Ubiquitin ligase RNF20 coordinates sequential adipose thermogenesis with brown and beige fat-specific substrates.

In mammals, brown adipose tissue (BAT) and inguinal white adipose tissue (iWAT) execute sequential thermogenesis to maintain body temperature during cold stimuli. BAT rapidly generates heat through brown adipocyte activation, and further iWAT gradually stimulates beige fat cell differentiation upon prolonged cold challenges. However, fat depot-specific regulatory mechanisms for thermogenic activation of two fat depots are poorly understood. Here, we demonstrate that E3 ubiquitin ligase RNF20 orchestrates adipose thermogenesis with BAT- and iWAT-specific substrates. Upon cold stimuli, BAT RNF20 is rapidly downregulated, resulting in GABPα protein elevation by controlling protein stability, which stimulates thermogenic gene expression. Accordingly, BAT-specific Rnf20 suppression potentiates BAT thermogenic activity via GABPα upregulation. Moreover, upon prolonged cold stimuli, iWAT RNF20 is gradually upregulated to promote de novo beige adipogenesis. Mechanistically, iWAT RNF20 mediates NCoR1 protein degradation, rather than GABPα, to activate PPARγ. Together, current findings propose fat depot-specific regulatory mechanisms for temporal activation of adipose thermogenesis.

Diagnosis of Primary Ciliary Dyskinesia via Whole Exome Sequencing and Histologic Findings.

PURPOSE: To assess the diagnostic potential of whole-exome sequencing (WES) and elucidate the clinical and genetic characteristics of primary ciliary dyskinesia (PCD) in the Korean population. MATERIALS AND METHODS: Forty-seven patients clinically suspected of having PCD were enrolled at a tertiary medical center. WES was performed in all patients, and seven patients received biopsy of cilia and transmission electron microscopy (TEM). RESULTS: Overall, PCD was diagnosed in 10 (21.3%) patients: eight by WES (8/47, 17%), four by TEM. Among patients diagnosed as PCD based on TEM results, two patients showed consistent results with WES and TEM of PCD (2/4, 50%). In addition, five patients, who were not included in the final PCD diagnosis group, had variants of unknown significance in PCD-related genes (5/47, 10.6%). The most frequent pathogenic (P)/likely pathogenic (LP) variants were detected in DNAH11 (n=4, 21.1%), DRC1 (n=4, 21.1%), and DNAH5 (n=4, 21.1%). Among the detected 17 P/LP variants in PCD-related genes in this study, 8 (47.1%) were identified as novel variants. Regarding the genotype-phenotype correlation in this study, the authors experienced severe PCD cases caused by the LP/P variants in MCIDAS, DRC1, and CCDC39. CONCLUSION: Through this study, we were able to confirm the value of WES as one of the diagnostic tools for PCD, which increases with TEM, rather than single gene tests. These results will prove useful to hospitals with limited access to PCD diagnostic testing but with relatively efficient in-house or outsourced access to genetic testing at a pre-symptomatic or early disease stage.

FAM3C in Cancer-Associated Adipocytes Promotes Breast Cancer Cell Survival and Metastasis.

Adipose tissue within the tumor microenvironment (TME) plays a critical role in supporting breast cancer progression. In this study, we identified FAM3 metabolism-regulating signaling molecule C (FAM3C) produced by cancer-associated adipocytes (CAA) as a key regulator of tumor progression. FAM3C overexpression in cultured adipocytes significantly reduced cell death in both adipocytes and cocultured breast cancer cells while suppressing markers of fibrosis. Conversely, FAM3C depletion in CAAs resulted in adipocyte-mesenchymal transition (AMT) and increased fibrosis within the TME. Adipocyte FAM3C expression was driven by TGFß signaling from breast cancer cells and was reduced upon treatment with a TGFß-neutralizing antibody. FAM3C knockdown in CAAs early in tumorigenesis in a genetically engineered mouse model of breast cancer significantly inhibited primary and metastatic tumor growth. Circulating FAM3C levels were elevated in patients with metastatic breast cancer compared with those with nonmetastatic breast cancer. These results suggest that therapeutic inhibition of FAM3C expression levels in CAAs during early tumor development could be a promising approach in the treatment of patients with breast cancer. SIGNIFICANCE: High FAM3C levels in cancer-associated adipocytes contribute to tumor-supportive niches and are tightly associated with metastatic growth, indicating that FAM3C inhibition could be beneficial for treating patients with breast cancer.

Unique adipose tissue invariant natural killer T cell subpopulations control adipocyte turnover in mice.

Adipose tissue invariant natural killer T (iNKT) cells are a crucial cell type for adipose tissue homeostasis in obese animals. However, heterogeneity of adipose iNKT cells and their function in adipocyte turnover are not thoroughly understood. Here, we investigate transcriptional heterogeneity in adipose iNKT cells and their hierarchy using single-cell RNA sequencing in lean and obese mice. We report that distinct subpopulations of adipose iNKT cells modulate adipose tissue homeostasis through adipocyte death and birth. We identify KLRG1+ iNKT cells as a unique iNKT cell subpopulation in adipose tissue. Adoptive transfer experiments showed that KLRG1+ iNKT cells are selectively generated within adipose tissue microenvironment and differentiate into a CX3CR1+ cytotoxic subpopulation in obese mice. In addition, CX3CR1+ iNKT cells specifically kill enlarged and inflamed adipocytes and recruit macrophages through CCL5. Furthermore, adipose iNKT17 cells have the potential to secrete AREG, and AREG is involved in stimulating adipose stem cell proliferation. Collectively, our data suggest that each adipose iNKT cell subpopulation plays key roles in the control of adipocyte turnover via interaction with adipocytes, adipose stem cells, and macrophages in adipose tissue.

Partial in vivo reprogramming enables injury-free intestinal regeneration via autonomous Ptgs1 induction.

Tissue regeneration after injury involves the dedifferentiation of somatic cells, a natural adaptive reprogramming that leads to the emergence of injury-responsive cells with fetal-like characteristics. However, there is no direct evidence that adaptive reprogramming involves a shared molecular mechanism with direct cellular reprogramming. Here, we induced dedifferentiation of intestinal epithelial cells using OSKM (Oct4, Sox2, Klf4, and c-Myc) in vivo. The OSKM-induced forced dedifferentiation showed similar molecular features of intestinal regeneration, including a transition from homeostatic cell types to injury-responsive-like cell types. These injury-responsive-like cells, sharing gene signatures of revival stem cells and atrophy-induced villus epithelial cells, actively assisted tissue regeneration following damage. In contrast to normal intestinal regeneration involving Ptgs2 induction, the OSKM promotes autonomous production of prostaglandin E2 via epithelial Ptgs1 expression. These results indicate prostaglandin synthesis is a common mechanism for intestinal regeneration but involves a different enzyme when partial reprogramming is applied to the intestinal epithelium.

Corrigendum: Genetic diagnosis of inborn errors of immunity using clinical exome sequencing.

[This corrects the article DOI: 10.3389/fimmu.2023.1178582.].

Effects of COVID-19 Stress on Healing Behavior in Residential Spaces.

This study aims to analyze the effects of COVID-19 stress on healing behavior in residential spaces. Based on the results, the study further examines the residential space as a healing environment according to space use behavior in the post-COVID era. A survey including 2101 Korean people was conducted. The COVID Stress Scale for Korean People (CSSK) was used to identify COVID-19 stress. In addition, using a literature review on healing environment factors and behavioral changes after the pandemic, survey questions were developed to assess changes in healing behavior. A frequency analysis was conducted for sociodemographic factors, and the relationship between COVID stress factors and healing behavior factors in residential spaces was examined using factor and correlation analyses. Multiple regression analysis was conducted to verify the effects of COVID stress factors on changes in healing behavior. The results revealed that COVID stress affected healing behavior in residential spaces; however, there were differences in healing behavior depending on the COVID stress factors. Fear of being infected positively affected infection prevention behavior in homes. Anger towards others negatively affected the establishment of elements that support various activities in the residential spaces and furniture arrangement. Stress from social distancing difficulties affected healing behavior in residential spaces but was not related to infection prevention behavior. Residential spaces can serve as healing spaces when people are provided with various spatial factors that support diverse types of behavior during a pandemic crisis.

A 3-dimensional evaluation of available retromolar space for the application of ramal plates.

INTRODUCTION: This study aimed to evaluate the available retromolar space for ramal plates in patients with Class I and III malocclusions and compare that space with and without third molars using cone-beam computed tomography. METHODS: Cone-beam computed tomography images of 30 patients (17 males, 13 females; mean age, 22.2 ± 4.5 years) with Class III malocclusion and 29 subjects (18 males, 11 females; mean age, 24.3 ± 3.7 years) with Class I malocclusion were analyzed. Available retromolar space at 4 axial levels of the second molar root and the volume of the retromolar bone were evaluated. Two-way repeated measures analysis of covariance (repeated measures analysis of covariance) was applied to compare the variables between Class I and III malocclusions and the presence of third molars. RESULTS: Patients with Class I and III relationships showed up to 12.7 mm of available retromolar space at 2 mm apical from the cementoenamel junction (CEJ). At 8 mm apical from CEJ, patients with Class III malocclusion had 11.1 mm of space, whereas those with a Class I relationship showed 9.8 mm of available space. When patients had third molars, the amount of available retromolar space was significantly greater in patients with a Class I and III relationship. However, patients with Class III malocclusion exhibited greater available retromolar space than those with a Class I relationship (P = 0.028). In addition, the bone volume was significantly greater in patients with Class III malocclusion than in patients with a Class I relationship and those with third molars than in those without them (P <0.001). CONCLUSIONS: Class I and III groups showed the availability of at least 10.0 mm of retromolar space 2 mm apical to the CEJ for molar distalization. Based on this information, it is suggested that clinicians consider available retromolar space for molar distalization in diagnosing and planning treatment for patients with Class I and III malocclusion.

Genetic diagnosis of inborn errors of immunity using clinical exome sequencing.

Inborn errors of immunity (IEI) include a variety of heterogeneous genetic disorders in which defects in the immune system lead to an increased susceptibility to infections and other complications. Accurate, prompt diagnosis of IEI is crucial for treatment plan and prognostication. In this study, clinical utility of clinical exome sequencing (CES) for diagnosis of IEI was evaluated. For 37 Korean patients with suspected symptoms, signs, or laboratory abnormalities associated with IEI, CES that covers 4,894 genes including genes related to IEI was performed. Their clinical diagnosis, clinical characteristics, family history of infection, and laboratory results, as well as detected variants, were reviewed. With CES, genetic diagnosis of IEI was made in 15 out of 37 patients (40.5%). Seventeen pathogenic variants were detected from IEI-related genes, BTK, UNC13D, STAT3, IL2RG, IL10RA, NRAS, SH2D1A, GATA2, TET2, PRF1, and UBA1, of which four variants were previously unreported. Among them, somatic causative variants were identified from GATA2, TET2, and UBA1. In addition, we identified two patients incidentally diagnosed IEI by CES, which was performed to diagnose other diseases of patients with unrecognized IEI. Taken together, these results demonstrate the utility of CES for the diagnosis of IEI, which contributes to accurate diagnosis and proper treatments.

High levels of intracellular endotrophin in adipocytes mediate COPII vesicle supplies to autophagosome to impair autophagic flux and contribute to systemic insulin resistance in obesity.

BACKGROUND AND AIMS: Extracellular matrix (ECM) homeostasis plays a crucial role in metabolic plasticity and endocrine function of adipose tissue. High levels of intracellular endotrophin, a cleavage peptide of type VI collagen alpha 3 chain (Col6a3), have been frequently observed in adipocyte in obesity and diabetes. However, how endotrophin intracellularly traffics and influences metabolic homeostasis in adipocyte remains unknown. Therefore, we aimed to investigate the trafficking of endotrophin and its metabolic effects in adipocytes depending on lean or obese condition. METHODS: We used doxycycline-inducible adipocyte-specific endotrophin overexpressed mice for a gain-of-function study and CRISPR-Cas9 system-based Col6a3-deficient mice for a loss-of-function study. Various molecular and biochemical techniques were employed to examine the effects of endotrophin on metabolic parameters. RESULTS: In adipocytes during obesity, the majority of endosomal endotrophin escapes lysosomal degradation and is released into the cytosol to mediate direct interactions between SEC13, a major component of coat protein complex II (COPII) vesicles, and autophagy-related 7 (ATG7), leading to the increased formation of autophagosomes. Autophagosome accumulation disrupts the balance of autophagic flux, resulting in adipocyte death, inflammation, and insulin resistance. These adverse metabolic effects were ameliorated by either suppressing ATG7 with siRNA ex vivo or neutralizing endotrophin with monoclonal antibodies in vivo. CONCLUSIONS: High levels of intracellular endotrophin-mediated autophagic flux impairment in adipocyte contribute to metabolic dysfunction such as apoptosis, inflammation, and insulin resistance in obesity.

Comparing Older Parents' and Adult Children's Fear of Falling and Perceptions of Age-Friendly Home Modification: An Integration of the Theories of Planned Behavior and Protection Motivation.

This study investigated how a fear of falling affects perceptions and behavioral intentions toward age-friendly home modification (AFHM) in older parents and adult children by integrating the theory of planned behavior (TPB) to explain AFHM decision-making processes and the protection motivation theory to explain the impact of a fear of falling on AFHM intention. The target population comprised older parents (≥75 years old) and adult children (45-64 years old) in Busan, South Korea (N = 600). The participants completed a self-administered questionnaire in March 2022. Independent t-test and path model analyses were conducted to compare primary constructs between older parents and adult children and analyze the relationships among a fear of falling, TPB components, and AFHM intention. Results showed that both groups had positive attitudes toward AFHM. However, adult children showed significantly higher rates of having a fear of falling, lower perceived behavioral control, and higher AFHM intention than older parents. The proposed research models were partially supported in the older-parent group and fully supported in the adult-children group. Adult children play a critical role in AFHM, along with older adults who are directly involved in an aging society. AFHM-supporting programs, including monetary and human-force assistance, education, related public advertisements, and an active AFHM market, should be expanded.

Association between face covering policies and the incidence of coronavirus disease 2019 in European countries.

OBJECTIVES: This study was conducted to determine the impact of the strengthening or relaxation of face covering mandates on the subsequent national case incidence of coronavirus disease 2019 (COVID-19) in Europe as the full vaccination rate was increasing. METHODS: European countries in which case incidence increased for 3 consecutive weeks were monitored and analyzed using COVID-19 incidence data shared by the World Health Organization (WHO). The epidemic trend of COVID-19 in Europe was compared with that of countries elsewhere in the world based on WHO weekly epidemiological reports from June 20 to October 30, 2021. In addition, this study provided insight into the impact of government mask mandates on COVID-19 incidence in Europe by measuring the index scores of those facial covering policies before and after mandate relaxation or strengthening. The effects of the vaccination rate and the speed of vaccination on COVID-19 incidence were also analyzed. RESULTS: The incidence of COVID-19 after the relaxation of face covering mandates was significantly higher than before relaxation. However, no significant difference was observed in vaccination rate between countries with increased and decreased incidence. Instead, rapid vaccination delayed the resurgence in incidence. CONCLUSIONS: The findings suggest that face covering policies in conjunction with rapid vaccination efforts are essential to help mitigate the spread of COVID-19.

Epigenetic regulation of Neuregulin 1 promotes breast cancer progression associated to hyperglycemia.

Hyperglycemia is a risk factor for breast cancer-related morbidity and mortality. Hyperglycemia induces Neuregulin 1 (Nrg1) overexpression in breast cancer, which subsequently promotes tumor progression. However, molecular mechanisms underlying hyperglycemia-induced Nrg1 overexpression remain poorly understood. Here, we show that hyperglycemia causes active histone modifications at the Nrg1 enhancer, forming enhanceosome complexes where recombination signal binding protein for immunoglobulin kappa J region (RBPJ), E1A binding protein p300 (P300), and SET domain containing 1 A (SETD1A) are recruited to upregulate Nrg1 expression. Deletions in RBPJ-binding sites causes hyperglycemia-controlled Nrg1 levels to be downregulated, resulting in decreased tumor growth in vitro and in vivo. Mice with modest-temporary hyperglycemia, induced by low-dose short-exposure streptozotocin, display accelerated tumor growth and lapatinib resistance, whereas combining lapatinib with N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S42 phenylglycine t-butyl ester (DAPT) ameliorates tumor growth under these modest hyperglycemic conditions by inhibiting NOTCH and EGFR superfamilies. NOTCH activity is correlated with NRG1 levels, and high NRG1 levels predicts poor outcomes, particularly in HER2-positive breast cancer patients. Our findings highlight the hyperglycemia-linked epigenetic modulation of NRG1 as a potential therapeutic strategy for treating breast cancer patients with diabetes.

DSCR1 deficiency ameliorates the Aß pathology of Alzheimer's disease by enhancing microglial activity.

Microglial phagocytosis and clearance are important for the removal of amyloid-ß (Aß) plaques in Alzheimer's disease (AD). Chronic exposure of microglia to Aß plaques leads to microglial metabolic dysfunction, and dysregulation of microglia can accelerate the deposition of Aß plaques and cause learning and memory impairment. Thus, regulating microglial Aß clearance is crucial for the development of therapeutics for AD-related dementia. Here, Down syndrome critical region 1 (DSCR1) deficiency ameliorated Aß plaque deposition in the 5xFAD mouse model of AD by altering microglial activity; however, the Aß synthesis pathway was not affected. DSCR1 deficiency improved spatial learning and memory impairment in 5xFAD mice. Furthermore, DSCR1-deficient microglia exhibited accelerated lysosomal degradation of Aß after phagocytosis, and BV2 cells with stable knockdown of DSCR1 demonstrated enhanced lysosomal activity. RNA-sequencing analysis showed that the transcriptional signatures associated with responses to IFN-γ were significantly up-regulated in DSCR1-knockdown BV2 cells treated with Aß. Our data strongly suggest that DSCR1 is a critical mediator of microglial degradation of amyloid plaques and a new potential microglial therapeutic target in AD.

The Efficacy of a Human-Ready miniMECP2 Gene Therapy in a Pre-Clinical Model of Rett Syndrome.

Inactivating mutations and the duplication of methyl-CpG binding protein 2 (MeCP2), respectively, mediate Rett syndrome (RTT) and MECP2 duplication syndrome. These disorders underscore the conceptual dose-dependent risk posed by MECP2 gene therapy for mosaic RTT patients. Recently, a miRNA-Responsive Autoregulatory Element (miRARE) mitigated the dose-dependent toxicity posed by self-complementary adeno-associated viral vector serotype 9 (AAV9) miniMECP2 gene therapy (scAAV9/miniMECP2-myc) in mice. Here, we report an efficacy assessment for the human-ready version of this regulated gene therapy (TSHA-102) in male Mecp2-/y knockout (KO) mice after intracerebroventricular (ICV) administration at postnatal day 2 (P2) and after intrathecal (IT) administration at P7, P14 (±immunosuppression), and P28 (±immunosuppression). We also report qPCR studies on KO mice treated at P7-P35; protein analyses in KO mice treated at P38; and a survival safety study in female adult Mecp2-/+ mice. In KO mice, TSHA-102 improved respiration, weight, and survival across multiple doses and treatment ages. TSHA-102 significantly improved the front average stance and swing times relative to the front average stride time after P14 administration of the highest dose for that treatment age. Viral genomic DNA and miniMECP2 mRNA were present in the CNS. MiniMeCP2 protein expression was higher in the KO spinal cord compared to the brain. In female mice, TSHA-102 permitted survivals that were similar to those of vehicle-treated controls. In all, these pivotal data helped to support the regulatory approval to initiate a clinical trial for TSHA-102 in RTT patients (clinical trial identifier number NCT05606614).

Dual-Functional Self-Attachable and Stretchable Interface for Universal Three-Dimensional Modular Electronics.

Stretchable electronics have become essential for custom-built electronics, self-assembling robotics, and wearable devices. Although many stretchable electronics contain integrated systems, they still limit bulky connection systems. We introduce a new dual-functioned self-attachable and stretchable interface (SASI), allowing a direct and instant interconnection between rigid and soft electronics. The SASI consists of a sticky and stretchable substrate and surface-embedded serpentine conductors with the single-sided polyimide fabricated using the embedded transfer process. The adhesion property of the SASI is controlled by the mixed elastomer ratio. The resulting sticky and conductive SASI can instantly adhere to a metal surface and create conductive paths. The SASI serpentine conductors exhibit high stretchability (∼290%) and provide self-attachable, re-attachable, and low-resistant electrical contacts (0.85 ohms in 0.25 mm2) between interfaces without pressure, heat, or extra solder. In addition, three-dimensional curved and modular electronics can be formed with the SASI by compiling functional blocks. SASI provides a novel strategy for assembling functional chips or modules for stretchable electronics, opening a path to onboard integrated electronics that are customizable by users for real-world stretchable electronics.

High density integration of stretchable inorganic thin film transistors with excellent performance and reliability.

Transistors with inorganic semiconductors have superior performance and reliability compared to organic transistors. However, they are unfavorable for building stretchable electronic products due to their brittle nature. Because of this drawback, they have mostly been placed on non-stretchable parts to avoid mechanical strain, burdening the deformable interconnects, which link these rigid parts, with the strain of the entire system. Integration density must therefore be sacrificed when stretchability is the first priority because the portion of stretchable wirings should be raised. In this study, we show high density integration of oxide thin film transistors having excellent performance and reliability by directly embedding the devices into stretchable serpentine strings to defeat such trade-off. The embedded transistors do not hide from deformation and endure strain up to 100% by themselves; thus, integration density can be enhanced without sacrificing the stretchability. We expect that our approach can create more compact stretchable electronics with high-end functionality than before.

Targeted erasure of DNA methylation by TET3 drives adipogenic reprogramming and differentiation.

DNA methylation is a crucial epigenetic modification in the establishment of cell-type-specific characteristics. However, how DNA methylation is selectively reprogrammed at adipocyte-specific loci during adipogenesis remains unclear. Here, we show that the transcription factor, C/EBPδ, and the DNA methylation eraser, TET3, cooperatively control adipocyte differentiation. We perform whole-genome bisulfite sequencing to explore the dynamics and regulatory mechanisms of DNA methylation in adipocyte differentiation. During adipogenesis, DNA methylation selectively decreases at adipocyte-specific loci carrying the C/EBP binding motif, which correlates with the activity of adipogenic promoters and enhancers. Mechanistically, we find that C/EBPδ recruits a DNA methylation eraser, TET3, to catalyse DNA demethylation at the C/EBP binding motif and stimulate the expression of key adipogenic genes. Ectopic expression of TET3 potentiates in vitro and in vivo adipocyte differentiation and recovers downregulated adipogenic potential, which is observed in aged mice and humans. Taken together, our study highlights how targeted reprogramming of DNA methylation through cooperative action of the transcription factor C/EBPδ, and the DNA methylation eraser TET3, controls adipocyte differentiation.

Common dental anomalies in Korean orthodontic patients: An update.

Objective: The aim of this study was 1) to investigate the prevalence and pattern of dental anomalies (DAs), 2) to compare DAs according to the type of malocclusion, and 3) to investigate the correlation between tooth impaction and other DAs in the Korean orthodontic population. Methods: A total of 3,240 orthodontic patients were classified as Class I, Class II, or Class III malocclusion groups. The presence and location of common DAs, including impaction, microdontia, agenesis, supernumerary tooth, transposition, and fusion, were identified by examining diagnostic records. Furthermore, samples were classified as Group 1 without impaction or Group 2 with impaction. The prevalence of other DAs concurrent with impaction was investigated and compared to Group 1. Results: Impaction was the most prevalent DA, followed by microdontia, agenesis, and supernumerary. Class I and Class III groups showed the same order of prevalence, but agenesis was more frequent than microdontia in the Class II group. The prevalence of the four DAs was lowest in the Class III group. Overall, 8.6% of patients were classified into Group 2. The incidence of DAs other than impaction and the prevalence of multiple concurrent DAs were significantly higher in Group 2. Impaction showed a significant relationship with supernumerary tooth, transposition, and fusion. Conclusions: The prevalence and pattern of DAs varied depending on the type of malocclusion. As there was a higher risk of other DAs in patients with impacted teeth, early detection of the impacted tooth and a detailed diagnosis of other possible DAs may be essential.