Abstract Background Sepsis is a fatal disease referring to the presence of a known or strongly suspected infection coupled with systemic and uncontrolled immune activation causing multiple organ failure. However, current knowledge of the role of lncRNAs in sepsis is still extremely limited. Methods We performed an in silico investigation of the gene coexpression pattern for the patients response to all-cause sepsis in consecutive intensive care unit (ICU) admissions. Sepsis coexpression gene modules were identified using WGCNA and enrichment analysis. lncRNAs were determined as sepsis biomarkers based on the interactions among lncRNAs and the identified modules. Results Twenty-three sepsis modules, including both differentially expressed modules and prognostic modules, were identified from the whole blood RNA expression profiling of sepsis patients. Five lncRNAs, FENDRR, MALAT1, TUG1, CRNDE, and ANCR, were detected as sepsis regulators based on the interactions among lncRNAs and the identified coexpression modules. Furthermore, we found that CRNDE and MALAT1 may act as miRNA sponges of sepsis related miRNAs to regulate the expression of sepsis modules. Ultimately, FENDRR, MALAT1, TUG1, and CRNDE were reannotated using three independent lncRNA expression datasets and validated as differentially expressed lncRNAs. Conclusion The procedure facilitates the identification of prognostic biomarkers and novel therapeutic strategies of sepsis. Our findings highlight the importance of transcriptome modularity and regulatory lncRNAs in the progress of sepsis.
Abstract Sepsis, caused by infections, sparks a dangerous bodily response. The transcriptional expression patterns of host responses aid in the diagnosis of sepsis, but the challenge lies in their limited generalization capabilities. To facilitate sepsis diagnosis, we present an updated version of single-cell Pair-wise Analysis of Gene Expression (scPAGE) using transfer learning method, scPAGE2, dedicated to data fusion between single-cell and bulk transcriptome. Compared to scPAGE, the upgrade to scPAGE2 featured ameliorated Differentially Expressed Gene Pairs (DEPs) for pretraining a model in single-cell transcriptome and retrained it using bulk transcriptome data to construct a sepsis diagnostic model, which effectively transferred cell-layer information from single-cell to bulk transcriptome. Seven datasets across three transcriptome platforms and fluorescence-activated cell sorting (FACS) were used for performance validation. The model involved four DEPs, showing robust performance across next-generation sequencing and microarray platforms, surpassing state-of-the-art models with an average AUROC of 0.947 and an average AUPRC of 0.987. Analysis of scRNA-seq data reveals higher cell proportions with JAM3-PIK3AP1 expression in sepsis monocytes, decreased ARG1-CCR7 in B and T cells. Elevated IRF6-HP in sepsis monocytes confirmed by both scRNA-seq and an independent cohort using FACS. Both the superior performance of the model and the in vitro validation of IRF6-HP in monocytes emphasize that scPAGE2 is effective and robust in the construction of sepsis diagnostic model. We additionally applied scPAGE2 to acute myeloid leukemia and demonstrated its superior classification performance. Overall, we provided a strategy to improve the generalizability of classification model that can be adapted to a broad range of clinical prediction scenarios.
Abstract Background With progress in tumor treatments, patient survival has been significantly extended; nevertheless, tumors and tumor treatments increase the risk of sepsis. Carrimycin may act as an immune-regulating treatment for tumor-related sepsis. We aimed to evaluate whether carrimycin regulates inflammation and immune function in tumor patients with sepsis. Methods We conducted a multicenter, randomized, placebo-controlled, double-blind clinical trial involving tumor patients with sepsis. The participant inclusion criteria were as follows: 1. age ≥ 18 and ≤ 75 years old; 2. condition consistent with sepsis 3.0 diagnostic criteria; 3. SOFA score of 2–13; and 4. patients with malignant tumors. Enrolled patients were assigned to either carrimycin treatment (400 mg/day) or placebo treatment (400 mg/day) orally once a day for 7 days. The primary outcome was immune-related indicators. Results A total of 120 patients were randomized, of whom 47 were assigned to receive carrimycin and 52 placebo. In immune and inflammation indicators, the HLA-DR and CD8 + T-cell levels showed promising trends, although there was no significant difference between the carrimycin and placebo groups ( P > 0.05). In the CD4 < 38.25 subgroup, the HLA-DR level of the carrimycin group was significantly better than that of the placebo group at 1 day after administration ( P = 0.023). In the CD8 < 25.195 subgroup, the degree of decrease in IL-8 in the carrimycin group was significantly higher than that in the placebo group at 1 ( P = 0.027) and 3 ( P = 0.034) days after administration. The CD8 + T-cell subset level of the carrimycin group was significantly better than that of the placebo group at 3 ( P = 0.027) and 5 ( P = 0.035) days after administration. The levels of SOFA, APACHE II, PCT and CRP were significantly reduced by carrimycin intervention. No serious adverse events were recorded. Conclusions In tumor patients with sepsis, especially those with immunocompromised function, carrimycin regulates the immune status by increasing the HLA-DR level and plays an anti-infective role to improve the severity of the disease but does not affect 28-day all-cause mortality. The trial was registered in the Chinese Clinical Trial Registry (http://www.chictr.org.cn) with the number ChiCTR2000032339 on April 26, 2020.
Fig. 1 PaO 2 /FiO 2 and PaO 2 improved in left lateral, right lateral, and prone position compared with supine position ( + P < 0.05 vs supine 0.5 h, * P < 0.05 vs supine 2.5 h, # P < 0.05 vs supine 4.5), and the improvement degree of PaO 2 /FiO 2 and PaO 2 in left and right lateral position was less than that of prone position ( & P < 0.05 vs supine prone 5 h, x P < 0.05 vs supine prone 6h)
Abstract Background Sepsis-associated acute kidney injury (SA-AKI) is a frequent complication in patients with sepsis and is associated with high mortality. Therefore, early recognition of SA-AKI is essential for administering supportive treatment and preventing further damage. This study aimed to identify and validate metabolite biomarkers of SA-AKI to assist in early clinical diagnosis. Methods Untargeted renal proteomic and metabolomic analyses were performed on the renal tissues of LPS-induced SA-AKI and sepsis mice. Glomerular filtration rate (GFR) monitoring technology was used to evaluate real-time renal function in mice. To elucidate the distinctive characteristics of SA-AKI, a multi-omics Spearman correlation network was constructed integrating core metabolites, proteins, and renal function. Subsequently, metabolomics analysis was used to explore the dynamic changes of core metabolites in the serum of SA-AKI mice at 0, 8, and 24 h. Finally, a clinical cohort (28 patients with SA-AKI vs. 28 patients with sepsis) serum quantitative metabolomic analysis was carried out to build a diagnostic model for SA-AKI via logistic regression (LR). Results Thirteen differential renal metabolites and 112 differential renal proteins were identified through a multi-omics study of SA-AKI mice. Subsequently, a multi-omics correlation network was constructed to highlight five core metabolites, i.e., 3-hydroxybutyric acid, 3-hydroxymethylglutaric acid, creatine, myristic acid, and inosine, the early changes of which were then observed via serum time series experiments of SA-AKI mice. The levels of 3-hydroxybutyric acid, 3-hydroxymethylglutaric acid, and creatine increased significantly at 24 h, myristic acid increased at 8 h, while inosine decreased at 8 h. Ultimately, based on the identified core metabolites, we recruited 56 patients and constructed a diagnostic model named IC3, using inosine, creatine, and 3-hydroxybutyric acid, to early identify SA-AKI (AUC = 0.90). Conclusions We proposed a blood metabolite model consisting of inosine, creatine, and 3-hydroxybutyric acid for the early screening of SA-AKI. Future studies will observe the performance of these metabolites in other clinical populations to evaluate their diagnostic role.
The novel Hsp90 inhibitor SNX-2112 showed broad antitumor activity. However, it was still necessary to optimize the therapeutic dosage of SNX-2112 applied on tumors to obtain effective therapy with minimal dose to reduce toxicity. We investigated the role of low-intensity US in promoting antitumorigenic effect of low doses of SNX-2112 on tongue squamous cell carcinoma.Cell viability was measured using CCK-8 assay or staining with Calcein AM/PI. Relative cumulative levels of SNX-2112 in cells were detected using high-performance liquid chromatography. The production of ROS was analyzed using fluorescence microscope and flow cytometer. Cellular apoptosis was detected using flow cytometer. The expression levels of proteins of the ERS-associated apoptosis signaling pathway were detected using Western blotting analysis. The efficacy and biosafety of SNX-2112 were also investigated in a mouse xenograft model.Low-intensity US combined with SNX-2112 exhibited significant antitumor effect, increased the absorption of SNX-2112 by cells even with a low dose, enhanced ROS generation and apoptosis. The combination regimen also inhibited the protein expression of Hsp90 and triggered apoptosis through endoplasmic reticulum stress (ERS) by enhancing PERK, CHOP and Bax protein levels, while downregulating the level of Bcl-2. Additionally, N-acetyl-L-cysteine (NAC), ROS scavenger, was able to reverse these results. Low-intensity US combined with SNX-2112 significantly inhibited tumor growth, prolonged survival of mice, decreased proliferation and promoted apoptosis with no visible damage or abnormalities in major organs in the mouse xenograft model with tongue squamous cell carcinoma.The antitumor effects of SNX-2112 were enhanced by low-intensity US. The most probable mechanism was that US sonoporation induced more SNX-2112 delivery to the cells and enhanced ROS production, triggering the ERS-associated apoptosis signaling pathway. Therefore, low-intensity US may increase the efficiency of conventional chemotherapy and reduce the dosage of SNX-2112 required and its side effects.
We aimed to identify and verify the key genes and lncRNAs associated with acute lung injury (ALI) and explore the pathogenesis of ALI. Research showed that lower expression of the lncRNA metastasis-associated lung carcinoma transcript 1 (MALAT1) alleviates lung injury induced by lipopolysaccharide (LPS). Nevertheless, the mechanisms of MALAT1 on cellular apoptosis remain unclear in LPS-stimulated ALI. We investigated the mechanism of MALAT1 in modulating the apoptosis of LPS-induced human pulmonary alveolar epithelial cells (HPAEpiC).Differentially expressed lncRNAs between the ALI samples and normal controls were identified using gene expression profiles. ALI-related genes were determined by the overlap of differentially expressed genes (DEGs), genes correlated with lung, genes correlated with key lncRNAs, and genes sharing significantly high proportions of microRNA targets with MALAT1. Quantitative real-time PCR (qPCR) was applied to detect the expression of MALAT1, microRNA (miR)-194-5p, and forkhead box P2 (FOXP2) mRNA in 1 μg/ml LPS-treated HPAEpiC. MALAT1 knockdown vectors, miR-194-5p inhibitors, and ov-FOXP2 were constructed and used to transfect HPAEpiC. The influence of MALAT1 knockdown on LPS-induced HPAEpiC proliferation and apoptosis via the miR-194-5p/FOXP2 axis was determined using Cell counting kit-8 (CCK-8) assay, flow cytometry, and Western blotting analysis, respectively. The interactions between MALAT1, miR-194-5p, and FOXP2 were verified using dual-luciferase reporter gene assay.We identified a key lncRNA (MALAT1) and three key genes (EYA1, WNT5A, and FOXP2) that are closely correlated with the pathogenesis of ALI. LPS stimulation promoted MALAT1 expression and apoptosis and also inhibited HPAEpiC viability. MALAT1 knockdown significantly improved viability and suppressed the apoptosis of LPS-stimulated HPAEpiC. Moreover, MALAT1 directly targeted miR-194-5p, a downregulated miRNA in LPS-stimulated HPAEpiC, when FOXP2 was overexpressed. MALAT1 knockdown led to the overexpression of miR-194-5p and restrained FOXP2 expression. Furthermore, inhibition of miR-194-5p exerted a rescue effect on MALAT1 knockdown of FOXP2, whereas the overexpression of FOXP2 reversed the effect of MALAT1 knockdown on viability and apoptosis of LPS-stimulated HPAEpiC.Our results demonstrated that MALAT1 knockdown alleviated HPAEpiC apoptosis by competitively binding to miR-194-5p and then elevating the inhibitory effect on its target FOXP2. These data provide a novel insight into the role of MALAT1 in the progression of ALI and potential diagnostic and therapeutic strategies for ALI patients.
Abstract Sepsis is a systemic inflammatory response that may be induced by trauma, infection, surgery, and burns. With the aim of discovering novel treatment targets for sepsis, this current study was conducted to investigate the effect and potential mechanism by which microRNA‐30a (miR‐30a) controls sepsis‐induced liver cell proliferation and apoptosis. Rat models of sepsis were established by applying the cecal ligation and puncture (CLP) method to simulate sepsis models. The binding site between miR‐30a and suppressor of cytokine signaling protein 1 (SOCS‐1) was determined by dual luciferase reporter gene assay. The gain‐of‐and‐loss‐of‐function experiments were applied to analyze the effects of miR‐30a and SOCS‐1 on liver cell proliferation and apoptosis of the established sepsis rat models. The expression of miR‐30a, SOCS‐1, Janus kinase 2 (JAK2), signal transducer and activator of transcription 3 (STAT3), Bcl‐2 associated X protein (Bax), B cell lymphoma‐2 (Bcl‐2), toll‐like receptor 4 (TLR4), and high‐mobility group box 1 (HMGB1), and the extent of JAK2 and STAT3 phosphorylation were all determined. Sepsis led to an elevation of miR‐30a and also a decline of SOCS‐1 in the liver cells. SOCS‐1 was negatively regulated by miR‐30a. Upregulated miR‐30a and downregulated SOCS‐1 increased the expression of JAK2, STAT3, Bax, TLR4, and HMGB1 as well as the extent of JAK2 and STAT3 phosphorylation whereas impeding the expression of SOCS‐1 and Bcl‐2. More important, either miR‐30a elevation or SOCS‐1 silencing suppressed liver cell proliferation and also promoted apoptosis. On the contrary, the inhibition of miR‐30a exhibited the opposite effects. Altogether, we come to the conclusion that miR‐30a inhibited the liver cell proliferation and promoted cell apoptosis by targeting and negatively regulating SOCS‐1 via the JAK/STAT signaling pathway in rats with sepsis.
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