Dysregulation of CREB5 Impairs Decidualization and Maternal-Fetal Interactions by Inhibiting Autophagy in Recurrent Spontaneous Abortion.

BACKGROUND: Clinically, recurrent spontaneous abortion (RSA) is a pregnancy illness that is difficult to treat. Impaired decidualization is a documented cause of RSA, but the etiology and mechanism are still unknown. cAMP-responsive element binding protein 5 (CREB5) is a member of the ATF/CREB family. CREB5 has been reported to be related to pathological pregnancy, but there are few related studies on this topic in patients with RSA, and the underlying mechanism is unclear. METHODS: We collected decidual tissues from RSA patients and healthy pregnant women to measure the expression level of CREB5, PRL, IGFBP1, ATG5, LC3B, and SQSTM/p62. Then, the changes in CREB5 expression and autophagy levels were measured in human endometrial stromal cells (hESCs) during decidualization. The expression levels of PRL and IGFBP1 were tested in sh-CREB5/ov-CREB5 hESCs after decidualization induction, and the autophagy level in sh-CREB5/ov-CREB5 hESCs was measured without decidualization induction. The decidualization ability of sh-CREB5 and ov-CREB5 hESCs treated with an autophagy inducer or inhibitor was measured. To investigate the effect of CREB5 in hESCs on the invasion and migration of HTR8/SVneo cells, we performed a coculture experiment. Finally, we examined the expression of CREB5 and autophagy key proteins in mouse decidual tissues by constructing an abortion mouse model. RESULTS: In our study, we found that the expression of CREB5 was unusually elevated in the uterine decidua of RSA patients, but the expression of PRL, IGFBP1, and autophagy were decreased. During the decidualization of hESCs, the expression of CREB5 gradually decreases in a time-dependent manner with increasing autophagy. Moreover, by knocking down or overexpressing CREB5 in hESCs, it was found that CREB5 can impair decidualization and reduce autophagy in hESCs. Furthermore, the damage caused by CREB5 in terms of decidualization can be reversed by the addition of an autophagy inducer (rapamycin). In addition, CREB5 can increase the secretion of proteins (IL-1ß and TGF-ß1) in hESCs to inhibit trophoblast invasion and migration. CONCLUSIONS: Our data support the supposition that CREB5 disturbs the decidualization of endometrial stromal cells and interactions at the maternal-fetal interface by inhibiting autophagy and that its abnormal upregulation and dysfunction may lead to RSA. It may function as a diagnostic and therapeutic target for RSA. Similarly, we found that in the spontaneous abortion mouse model, the expression of CREB5 in the decidual tissue of the abortion group was significantly increased, and autophagy was decreased.

Metabolic reprogramming based on RNA sequencing of gemcitabine-resistant cells reveals the FASN gene as a therapeutic for bladder cancer.

Bladder cancer (BLCA) is the most frequent malignant tumor of the genitourinary system. Postoperative chemotherapy drug perfusion and chemotherapy are important means for the treatment of BLCA. However, once drug resistance occurs, BLCA develops rapidly after recurrence. BLCA cells rely on unique metabolic rewriting to maintain their growth and proliferation. However, the relationship between the metabolic pattern changes and drug resistance in BLCA is unclear. At present, this problem lacks systematic research. In our research, we identified and analyzed resistance- and metabolism-related differentially expressed genes (RM-DEGs) based on RNA sequencing of a gemcitabine-resistant BLCA cell line and metabolic-related genes (MRGs). Then, we established a drug resistance- and metabolism-related model (RM-RM) through regression analysis to predict the overall survival of BLCA. We also confirmed that RM-RM had a significant correlation with tumor metabolism, gene mutations, tumor microenvironment, and adverse drug reactions. Patients with a high drug resistance- and metabolism-related risk score (RM-RS) showed more active lipid synthesis than those with a low RM-RS. Further in vitro and in vivo studies were implemented using Fatty Acid Synthase (FASN), a representative gene, which promotes gemcitabine resistance, and its inhibitor (TVB-3166) that can reverse this resistance effect.

In-hospital mortality and length of hospital stay in infants requiring tracheostomy with bronchopulmonary dysplasia.

PURPOSE: To investigate in-hospital mortality and hospital length of stay (LOS) in infants requiring tracheostomy with bronchopulmonary dysplasia (BPD). METHODS: We explored the correlation between tracheostomy with in-hospital mortality and LOS in infant patients hospitalized with BPD, using the data from Nationwide Inpatient Sample between 2008 and 2017 in the United States. In-hospital mortality and LOS was compared in patients who underwent tracheostomy with those patients who did not after propensity-score matching. RESULTS: A total of 10,262 children ≤2 years old hospitalized with BPD, 847 (8%) underwent tracheostomy, and 821 patients underwent tracheostomy were matched with 1602 patients without tracheostomy. Tracheostomy group was correlated with higher in-hospital mortality(OR(95%CI):2.98(2.25-3.95)) and prolonged LOS(absolute difference(95%CI):97.0(85.6-108.4)). CONCLUSIONS: Tracheostomy was correlated with increased in-hospital mortality and prolonged LOS. Such information may contribute to better decision-making process between clinicians and parents regarding tracheostomy to manage parent expectations, as well as better interdisciplinary teamwork.

Diabetes risk prediction model based on community follow-up data using machine learning.

Diabetes is a chronic metabolic disease characterized by hyperglycemia, the follow-up management of diabetes patients is mostly in the community, but the relationship between key lifestyle indicators in community follow-up and the risk of diabetes is unclear. In order to explore the association between key life characteristic indicators of community follow-up and the risk of diabetes, 252,176 follow-up records of people with diabetes patients from 2016 to 2023 were obtained from Haizhu District, Guangzhou. According to the follow-up data, the key life characteristic indicators that affect diabetes are determined, and the optimal feature subset is obtained through feature selection technology to accurately assess the risk of diabetes. A diabetes risk assessment model based on a random forest classifier was designed, which used optimal feature parameter selection and algorithm model comparison, with an accuracy of 91.24% and an AUC corresponding to the ROC curve of 97%. In order to improve the applicability of the model in clinical and real life, a diabetes risk score card was designed and tested using the original data, the accuracy was 95.15%, and the model reliability was high. The diabetes risk prediction model based on community follow-up big data mining can be used for large-scale risk screening and early warning by community doctors based on patient follow-up data, further promoting diabetes prevention and control strategies, and can also be used for wearable devices or intelligent biosensors for individual patient self examination, in order to improve lifestyle and reduce risk factor levels.

Reconstitution of morphogen shuttling circuits.

Developing tissues form spatial patterns by establishing concentration gradients of diffusible signaling proteins called morphogens. The bone morphogenetic protein (BMP) morphogen pathway uses a family of extracellular modulators to reshape signaling gradients by actively "shuttling" ligands to different locations. It has remained unclear what circuits are sufficient to enable shuttling, what other patterns they can generate, and whether shuttling is evolutionarily conserved. Here, using a synthetic, bottom-up approach, we compared the spatiotemporal dynamics of different extracellular circuits. Three proteins-Chordin, Twsg, and the BMP-1 protease-successfully displaced gradients by shuttling ligands away from the site of production. A mathematical model explained the different spatial dynamics of this and other circuits. Last, combining mammalian and Drosophila components in the same system suggests that shuttling is a conserved capability. Together, these results reveal principles through which extracellular circuits control the spatiotemporal dynamics of morphogen signaling.

Updated skin transcriptomic atlas depicted by reciprocal contribution of single-nucleus RNA sequencing and single-cell RNA sequencing.

BACKGROUND: Single-cell RNA sequencing (scRNA-seq) has advanced our understanding of skin biology, but its utility is restricted by the requirement of fresh samples, inadequate dissociation-induced cell loss or death, and activation during tissue digestion. Single-nucleus RNA sequencing (snRNA-seq) can use frozen, hard-to-dissociate materials, which might be a promising method to circumvent the limitations of scRNA-seq for the skin tissue. OBJECTIVE: To profile skin cells using snRNA-seq in parallel with scRNA-seq. METHODS: We performed snRNA-seq in parallel with scRNA-seq for the bisected skin sample of one person and integrated previously published scRNA-seq data for analysis. We comparatively analyzed the differences in cell proportions and gene expression between the two methods. The differentiation trajectories of keratinocytes and fibroblasts were analyzed by Slingshot analysis. RESULTS: snRNA-seq was less susceptible to contamination from mitochondrial and ribosomal RNA, and exhibited a greater capacity to detect transcription factors. snRNA-seq identified more spatially and functionally relevant keratinocyte clusters that constitute cell trajectories with expected differentiation dynamics. Novel markers, e.g., LYPD3, EMP2, and CSTB, were revealed for different differentiation stages of keratinocytes, and NFIB and GRHL1 were identified as transcription factors involving in the proliferation and functional differentiation of keratinocytes. Fibroblasts were found in a state of activation in scRNA-seq. And scRNA-seq detected a greater number of immune cells. CONCLUSIONS: We generated an updated atlas of the skin transcriptome based on the reciprocal contribution of scRNA-seq and snRNA-seq.

Peripheral blood mononuclear cell- transcriptome signatures of atopic dermatitis and prediction for the efficacy of dupilumab.

BACKGROUND: Few studies have explored transcriptome of the peripheral blood mononuclear cells (PBMCs) of atopic dermatitis (AD). Parameters for prediction of the efficacy of dupilumab in AD remain obscure. OBJECTIVE: To explore transcriptome signature of the PBMCs from Chinese AD patients and the usage in predication for the efficacy of dupilumab. METHODS: A total of 56 moderate-to-severe adult AD patients were enrolled and followed up for 16 week-dupilumab treatment. PBMCs samples were collected at baseline and 16 weeks after dupilumab treatment. Thirty-five patients were subjected to RNA-sequencing. Weighted gene co-expression network analysis (WGCNA) was used to find genes for prediction of dupilumab efficacy, which was validated in the rest 21 AD patients. Another 30 healthy individuals were enrolled and subjected to RNA-sequencing as healthy controls. RESULTS: Upregulation of the T helper (Th) 2/Th22 pathway, Th17 antimicrobial genes, and natural T-regulatory cell abundance in the PBMCs of AD cases was observed, whereas TGF-ß signaling and NK-cell signaling were decreased. Dupilumab treatment reversed the increase in the expression of Th2 cytokine receptors. WGCNA identified two immune-related modules that were correlated significantly with the efficacy of dupilumab. Hub gene MAP2K3 and UBE2L3 of these two modules demonstrated potential predictive ability for efficacy in the RNA-sequencing group by Spearman correlation, ROC analysis, and regression analysis, which was further validated in additional 21 AD cases. CONCLUSION: We firstly revealed the molecular phenotype of PBMCs in Chinese patients with AD, and uncovered two molecules that might be useful for prediction of the efficacy of dupilumab.

A prediction model identifying glycolysis signature as therapeutic target for psoriasis.

Background: The hyperproliferation featured with upregulated glycolysis is a hallmark of psoriasis. However, molecular difference of keratinocyte glycolysis amongst varied pathologic states in psoriasis remain elusive. Objectives: To characterize glycolysis status of psoriatic skin and assess the potential of glycolysis score for therapeutic decision. Methods: We analyzed 345414 cells collected from different cohorts of single-cell RNA seq database. A new method, Scissor, was used to integrate the phenotypes in GSE11903 to guide single-cell data analysis, allowing identification of responder subpopulations. AUCell algorithm was performed to evaluate the glycolysis status of single cell. Glycolysis signature was used for further ordering in trajectory analysis. The signature model was built with logistic regression analysis and validated using external datasets. Results: Keratinocytes (KCs) expressing SLC2A1 and LDH1 were identified as a novel glycolysis-related subpopulation. Scissor+ cells and Scissor- cells were defined as response and non-response phenotypes. In Scissor+ SLC2A1+ LDH1+ KCs, ATP synthesis pathway was activated, especially, the glycolysis pathway being intriguing. Based on the glycolysis signature, keratinocyte differentiation was decomposed into a three-phase trajectory of normal, non-lesional, and lesional psoriatic cells. The area under the curve (AUC) and Brier score (BS) were used to estimate the performance of the glycolysis signature in distinguishing response and non-response samples in GSE69967 (AUC =0.786, BS =17.7) and GSE85034 (AUC=0.849, BS=11.1). Furthermore, Decision Curve Analysis suggested that the glycolysis score was clinically practicable. Conclusion: We demonstrated a novel glycolysis-related subpopulation of KCs, identified 12-glycolysis signature, and validated its promising predictive efficacy of treatment effectiveness.

Temporal trend and factors associated with post-endoscopic retrograde cholangiopancreatography pancreatitis in children in the USA: a nationwide, retrospective cohort study.

Pancreatitis is the most common adverse event following endoscopic retrograde cholangiopancreatography (ERCP). Meanwhile, the national temporal trend of post-ERCP pancreatitis (PEP) in children remains to be reported. The purpose of this study is to investigate the temporal trend and factors associated with PEP in children. We conducted a nationwide study using data from the National Inpatient Sample database during 2008-2017 and included all patients aged ≤ 18 years who underwent ERCP. The primary outcomes were temporal trends and factors associated with PEP. The secondary outcomes were in-hospital mortality, total charges (TC), and total length of stay (LOS). A total of 45,268 hospitalized pediatric patients who underwent ERCP were analyzed; of whom, 2043 (4.5%) were diagnosed with PEP. The prevalence of PEP decreased from 5.0% in 2008 to 4.6% in 2017 (P = 0.0002). In multivariable logistic analysis, adjusted risk factors of PEP were hospitals located in the West (aOR 2.09, 95% CI 1.36-3.20; P < .0001), bile duct stent insertion (aOR 1.49, 95% CI, 1.08-2.05; P = 0.0040), and end-stage renal disease (aOR 8.05, 95% CI 1.66-39.16; P = 0.0098). Adjusted protective factors of PEP were increasing age (aOR 0.95, 95% CI 0.92-0.98; P = 0.0014) and hospitals located in the South (aOR 0.53, 95% CI 0.30-0.94; P < .0001). In-hospital mortality, TC, and LOS were higher in patients with PEP than those without PEP. CONCLUSION: This study shows a decreasing national trend over time and identifies multiple protective and risk factors for pediatric PEP. Endoscopists can use the insights from this study to evaluate relevant factors before performing ERCP in children to prevent PEP and reduce the medical-care burden. WHAT IS KNOWN: • Although ERCP has become indispensable procedure in children as they are in adults, education and training programs for ERCP in children are underdeveloped in many countries. • PEP is the most common and most serious adverse event following ERCP. Research on PEP in adults showed rising hospital admission and mortality rates associated with PEP in the USA. WHAT IS NEW: • The national temporal trend of PEP among pediatric patients in the USA was decreasing from 2008 to 2017. • Older age was a protective factor for PEP in children, while end-stage renal disease and stent insertion into the bile duct were risk factors.

Dysregulated lipidome of sebum in patients with atopic dermatitis.

BACKGROUND: Lipids are the major components of skin barrier, mainly produced by keratinocytes and sebaceous glands. Previous studies on barrier dysfunction of atopic dermatitis (AD) mainly focus on the lipids from keratinocytes, whereas the role of sebaceous gland-derived lipids in AD has long been underrecognized. METHODS: The sebum secreted on the skin surface of AD patients was measured using the Delfin Sebum Scale. Sebum was collected using Sebutape patches and subjected for liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis. Multivariate data analysis was applied to explore the relationship among the lipidome, clinical features, and sebaceous gland-related molecules. RESULTS: The amount of sebum secreted from sebaceous glands was decreased in AD patients and was negatively correlated with the barrier function and disease severity. LC-MS/MS revealed the lipidome of sebum, which clustered distinctly between AD patients and healthy individuals. Among the differential lipid subclasses, triglycerides (TG) were exclusively decreased in AD patients and correlated with disease severity. The first principal component scores of AD patients, which represented the main signature of the lipidome, were positively correlated with the SCORAD scores and were significantly different across the patient groups with differential clinical symptoms such as skin dryness and pruritus. Further analysis on the previously published transcriptome data revealed aberrant expression of lipid metabolism-related genes in non-lesional skin of AD patients, which was associated with skin inflammation and barrier dysfunction and mainly derived from inner root sheath keratinocytes and sebaceous gland cells. CONCLUSION: Atopic dermatitis patients demonstrated a deviated lipidome of sebum and aberrant lipid metabolism in sebaceous glands, indicating a possible role of lipids from sebaceous glands in the pathogenesis of AD.

Deletion of Insulin-like growth factor II mRNA-binding protein 3 participates in the pathogenesis of recurrent spontaneous abortion by inhibiting IL-10 secretion and inducing M1 polarization.

Insulin-like growth factor II mRNA-binding protein 3 (IGF2BP3) has been proved to affect trophoblast function and embryonic development, but its role and potential mechanism in recurrent spontaneous abortion (RSA) are not clear. RSA is a complex reproductive disease, causing physical and mental damage to patients. In recent years, many studies have found that immune microenvironment is vital to maintain successful pregnancy in the maternal fetal interface. Therefore, this study aims to explore the role of IGF2BP3 in affecting macrophage polarization and its possible mechanism. In this article, we found that IGF2BP3 expression was decreased in placental villous samples of human and RSA mouse model, and knockdown of IGF2BP3 in HTR8/SVneo cells promotes M1 Mφ polarization. Combining with RNA sequencing analysis, we found that IGF2BP3 may regulate the Mφ polarization by affecting the expression of trophoblast cytokines, especially IL-10 secretion. Further mechanistic studies showed that knockdown of IGF2BP3 decreased expression of IL-10 by activating NF-κB pathway. Moreover, we found that M2 Mφ promote trophoblast invasion not IGF2BP3 dependent. Our study reveals the interaction between trophoblast cells and macrophages at the maternal-fetal interface of RSA patients, and will provide theoretical guidance for its diagnosis and treatment of RSA patients.

IL-17D-induced inhibition of DDX5 expression in keratinocytes amplifies IL-36R-mediated skin inflammation.

Aberrant RNA splicing in keratinocytes drives inflammatory skin disorders. In the present study, we found that the RNA helicase DDX5 was downregulated in keratinocytes from the inflammatory skin lesions in patients with atopic dermatitis and psoriasis, and that mice with keratinocyte-specific deletion of Ddx5 (Ddx5∆KC) were more susceptible to cutaneous inflammation. Inhibition of DDX5 expression in keratinocytes was induced by the cytokine interleukin (IL)-17D through activation of the CD93-p38 MAPK-AKT-SMAD2/3 signaling pathway and led to pre-messenger RNA splicing events that favored the production of membrane-bound, intact IL-36 receptor (IL-36R) at the expense of soluble IL-36R (sIL-36R) and to the selective amplification of IL-36R-mediated inflammatory responses and cutaneous inflammation. Restoration of sIL-36R in Ddx5∆KC mice with experimental atopic dermatitis or psoriasis suppressed skin inflammation and alleviated the disease phenotypes. These findings indicate that IL-17D modulation of DDX5 expression controls inflammation in keratinocytes during inflammatory skin diseases.

A dysregulated sebum-microbial metabolite-IL-33 axis initiates skin inflammation in atopic dermatitis.

Microbial dysbiosis in the skin has been implicated in the pathogenesis of atopic dermatitis (AD); however, whether and how changes in the skin microbiome initiate skin inflammation, or vice versa, remains poorly understood. Here, we report that the levels of sebum and its microbial metabolite, propionate, were lower on the skin surface of AD patients compared with those of healthy individuals. Topical propionate application attenuated skin inflammation in mice with MC903-induced AD-like dermatitis by inhibiting IL-33 production in keratinocytes, an effect that was mediated through inhibition of HDAC and regulation of the AhR signaling pathway. Mice lacking sebum spontaneously developed AD-like dermatitis, which was improved by topical propionate application. A proof-of-concept clinical study further demonstrated the beneficial therapeutic effects of topical propionate application in AD patients. In summary, we have uncovered that the dysregulated sebum-microbial metabolite-IL-33 axis might play an initiating role in AD-related skin inflammation, thereby highlighting novel therapeutic strategies for the treatment of AD.

YY1/ITGA3 pathway may affect trophoblastic cells migration and invasion ability.

Recurrent spontaneous abortion (RSA) is a disturbing pregnancy disorder experienced by ~2.5% of women attempting to conceive. The pathogenesis of RSA is still unclear. Previous findings revealed that transcription factor YIN-YANG 1(YY1) was related to the pathogenesis of RSA by influence trophoblastic cell invasion ability. Present study aimed to investigate more specific molecular mechanism of YY1 playing in trophoblastic cells. In our research, RNA-seq and Chip-seq were used to find significant changed genes between si-YY1(Knock down of YY1) HTR-8/SVneo cells(n = 3) and HTR-8/SVneo cells(n = 3). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis results suggested that Integrins related pathway maybe necessary to biological functions of trophoblastic cells. Chip-seq dataset analysis results predict YY1 can regulate ITGA3/7 expression by binding to the promoter region of ITGA3/7. Furthermore, results from chip experiment, RT-PCR, Dual-luciferase reporter gene assay showed that YY1 was able to bind to the promoter region of ITGA3 and regulate ITGA3 mRNA and protein expression. However, ITGA7 could not be significant influenced by YY1. Besides, gene silencing experiment, Western blot and Immunofluorescence assay confirmed that both YY1 and ITGA3 can accelerate phosphorylation focal adhesion kinase and affect cytoskeleton formation in HTR-8/SVneo cells. In conclusion, YY1/ITGA3 play a critical role in trophoblast invasion ability by regulating cytoskeleton formation.

Fertility Intention Among Chinese Reproductive Couples During the COVID-19 Outbreak: A Cross-Sectional Study.

A decline in the fertility rate has been observed worldwide, which hampers social development severely. Given the impacts of COVID-19 on individuals and society, it is of great significance to investigate the fertility intention of reproductive couples under COVID-19. The convenience sampling method was used to obtain our study sample. The self-administered questionnaire included the following components: sociodemographic characteristics (age, residence, education, occupation, characteristics of the couples, and annual household income), reproductive history (parity, number of children, child gender, and duration of preparing pregnancy), and attitudes toward COVID-19, was distributed online via an applet of WeChat. The results showed that among 4,133 valid questionnaires, 1,091 had fertility intention before COVID-19, whereas 3,042 did not, indicating a fertility intention rate of 26.4% among participating couples. Of the 1,091 couples who had fertility intention before COVID-19, 520 (47.7%) were affected by the outbreak, whereas 571 (52.3%) were not. By multivariable logistic regression analysis, we further found that couples living in Hubei Province, the epicenter in China (OR 2.20, 95% CI 1.35-3.60), and couples who prepared for pregnancy longer before COVID-19 (OR 1.19, 95% CI 1.06-1.33) were more likely to change their fertility intention under the pandemic. In addition, most of the participants reported their fertility intention was affected by the inconvenience of seeking medical service under COVID-19. Therefore, more forms of medical services to provide convenience for patients might be effective ways to reverse the declined fertility intention rate in facing COVID-19.

TNF-α/anti-TNF-α drugs and its effect on pregnancy outcomes.

Pregnancy is a complex biological process. The establishment and maintenance of foetal-maternal interface are pivotal events. Decidual immune cells and inflammatory cytokines play indispensable roles in the foetal-maternal interface. The disfunction of decidual immune cells leads to adverse pregnancy outcome. Tumour necrosis factor (TNF)-α, a common inflammatory cytokine, has critical roles in different stages of normal pregnancy process. However, the relationship between the disorder of TNF-α and adverse pregnancy outcomes, including preeclampsia (PE), intrauterine growth restriction (IUGR), spontaneous abortion (SA), preterm birth and so on, is still indefinite. In this review, we thoroughly reviewed the effect of TNF-α disorder on pathological conditions. Moreover, we summarized the reports about the adverse pregnancy outcomes (PE, IUGR, SA and preterm birth) of using anti-TNF-α drugs (infliximab, etanercept and adalimumab, certolizumab and golimumab) currently in the clinical studies. Overall, IUGR, SA and preterm birth are the most common adverse pregnancy outcomes of anti-TNF-α drugs. Our review may provide insight for the immunological treatment of pregnancy-related complication, and help practitioners make informed decisions based on the current evidences.

The Mechanism of Insulin-Like Growth Factor II mRNA-Binging Protein 3 Induce Decidualization and Maternal-Fetal Interface Cross Talk by TGF-ß1 in Recurrent Spontaneous Abortion.

Recurrent spontaneous abortion (RSA) is defined as the loss of two or more consecutive intrauterine pregnancies that are clinically established early in pregnancy. To date, the etiology and underlying mechanisms of RSA remain unclear. It is widely thought that the impairment of decidualization is inclined to induce subsequent pregnancy failure and leads to the dysregulation of extra-villous trophoblast invasion and proliferation through maternal-fetal cross talk. However, the mechanism of decidualization in RSA has yet to be understood. In our study, we demonstrate that decidual samples from RSA patients have significantly higher insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3) and lower TGF-ß1 levels compared to healthy controls. In addition, the overexpression of IGF2BP3 in human endometrial stromal cells (hESCs) can lead to the impairment of decidualization in vitro-induced model and the abnormal cell cycle regulation. Furthermore, TGF-ß1 and MMP9 levels were greatly increased after decidualization, whereas IGF2BP3 overexpression inhibited endometrial mesenchymal decidualization by downregulating TGF-ß1, impeding maternal-fetal interface cytokine cross talk, and limiting the ability of trophoblast invasion. In conclusion, our investigation first demonstrates that abnormal elevation of IGF2BP3 in the pregnant endometrium leads to the impairment of decidualization and abnormal trophoblast invasion, thereby predisposing individuals to RSA.

Construction and Validation of a Ferroptosis-Related Prognostic Signature for Melanoma Based on Single-Cell RNA Sequencing.

Melanoma, the deadliest type of skin cancer, is on the rise globally. The generally poor prognosis makes melanoma still an enormous public health problem. Ferroptosis is a newly emerging form of iron-dependent regulated cell death, which has been implicated in the development and treatment of several tumors. However, whether there is a connection between ferroptosis-related genes and the prognosis of melanoma patients remains an enigma. In the present study, we identified a ferroptosis-related genes signature to predict the prognosis of melanoma patients by analyzing single-cell RNA-sequencing data from the Gene Expression Omnibus (GEO). Single-cell trajectory analysis was performed to explore malignant differentiation. CellChat was used to investigate intercellular communications in melanoma. Collectively, a novel four-gene signature (CP, MAP1LC3A, transferrin, and TP53) was constructed for prognosis prediction. COX proportional hazards regression analysis showed that the established ferroptosis-associated risk model was an independent prognostic predictor for melanoma patients (HR = 2.3293; 95%CI 1.1528-4.706) (p < 0.018). Patients with low-risk scores had significantly better overall survival (OS) than those with high-risk scores in The Cancer Genome Atlas, GSE59455, and GSE22153 dataset (p = 0.0015, p = 0.031, p = 0.077). Furthermore, the gene expression level of the four genes were verified in multistrain melanoma cell lines and normal human epidermal melanocytes (NHEM). The protein expression level of the four genes in clinical samples were further verified in the Human Protein Atlas (HPA) databases. Taken together, our study identified the prognostic significance of the ferroptosis-related genes in melanoma and developed a novel four-gene prognostic signature, which may shed light on the prognostic assessment and clinical decision making for melanoma patients.

Protease-controlled secretion and display of intercellular signals.

To program intercellular communication for biomedicine, it is crucial to regulate the secretion and surface display of signaling proteins. If such regulations are at the protein level, there are additional advantages, including compact delivery and direct interactions with endogenous signaling pathways. Here we create a modular, generalizable design called Retained Endoplasmic Cleavable Secretion (RELEASE), with engineered proteins retained in the endoplasmic reticulum and displayed/secreted in response to specific proteases. The design allows functional regulation of multiple synthetic and natural proteins by synthetic protease circuits to realize diverse signal processing capabilities, including logic operation and threshold tuning. By linking RELEASE to additional sensing and processing circuits, we can achieve elevated protein secretion in response to "undruggable" oncogene KRAS mutants. RELEASE should enable the local, programmable delivery of intercellular cues for a broad variety of fields such as neurobiology, cancer immunotherapy and cell transplantation.