of the biology of CMS4 colorectal cancer is yielding potential biomarkers and novel treatment approaches. This overview aims to provide insights into the clinico-biological characteristics of the CMS4 subgroup, the molecular pathways driving this subtype, and available diagnostic options. We also emphasize the therapeutic challenges associated with this subtype, offering potential explanations. Finally, we summarize the current tailored treatments for CMS4 colorectal cancer emerging from fundamental and preclinical studies.

Informal coercion in inpatient mental healthcare: a scoping review protocol.

INTRODUCTION: Comprehending informal coercion, which encompasses a wide range of phenomena characterised by subtle and non-legalised pressures, can be complex. Its use is underestimated within the continuum of coercion in mental health, although its application may have a negative impact on the persons involved. A better understanding of informal coercion is crucial for improving mental healthcare and informing future research. This scoping review aims to explore the nature, extent and consequences of informal coercion in mental health hospitalisation to clarify this phenomenon, establish its boundaries more clearly and identify knowledge gaps. METHODS AND ANALYSIS: Following the methodological framework from the Joanna Briggs Institute, this scoping review will encompass 10 key steps. Literature searches will be conducted in electronic databases, including CINAHL, PubMed, MEDLINE, EMBASE, Web of Science, PsycINFO, and ProQuest Dissertation and Theses. Then, a search in grey literature sources (Open Grey, Grey Guide), psychiatric and mental health journals, government agencies and among the references of selected studies will be conducted. The research will include all literature focusing on informal coercion with inpatients aged 18 and above. Data will be extracted and analysed descriptively, mapping the available knowledge and identifying thematic patterns. The quality of included studies will be assessed using appropriate appraisal tools. An exploratory search was conducted in November 2023 and will be updated in December 2023 when the selection of relevant evidence will begin. ETHICS AND DISSEMINATION: Ethical approval is not required as this study involves the analysis of existing published literature. The findings will be disseminated through a peer-reviewed publication and presentations at relevant conferences. They will be shared with people living with mental disorders and professionals working in mental healthcare.