Gene silencing dynamics are modulated by transiently active regulatory elements.

Transcriptional enhancers have been extensively characterized, but cis-regulatory elements involved in acute gene repression have received less attention. Transcription factor GATA1 promotes erythroid differentiation by activating and repressing distinct gene sets. Here, we study the mechanism by which GATA1 silences the proliferative gene Kit during murine erythroid cell maturation and define stages from initial loss of activation to heterochromatinization. We find that GATA1 inactivates a potent upstream enhancer but concomitantly creates a discrete intronic regulatory region marked by H3K27ac, short noncoding RNAs, and de novo chromatin looping. This enhancer-like element forms transiently and serves to delay Kit silencing. The element is ultimately erased via the FOG1/NuRD deacetylase complex, as revealed by the study of a disease-associated GATA1 variant. Hence, regulatory sites can be self-limiting by dynamic co-factor usage. Genome-wide analyses across cell types and species uncover transiently active elements at numerous genes during repression, suggesting that modulation of silencing kinetics is widespread.

Interspecies regulatory landscapes and elements revealed by novel joint systematic integration of human and mouse blood cell epigenomes.

Knowledge of locations and activities of cis-regulatory elements (CREs) is needed to decipher basic mechanisms of gene regulation and to understand the impact of genetic variants on complex traits. Previous studies identified candidate CREs (cCREs) using epigenetic features in one species, making comparisons difficult between species. In contrast, we conducted an interspecies study defining epigenetic states and identifying cCREs in blood cell types to generate regulatory maps that are comparable between species, using integrative modeling of eight epigenetic features jointly in human and mouse in our Validated Systematic Integration (VISION) Project. The resulting catalogs of cCREs are useful resources for further studies of gene regulation in blood cells, indicated by high overlap with known functional elements and strong enrichment for human genetic variants associated with blood cell phenotypes. The contribution of each epigenetic state in cCREs to gene regulation, inferred from a multivariate regression, was used to estimate epigenetic state regulatory potential (esRP) scores for each cCRE in each cell type, which were used to categorize dynamic changes in cCREs. Groups of cCREs displaying similar patterns of regulatory activity in human and mouse cell types, obtained by joint clustering on esRP scores, harbor distinctive transcription factor binding motifs that are similar between species. An interspecies comparison of cCREs revealed both conserved and species-specific patterns of epigenetic evolution. Finally, we show that comparisons of the epigenetic landscape between species can reveal elements with similar roles in regulation, even in the absence of genomic sequence alignment.

Up-regulation of BTN3A1 on CD14+ cells promotes Vγ9Vδ2 T cell activation in psoriasis.

Vγ9Vδ2 T cells play an important role in the development and progression of psoriasis vulgaris (PV), but how they promote skin inflammation and the molecular mechanisms underlying Vγ9Vδ2 T cell dysfunction are poorly understood. Here, we show that circulating Vγ9Vδ2 T cells are decreased and exhibit enhanced proliferation and increased production of IFN-γ and TNF-α in PV patients. Monocytes from PV patients express higher levels of the phosphoantigen sensor butyrophilin 3A1 (BTN3A1) than monocytes from healthy controls. Blockade of BTN3A1 suppresses Vγ9Vδ2 T cell activation and abolishes the difference in Vγ9Vδ2 T cell activation between PV patients and healthy controls. The CD14+ cells in PV skin lesions highly express BTN3A1 and juxtapose to Vδ2 T cells. In addition, IFN-γ induces the up-regulation of BTN3A1 on monocytes. Collectively, our results demonstrate a crucial role of BTN3A1 on monocytes in regulating Vγ9Vδ2 T cell activation and highlight BTN3A1 as a potential therapeutic target for psoriasis.

FeP-Based Nanotheranostic Platform for Enhanced Phototherapy/Ferroptosis/Chemodynamic Therapy.

Ferroptosis is an iron-dependent and lipid peroxides (LPO)-overloaded programmed damage cell death, induced by glutathione (GSH) depletion and glutathione peroxide 4 (GPX4) inactivation. However, the inadequacy of endogenous iron and reactive oxygen species (ROS) restricts the efficacy of ferroptosis. To overcome this obstacle, a near-infrared photo-responsive FeP@PEG NPs is fabricated. Exogenous iron pool can enhance the effect of ferroptosis via the depletion of GSH and further regulate GPX4 inactivation. Generation of ·OH derived from the Fenton reaction is proved by increased accumulation of lipid peroxides. The heat generated by photothermal therapy and ROS generated by photodynamic therapy can enhance cell apoptosis under near-infrared (NIR-808 nm) irradiation, as evidenced by mitochondrial dysfunction and further accumulation of lipid peroxide content. FeP@PEG NPs can significantly inhibit the growth of several types of cancer cells in vitro and in vivo, which is validated by theoretical and experimental results. Meanwhile, FeP@PEG NPs show excellent T2-weighted magnetic resonance imaging (MRI) property. In summary, the FeP-based nanotheranostic platform for enhanced phototherapy/ferroptosis/chemodynamic therapy provides a reliable opportunity for clinical cancer theranostics.

Unlinking an lncRNA from Its Associated cis Element.

Long non-coding (lnc) RNAs can regulate gene expression and protein functions. However, the proportion of lncRNAs with biological activities among the thousands expressed in mammalian cells is controversial. We studied Lockd (lncRNA downstream of Cdkn1b), a 434-nt polyadenylated lncRNA originating 4 kb 3' to the Cdkn1b gene. Deletion of the 25-kb Lockd locus reduced Cdkn1b transcription by approximately 70% in an erythroid cell line. In contrast, homozygous insertion of a polyadenylation cassette 80 bp downstream of the Lockd transcription start site reduced the entire lncRNA transcript level by >90% with no effect on Cdkn1b transcription. The Lockd promoter contains a DNase-hypersensitive site, binds numerous transcription factors, and physically associates with the Cdkn1b promoter in chromosomal conformation capture studies. Therefore, the Lockd gene positively regulates Cdkn1b transcription through an enhancer-like cis element, whereas the lncRNA itself is dispensable, which may be the case for other lncRNAs.

Establishment of a Mouse Model for Porokeratosis Due to Mevalonate Diphosphate Decarboxylase Deficiency.

INTRODUCTION: Porokeratosis (PK) is an autoinflammatory keratinization disease (AIKD) characterized by circular or annular skin lesions with a hyperkeratotic rim, pathologically shown as the cornoid lamella. Four genes that cause PK are associated with the mevalonate (MV) pathway. In Chinese PK patients, mevalonate diphosphate decarboxylase (MVD) is the most common causative gene. The lack of an animal model has greatly limited research on PK pathogenesis. MATERIALS AND METHODS: In this research, we constructed K14-CreERT2-Mvdfl/fl mice using the Cre-LoxP system to create a mouse model for in-depth studies of PK. The Epidermal Mvd gene was knocked out by intraperitoneal injection of Tamoxifen (TAM). Pathology, immunohistochemistry, RNA-seq, and Western Blot analysis were performed. RESULTS: Skin lesions appeared following Mvd deficiency, and pathological examination revealed the characteristic cornoid lamella, as well as cutaneous inflammation. Furthermore, we observed elevated levels of IL-17A and IL-1ß, and a decreased Loricrin level in epidermal Mvd-deficient mice. Compared with the wild-type (WT) group, Mvd deficiency activated the expression of lipid metabolism-related proteins. CONCLUSION: We developed the first mouse model for PK research, enabling further studies on disease development and treatment approaches.

Direct modulation bandwidth enhancement of uncooled DFB laser operating over a wide temperature range based on groove-in-trench waveguide structure.

Uncooled direct modulation DFB laser offers high speed transmission rate over a wide temperature range with high reliability and low cost, making it a cost-effective light source choice for 5G fronthaul and data center applications. However, a significant 3dB bandwidth decrease can be observed in high temperature for conventional DFB lasers. We present an uncooled DFB laser operating up to 85°C with extended direct modulation bandwidth and high reliability based on a novel groove-in-trench ridge waveguide structure, where two narrow grooves penetrating the active layer are etched symmetrically in the two conventional trenches by deep wet etching, respectively. By optimizing the distance between the groove and the mesa stripe, we obtain a 3dB bandwidth of 15.3 GHz at 85°C, which is a 3.7 GHz improvement compared with the conventional ridge waveguide DFB laser. Transmissions of 25 Gb/s NRZ signal at 25°C and 85°C with clear eye openings have been demonstrated. It also achieves 25 Gb/s transmission over 10 km optical fiber with a low power penalty of 0.5 dB for a bit error rate of 10-12 at 85°C. In addition, the result of 2000-hour aging test shows that the proposed groove-in-trench structure DFB lasers have the same excellent reliability as the conventional ones.

Experiment demonstration of high speed 1.3 µm grating assisted surface-emitting DFB lasers.

Surface emitting lasers are attractive light sources for silicon integrated photonic circuits. High speed direct operation is of great importance for these lasers in high capacity and low cost on-chip communication system. Here, we demonstrate a 1.3 µm surface emitting ridge-waveguide distributed feedback (DFB) laser with second order grating and λ/4 phase shift grating, which can achieve a 24 Gb/s operation over a wide temperature. The fabricated lasers can achieve low threshold current as 6.8 mA, and 12.5 mA at 20, and 70°C, respectively. Stable single mode operation has been observed with high side mode suppression ratio (SMSR) > 40 dB at all temperatures (20-70 °C). Meanwhile, the surface emitting optical power can reach 1.7 mW at high temperature as 70 °C. 3 dB bandwidth of small signal response is 21 GHz and 12 GHz at 20 °C and 70 °C respectively. The far-field divergence angle of surface emitting beam is 13.4°×20.2° of 10 µm length second order grating coupler. The proposed laser may have great advantages of single mode, high speed modulation and good temperature tolerance. In addition, compared with conventional DFB lasers, the surface emitting DFB laser has no additional manufacturing process, which is simple to fabricate and easy to integrate with silicon platform.

Activation of Autophagy Induces Monocrotaline-Induced Pulmonary Arterial Hypertension by FOXM1-Mediated FAK Phosphorylation.

PURPOSE: It has been shown that activation of autophagy promotes the development of pulmonary arterial hypertension (PAH). Meanwhile, forkhead box M1 (FOXM1) has been found to induce autophagy in several types of cancer. However, it is still unclear whether FOXM1 mediates autophagy activation in PAH, and detailed mechanisms responsible for these processes are indefinite. METHOD: PAH was induced by a single intraperitoneal injection of monocrotaline (MCT) to rats. The right ventricle systolic pressure (RVSP), right ventricular hypertrophy index (RVHI), percentage of medial wall thickness (%MT), α-smooth muscle actin (α-SMA) staining, and Ki67 staining were performed to evaluate the development of PAH. The protein levels of FOXM1, phospho-focal adhesion kinase (p-FAK), FAK, and LC3B were determined by immunoblotting or immunohistochemistry. RESULTS: FOXM1 protein level and FAK activity were significantly increased in MCT-induced PAH rats, this was accompanied with the activation of autophagy. Pharmacological inhibition of FOXM1 or FAK suppressed MCT-induced autophagy activation, decreased RVSP, RVHI and %MT in MCT-induced PAH rats, and inhibited the proliferation of pulmonary arterial smooth muscle cells and pulmonary vessel muscularization in MCT-induced PAH rats. CONCLUSION: FOXM1 promotes the development of PAH by inducing FAK phosphorylation and subsequent activation of autophagy in MCT-treated rats.

Factors affecting physicians using mobile health applications: an empirical study.

BACKGROUND: Mobile health applications (mHealth apps) have created innovative service channels for patients with chronic diseases. These innovative service channels require physicians to actively use mHealth apps. However, few studies investigate physicians' participation in mHealth apps. OBJECTIVE: This study aims to empirically explore factors affecting physicians' usage behaviors of mHealth apps. Based on the extended Unified Theory of Acceptance and Use of Technology (UTAUT2) and mHealth apps features, we propose a research model including altruism, cognitive trust, and online ratings. METHODS: We collected data from physicians who have used mHealth apps and conducted a factor analysis to verify the convergence and discriminative effects. We used a hierarchical regression method to test the path coefficients and statistical significance of our research model. In addition, we adopted bootstrapping approach and further analyzed the mediating effects of behavioral intention between all antecedent variables and physicians' usage behavior. Finally, we conducted three robustness analyses to test the validity of results and tested the constructs to verify the common method bias. RESULTS: Our results support the effects of performance expectancy, effort expectancy, social influence, and altruism on the behavioral intentions of physicians using mHealth apps. Moreover, facilitating conditions and habits positively affect physicians using mHealth apps through the mediating effort of behavioral intention. Physicians' cognitive trust and online rating have significant effects on their usage behaviors through the mediating efforts of behavioral intention. CONCLUSIONS: This study contributes to the existing literature on UTAUT2 extension of physicians' acceptance of mHealth apps by adding altruism, cognitive trust, and online ratings. The results of this study provide a novel perspective in understanding the factors affecting physicians' usage behaviors on mHealth apps in China and provide such apps' managers with an insight into the promotion of physicians' active acceptance and usage behaviors.