摘要：Targeted protein degradation is a pharmacological modality that is based on the induced proximity of an E3 ubiquitin ligase and a target protein to promote target ubiquitination and proteasomal degradation. This has been achieved either via proteolysis-targeting chimeras (PROTACs)—bifunctional compounds composed of two separate moieties that individually bind the target and E3 ligase, or via molecular glues that monovalently bind either the ligase or the target1,2,3,4. Here, using orthogonal genetic screening, biophysical characterization and structural reconstitution, we investigate the mechanism of action of bifunctional degraders of BRD2 and BRD4, termed intramolecular bivalent glues (IBGs), and find that instead of connecting target and ligase in trans as PROTACs do, they simultaneously engage and connect two adjacent domains of the target protein in cis. This conformational change ‘glues’ BRD4 to the E3 ligases DCAF11 or DCAF16, leveraging intrinsic target–ligase affinities that do not translate to BRD4 degradation in the absence of compound. Structural insights into the ternary BRD4–IBG1–DCAF16 complex guided the rational design of improved degraders of low picomolar potency. We thus introduce a new modality in targeted protein degradation, which works by bridging protein domains in cis to enhance surface complementarity with E3 ligases for productive ubiquitination and degradation.

摘要：本研究评估FAK抑制剂（VS-6063）与PI3K/mTOR抑制剂（PF-04691502）联合用于宫颈癌和胰腺癌治疗的效果。研究发现两种抑制剂联合可协同抑制肿瘤细胞生长，通过促进细胞凋亡和抑制有丝分裂发挥作用。利用Promega RealTime-Glo™ MT Cell Viability Assay连续监测72小时细胞活力变化，证实联合用药较单药具有更强的细胞毒作用。进一步研究表明PI3K抑制可诱导多个RTK活化并继发激活FAK，而FAK抑制剂可有效阻断这一补偿机制，从而增强抗肿瘤效应。

摘要：研究发现ADAM17抑制剂GW280264X可显著增强PARP抑制剂Olaparib对卵巢癌的杀伤效果。在2D细胞模型、3D肿瘤球模型以及患者来源原代细胞中均观察到细胞活性下降、Caspase活化增强及细胞毒性增加。研究利用RealTime-Glo™ MT对OVCAR-8肿瘤球进行动态活力检测，证实ADAM17抑制可显著降低Olaparib IC50并产生协同效应，为克服PARPi耐药提供了新的联合治疗策略。

摘要：建立包含15个头颈癌患者来源类器官（PDTO）的Head and Neck Organoid Biobank（HNOB），系统研究TP53突变和HPV16感染对放疗反应的影响。研究开发基于Promega RealTime-Glo™ MT的3D动态放疗响应（Radioresponse）检测体系，可连续监测类器官放疗后144小时的活性变化，并利用GR metrics评估放疗敏感性。结果显示类器官放疗反应具有显著患者异质性，并与患者临床复发风险相关。TP53缺失促进生长但不足以导致放疗耐受，而HPV16 E6/E7表达可增强放疗敏感性并诱导G2期阻滞。该研究支持类器官结合RealTime-Glo™ MT作为精准放疗预测平台。

摘要：研究利用CyTOF单细胞蛋白组学系统解析卵巢高级别浆液性癌（HGSC）对卡铂治疗的DNA损伤应答（DDR）。通过分析数百万单细胞，鉴定出8个DDR蛋白模块，其中由pH2AX、pATM、pCHK1/2、pRPA等构成的Module 6与卡铂敏感性显著相关，而耐药细胞则激活更复杂的DDR网络。研究采用Promega RealTime-Glo™ MT Cell Viability Assay测定卡铂、Talazoparib、Rucaparib及联合治疗的IC50和细胞活性，支持Module 6作为预测铂类敏感性的潜在功能性生物标志物。

摘要：Talazoparib显著增强INO对ALL细胞的杀伤作用，通过RealTime-Glo™ MT评估细胞活性、IC50和协同效应。

摘要：发现同时抑制GSH和核苷酸合成可在GBM中产生合成致死作用，采用RealTime-Glo™ MT评估药物敏感性和联合效果。

摘要：建立临床相关复发GBM模型，通过RealTime-Glo™ MT和生物发光技术筛选复发肿瘤的特异性治疗脆弱性。

摘要：构建SVF-GelMA 3D生物打印模型用于胃肠组织再生，通过RealTime-Glo™ MT监测细胞活性，证实SVF促进组织修复和血管生成。

摘要：Inflammasome sensors detect pathogen- and danger-associated molecular patterns and promote inflammation and pyroptosis1. NLRP1 was the first inflammasome sensor to be described, and its hyperactivation is linked to autoinflammatory disease and cancer2-6. However, the mechanism underlying the activation and regulation of NLRP1 has not been clearly elucidated4,7,8. Here we identify ubiquitously expressed endogenous thioredoxin (TRX) as a binder of NLRP1 and a suppressor of the NLRP1 inflammasome. The cryo-electron microscopy structure of human NLRP1 shows NLRP1 bound to Spodoptera frugiperda TRX. Mutagenesis studies of NLRP1 and human TRX show that TRX in the oxidized form binds to the nucleotide-binding domain subdomain of NLRP1. This observation highlights the crucial role of redox-active cysteines of TRX in NLRP1 binding. Cellular assays reveal that TRX suppresses NLRP1 inflammasome activation and thus negatively regulates NLRP1. Our data identify the TRX system as an intrinsic checkpoint for innate immunity and provide opportunities for future therapeutic intervention in NLRP1 inflammasome activation targeting this system.