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Cellular and Molecular Gastroenterology... May 2024Type 2 innate lymphoid cells (ILC2s) and interleukin-13 (IL-13) promote the onset of spasmolytic polypeptide-expressing metaplasia (SPEM) cells. However, little is known...
BACKGROUND & AIMS
Type 2 innate lymphoid cells (ILC2s) and interleukin-13 (IL-13) promote the onset of spasmolytic polypeptide-expressing metaplasia (SPEM) cells. However, little is known about molecular effects of IL-13 in SPEM cells. We now sought to establish a reliable organoid model, Meta1 gastroids, to model SPEM cells in vitro. We evaluated cellular and molecular effects of ILC2s and IL-13 on maturation and proliferation of SPEM cells.
METHODS
We performed single-cell RNA sequencing to characterize Meta1 gastroids, which were derived from stomachs of Mist1-Kras transgenic mice that displayed pyloric metaplasia. Cell sorting was used to isolate activated ILC2s from stomachs of IL-13-tdTomato reporter mice treated with L635. Three-dimensional co-culture was used to determine the effects of ILC2s on Meta1 gastroids. Mouse normal or metaplastic (Meta1) and human metaplastic gastroids were cultured with IL-13 to evaluate cell responses. Air-Liquid Interface culture was performed to test long-term culture effects of IL-13. In silico analysis determined possible STAT6-binding sites in gene promoter regions. STAT6 inhibition was performed to corroborate STAT6 role in SPEM cells maturation.
RESULTS
Meta1 gastroids showed the characteristics of SPEM cell lineages in vitro even after several passages. We demonstrated that co-culture with ILC2s or IL-13 treatment can induce phosphorylation of STAT6 in Meta1 and normal gastroids and promote the maturation and proliferation of SPEM cell lineages. IL-13 upregulated expression of mucin-related proteins in human metaplastic gastroids. Inhibition of STAT6 blocked SPEM-related gene expression in Meta1 gastroids and maturation of SPEM in both normal and Meta1 gastroids.
CONCLUSIONS
IL-13 promotes the maturation and proliferation of SPEM cells consistent with gastric mucosal regeneration.
PubMed: 38815928
DOI: 10.1016/j.jcmgh.2024.101366 -
The Journal of Biological Chemistry May 2024The metastasis suppressor, N-myc downstream regulated gene-1 (NDRG1), inhibits pro-oncogenic signaling in pancreatic cancer (PC). This investigation dissected a novel...
The metastasis suppressor, N-myc downstream regulated gene-1 (NDRG1), inhibits pro-oncogenic signaling in pancreatic cancer (PC). This investigation dissected a novel mechanism induced by NDRG1 on WNT/β-catenin signaling in multiple PC cell-types. NDRG1 overexpression decreased β-catenin and down-regulated glycogen synthase kinase-3β (GSK-3β) protein levels and its activation. However, β-catenin phosphorylation at Ser33, Ser37, and Thr41 that are classically induced by GSK-3β were significantly increased after NDRG1 overexpression, suggesting a GSK-3β-independent mechanism. Intriguingly, NDRG1 overexpression up-regulated protein kinase Cα (PKCα), with PKCα silencing preventing β-catenin phosphorylation at Ser33, Ser37, and Thr41, and decreasing β-catenin expression. Further, NDRG1 and PKCα were demonstrated to associate, with PKCα stabilization occurring after NDRG1 overexpression. In fact, PKCα half-life increased from 1.5 ± 0.8 h (3) in control cells to 11.0 ± 2.5 h (3) after NDRG1 overexpression. Thus, NDRG1 overexpression leads to the association of NDRG1 with PKCα and PKCα stabilization, resulting in β-catenin phosphorylation at Ser33, Ser37, and Thr41. In fact, the association between PKCα, NDRG1, and β-catenin was identified, with the formation of a potential metabolon that promotes the latter β-catenin phosphorylation. This anti-oncogenic activity of NDRG1 was multi-modal, with the above mechanism accompanied by the down-regulation of the nucleo-cytoplasmic shuttling protein, p21-activated kinase 4 (PAK4), that is involved in β-catenin nuclear translocation, inhibition of AKT phosphorylation (Ser473), and decreased β-catenin phosphorylation at Ser552 that suppresses its transcriptional activity. These mechanisms of NDRG1 activity are important to dissect to understand the marked anti-cancer efficacy of NDRG1-inducing thiosemicarbazones that up-regulate PKCα and inhibit WNT signaling.
PubMed: 38815861
DOI: 10.1016/j.jbc.2024.107417 -
Kidney & Blood Pressure Research May 2024Background Chronic kidney disease (CKD) is a progressive systemic condition characterized by numerous complications. Among these, alterations in skeletal muscle... (Review)
Review
Background Chronic kidney disease (CKD) is a progressive systemic condition characterized by numerous complications. Among these, alterations in skeletal muscle physiology, such as sarcopenia, are particularly significant, as they are associated with poor outcomes and reduced quality of life. Summary Various interventions, including pharmacological approaches and lifestyle modifications have been investigated to slow CKD progression and prevent or treat its complications. Physical exercise, in particular, has emerged as a promising intervention with multiple beneficial effects. These include improvements in physical functioning, increased muscle mass, modulation of metabolic abnormalities, and reduced cardiovascular risk. However, the pathophysiology of physical exercise in patients with kidney disease is complex and remains only partially understood. A crucial advancement in understanding this phenomenon has been the identification of myokines-molecules expressed and released by skeletal muscle in response to physical activity. These myokines can exert both paracrine and systemic effects, influencing not only skeletal muscle physiology but also other processes such as energy metabolism and lipid regulation. Key Messages The interplay among skeletal muscle, physical activity, and myokines may act as a pivotal regulator in various physiological processes, including aging, as well as in pathological conditions like cachexia and sarcopenia, frequently observed in CKD patients at different stages, including patients on dialysis. Despite the potential importance of this relationship, only a limited number of studies have explored the relationship between exercise and myokine, and the effect of this interaction on experimental models or individuals with kidney disease. In the following sections, we review and discuss this topic.
PubMed: 38815556
DOI: 10.1159/000539489 -
Redox Biology May 2024Hypoxia-inducible factor 1 alpha (HIF-1α) is a major molecular mediator of the hypoxic response. In the endometrium, local hypoxic conditions induced by hormonal... (Review)
Review
Hypoxia-inducible factor 1 alpha (HIF-1α) is a major molecular mediator of the hypoxic response. In the endometrium, local hypoxic conditions induced by hormonal fluctuations and endometrial vascular remodeling contribute to the production of HIF-1α, which plays an indispensable role in a series of physiological activities, such as menstruation and metamorphosis. The sensitive regulation of HIF-1α maintains the cellular viability and regenerative capacity of the endometrium against cellular stresses induced by hypoxia and excess reactive oxygen species. In contrast, abnormal HIF-1α levels exacerbate the development of various endometrial pathologies. This knowledge opens important possibilities for the development of promising HIF-1α-centered strategies to ameliorate endometrial disease. Nonetheless, additional efforts are required to elucidate the regulatory network of endometrial HIF-1α and promote the applications of HIF-1α-centered strategies in the human endometrium. Here, we summarize the role of the HIF-1α-mediated pathway in endometrial physiology and pathology, highlight the latest HIF-1α-centered strategies for treating endometrial diseases, and improve endometrial receptivity.
PubMed: 38815332
DOI: 10.1016/j.redox.2024.103205 -
The Oncologist May 2024The National Cancer Institute-Children's Oncology Group Pediatric Molecular Analysis for Therapy Choice (MATCH) precision oncology platform trial enrolled children aged...
BACKGROUND
The National Cancer Institute-Children's Oncology Group Pediatric Molecular Analysis for Therapy Choice (MATCH) precision oncology platform trial enrolled children aged 1-21 years with treatment-refractory solid tumors and predefined actionable genetic alterations. Patients with tumors harboring alterations in DNA damage repair (DDR) genes were assigned to receive olaparib.
METHODS
Tumor and blood samples were submitted for centralized molecular testing. Tumor and germline sequencing were conducted in parallel. Olaparib was given twice daily for 28-day cycles starting at a dose 30% lower than the adult recommended phase 2 dose (RP2D). The primary endpoint was the objective response.
RESULTS
Eighteen patients matched (1.5% of those screened) based on the presence of a deleterious gene alteration in BRCA1/2, RAD51C/D, or ATM detected by tumor sequencing without germline subtraction or analysis of loss of heterozygosity (LOH). Eleven (61%) harbored a germline mutation, with only one exhibiting LOH. Six patients enrolled and received the olaparib starting dose of 135 mg/m2/dose. Two participants were fully evaluable; 4 were inevaluable because <85% of the prescribed dose was administered during cycle 1. There were no dose-limiting toxicities or responses. Minimal hematologic toxicity was observed.
CONCLUSION
Most DDR gene alterations detected in Pediatric MATCH were germline, monoallelic, and unlikely to confer homologous recombination deficiency predicting sensitivity to olaparib monotherapy. The study closed due to poor accrual.
CLINICALTRIALS.GOV IDENTIFIER
NCT03233204. IRB approved: initial July 24, 2017.
PubMed: 38815151
DOI: 10.1093/oncolo/oyae096 -
PloS One 2024After its emergence in China, the coronavirus SARS-CoV-2 has swept the world, leading to global health crises with millions of deaths. COVID-19 clinical manifestations...
BACKGROUND
After its emergence in China, the coronavirus SARS-CoV-2 has swept the world, leading to global health crises with millions of deaths. COVID-19 clinical manifestations differ in severity, ranging from mild symptoms to severe disease. Although perturbation of metabolism has been reported as a part of the host response to COVID-19 infection, scarce data exist that describe stage-specific changes in host metabolites during the infection and how this could stratify patients based on severity.
METHODS
Given this knowledge gap, we performed targeted metabolomics profiling and then used machine learning models and biostatistics to characterize the alteration patterns of 50 metabolites and 17 blood parameters measured in a cohort of 295 human subjects. They were categorized into healthy controls, non-severe, severe and critical groups with their outcomes. Subject's demographic and clinical data were also used in the analyses to provide more robust predictive models.
RESULTS
The non-severe and severe COVID-19 patients experienced the strongest changes in metabolite repertoire, whereas less intense changes occur during the critical phase. Panels of 15, 14, 2 and 2 key metabolites were identified as predictors for non-severe, severe, critical and dead patients, respectively. Specifically, arginine and malonyl methylmalonyl succinylcarnitine were significant biomarkers for the onset of COVID-19 infection and tauroursodeoxycholic acid were potential biomarkers for disease progression. Measuring blood parameters enhanced the predictive power of metabolic signatures during critical illness.
CONCLUSIONS
Metabolomic signatures are distinctive for each stage of COVID-19 infection. This has great translation potential as it opens new therapeutic and diagnostic prospective based on key metabolites.
Topics: Humans; COVID-19; Machine Learning; Male; Female; Biomarkers; Middle Aged; Metabolomics; Adult; Severity of Illness Index; SARS-CoV-2; Aged; Metabolome
PubMed: 38814977
DOI: 10.1371/journal.pone.0302977 -
Psychiatrike = Psychiatriki May 2024According to international experience, the conditions for the successful outcome of a psychiatric reform are the following: (a) Existence of political will (supporting a...
According to international experience, the conditions for the successful outcome of a psychiatric reform are the following: (a) Existence of political will (supporting a national plan with assessment, monitoring, and corrective intervention procedures for structural dysfunctions, etc.). (b) Strong mental health leadership (executive expertise and skills that advance the public health agenda). (c) Challenging the dominance of the biomedical model in therapeutic practice through the promotion of holistic care practices, evidence-based innovative actions, collaborative care, the promotion of recovery culture, and the and the use of innovative digital tools. (d) Ensuring necessary resources over time, so that resources from the transition of the asylum model to a model of sectorial community mental health services "follow" the patient. (e) Strengthening the participation of service recipients and their families in decision-making processes and evaluation of care quality. (f) Practices based on ethical principles (value-based practice) and not only on the always necessary documentation (evidence-based practice).1- 4 Convergent evidence from the "ex post" evaluation of the implementation of the national plan Psychargos 2000-20095 and from the recent rapid assessment of the psychiatric reform by the Ministry of Health and the WHO Athens office (SWOT analysis)6 indicates "serious fragmentation of services, an uncoordinated system that often results in inappropriate service provision, a lack of epidemiological studies and studies concerning the local needs of specific populations, uneven development of services between different regions of the country, a large number of specialized professionals with significant deficits in community psychiatry expertise, a lack of personnel in supportive roles, significant gaps in specialized services (for individuals with autism spectrum disorders, intellectual disabilities, eating disorders, old and new addictions, and community forensic psychiatry services)". We would also like to highlight lack of coordination and collaboration among different mental health service systems (public primary and secondary service providers, NGOs, municipal services, mental health services of the armed forces, private sector), complete absence of systematic evaluation and monitoring (lack of quality of care indicators, clinical outcomes, epidemiological profile of each service), lack of quality assurance mechanisms and clinical management systems, insufficient number of beds mainly for acute cases, unclear protocols for discharge issuance and ensuring continuity of care, deficient budget for Mental Health in relation to the overall healthcare expenditure (currently 3.3%), and finally, one of the highest rates of involuntary hospitalizations in Europe, which is linked to serious issues concerning the protection of the rights of service users. After the pandemic and the emergence of the silent but expected mental health pandemic, WHO, EU, and the Greek Ministry of Health emphasized the need to adopt a public mental health agenda with an emphasis on community psychiatry in order to address both the old structural dysfunctions and inadequacies of psychiatric reform (regulation 815/1984, Leros I-Leros II plan, Psychargos A & B, incomplete implementation of laws 2071/1992 & 2716/1999, incomplete deinstitutionalization of the remaining psychiatric hospitals). However, it is time to reflect that it is not possible to talk today about the need to update and implement a new national plan to upgrade mental health in the country without answering basic questions, both old and new, about the wider context of its implementation. The transformation of the deficient psychiatric care in the country cannot be completed without the urgent restructuring of the National Health System7 and the reform of the Greek welfare state itself, which is also characterized by irrationality, inequalities, bureaucratic inefficiency, and fragmentation.8 As we should have learned from the bankruptcy and the prolonged economic, social, and cultural crisis in our country, reforms usually pay off in the long term, while the time horizon of the applied policies is narrow and usually reaching the next election. The fact is that in any reform effort, including psychiatry, the political system does not demonstrate the ability to promote transparency, evaluation, stable rules of regulation, reference to a universally applicable legal and institutional framework, the limitation of clientelism and guild resistances. From this point of view, it is necessary to give meaning in the context of Greek psychiatric reform to the professional burnout of the National Health System workers, the lack of motivation and vision, the intrusion into the NGO space by new entities without any connection to the culture of psychiatry reform, the guild resistances of all relevant specialties, the selective use of psychotherapeutic techniques, as trends of discrediting the relief of social and psychological suffering in the field of public mental health. There is an urgent need to understand new pathologies (narcissistic disorders, new forms of addiction, eating disorders, "pathology of emptiness", adolescent delinquency and suicide, psychosomatic manifestations due to high stress, pathology of fluid social ties, deficient socialization of young people "outside of their algorithms") through a solid and coherent analysis of the toxic postmodernity culture. In addition to the social determinants of mental health,9 it is necessary in clinical work to also assess the psychological factors, such as uncertainty, conflict, loss of control, and incomplete information, that burden human health.10 In order to reduce the gap between declarations and real life, there is an urgent need to overcome the blind spots of psychiatric reform in the country by establishing internal and external evaluation processes, training young professionals in holistic care and community networking and communication skills, retraining leaders for organizational change, and strengthening the participation of service users in the context of deepening democracy in mental health. As mental health professionals, the object of our work in the community should be the reconstruction of meaning and the fragile or non-existent social bond in subjects who have been cut off from any possible production of meaning and participation in their history. Why should our therapeutic responses be stereotypically repetitive in the face of these complex, radical changes in the meta-context and the new demands of our patients? After all, as the philosopher Ernst Bloch puts it, utopia is "that which does not exist yet.".
PubMed: 38814270
DOI: 10.22365/jpsych.2024.009 -
Microbiology Spectrum May 2024is the leading cause of bacterial sexually transmitted infections in the USA and of preventable blindness worldwide. This obligate intracellular pathogen replicates...
is the leading cause of bacterial sexually transmitted infections in the USA and of preventable blindness worldwide. This obligate intracellular pathogen replicates within a membrane-bound inclusion, but how it acquires nutrients from the host while avoiding detection by the innate immune system is incompletely understood. accomplishes this in part through the translocation of a unique set of effectors into the inclusion membrane, the lusion membrane proteins (Incs). Incs are ideally positioned at the host-pathogen interface to reprogram host signaling by redirecting proteins or organelles to the inclusion. Using a combination of co-affinity purification, immunofluorescence confocal imaging, and proteomics, we characterize the interaction between an early-expressed Inc of unknown function, Tri1, and tumor necrosis factor receptor-associated factor 7 (TRAF7). TRAF7 is a multi-domain protein with a RING finger ubiquitin ligase domain and a C-terminal WD40 domain. TRAF7 regulates several innate immune signaling pathways associated with infection and is mutated in a subset of tumors. We demonstrate that Tri1 and TRAF7 specifically interact during infection and that TRAF7 is recruited to the inclusion. We further show that the predicted coiled-coil domain of Tri1 is necessary to interact with the TRAF7 WD40 domain. Finally, we demonstrate that Tri1 displaces the native TRAF7 binding partners, mitogen-activated protein kinase kinase kinase 2 (MEKK2), and MEKK3. Together, our results suggest that by displacing TRAF7 native binding partners, Tri1 has the capacity to alter TRAF7 signaling during infection.IMPORTANCE is the leading cause of bacterial sexually transmitted infections in the USA and preventable blindness worldwide. Although easily treated with antibiotics, the vast majority of infections are asymptomatic and therefore go untreated, leading to infertility and blindness. This obligate intracellular pathogen evades the immune response, which contributes to these outcomes. Here, we characterize the interaction between a -secreted effector, Tri1, and a host protein involved in innate immune signaling, TRAF7. We identified host proteins that bind to TRAF7 and demonstrated that Tri1 can displace these proteins upon binding to TRAF7. Remarkably, the region of TRAF7 to which these host proteins bind is often mutated in a subset of human tumors. Our work suggests a mechanism by which Tri1 may alter TRAF7 signaling and has implications not only in the pathogenesis of infections but also in understanding the role of TRAF7 in cancer.
PubMed: 38814079
DOI: 10.1128/spectrum.00453-24 -
Haematologica May 2024Whereas immunotherapies have revolutionized the treatment of different solid and hematological cancers, their efficacy in nodal peripheral T-cell lymphomas (PTCLs) is...
Whereas immunotherapies have revolutionized the treatment of different solid and hematological cancers, their efficacy in nodal peripheral T-cell lymphomas (PTCLs) is limited, due to a lack of understanding of the immune response they trigger. To fully characterize the immune tumor microenvironment (TME) of PTCLs, we performed spectral flow cytometry analyses on 11 angioimmunoblastic T-cell lymphomas (AITL), 7 PTCL, not otherwise specified (PTCL, NOS) lymph node samples, and 10 non-tumoral control samples. The PTCL TME contained a larger proportion of regulatory T cells and exhausted CD8+ T cells, with enriched expression of druggable immune checkpoints. Interestingly, CD39 expression was up-regulated at the surface of most immune cells, and a multi-immunofluorescence analyses on a retrospective cohort of 43 AITL patients demonstrated a significant association between high CD39 expression by T cells and poor patient prognosis. Together, our study unravels the complex TME of nodal PTCLs, identifies targetable immune checkpoints, and highlights CD39 as a novel prognostic factor.
PubMed: 38813724
DOI: 10.3324/haematol.2023.284448 -
Turkish Journal of Medical Sciences 2023Heavy-ion irradiation seriously perturbs cellular homeostasis and thus damages cells. Vascular endothelial cells (ECs) play an important role in the pathological process...
BACKGROUND/AIM
Heavy-ion irradiation seriously perturbs cellular homeostasis and thus damages cells. Vascular endothelial cells (ECs) play an important role in the pathological process of radiation damage. Protecting ECs from heavy-ion radiation is of great significance in the radioprotection of normal tissues. In this study, the radioprotective effect of β-D-glucan (BG) derived from on human umbilical vein endothelial cell (EA.hy926) cytotoxicity produced by carbon-ion irradiation was examined and the probable mechanism was established.
MATERIALS AND METHODS
EA.hy926 cells were divided into seven groups: a control group; 1, 2, or 4 Gy radiation; and 10 μg/mL BG pretreatment for 24 h before 1, 2, or 4 Gy irradiation. Cell survival was assessed by colony formation assay. Cell cycles, apoptosis, DNA damage, and reactive oxygen species (ROS) levels were measured through flow cytometry. The level of malondialdehyde and antioxidant enzyme activities were analyzed using assay kits. The activation of NF-κB was analyzed using western blotting and a transcription factor assay kit. The expression of downstream target genes was detected by western blotting.
RESULTS
BG pretreatment significantly increased the survival of irradiated cells, improved cell cycle progression, and decreased DNA damage and apoptosis. The levels of ROS and malondialdehyde were also decreased by BG. Further study indicated that BG increased the antioxidant enzyme activities, activated Src, and promoted NF-κB activation, especially for the p65, p50, and RelB subunits. The activated NF-κB upregulated the expression of antioxidant protein MnSOD, DNA damage-response and repair-related proteins BRCA2 and Hsp90α, and antiapoptotic protein Bcl-2.
CONCLUSION
Our results demonstrated that BG protects EA.hy926 cells from high linear-energy-transfer carbon-ion irradiation damage through the upregulation of prosurvival signaling triggered by the interaction of BG with its receptor. This confirms that BG is a promising radioprotective agent for heavy-ion exposure.
Topics: Humans; NF-kappa B; Human Umbilical Vein Endothelial Cells; Apoptosis; Cell Survival; DNA Damage; Reactive Oxygen Species; beta-Glucans; Radiation-Protective Agents
PubMed: 38813508
DOI: 10.55730/1300-0144.5731