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关键词： CRISPR screening   cytokine receptors   cytokine signaling   interleukin-15   natural killer cells   post-translational modifications  

摘要： Interleukin-15 receptor (IL-15R) agonists induce anti-tumor immunity in pre-clinical models. However, dose-limiting toxicity has hampered their clinical development. We performed genome-wide CRISPR screens to reveal the complete IL-15R signaling mechanism in natural killer (NK) cells and discovered that ubiquitin-dependent IL-15R degradation is the dominant mechanism restraining IL-15R signaling. Key hits included the NEDD8 E2-conjugating enzyme UBE2F, the ubiquitin E3-ligase ARIH2, and Cullin-5 RING E3 ligase (CRL5) members. We found that UBE2F was required for neddylation and activation of CUL5, whereas ARIH2 contributed to CRL5-mediated IL-15RB degradation. Ablation of ARIH2 or UBE2F increased IL-15RB surface expression and enhanced signaling, resulting in proinflammatory cytokine production and augmented natural and CAR-mediated cytotoxicity. In mice lacking Arih2, Rnf7, or Ube2f, we observed that the IL-15R hyperresponsive NK cells exhibited superior in vivo anti-tumor immunity against primary and disseminated metastatic tumors. Thus, we have identified the enzymes UBE2F and ARIH2 as tractable immunotherapy drug targets.

关键词： 肺肿瘤   肺炎克雷伯菌   肿瘤微环境   髓源性抑制细胞   CD8   T淋巴细胞   肿瘤进展   免疫耐受   C57BL小鼠  

摘要： 目的 明确肿瘤内肺炎克雷伯菌（KP）对肺癌进展的影响，并初步探究可能的机制。方法构建小鼠Lewis肺癌皮下瘤模型，通过瘤内注射的方法使KP定植肿瘤组织。将小鼠随机分为对照组（Vehicle组，n=8）和实验组（KP组，n=8），Vehicle组小鼠瘤内注射磷酸缓冲盐溶液。利用qRT-PCR和FISH实验验证定植效果，并观察KP对肿瘤生长的影响。动物实验结束后取材，借助流式细胞术、免疫荧光染色检测小鼠肿瘤组织中各亚群免疫细胞的数量和功能，利用转录组测序、qRT-PCR、ELISA检测小鼠肿瘤组织中基因或蛋白表达的改变。结果与Vehicle组相比，KP组小鼠皮下瘤的体积更大（P<0.01），质量更重（P<0.05）。与Vehicle组相比，KP组小鼠肿瘤组织中CD8+T细胞比例降低（P<0.05）、Perforin+CD8+T细胞比例降低（P<0.05），CD4+T细胞比例降低（P<0.01），多形核髓源性抑制细胞比例升高（P<0.01），巨噬细胞及单核细胞样髓源性抑制细胞比例未见明显改变。与Vehicle组相比，KP组小鼠肿瘤组织中CD274、ARG2、ARG1、NOS2、IL-6、IL-1β、CXCL2、CXCL5基因表达上调（P<0.05）。与Vehicle组相比，KP组小鼠肿瘤组织中IL-1β含量增加（P<0.05）。结论肿瘤内定植的KP可通过募集髓源性抑制细胞抑制CD8+T细胞的数量和功能，重塑免疫微环境，进而促进肺癌进展。

关键词： Breast cancer   Monopolar spindle kinase I   Radiosensitization   Radiotherapy   Type I interferon  

摘要： Triple negative breast cancer (TNBC) is an aggressive breast cancer subtype that disproportionately impacts Black women and has limited effective therapeutic options. Consequently, there is an urgent need to develop novel approaches for the treatment of TNBC. Previously, we identified monopolar spindle kinase I (Mps1 or TTK), which is upregulated in TNBC patients after radiotherapy, as a potential therapeutic target. We found that TTK inhibition sensitizes human TNBC to radiotherapy (RT) both in vitro and in vivo; however, these studies were performed in immunodeficient models. Here, we extended those studies into syngeneic murine models of TNBC using two TTK inhibitors: empesertib and the novel TTK inhibitor CFI-402257 (also known as luvixasertib) that was recently granted FDA fast track approval in breast cancer. These studies demonstrate that TTK inhibition radiosensitizes syngeneic murine models of TNBC and increases the production of micronuclei and aneuploidy. Mechanistic studies demonstrate that TTK inhibition and RT alter the tumor immune microenvironment of TNBC by modifying the production of antitumoral type I interferon (T1IFN). In vivo, TTK inhibition sensitizes syngeneic models of TNBC to RT. Furthermore, combining TTK inhibition and RT also potentiates T1IFN signaling, suggesting that combination treatment may induce antitumoral immunity in immunocompetent models. Taken together, these studies demonstrate that TTK inhibition enhances radiosensitivity and TTK inhibition with RT modulates the immune landscape of TNBC. Collectively, this combination may represent a novel therapeutic strategy to improve outcomes for patients with TNBC by both direct tumor cytotoxicity and by promoting an immune-responsive environment.

关键词： Autophagy   Hematology   Metabolism   Mitochondria   Platelets  

摘要： Platelets play a dual role in hemostasis and inflammation-associated thrombosis and hemorrhage. While the mechanisms linking inflammation to platelet dysfunction remain poorly understood, our previous work demonstrated that TNFα alters mitochondrial mass, platelet activation, and autophagy-related pathways in megakaryocytes. Here, we hypothesized that TNFα impairs platelet function by disrupting autophagy, a process critical for mitochondrial health and cellular metabolism. Using human and murine models of TNFα-driven diseases, including myeloproliferative neoplasms and rheumatoid arthritis, we found that TNFα downregulates STX17, a key mediator of autophagosome-lysosome fusion. This disruption inhibited autophagy, leading to the accumulation of dysfunctional mitochondria and reduced mitochondrial respiration. These metabolic alterations compromised platelet-driven clot contraction, a process linked to thrombotic and hemorrhagic complications. Our findings reveal a mechanism by which TNFα disrupts hemostasis through autophagy inhibition, highlighting TNFα as a critical regulator of platelet metabolism and function. This study provides new insights into inflammation-associated pathologies and suggests autophagy-targeting strategies as potential therapeutic avenues to restore hemostatic balance.

关键词： Extracellular vesicles   IL-3   Th2 cells   allergic   asthma   eosinophils   lung   type 2 inflammation  

摘要： BACKGROUND:Extracellular vesicles (EVs) mediate intercellular communication during immune responses. EVs are abundant in respiratory biofluids, and the composition of EVs in the lung changes during inflammation. OBJECTIVE:We aimed to quantify the contribution of T cells to airway EVs in eosinophilic lung inflammation and ascertain their function during a type 2 inflammatory response. METHODS:Genetic membrane tagging was combined with single vesicle flow cytometry to quantify T cell EVs in the airways of mice challenged with ovalbumin or house dust mite. EVs were purified from mouse T helper type 2 (Th2) cell cultures and their functions on eosinophils assessed by flow cytometry and RNA sequencing. Th2 cell EVs were instilled into the lungs of mice to determine effects on lung eosinophilia. Finally, the function of an EV protein cargo was tested using inhibitors and blocking antibodies. RESULTS:T cell EVs are increased in the airways of mice after ovalbumin or house dust mite induced inflammation. EVs secreted by Th2 cells inhibit apoptosis and induce activating pathways in eosinophils in vitro. This effect depends on re-stimulation through the T cell receptor. Th2 cell EVs prolong eosinophilia in vivo during acute eosinophilic inflammation. Th2 cell EVs carry the cytokine IL-3 as a surface cargo, which inhibits apoptosis by activating Jak1/2-dependent pro-survival programs in eosinophils. CONCLUSION:Th2 cell EVs promote eosinophil survival through the EV cargo IL-3, supporting a role for EVs as vehicles of cytokine-based communication in lung inflammation.

关键词： 肝纤维化   JAK-STAT通路   细胞因子  

摘要： 肝纤维化是指因各类损伤因素致使肝脏受损，进而引发肝细胞水肿、变性、坏死等情况，同时伴有纤维组织增生。这一过程是肝脏受到损伤后的一种疤痕修复反应，最终可能发展为肝硬化。近年来，肝纤维化信号转导通路的深入研究为抗肝纤维化药物的研发提供了重要依据。JAK-STAT信号转导通路作为常见的细胞信号传导机制，研究表明其在调控肝纤维化疾病中发挥着重要作用。在肝脏中JAK-STAT信号通路可被多种细胞因子和生长激素激活，在抗病毒防御、急性期反应、肝损伤、修复和炎症中发挥关键作用。本文综述了JAK-STAT信号通路在肝纤维化中的研究进展。

关键词： IL-33   immunotherapy   pancreatic ductal adenocarcinoma   prognosis   regulatory T cells   tumor microenvironment  

摘要： BACKGROUND:Interleukin-33 (IL-33), a member of the IL-1 cytokine family, is constitutively expressed in barrier cells such as endothelial cells and fibroblasts. While the expression of IL-33 in regulatory T cells (Tregs) has been previously reported, its clinical significance in pancreatic ductal adenocarcinoma (PDAC) remains unclear. This study aims to investigate the clinical relevance and biological role of IL-33+ Tregs in PDAC. METHODS:Infiltration of IL-33+ Tregs was assessed by immunohistochemistry in 215 patients from our institute. The correlation between IL-33+ Tregs and clinical characteristics was analyzed. Additionally, the functional status of cytotoxic T cells in relation to IL-33+ Treg infiltration was examined. The impact of IL-33+ Tregs on the tumor microenvironment (TME) was further evaluated both in silico and in vitro. RESULTS:IL-33+ Tregs infiltration was confirmed in PDAC tissues, and its abundance was positively associated with microvascular invasion, perineural invasion, and elevated serum CA19-9 levels. Patients with higher tumor-infiltrating IL-33+ Tregs demonstrated poorer overall survival (OS) and recurrence-free survival (RFS) compared to those with lower infiltration levels. Multivariate analysis confirmed IL-33+ Tregs as an independent prognostic factor for both OS and RFS, with improved survival prediction when combined with tumor differentiation. Subgroup analyses indicated that serum CA19-9 was not a useful risk stratification tool in patients with high IL-33+ Treg infiltration, and these patients showed limited survival benefit from adjuvant chemotherapy. Furthermore, increased IL-33+ Treg infiltration was associated with more pronounced immunosuppressive TME, marked by a reduction in cytotoxic phenotypes and an upregulation of exhausted markers on CD8+ T cells. CONCLUSION:Our findings identify IL-33+ Tregs as a novel subtype of Tregs, with strong prognostic value for survival risk stratification and therapeutic response prediction

关键词： BET bromodomains   Bromodomain‐containing protein 4 (BRD4)   Diabetes   Epigenetics   Gene regulation   Inflammation   NF‐κB transcription factor   Small molecule   Transcription   β-cell  

摘要： Cytokine-stimulated transcription of NF-κB target genes is linked to the development of multiple inflammatory and autoimmune diseases. Inhibitors of bromodomain and extraterminal domain (BET) epigenetic reader proteins attenuate inflammatory gene transcription and delay the onset of several inflammatory diseases, including autoimmune diabetes. Our previous studies showed that BET bromodomain inhibitors disrupt the interaction between BET family member BRD4 and NF-κB transcription factor p65 in β-cells, thus attenuating cytokine-stimulated NF-κB-dependent gene and functional changes. However, the role of NF-κB in developing inflammatory disease is controversial, as NF-κB inhibition can promote disease progression in some contexts. NF-κB target genes play both physiological and pathophysiological roles in regulating the cellular response to cytokines. Here, using cytokine-stimulated pancreatic β-cells as an inflammatory disease model, we show that NF-κB-dependent gene products that participate in inflammation are sensitive to BET bromodomain inhibition. In contrast, gene products that maintain cellular homeostasis or protect β-cells from stressors are largely insensitive to BET bromodomain inhibition. These studies define a novel and selective role for BET bromodomain-containing proteins in regulating inflammatory gene activation.

关键词： Head and Neck Cancer   Immune Checkpoint Inhibitor   Immunotherapy   JAK-STAT   Macrophage  

摘要： BACKGROUND:Immune checkpoint blockade therapy has shown limited efficacy in head and neck squamous cell carcinoma (HNSCC). Sialic acid binding immunoglobulin-like lectin (Siglec)-15 has been identified as a novel immune evasion biomarker, while the role of Siglec-10 in the specific immune suppressive tumor microenvironment remains largely unknown. METHODS:Immunohistochemical assays were employed to investigate the correlation of the expressions of Siglec-10 and Siglec-15 with the clinicopathological features as well as the prognosis of immunotherapy in patients with HNSCC. The Gene Expression Omnibus datasets were used to identify the upstream transcriptional regulators of in tumor-associated macrophages (TAMs) and the downstream biological functions it mediates. These findings were then validated through in vitro and in vivo experiments. The impact of deficiency on the efficacy of immunotherapy and the activation of CD8+T cells was analyzed in mouse HNSCC tumor-bearing models. RESULTS:The expression of Siglec-G/10, rather than that of Siglec-15, was positively correlated with immune suppressive marker programmed death-ligand 1 (PD-L1) expression and was associated with cervical lymph node metastasis, poorer pathologic stage, and lower sensitivity to immunotherapy. deficiency rescued the immune suppressive tumor microenvironment, as evidenced by decreased TAM-associated phenotype and increased CD8+T cell infiltration and activation, which inhibited tumor growth significantly. Single-cell sequence and transcription factor prediction revealed that signal transducer and activator of transcription 6 (STAT6) could induce Siglec-G/10 transcription. Interleukin (IL)-4 could upregulate Siglec-G/10 expression significantly via STAT6 activation, as proved by overexpression and inhibition of STAT6. Signal transduction mechanism revealed that Siglec-G/10 could promote TAM differentiation and activation via increasing HIF1α (hypoxia-inducible factor 1α) expression. Furthermor

关键词： ABO血型   A抗原表达减弱   基因测序  

摘要： 目的：对ABO血型A抗原表达减弱者进行基因测序分析，探讨其分子基础。方法：采用盐水试管凝集法进行ABO血型正反检测，采用吸收放散试验进行红细胞上弱抗原检测，采用聚合酶链反应直接测序技术、Sanger测序法分别进行ABO基因分型和测序，采用Py MOL软件进行蛋白3D结构模型分析。结果：30例红细胞与抗A发生弱凝集或不凝集反应，与抗-A1不凝集；血清与B型红细胞发生4+凝集或不凝集反应，与O型红细胞发生不凝集反应；其中2例吸收放散试验阳性。血清学结果疑示Aw亚型9例、Aw B 15例、A3亚型3例、Ael亚型2例、Ax亚型1例，基因测序结果分别为2例ABO*A1.02/ABO*O.01.01、1例ABO*A1.01/ABO*O.01.01、1例ABO*A1.02/ABO*O.01.13、3例ABO*A2.05/ABO*O.01.01、1例ABO*A2.05/ABO*O.01.13、5例ABO*A2.05/ABO*B.01、1例ABO*A2.08/ABO*O.01.02、3例ABO*A2.08/ABO*B.01、1例ABO*B(A).02/ABO*O.01.02、2例ABO*B(A).02/ABO*O.01.01、1例ABO*B(A).04/ABO*A1.01、2例ABO*B(A).04/ABO*O.01.02、1例ABO*B(A).04/ABO*O.01.01、1例ABO*A3.07/ABO*0.01.02、2例ABO*A3.07/ABO*0.01.01、1例ABO*Ael.06/ABO*O.01.02、1例ABO*O.01.02/ABO*0.01.02、1例ABO*Ax.03/ABO*0.01.02。结论：A抗原表达减弱者基因存在一定异质性，基因型以A205亚型多见。