Purpose: This study aimed to determine whether structural magnetic resonance imaging (MRI) can accurately differentiate between distensible orbital venous malformations (OVMs) and nondistensible OVMs. Methods: Fifty-one patients with nondistensible OVMs and 65 patients with distensible OVMs who underwent conventional MRI, dynamic contrast-enhanced (DCE) MRI, and diffusion-weighted imaging (DWI) MRI examinations were evaluated. The time-intensity curve (TIC) and apparent diffusion coefficient were calculated. Statistical analysis, including univariate and multivariate analysis, was performed. Receiver operating characteristic curve analysis and logistic regression analysis were used to assess the diagnostic ability of single and combined metrics. Results: In conventional MRI, nondistensible OVMs were more likely to present an enlarged orbital cavity (P = 0.032), a cystic component (P = 0.023), and a fluid-fluid level (P = 0.019), whereas distensible OVMs tended to exhibit a peak-like leading edge (P < 0.001). In DCE MRI, nondistensible OVM was more likely to present with a persistent TIC pattern and regional enhancement pattern, while distensible OVM was more likely to show a plateau TIC pattern (P < 0.001) and had a parallel proportion with regional and global enhancement patterns (P = 0.002). The combination of multiparametric MRI features, including an enlarged orbital cavity, a peak-like leading edge, and the type of TIC curve, demonstrated a high diagnostic efficiency (area under the curve = 0.869) in distinguishing between the two groups. Notably, the peak-like leading edge revealed a diagnostic specificity as high as 0.902. Conclusions: A combination of conventional MRI, DCE MRI, and DWI MRI yields high sensitivity and specificity in differentiating distensible OVMs from nondistensible OVMs.

AIM: To explore the prognostic factors for lacrimal gland adenoid cystic carcinoma (LGACC) in Chinese patients. METHODS: Clinical and histopathological data were reviewed in patients with pathologically confirmed LGACC. Local recurrence, metastasis, and disease-specific death were the main outcome measures. Univariate and multivariate analyses were performed by the Kaplan-Meier method and a Cox proportional hazard model. RESULTS: This retrospective cohort study included 45 patients with pathologically confirmed LGACC between January 2008 and June 2022. Tumor (T) classification (P=0.005), nodal metastasis (N) classification (P=0.018) and positive margin (P=0.008) were independent risk factors of recurrence; T (P=0.013) and N (P=0.003) classification and the basaloid tumor type (P=0.032) were independent risk factors for metastasis; T classification (P<0.001) was an independent factor of death of disease. In the further analysis, the durations from first surgery to radiotherapy is correlated with metastatic risk in LGACC patients with basaloid component (P=0.022). CONCLUSION: Histological subtype should be emphasized when evaluating prognosis and guiding treatment. Timely radiotherapy may reduce the risk of metastasis in patients with basaloid component.

Cancer-associated fibroblasts (CAFs) exhibit notable versatility, plasticity, and robustness, actively participating in cancer progression through intricate interactions within the tumor microenvironment (TME). N6-methyladenosine (m6A) modification is the most prevalent modification in eukaryotic mRNA, playing essential roles in mRNA metabolism and various biological processes. Howbeit, the precise involvement of m6A in CAF activation remains enigmatic. In this study, we revealed that the m6A demethylase FTO supports CAF-mediated angiogenesis through activation of EGR1 and VEGFA in conjunctival melanoma (CoM). First, single-cell transcriptome analysis revealed that FTO was specifically upregulated in the CAF population, thereby contributing to the hypo-m6A status in the TME of CoM. Moreover, CAFs of CoM displayed extensive proangiogenic potential, which was largely compromised by FTO inhibition, both in vitro and in vivo. By employing multi-omics analysis, we showed that FTO effectively eliminates the m6A modifications of VEGFA and EGR1. This process subsequently disrupts the YTHDF2-dependent mRNA decay pathway, resulting in increased mRNA stability and upregulated expression of these molecules. Collectively, our findings initially indicate that the upregulation of FTO plays a pivotal role in tumor development by promoting CAF-mediated angiogenesis. Therapeutically, targeting FTO may show promise as a potential antiangiogenic strategy to optimize cancer treatment.

Consumption of ultra-processed foods (UPF) has been associated with increased risks of various age-related diseases. However, the potential association between UPF consumption and age-related eye diseases (AREDs) remains unclear. We aim to assess the associations between consumption of UPF and risk of AREDs including age-related macular degeneration (AMD), cataract and glaucoma. We included 156,232 individuals aged 50 or older, who were free from AREDs from UK biobank study. Dietary intake data were collected using 24-h dietary assessments. UPF is defined according to the NOVA classification, and all participants are divided into four quartiles based on the weight proportion (

Optic pathway gliomas (OPGs) are most predominant pilocytic astrocytomas, which are typically diagnosed within the first decade of life. The majority of affected children with OPGs also present with neurofibromatosis type 1 (NF1), the most common tumor predisposition syndrome. OPGs in individuals with NF1 primarily affect the optic pathway and lead to visual disturbance. However, it is challenging to assess risk in asymptomatic patients without valid biomarkers. On the other hand, for symptomatic patients, there is still no effective treatment to prevent or recover vision loss. Therefore, this review summarizes current knowledge regarding the pathogenesis of NF1-associated OPGs (NF1-OPGs) from preclinical studies to seek potential prognostic markers and therapeutic targets. First, the loss of the NF1 gene activates 3 distinct Ras effector pathways, including the PI3K/AKT/mTOR pathway, the MEK/ERK pathway, and the cAMP pathway, which mediate glioma tumorigenesis. Meanwhile, non-neoplastic cells from the tumor microenvironment (microglia, T cells, neurons, etc.) also contribute to gliomagenesis via various soluble factors. Subsequently, we investigated potential genetic risk factors, molecularly targeted therapies, and neuroprotective strategies for tumor prevention and vision recovery. Last, potential directions and promising preclinical models of NF1-OPGs are presented for further research. On the whole, NF1-OPGs develop as a result of the interaction between glioma cells and the tumor microenvironment. Developing effective treatments require a better understanding of tumor molecular characteristics, as well as multistage interventions targeting both neoplastic cells and non-neoplastic cells.

Purpose To investigate the association of metabolism-related proteins and clinicopathological features with poor prognosis in lacrimal gland adenoid cystic carcinoma (LGACC). Methods Clinicopathological data for 39 Chinese patients with LGACC enrolled were retrospectively analysed. Disease progression included death, recurrence, further nodal metastasis, and distant metastasis. Expression of ASCT2 and GLS1 were evaluated by immunohistochemistry. Kaplan–Meier survival curves and Cox proportional hazards regression models were used for risk factor analyses. Results At the end of follow-up, 14 patients (35.9%) developed local recurrence, 13 patients (33.3%) developed distant metastasis, 3 patients (7.7%) developed lymph node metastasis, and 9 patients (23.1%) died. Among the 13 patients who developed distant metastasis, lung metastasis was observed in 8 patients (61.5%), the brain in 8 patients (61.5%), and bone in 1 patient (7.7%). ASCT2 was expressed in 16 (57.14%) cases, while GLS1 had high expression in 19 (67.9%) cases. Advanced T category (≥T3), bone erosion, basaloid subtype, and ASCT2 (-) were associated with disease progression. Basaloid subtype was an independent risk factor for local recurrence ( P = 0.028; HR, 12.12; 95% CI, 1.3–111.5). ASCT2(-) was an independent risk factor for distant metastasis ( P = 0.016; HR, 14.46; 95% CI, 1.6–127.5) and was associated with basaloid subtype ( P = 0.019). Conclusions For LGACC, ≥T3 category, basaloid subtype, and bone erosion were high-risk predictors. ASCT2(-) was an independent risk factor for distant metastasis, which suggested that it could be a potential biomarker for LGACC.

Adoptive immunotherapy in the T cell landscape exhibits efficacy in cancer treatment. Over the past few decades, genetically modified T cells, particularly chimeric antigen receptor T cells, have enabled remarkable strides in the treatment of hematological malignancies. Besides, extensive exploration of multiple antigens for the treatment of solid tumors has led to clinical interest in the potential of T cells expressing the engineered T cell receptor (TCR). TCR-T cells possess the capacity to recognize intracellular antigen families and maintain the intrinsic properties of TCRs in terms of affinity to target epitopes and signal transduction. Recent research has provided critical insight into their capability and therapeutic targets for multiple refractory solid tumors, but also exposes some challenges for durable efficacy. In this review, we describe the screening and identification of available tumor antigens, and the acquisition and optimization of TCRs for TCR-T cell therapy. Furthermore, we summarize the complete flow from laboratory to clinical applications of TCR-T cells. Last, we emerge future prospects for improving therapeutic efficacy in cancer world with combination therapies or TCR-T derived products. In conclusion, this review depicts our current understanding of TCR-T cell therapy in solid neoplasms, and provides new perspectives for expanding its clinical applications and improving therapeutic efficacy.

Purpose:To investigate the correlation between apparent diffusion coefficient (ADC) histograms and high-risk clinicopathologic features related to uveal melanoma (UM) prognosis. Methods:This retrospective study included 53 patients with UM who underwent diffusion-weighted imaging (DWI) between August 2015 and March 2024. Axial DWI was performed with a single-shot spin-echo echo-planar imaging sequence. ADC histogram parameters of ADCmean, ADC50%, interquartile range (IQR), skewness, kurtosis, and entropy were obtained from DWI. The relationships between histogram parameters and high-risk clinicopathological characteristics including tumor size, preoperative retinal detachment, histological subtypes, Ki-67 index, and chromosome status, were analyzed by Spearman correlation analysis, Mann-Whitney U test, or Kruskal-Wallis test. Results:A total of 53 patients (mean ± SD age, 55 ± 15 years; 22 men) were evaluated. The largest basal diameter (LBD) was correlated with kurtosis (r = 0.311, P = 0.024). Tumor prominence (TP) was correlated with entropy (r = 0.581, P < 0.001) and kurtosis (r = 0.273, P = 0.048). Additionally, significant correlations were identified between the Ki-67 index and ADCmean (r = -0.444, P = 0.005), ADC50% (r = -0.487, P = 0.002), and skewness (r = 0.394, P = 0.014). Finally, entropy was correlated with monosomy 3 (r = 0.541, P = 0.017). Conclusions:The ADC histograms provided valuable insights into high-risk clinicopathologic features of UM and hold promise in the early prediction of UM prognosis.

BACKGROUND:ARID1A, a subunit of the SWI/SNF chromatin remodeling complex, is thought to play a significant role both in tumor suppression and tumor initiation, which is highly dependent upon context. Previous studies have suggested that ARID1A deficiency may contribute to cancer development. The specific mechanisms of whether ARID1A loss affects tumorigenesis by RNA editing remain unclear.RESULTS:Our findings indicate that the deficiency of ARID1A leads to an increase in RNA editing levels and alterations in RNA editing categories mediated by adenosine deaminases acting on RNA 1 (ADAR1). ADAR1 edits the CDK13 gene at two previously unidentified sites, namely Q113R and K117R. Given the crucial role of CDK13 as a cyclin-dependent kinase, we further observed that ADAR1 deficiency results in changes in the cell cycle. Importantly, the sensitivity of ARID1A-deficient tumor cells to SR-4835, a CDK12/CDK13 inhibitor, suggests a promising therapeutic approach for individuals with ARID1A-mutant tumors. Knockdown of ADAR1 restored the sensitivity of ARID1A deficient cells to SR-4835 treatment.CONCLUSIONS:ARID1A deficiency promotes RNA editing of CDK13 by regulating ADAR1.

Objective This study aims to delineate the key characteristics of eyelid neoplasms in central and southern coastal China and to explore potential factors affecting tumour development. Methods This was a retrospective, multicentre observational study. In total, 1302 cases of eyelid neoplasms diagnosed in 2 tertiary hospitals were reviewed from January 2013 to May 2023. Histogenesis, pathologic diagnosis, and potential risk factors were investigated, and the findings were compared with data from various regions and countries. Age- and sex-specific incidence rates, time trends, distributions across age groups, and the relationship with the Sociodemographic Index, latitude, and altitude were calculated. Results There were 1162 benign (89.2%) and 140 malignant (10.8%) cases, representing 63 pathologic types. The median age of benign cases (50 years) was significantly lower than that of malignant cases (72 years) (P < 0.05). The most common benign and malignant lesions were intradermal nevus (28.83%) and basal cell carcinoma (BCC) (74.29%). A relatively younger median age (68 years) for sebaceous gland carcinoma (SGC) was observed, with a male preponderance (sex ratio 2:1). Analyses of the proportions of BCC, SGC, and squamous cell carcinoma in eyelid malignancies revealed significant regression equations (P < 0.05). Conclusions Most eyelid neoplasms are of epithelial and melanocytic origin, with benign tumours being predominant. The regions with greater Sociodemographic Index exhibit a greater incidence of BCC and a lower incidence of squamous cell carcinoma and SGC. BCC appears to have a greater incidence in regions located at higher latitudes.

Purpose: Ocular melanoma is a common primary malignant ocular tumor in adults with limited effective treatments. Epigenetic regulation plays an important role in tumor development. The switching/sucrose nonfermentation (SWI/SNF) chromatin remodeling complex and bromodomain and extraterminal domain family proteins are epigenetic regulators involved in several cancers. We aimed to screen a candidate small molecule inhibitor targeting these regulators and investigate its effect and mechanism in ocular melanoma. Methods: We observed phenotypes caused by knockdown of the corresponding gene and synergistic effects with BRD inhibitor treatment and SWI/SNF complex knockdown. The effect of JQ-1 on ocular melanoma cell cycle and apoptosis was analyzed with flow cytometry. Via RNA sequencing, we also explored the mechanism of BRD4. Results: The best tumor inhibitory effect was observed for the BRD4 inhibitor (JQ-1), although there were no statistically obvious changes in the shBRD4 and shBRD9 groups. Interestingly, the inhibitory effect of JQ-1 was decrease in the shBRD4 group. JQ-1 inhibits the growth of melanoma in various cell lines and in tumor-bearing mice. We found 17 of these 28 common differentially expressed genes were downregulated after MEL270 and MEL290 cells treated with JQ-1. Four of these 17 genes, TP53I11, SH2D5, SEMA5A, and MDGA1, were positively correlated with BRD4. In TCGA database, low expression of TP53I11, SH2D5, SEMA5A, and MDGA1 improved the overall survival rate of patients. Furthermore, the disease-free survival rate was increased in the groups with low expression of TP53I11, SH2D5, and SEMA5A. Conclusions: JQ-1 may act downstream of BRD4 and suppress ocular melanoma growth by inducing G1 cell cycle arrest.

BACKGROUND/AIMS:Extranodal marginal zone lymphoma of ocular adnexa (OA-EMZL) is the most frequent type of ocular adnexal lymphomas, with a high rate of disease recurrence. Precise patient stratification based on disease recurrence is understudied. This study aims to identify risk factors of distant recurrence (DR) and local recurrence (LR) to construct a prognostic model optimising rapid decision of therapeutic strategies. METHODS:A total of 104 patients diagnosed with OA-EMZL between January 2011 and February 2020 were enrolled. Propensity score matching was performed for DR and LR groups. A nomogram was generated using a multivariate Cox proportional hazards model. RESULTS:After matching, different independent risk factors of DR and LR were identified. Monocyte percentage (p=0.015) and M category >0 (p=0.043) were significant independent risk factors of DR. Epiphora (p<0.001) was the significant independent risk factor of LR. Three factors (monocyte percentage, M category >0, age >60) were integrated into the nomogram to predict the risk of DR. It had a relatively better discriminative ability for distant recurrence-free survival (C-index: 3-year, 0.784; 6-year, 0.801) than IPI score (C-index: 3-year, 0.663; 6-year, 0.673) in the cohort of all patients. CONCLUSION:Our analyses suggested DR and LR as two distinct prognostic events, and additionally identified novel risk factors of them. The nomogram may serve as a practical tool for the prognostic estimation and rapid decision of therapeutic strategies for patients with OA-EMZL.

Metabolic reprogramming of amino acids has been increasingly recognized to initiate and fuel tumorigenesis and survival. Therefore, there is emerging interest in the application of amino acid metabolic strategies in antitumor therapy. Tremendous efforts have been made to develop amino acid metabolic node interventions such as amino acid antagonists and targeting amino acid transporters, key enzymes of amino acid metabolism, and common downstream pathways of amino acid metabolism. In addition to playing an essential role in sustaining tumor growth, new technologies and studies has revealed amino acid metabolic reprograming to have wide implications in the regulation of antitumor immune responses. Specifically, extensive crosstalk between amino acid metabolism and T cell immunity has been reported. Tumor cells can inhibit T cell immunity by depleting amino acids in the microenvironment through nutrient competition, and toxic metabolites of amino acids can also inhibit T cell function. In addition, amino acids can interfere with T cells by regulating glucose and lipid metabolism. This crucial crosstalk inspires the exploitation of novel strategies of immunotherapy enhancement and combination, owing to the unprecedented benefits of immunotherapy and the limited population it can benefit. Herein, we review recent findings related to the crosstalk between amino acid metabolism and T cell immunity. We also describe possible approaches to intervene in amino acid metabolic pathways by targeting various signaling nodes. Novel efforts to combine with and unleash potential immunotherapy are also discussed. Hopefully, some strategies that take the lead in the pipeline may soon be used for the common good.

Abstract Objective Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. Recently, RNA modifications have received increasing attention and have been shown to regulate tumorigenesis and immune response. However, the specific roles of N7-methylguanosine (m7G) on the prognosis, tumour immune microenvironment and immunotherapy in UM remain poorly understood. Methods Gene expression data and clinical data of 80 and 28 UM cases were obtained from The Cancer Genome Atlas and Gene Expression Omnibus databases, respectively. Of the 26 m7G regulators, eight prognosis-related m7G regulators were identified and analyzed for their correlations with survival outcomes, immune cell infiltration and immunotherapy sensitivity. Single-cell data were downloaded and used to validate the changes in the immune microenvironment. Results A total of eight prognostic m7G regulators––EIF3D, EIF4E2, EIF4E3, EIF4G3, NUDT16, NUDT16L1, SNUPN and WDR4––were identified to construct a m7G-related risk model. Patients were divided into high- and low-risk groups based on this model. The high-risk group was characterized by a poorer prognosis and higher infiltration abundance of immunosuppressed CD8 T cells. Immune cell depletion markers were significantly overexpressed in high-risk patients and predicted better response to immunotherapy. Additionally, we found that m7G regulators were most closely related to the function of the monocyte/macrophage. Conclusions We constructed a risk model comprising eight m7G regulators with good predictive power for prognosis and immunotherapy sensitivity. We also found that m7G regulators are associated with monocyte/macrophage immunity in UM. These novel findings may provide new insights into m7G regulators as prognostic and immunotherapeutic biomarkers for UM and therapies targeting m7G regulators.

With integrative molecular profiling and case-control profiling, we identified immunoglobulin M (IgM) as a tumour-derived serum biomarker for ocular adnexal extranodal marginal zone lymphoma (OA-EMZL) and further constructed a preoperative diagnostic model. OA-EMZL is the commonest subtype of ocular adnexal lymphomas (OALs),1 and the second common subtype of extranodal marginal zone lymphomas (EMZLs).2 It has similar clinical features and distinct treatment strategies compared with other orbital space-occupying lesions.3 Few tools are designed for preoperative differential diagnosis of OA-EMZL.3 A definite diagnosis of OA-EMZL requires a biopsy and a complicated pathological examination. Together, these make the efficient differential diagnosis of OA-EMZL an issue in clinical practice. Thus, we aim to find a biomarker for OA-EMZL to facilitate preoperative decision-making. Herein, we investigated serum proteome and tissue transcriptome in 83 samples to identify potential tumour-derived diagnostic serum biomarkers (Figure 1A–C and Tables S1 and S2).4-7 Differentially expressed proteins (DEPs) detected by serum proteome are not identically secreted by tumour tissues. Based on previous research,4 we hypothesized that tumour-derived biomarkers should be positively correlated in abundance between omics (Figure 1D), and concordantly differentially expressed in both omics (Figure 1E and Table S3). Immunoglobulin Heavy Constant Mu (IGHM) was the highest correlated gene (R = 0.56, p < 0.001), and the most dysregulated DEP in the proteome (Log2(foldchange) = 2.64, p < 0.001) (Figure 1D,E). The dysregulation of IGHM was consistent across subgroups in both proteome and transcriptome (Figure 1G). IGHM encodes an indispensable constant region of the IgM heavy chain. EMZL originates from marginal zone B-cell,8 which is the main producer of secreted IgM.9 Immunofluorescent staining suggested IgM was expressed in neoplastic cells instead of bystanders (Figure S1). The malignant B cells exhibit monotypic immunoglobulin expression with light chain restriction. We developed an “absolute κ/λ score” to quantify monoclonality in multi-omics data. The score calculated by transcriptomic and proteomic data were both significantly higher in EMZLs compared with nonmalignant diseases (Figure S2A,B) and significantly correlated in EMZLs (Figure S2C). The serum score was highly correlated to the serum IGHM in EMZLs instead of controls (Figure S2D,E). This evidence supports the tumour-derived nature of serum IgM and further attributes the elevated IgM to the neoplastic cells. We investigated 205 patients with pathologically confirmed diagnoses who underwent serum immunoglobulin examination in a case-control analysis (Table S4). Patients with OAL apart from EMZL, lymphoid hyperplasia and chronic orbital inflammation (idiopathic orbital inflammation, IgG4-related disease, Granuloma, Mikulicz disease, Sarcoidosis disease, Kimura disease, Sjögren syndrome and Amyloidosis) were enrolled as controls. Serum IgM was significantly higher in EMZL compared with other subgroups and diagnoses (Figure 2A and Figure S3A). Among immunoglobulins, IgM exhibited a strong diagnostic potential in differentiating EMZLs from controls (Figure 2B). It also consistently distinguished EMZLs from subgroups and diagnoses (Figure S3B). Multivariate logistic regression indicated that serum IgM (p < 0.001) was the independent predictor for EMZL (Table S5). Paired IgM concentrations were available in 70 patients at diagnosis and after treatment. OA-EMZL patients exhibited a significantly decreased serum IgM after initial treatment (Figure 2C). There was no significant correlation between serum IgM and clinicopathological features of EMZL patients (Figure 2D,F). The results indicated the consistency of serum IgM in EMZLs. Furthermore, we evaluated potential confounders based on the previous report.10 Different genders exhibited a similar serum IgM concentration in all subgroups (Figure S4A). Spearman correlation analysis showed a significant negative correlation between serum IgM and age in control and chronic inflammation groups (Figure 2D). Restricted cubic splines (RCS) analysis suggested serum IgM as a linear variable (Figure S4B), and age as a non-linear variable (p for non-linearity = 0.002) (Figure S5A). Thus, we defined an optimal cutoff for age at diagnosis as 47 years based on receiver operating characteristic (ROC) analysis and converted it into a category variable (Figure S5B). To simplify the usage of serum IgM concentration in the clinical context, we conducted propensity score matching (PSM) to minimize the potential bias and further converted IgM into an ordinal variable (Figure 3A and Table S6). After PSM, serum IgM consistently distinguished EMZLs from controls (Figure 3B). IgM was fitted consistently as a linear variable (Figure 3C). The cutoff value of serum IgM concentration at 1.46 g/L (Specificity 0.897; Sensitivity 0.559) was determined by ROC; 0.88 g/L (Specificity 0.560; Sensitivity 0.853) was determined by the minimal Akaike information criterion (AIC) value of RCS (Table S7). The ordinal IgM exhibited a comparable area under the curve (AUC) compared with the continuous (Figure 3D,E). Serum IgM solely is not sufficient for the preliminary diagnosis based on its AUC. Thus, we further constructed a diagnostic model. In the validation cohort, multivariate analysis showed IgM between 0.88–1.46 g/L (p = 0.017), IgM ≥1.46 g/L (p < 0.001), and age at diagnosis < 47 (p = 0.008) were independent predictors of EMZL (Table S8). Lacrimal gland involvement was included in the equation based on AIC. A nomogram entitled preliminary EMZL diagnostic model (preEM) was constructed based on the results (Figure 4A). Compared with continuous or ordinal IgM, preEM exhibited relatively higher discriminative ability in both derivation and validation sets (Figure 4B,C). The calibration plots exhibited a great agreement between nomogram predictions and actual observations (Figure 4D,E). Previous findings have established the diagnostic application of diffusion-weighted magnetic resonance imaging (MRI) for OAL.3 Herein, we identified preEM as a practical preoperative diagnostic model for its major subtype, OA-EMZL. The combination of these two might provide a complete diagnostic process for orbital lymphoproliferative diseases and chronic inflammation. Although EMZLs exhibited a high IgM, the results suggested an intertumoral heterogeneity (Figures 1G and 2A). We performed a differential expression analysis based on transcriptomic data between IgM-high and IgM-low EMZLs to investigate potential mechanisms (Figure S6A). The result suggested that activated GPCR, WNT and interleukin 10-related genesets were associated with higher IgM (Figure S6B,C). Further study is required to decipher this heterogeneity and the limitation of IgM for diagnosis. Serum IgM is a tumour-derived biomarker used to differentiate OA-EMZL from idiopathic orbital inflammation, IgG4 related disease and orbital lymphoproliferative disorders. The preEM constructed based on ordinal serum IgM, age at diagnosis, and lacrimal gland involvement is a practical preoperative diagnostic tool for OA-EMZL. Our proteomics and mass spectrometry work were performed at the Proteomics Platform of Core Facility of Basic Medical Sciences, Shanghai Jiao Tong University School of Medicine (SJTU-SM). We are grateful to our collaborating statistician Chang Cui and pathologists Cen Qiu, Gaoxing Zheng, and Yaoxing Guo for their constructive advice and valuable work. The authors declare no conflict of interest. This work was supported by the Project of Biobank of Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine. (grant number YBKA201907; YBKA202208). The funding organization had no role in the design or conduct of this research. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Abstract Background Lacrimal gland adenoid cystic carcinoma is the most common lacrimal gland malignant epithelial tumour, characterized by high rates of relapse and distant spread. As few studies have reported pathological biomarkers for LGACC, we aimed to identify biomarkers that can predict the recurrence and metastasis of LGACC. Methods Twenty-eight patients diagnosed with LGACC between 2008 and 2022 were included. The level of expression of ASCT2, GLS1, MCT4, and SHMT1 in patients with LGACC was evaluated by immunohistochemistry and the concordance was calculated. Patients were divided into negative and positive groups according to the staining results of ASCT2, and high or low expression groups according to the staining scores of GLS1, MCT4, and SHMT1. The recurrence-free survival, metastasis-free survival, and overall survival were analysed and compared between groups. Results The study participants comprised 16 males and 12 females, with a median age of 52.4 years (range: 30–77 years). During a 34.3-month median follow-up (range: 0.4–125.1 months), 7 (25%) patients suffered recurrence, 8 (28.6%) patients had metastasis, and 4 (14.3%) patients died due to LGACC. Patients with negative ASCT2 expression had a significantly shorter recurrence-free survival (p = 0.007), metastasis-free survival (p = 0.033), and poorer overall survival (p = 0.028). Multivariate analysis showed that ASCT2 was an independent risk factor for disease recurrence (HR = 0.1, 95% CI: 0.012–0.88, p = 0.038). Conclusions ASCT2 negative expression is a predictor to identify patients with poor recurrence-free survival, metastasis-free survival, and overall survival.

Ocular adnexal extranodal marginal zone lymphoma (OA-EMZL) is the most frequent subtype of ocular adnexal lymphoma, with a high propensity for recurrence. Distant recurrence (DR) as an essential prognostic event has unique clinical risk factors, but whether distinct molecular features exist remains poorly understood. Here, we identified potential biomarkers using proteomic analysis of 27 OA-EMZL samples. The MYC-targeted genes PCNA, MCM6, and MCM4 were identified as candidates. MYC-targeted genes were further identified as the most significantly activated gene set in patients with DR. The candidate genes were verified in samples from 11 patients with DR and 33 matched controls using immunohistochemistry. The 3-year and 5-year AUC values of MCM6 (0.699 and 0.757) were higher than those of Ki-67 (0.532 and 0.592). High expressions of MCM6 and MCM4 were significantly associated with shorter distant recurrence–free survival (Log-rank p = 0.017, Log-rank p = 0.0053). Multivariate Cox regression identified MCM6 expression as an independent risk factor for DR (HR, 6.86; 95

Venous malformation (VM) and cavernous venous malformation (CVM) are two types of vascular malformations. Even if the two diseases are similar in appearance and imaging, the distinct cellular components and signaling pathways between them might help distinguish the two from a molecular perspective. Here, we performed single-cell profiling of 35,245 cells from two VM samples and three CVM samples, with a focus on endothelial cells (ECs), smooth muscle cells (SMCs) and immune microenvironment (IME). Clustering analysis based on differential gene expression unveiled 11 specific cell types, and determined CVM had more SMCs. Re-clustering of ECs and SMCs indicated CVM was dominated by arterial components, while VM is dominated by venous components. Gene set variation analysis suggested the activation of inflammation-related pathways in VM ECs, and upregulation of myogenesis pathway in CVM SMCs. In IME analysis, immune cells were identified to accounted for nearly 30% of the total cell number, including macrophages, monocytes, NK cells, T cells and B cells. Notably, more macrophages and monocytes were discovered in VM, indicating innate immune responses might be more closely related to VM pathogenesis. In addition, angiogenesis pathway was highlighted among the significant pathways of macrophages & monocytes between CVM and VM. In VM, VEGFA was highly expressed in macrophages & monocytes, while its receptors were all abundantly present in ECs. The close interaction of VEGFA on macrophages with its receptors on ECs was also predicted by CellPhoneDB analysis. Our results document cellular composition, significant pathways, and critical IME in CVM and VM development.

Background Ocular adnexal B-cell lymphoma (OABL) is a rare subtype of non-Hodgkin lymphoma. The molecular characteristics of OABL remain poorly understood. We performed an integrated study to investigate the proteotranscriptome landscape and identify novel molecular characteristics and biomarkers of OABL. Methods Integrated quantitative proteome and transcriptome were performed on 40 OABL 12 idiopathic orbital inflammation, 6 reactive lymphoid hyperplasia, and 13 aesthetic orbital plastic surgery specimens. Complete clinicopathologic and prognostic data of the patients were recorded. Results We identified high global protein-mRNA concordance as a novel characteristic of OABL. High concordance was related to OABL recurrence. By integrated expression profile, motif enrichment and trend analysis, we found that alternative splicing is inflammation-independently dysregulated in OABL. After portraying the aberrant alternative splicing event landscape, we demonstrated the oncogenic role of ADAR, a core splicing regulator that regulates the splicing of Rho GTPase and cell cycle members. We found that ADAR regulates cell proliferation and Rho GTPase inhibitor sensitivity of lymphoma. We identified DNAJC9 as a potential biomarker for OABL in proteomic analyses. Immunohistochemistry and immunofluorescent staining showed the nuclear staining of DNAJC9 was significantly higher in extranodal marginal zone lymphomas compared with inflammation specimens. Conclusions These results provide an integrated gene expression profiling and demonstrate that high global protein-mRNA concordance is a prognosis-related molecular characteristic of OABL. We portray the alternative splicing events landscape of OABL, and reveal the oncogenic role of ADAR. We identified strong nuclear staining of DNAJC9 as a promising pathology diagnostic biomarker for extranodal marginal zone lymphomas.

The blepharophimosis–ptosis–epicanthus inversus syndrome (BPES) is a rare autosomal dominant disease mainly caused by FOXL2 variants. This genetic disorder is usually characterized by eyelid malformation and ovarian dysfunction. However, no reliable genotype/phenotype correlations have been established considering the ovarian phenotype. Here, we detected 15 FOXL2 variants including nine novel ones from 7 families and 8 sporadic cases, which expanded the spectrum of FOXL2 variants and identified a potential clinical cause. Functional studies, with respect to the effect of FOXL2 on the StAR promoter, showed that non-sense variants that lead to protein truncation before the polyalanine tract and missense variants [c.307C > T; p.(Arg103Cys), c.311A > C; p.(His104Pro), c.320G > A; p.(Ser107Asn), and c.335T > A; p.(Phe112Tyr)] within the central portion of the FOXL2 forkhead domain significantly affect its suppressor activity. Such changes may explain the mechanism underlying a more severe phenotype, more likely to result in BPES type I. Furthermore, the missenses variants c.307C > T; p.(Arg103Cys), c.311A > C; p.(His104Pro), and c.320G > A; p.(Ser107Asn) were not able to transactivate OSR2, which is consistent with the eyelid malformation in these patients. The results from our cohort have expanded the spectrum of FOXL2 variants and have provided insights into genotype/phenotype correlations.