Identification of systemic biomarkers and potential drug targets for age-related macular degeneration.

Purpose: Since age-related macular degeneration (AMD) is tightly associated with aging and cellular senescence, objective of this study was to investigate the association between plasma levels of senescence-related proteins (SRPs) and risk of AMD. Design: The whole study was based on two-sample Mendelian randomization (MR) analysis. Methods: For MR analysis, the primary approach for MR analysis was the inverse-variance weighted (IVW) method and the heterogeneity and pleiotropy of results were tested. The instrumental single-nucleotide polymorphisms (SNPs) associated with 110 SRPs were filtered and selected from a large genome-wide association study (GWAS) for plasma proteome involving 35,559 participants. The GWAS data of AMD was obtained from FinnGen consortium (6,157 AMD cases and 288,237 controls) and further validated by using data from UK Biobank consortium (3,553 AMD cases and 147,089 controls). Results: The MR results at both discovery and validation stages supported the causality (IVW-P < 0.00045) between plasma levels of 4 SRPs (C3b, CTNNB1, CCL1, and CCL3L1) and the risk of AMD and supported potential causality (IVW-P < 0.05) between other 10 SRPs and risk of AMD. No heterogeneity or pleiotropy in these results was detected. Conclusion: Our findings supported that high plasma levels of C3b, CTNNB1, CCL1, and CCL3L1 were associated with increased risk of AMD, thereby highlighting the role of systemic inflammation in AMD pathogenesis and providing the rationale for developing new preventative and therapeutic strategies.

Sleep duration and age-related macular degeneration: a cross-sectional and Mendelian randomization study.

Purpose: To investigate the association between sleep duration and age-related macular degeneration (AMD). Design: Cross-sectional study, bidirectional two-sample Mendelian randomization (MR). For cross-sectional analysis, we used survey data of 5,481 participants aged ≥40 years from the 2005 to 2008 National Health and Nutrition Examination Survey (NHANES). For MR analysis, we used sleep- and AMD-associated genome-wide association studies (GWAS) data involving large populations. Methods: The association between sleep duration and AMD was assessed using logistic regression models. For MR analysis, the primary approach for MR analysis was the inverse-variance weighted (IVW) method. Results: In cross-sectional analysis, after adjusting for multiple covariates, short sleep duration (SSD) was found to be associated with increased risk of early AMD [odds ratio (OR) = 1.364, P = 0.036). MR analysis supported the results of cross-sectional analysis: SSD increases the risk of early AMD (ß = 0.102, IVW-P = 0.003). Conclusion: Our findings provide the evidence supporting the association between sleep deficiency and higher risk of AMD. Further studies are required to confirm our findings and elucidate the mechanisms underlying this association.

Single-cell sequencing reveals an important role of SPP1 and microglial activation in age-related macular degeneration.

Purpose: To investigate the role of senescence-related cytokines (SRCs) in the pathophysiology of age-related macular degeneration (AMD). Design: The whole study is based on single-cell and bulk tissue transcriptomic analysis of the human neuroretinas with or without AMD. The transcriptomic data of human neuroretinas was obtained from Gene-Expression Omnibus (GEO) database. Methods: For single-cell transcriptomic analysis, the gene expression matrix goes through quality control (QC) filtering, being normalized, scaled and integrated for downstream analysis. The further analyses were performed using Seurat R package and CellChat R package. After cell type annotation, the expression of phenotype and functional markers of microglia was investigated and cell-cell communication analysis was performed. For bulk tissue transcriptomic analysis, GSE29801 dataset contains the transcriptomic data of human macular neuroretina (n = 118) from control group and AMD patients. The expression of SPP1 in control and AMD subtypes were compared by Student's t-test. In addition, the AMD macular neuroretina were classified into SPP1-low and SPP1-high groups according to the expression level of SPP1. The differentially expressed genes between these two groups were subsequently identified and the pathway enrichment analysis for these genes was further conducted. Results: Secreted phosphoprotein 1, as an SRC, was revealed to be highly expressed in microglia of AMD neuroretina and the SPP1-receptor signaling was highly activated in AMD neuroretina. In addition, SPP1 signaling was associated with the pro-inflammatory phenotype and phagocytic state of microglia. SPP1 expression was elevated in macular neuroretina with late dry and wet AMD and the inflammatory pathways were found to be activated in SPP1-high AMD macular neuroretina. Conclusion: Our findings indicated that SPP1 and microglial activation might play an important role in the pathophysiology of AMD. Therefore, SPP1 might serve as a potential therapeutic target for AMD. More in vitro and in vivo studies are required to confirm the results and the therapeutic effect of SPP1-targeting strategy.

Two Pyroptosis-Related Subtypes with Distinct Immune Microenvironment Characteristics and a Novel Signature for Predicting Immunotherapy Response and Prognosis in Uveal Melanoma.

PURPOSE: Uveal melanoma (UM) is the most common primary intraocular malignant tumor in adults with poor prognosis. Pyroptosis is a well-known form of programmed cell death. However, pyroptosis has not been sufficiently discussed in UM. This study aims to explore the expression patterns of pyroptosis-related genes (PRGs) and their relationship with tumor microenvironment (TME) characteristics and prognosis in UM. METHODS: In this study, unsupervised clustering analysis was performed based on the expression of 10 PRGs to identify pyroptotic subtypes. TME characteristics were evaluated by using the ssGSEA, ESTIMATE and CIBERSORT algorithms. In addition, a related scoring model (PRscore) was established to quantify pyroptotic patterns of individual UM patients via principal component analysis. The correlation between PRscore and TME characteristics was assessed by Spearman analysis. Furthermore, univariate and multivariate Cox regression analysis were performed to identify whether the PRscore can be an independent factor for predicting the overall survival (OS) of UM patients. RESULTS: The results revealed that there were two distinct pyroptotic patterns with different TME characteristics in UM. PRscore was found to be associated with TME characteristics and patient prognosis. In addition, combined with the clinical characteristics, the PRscore was found to be an independent factor for predicting the OS of UM patients. Furthermore, PRscore might be a useful tool for predicting the response to immunotherapy in UM patients. CONCLUSIONS: The status of pyroptosis was associated with TME characteristics in UM. In addition, evaluating the pyroptotic pattern (Prscore) would help us to predict the prognosis and immunotherapy response of individual UM patient. Furthermore, our results may offer novel insights into the development of a promising strategy for treating UM, i.e. the combination of chemodrugs targetting the induction of pyroptosis and immune checkpoint inhibitors (ICIs).

Integrative analysis identifies key genes related to metastasis and a robust gene-based prognostic signature in uveal melanoma.

PURPOSE: Uveal melanoma (UM) is an aggressive intraocular malignancy, leading to systemic metastasis in half of the patients. However, the mechanism of the high metastatic rate remains unclear. This study aimed to identify key genes related to metastasis and construct a gene-based signature for better prognosis prediction of UM patients. METHODS: Weighted gene co-expression network analysis (WGCNA) was used to identify the co-expression of genes primarily associated with metastasis of UM. Univariate, Lasso-penalized and multivariate Cox regression analyses were performed to establish a prognostic signature for UM patients. RESULTS: The tan and greenyellow modules were significantly associated with the metastasis of UM patients. Significant genes related to the overall survival (OS) in these two modules were then identified. Additionally, an OS-predicting signature was established. The UM patients were divided into a low- or high-risk group. The Kaplan-Meier curve indicated that high-risk patients had poorer OS than low-risk patients. The receiver operating curve (ROC) was used to validate the stability and accuracy of the final five-gene signature. Based on the signature and clinical traits of UM patients, a nomogram was established to serve in clinical practice. CONCLUSIONS: We identified key genes involved in the metastasis of UM. A robust five-gene-based prognostic signature was constructed and validated. In addition, the gene signature-based nomogram was created that can optimize the prognosis prediction and identify possible factors causing the poor prognosis of high-risk UM patients.

Identification of survival-related genes and a novel gene-based prognostic signature involving the tumor microenvironment of uveal melanoma.

Uveal melanoma (UM) is the most common primary intraocular malignant tumor in adults and almost fifty percent of patients subsequently develop systemic metastases usually involving the liver. The tumor microenvironment (TME) is crucial to the initiation and progression of tumors. In the present study, we comprehensively evaluated the TME of primary UM samples from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) database by using several bioinformatic algorithms. The prognostic value of immune score and infiltrating immune cells in the TME were evaluated. Differentially expressed genes between the low- and high-immune score groups were also identified. The majority of tumor-infiltrating lymphocytes in UM have been determined to be activated CD8 + T cells. Therefore, weighted gene co-expression network analysis (WGCNA) was performed to identify the co-expression modules and genes significantly associated with the level of infiltrating CD8 + T cells in UM. Survival-related genes involved in the TME were identified by univariate Cox regression analysis. Furthermore, an eight-gene-based prognostic signature was established in the training dataset TCGA-UM via Lasso-penalized and multivariate Cox regression analyses. The predictive value of this signature was validated in two testing datasets. Besides, a nomogram was established to serve clinical practice. Moreover, hub genes involved in the infiltrating CD8 + T cells were identified and a potential targeted therapy for preventing metastasis of UM was proposed based on the results. In summary, our results provided a robust gene-based prognostic signature for predicting prognosis of UM patients and proposed a potential targeted therapy for preventing UM metastasis.

Identification of the key genes and pathways involved in B cells in primary Sjögren' s syndrome.

Primary Sjögren' s syndrome (pSS) is a relatively common autoimmune disease, which mainly involves the exocrine glands, causing dry eye, dryness of mouth, fatigue and pain in the joints, thus severely affecting the normal lives of patients. B cell populations are considered to play an important role in their pathogenesis and pSS patients are generally characterized by exhibiting biological signs of B cell activation. Moreover, another important characterized change in the peripheral blood of pSS patients is found to be the decreased number of circulating memory B cells. However, the mechanisms underlying the B cell activation and the decreased level of circulating memory B cells in pSS patients are still unclear. Therefore, we identified key genes and pathways involved in B cells in pSS through a combination of several bioinformatic approaches including Cell-type Identification By Estimating Relative Subsets Of RNA Transcripts (CIBERSORT) and weighted gene co-expression network analysis (WGCNA) using gene expression data of pSS patients and controls from an open database Gene Expression Omnibus (GEO). The results may provide some novel insights into the pathogenesis of pSS. Moreover, we constructed and validated a diagnostic model for pSS by using the expression patterns of these key genes, which may assist clinicians in diagnosing pSS.

Thermoresponsive GenisteinNLC-dexamethasone-moxifloxacin multi drug delivery system in lens capsule bag to prevent complications after cataract surgery.

Cataract surgery is the most common intraocular procedure. To decrease postsurgical inflammation, prevent infection and reduce the incidence of secondary cataract, we built a temperature-sensitive drug delivery system carrying dexamethasone, moxifloxacin and genistein nanostructured lipid carrier (GenNLC) modified by mPEG-PLA based on F127/F68 as hydrogel. Characterizations and release profiles of the drug delivery system were studied. In vitro functions were detected by CCK-8 test, immunofluorescence, wound-healing assay, real time-PCR and western blotting. The size of GenNLCs was 39.47 ± 0.69 nm in average with surface charges of - 4.32 ± 0.84 mV. The hydrogel gelation temperature and time were 32 °C, 20 s with a viscosity, hardness, adhesiveness and stringiness of 6.135 Pa.s, 54.0 g, 22.0 g, and 3.24 mm, respectively. Transmittance of the gel-release medium was above 90% (93.44 ± 0.33% to 100%) at range of 430 nm to 800 nm. Moxifloxacin released completely within 10 days. Fifty percent of dexamethasone released at a constant rate in the first week, and then released sustainably with a tapering down rate until day 30. Genistein released slowly but persistently with a cumulative release of 63% at day 40. The thermoresponsive hydrogel inhibited the proliferation, migration and epithelial-mesenchymal transition of SRA 01/04 cells, which were confirmed by testing CCK-8, wound-healing assay, western blot, real time-PCR (RT-PCR) and immunofluorescence. These results support this intracameral thermoresponsive in situ multi-drug delivery system with programmed release amounts and release profiles to cut down the need of eye drops for preventing inflammation or infection and to reduce posterior capsular opacification following cataract surgery.