原创 李娅妮 华西医院耳鼻喉科

华西耳鼻喉前沿学术速递——文献导读（第64期）

【Nature Cancer】2024年9-12月刊论文导读

期刊介绍：

《Nature Cancer》旨在发表涵盖癌症研究全领域最重要的进展，涉及生命科学、物理科学、应用科学和社会科学，涵盖基础临床前研究、转化医学研究和临床研究。该期刊视角广阔，涵盖所有癌症研究领域，提供关于癌症生物学和遗传学的新见解、开发与提供诊断和治疗的新方法，以及理解癌症对全球社会影响的新方式，确保发表的工作能够覆盖最广泛的读者群体。在行业领域中学术影响力很大，属于国际一流期刊。属于JCR Q1，中科院1区SCI期刊，影响因子指数23.5。

2024 年 9 月，第 5 卷第 9 期，共发表14篇文章，其中2篇News & Views；3篇Research Briefings；1篇Reviews；8篇Research.

1. A deep-learning framework to predict cancer treatment response from histopathology images through imputed transcriptomics

通过估算转录组学组织病理学图像预测癌症治疗反应的深度学习框架

澳大利亚国立大学与美国国家癌症研究所共同合作

Advances in artificial intelligence have paved the way for leveraging hematoxylin and eosin-stained tumor slides for precision oncology. We present ENLIGHT–DeepPT, an indirect two-step approach consisting of (1) DeepPT, a deep-learning framework that predicts genome-wide tumor mRNA expression from slides, and (2) ENLIGHT, which predicts response to targeted and immune therapies from the inferred expression values. We show that DeepPT successfully predicts transcriptomics in all 16 The Cancer Genome Atlas cohorts tested and generalizes well to two independent datasets. ENLIGHT–DeepPT successfully predicts true responders in five independent patient cohorts involving four different treatments spanning six cancer types, with an overall odds ratio of 2.28 and a 39.5% increased response rate among predicted responders versus the baseline rate. Notably, its prediction accuracy, obtained without any training on the treatment data, is comparable to that achieved by directly predicting the response from the images, which requires specific training on the treatment evaluation cohorts.

人工智能的进步为利用苏木精和伊红染色肿瘤切片实现精准肿瘤学铺平了道路。文章中，研究团队提出了ENLIGHT-DeepPT——一种间接的两步法框架，其包含：(1) DeepPT（一种通过病理切片预测全基因组肿瘤mRNA表达的深度学习框架），以及(2) ENLIGHT（基于推断的表达值预测靶向治疗和免疫治疗应答的系统）。研究表明，DeepPT成功预测了癌症基因组图谱（TCGA）全部16个测试队列的转录组数据，并在两个独立数据集上展现出良好的泛化能力。ENLIGHT-DeepPT在涉及4种不同治疗方案、覆盖6种癌症类型的5个独立患者队列中，成功预测了真实治疗应答者，整体优势比达2.28，预测应答者的应答率较基线水平提高了39.5%。值得注意的是，该方法在未经任何治疗数据训练的情况下，其预测准确度与直接通过图像预测治疗反应（需在治疗评估队列进行专门训练）的模型相当。

2. Single-cell multiomic dissection of response and resistance to chimeric antigen receptor T cells against BCMA in relapsed multiple myeloma

针对复发型多发性骨髓瘤BCMA靶向嵌合抗原受体T细胞疗法应答与耐药机制的单细胞多组学解析

莱比锡大学医院

Markers that predict response and resistance to chimeric antigen receptor (CAR) T cells in relapsed/refractory multiple myeloma are currently missing. We subjected mononuclear cells isolated from peripheral blood and bone marrow before and after the application of approved B cell maturation antigen-directed CAR T cells to single-cell multiomic analyses to identify markers associated with resistance and early relapse. Differences between responders and nonresponders were identified at the time of leukapheresis. Nonresponders showed an immunosuppressive microenvironment characterized by increased numbers of monocytes expressing the immune checkpoint molecule CD39 and suppressed CD8+ T cell and natural killer cell function. Analysis of CAR T cells showed cytotoxic and exhausted phenotypes in hyperexpanded clones compared to low/intermediate expanded clones. We identified potential immunotherapy targets on CAR T cells, like PD1, to improve their functionality and durability. Our work provides evidence that an immunosuppressive microenvironment causes resistance to CAR T cell therapies in multiple myeloma.

预测复发/难治性多发性骨髓瘤患者对嵌合抗原受体（CAR）T细胞疗法应答与耐药性的生物标志物目前尚不明确。为识别与耐药和早期复发相关的标志物，该研究对应用已获批的靶向B细胞成熟抗原的CAR T细胞治疗前后，从外周血和骨髓中分离的单核细胞进行了单细胞多组学分析。结果显示，在白细胞分离术阶段，反应者和无反应者之间已显示出差异。无反应者表现出一种免疫抑制性微环境，其特征为表达免疫检查点分子CD39的单核细胞数量增加，并且CD8+ T细胞和自然杀伤细胞功能受抑制。对CAR T细胞的分析显示，与低/中度扩增克隆相比，高度扩增克隆呈现出细胞毒性表型和耗竭表型共存的特征。该研究还鉴定了CAR T细胞（如PD1）上的潜在免疫治疗靶点，以改善其功能和持久性。该研究提供了证据，证明免疫抑制性微环境是导致多发性骨髓瘤对CAR T细胞治疗产生耐药性的原因。

3. Tumor evolution metrics predict recurrence beyond 10 years in locally advanced prostate cancer

肿瘤演变指标预测局部晚期前列腺癌10年以上的复发风险

英国癌症研究所的科研团队

Cancer evolution lays the groundwork for predictive oncology. Testing evolutionary metrics requires quantitative measurements in controlled clinical trials. We mapped genomic intratumor heterogeneity in locally advanced prostate cancer using 642 samples from 114 individuals enrolled in clinical trials with a 12-year median follow-up. We concomitantly assessed morphological heterogeneity using deep learning in 1,923 histological sections from 250 individuals. Genetic and morphological (Gleason) diversity were independent predictors of recurrence (hazard ratio (HR) = 3.12 and 95% confidence interval (95% CI) = 1.34–7.3; HR = 2.24 and 95% CI = 1.28–3.92). Combined, they identified a group with half the median time to recurrence. Spatial segregation of clones was also an independent marker of recurrence (HR = 2.3 and 95% CI = 1.11–4.8). We identified copy number changes associated with Gleason grade and found that chromosome 6p loss correlated with reduced immune infiltration. Matched profiling of relapse, decades after diagnosis, confirmed that genomic instability is a driving force in prostate cancer progression. This study shows that combining genomics with artificial intelligence-aided histopathology leads to the identification of clinical biomarkers of evolution.

癌症演变为预测肿瘤学奠定了基础。验证进化指标需要通过对照临床试的定量检测。该研究使用来自114名参与临床试验受试者的642份样本（中位随访时间12年），绘制了局部晚期前列腺癌的基因组瘤内异质性图谱。该研究还使用深度学习评估了250名个体的1923个组织学切片的形态学异质性。结果显示，基因组和形态学（Gleason评分）多样性是独立的复发预测指标（风险比HR= 3.12，95%置信区间（CI）= 1.34–7.3；HR = 2.24，95% CI = 1.28–3.92）。联合分析可将中位无复发生存时间缩短一半的患者群体有效区分。克隆的空间分离也是复发的独立标志（HR = 2.3，95% CI = 1.11–4.8）。该研究发现与Gleason分级相关的拷贝数变化，其中6号染色体短臂丢失与免疫浸润减少存在显著相关性。对确诊数十年后复发样本的匹配分析证实了基因组不稳定性是前列腺癌进展的核心驱动因素。该研究表明，将基因组学与人工智能辅助的组织病理学相结合，可以识别出肿瘤演变的临床生物标志物。

4. Co-targeting SOS1 enhances the antitumor effects of KRASG12C inhibitors by addressing intrinsic and acquired resistance

共靶向SOS1通过解决内源性和获得性耐药增强KRASG12C抑制剂的抗肿瘤作用

奥地利勃林格殷格翰维也纳区域中心的科研团队

Combination approaches are needed to strengthen and extend the clinical response to KRASG12C inhibitors (KRASG12Ci). Here, we assessed the antitumor responses of KRASG12C mutant lung and colorectal cancer models to combination treatment with a SOS1 inhibitor (SOS1i), BI-3406, plus the KRASG12C inhibitor, adagrasib. We found that responses to BI-3406 plus adagrasib were stronger than to adagrasib alone, comparable to adagrasib with SHP2 (SHP2i) or EGFR inhibitors and correlated with stronger suppression of RAS-MAPK signaling. BI-3406 plus adagrasib treatment also delayed the emergence of acquired resistance and elicited antitumor responses from adagrasib-resistant models. Resistance to KRASG12Ci seemed to be driven by upregulation of MRAS activity, which both SOS1i and SHP2i were found to potently inhibit. Knockdown of SHOC2, a MRAS complex partner, partially restored response to KRASG12Ci treatment. These results suggest KRASG12C plus SOS1i to be a promising strategy for treating both KRASG12Ci naive and relapsed KRASG12C-mutant tumors.

为增强和延长对 KRASG12C抑制剂 (KRASG12Ci) 的临床疗效，联合治疗策略势在必行。这项研究评估了 KRASG12C突变型肺癌和结直肠癌模型对 SOS1 抑制剂 (SOS1i) BI-3406 和 联合KRASG12C抑制剂adagrasib（阿达格拉西布）的抗肿瘤反应。研究发现，BI -3406 联合adagrasib的疗效显著优于adagrasib单药治疗，且与adagrasib联合 SHP2 (SHP2i) 或 EGFR 抑制剂的疗效相当，同时显示出对RAS-MAPK信号传导更强的抑制作用。BI-3406联合adagrasib 治疗还可延缓了获得性耐药性的出现，并在adagrasib耐药模型中产生了抗肿瘤反应。对 KRASG12Ci 的耐药性可能是由 MRAS 活性上调引起的，SOS1i 和 SHP2i 均被发现能有效抑制 MRAS 活性。敲除MRAS 复合物伴侣SHOC2可部分恢复对 KRASG12Ci 治疗的反应。这些结果表明， KRASG12C联合 SOS1i 是一种在KRASG12Ci初治和复发性KRASG12C突变肿瘤中具有良好前景的治疗策略。

5. Vascular heterogeneity of tight junction Claudins guides organotropic metastasis

紧密连接蛋白的血管异质性引导了乳腺癌器官趋向性转移

美国德克萨斯大学MD Anderson Cancer Center（MD安德森癌症中心）Raghu Kalluri团队

Carcinomas are associated with metastasis to specific organs while sparing others. Breast cancer presents with lung metastasis but rarely kidney metastasis. Using this difference as an example, we queried the mechanism(s) behind the proclivity for organ-specific metastasis. We used spontaneous and implant models of metastatic mammary carcinoma coupled with inflammatory tissue fibrosis, single-cell sequencing analyses and functional studies to unravel the causal determinants of organ-specific metastasis. Here we show that lung metastasis is facilitated by angiopoietin 2 (Ang2)-mediated suppression of lung-specific endothelial tight junction protein Claudin 5, which is augmented by the inflammatory fibrotic microenvironment and prevented by anti-Ang2 blocking antibodies, while kidney metastasis is prevented by non-Ang2-responsive Claudins 2 and 10. Suppression of Claudins 2 and 10 was sufficient to induce the emergence of kidney metastasis. This study illustrates the influence of organ-specific vascular heterogeneity in determining organotropic metastasis, independent of cancer cell-intrinsic mechanisms.

癌症转移具有器官特异性倾向，例如乳腺癌通常转移至肺部，但很少转移至肾脏。本研究以这一差异为切入点，探究器官特异性转移的内在机制。通过自发性和植入性转移性乳腺癌模型，结合炎症性组织纤维化模型、单细胞测序分析以及功能性研究，该研究揭示了器官特异性转移的决定因素。研究发现，血管生成素 2 (Ang2) 介导的抑制作用降低了肺部特异性内皮紧密连接蛋白 Claudin 5 的表达，这种作用促进了肺转移，并受到炎症性纤维化微环境的增强。而通过使用抗 Ang2 阻断抗体可以有效阻止这一过程。相比之下，肾脏的转移受到对 Ang2 不敏感的 Claudins 2 和 Claudins 10 的保护。进一步研究表明，抑制 Claudins 2 和 Claudins 10 的表达即可诱导肾转移的发生。本研究表明，器官特异性的血管异质性在决定器官选择性转移中起着关键作用，这一机制与癌细胞的内在特性无关。

6. A targetable type III immune response with increase of IL-17A expressing CD4+ T cells is associated with immunotherapy-induced toxicity in melanoma

IL-17A 表达的 CD4+ T 细胞介导的可靶向 III 型免疫反应与黑色素瘤免疫治疗相关毒性有关

瑞士苏黎世大学研究团队

Immune checkpoint inhibitors are standard-of-care for the treatment of advanced melanoma, but their use is limited by immune-related adverse events. Proteomic analyses and multiplex cytokine and chemokine assays from serum at baseline and at the adverse event onset indicated aberrant T cell activity with differential expression of type I and III immune signatures. This was in line with the finding of an increase in the proportion of CD4+ T cells with IL-17A expression at the adverse event onset in the peripheral blood using flow cytometry. Multiplex immunohistochemistry and spatial transcriptomics on immunotherapy-induced skin rash and colitis showed an increase in the proportion of CD4+ T cells with IL-17A expression. Anti-IL-17A was administered in two patients with mild myocarditis, colitis and skin rash with resolution of the adverse events. This study highlights the potential role of type III CD4+ T cells in adverse event development and provides proof-of-principle evidence for a clinical trial using anti-IL-17A for treating adverse events.

免疫检查点抑制剂是治疗晚期黑色素瘤的标准疗法，但其应用受到免疫相关不良事件的限制。通过对基线和不良事件发生时血清的蛋白质组学分析及多重细胞因子和趋化因子检测，发现T细胞活性异常，并伴随I型和III型免疫特征的差异性表达。这一结果与流式细胞术检测到的不良事件发生时外周血中IL-17A表达的CD4+ T细胞比例增加的结果一致。针对免疫治疗引起的皮疹和结肠炎的多重免疫组化和空间转录组学分析同样显示IL-17A表达的CD4+ T细胞比例增加。针对两例轻度心肌炎、结肠炎和皮疹的患者，使用抗IL-17A治疗后，不良事件得以缓解。本研究揭示了III型CD4+ T细胞在不良事件发展中的潜在作用，并为利用抗IL-17A治疗不良事件的临床试验提供了原理验证证据。

7. Integrative single-cell analysis of human colorectal cancer reveals patient stratification with distinct immune evasion mechanisms

对人类结直肠癌的整合单细胞分析揭示了患者分层具有不同的免疫逃避机制

北京大学张泽民院士团队

The tumor microenvironment (TME) considerably influences colorectal cancer (CRC) progression, therapeutic response and clinical outcome, but studies of interindividual heterogeneities of the TME in CRC are lacking. Here, by integrating human colorectal single-cell transcriptomic data from approximately 200 donors, we comprehensively characterized transcriptional remodeling in the TME compared to noncancer tissues and identified a rare tumor-specific subset of endothelial cells with T cell recruitment potential. The large sample size enabled us to stratify patients based on their TME heterogeneity, revealing divergent TME subtypes in which cancer cells exploit different immune evasion mechanisms. Additionally, by associating single-cell transcriptional profiling with risk genes identified by genome-wide association studies, we determined that stromal cells are major effector cell types in CRC genetic susceptibility. In summary, our results provide valuable insights into CRC pathogenesis and might help with the development of personalized immune therapies.

肿瘤微环境（TME）对结直肠癌（CRC）的进展、治疗反应和临床结果有显著影响，但缺乏关于CRC中TME个体间异质性的研究。在本研究中，研究者通过整合来自约200例供体的结直肠癌单细胞转录组数据，全面表征了TME相对于非癌组织的转录重塑特征，并鉴定出一类具有T细胞募集潜力的罕见肿瘤特异性内皮细胞亚群。大样本量使研究者能基于TME的异质性对患者进行分型，揭示了不同的TME亚型，其中癌细胞利用不同的免疫逃逸机制。此外，通过将单细胞转录特征与全基因组关联研究鉴定的风险基因相关联，研究者确定基质细胞是CRC遗传易感性的主要效应细胞类型。总之，本研究结果为CRC的发病机制提供了重要见解，并可能有助于个性化免疫疗法的开发。

Integrative proteogenomic profiling of high-risk prostate cancer samples from Chinese patients indicates metabolic vulnerabilities and diagnostic biomarkers

中国高危前列腺癌患者的整合蛋白质基因组学分析揭示代谢脆弱性与诊断生物标志物

中国科学院上海药物研究所谭敏佳课题组、中科中山药物创新研究院徐骏宇课题组，联合中国科学院营养与健康研究所秦骏课题组、上海交通大学附属仁济医院薛蔚/董柏君团队、上海交通大学生命科学技术学院李婧课题组

Prostate cancer (PCa) exhibits significant geoethnic disparities as reflected by distinct variations in the cancer genome and disease progression. Here, we perform a comprehensive proteogenomic characterization of localized high-risk PCa utilizing paired tumors and nearby tissues from 125 Chinese male patients, with the primary objectives of identifying potential biomarkers, unraveling critical oncogenic events and delineating molecular subtypes with poor prognosis. Our integrated analysis highlights the utility of GOLM1 as a noninvasive serum biomarker. Phosphoproteomics analysis reveals the crucial role of Ser331 phosphorylation on FOXA1 in regulating FOXA1-AR-dependent cistrome. Notably, our proteomic profiling identifies three distinct subtypes, with metabolic immune-desert tumors (S-III) emerging as a particularly aggressive subtype linked to poor prognosis and BCAT2 catabolism-driven PCa progression. In summary, our study provides a comprehensive resource detailing the unique proteomic and phosphoproteomic characteristics of PCa molecular pathogenesis and offering valuable insights for the development of diagnostic and therapeutic strategies.

前列腺癌（PCa）表现出显著的地缘种族差异，这体现在癌症基因组和疾病进程的显著变化上。在该研究中，利用来自125名中国男性患者的配对肿瘤和癌旁组织样本，对局部高危前列腺癌进行了全面的蛋白质基因组学分析，旨在识别潜在的生物标志物、揭示关键致癌事件，并界定预后不良的分子亚型。综合分析显示，GOLM1 可作为一种非侵入性的血清生物标志物。磷酸化蛋白质组学分析进一步揭示了 FOXA1 蛋白Ser331 磷酸化在调控 FOXA1-AR 依赖性染色质调控网络中的重要作用。值得注意的是，蛋白质组学分析识别了三种分子亚型，其中代谢免疫凋亡肿瘤（S-III）是一种特别侵袭性的亚型，与预后不良以及由 BCAT2 分解代谢驱动的PCa进展密切相关。总之，该研究提供了详细阐释PCa分子发病机制的独特蛋白质组和磷酸化蛋白质组特征的宝贵资源，为诊断和治疗策略的开发提供了重要的理论依据和实践指导。

2024 年 10 月，第 5 卷第 10 期，共发表12篇文章，其中2篇News & Views；2篇Research Briefings；8篇Research.

1. Clinical response and pathway-specific correlates following TIGIT–LAG3 blockade in myeloma: the MyCheckpoint randomized clinical trial

多发性骨髓瘤中TIGIT-LAG3联合阻断后的临床反应与通路特异性相关性：MyCheckpoint随机临床试验

美国伊坎医学院Tisch 癌症研究所和埃默里大学 Winship 癌症研究所牵头的骨髓瘤多学科研究团队

Persons with myeloma were randomized to receive an anti-TIGIT (T cell immunoreceptor) or anti-LAG3 (lymphocyte activation gene) antibody followed by combination with pomalidomide and dexamethasone (NCT04150965). Primary and secondary endpoints were safety and efficacy, respectively. Therapy was well tolerated without dose-limiting toxicity. Durable clinical responses were observed in both the anti-TIGIT(three of six participants) and the anti-LAG3 (two of six participants) arms. Anti-LAG3 responders had higher naive cluster of differentiation 4 (CD4)-positive T cells and lower programmed cell death protein 1-positive effector T cells. Anti-TIGIT responders had higher CD226 expression, natural killer cell activation and lower CD112 expression. These data demonstrate the clinical activity of TIGIT–LAG3 blockade and identify pathway-specific response correlates in myeloma.

多发性骨髓瘤患者被随机分配接受抗 TIGIT（T 细胞免疫受体）或抗 LAG3（淋巴细胞活化基因）抗体治疗，随后联合泊马度胺和地塞米松（临床试验编号NCT04150965）。主要终点和次要终点分别为安全性和有效性。治疗耐受性良好，未观察到剂量限制性毒性。在抗 TIGIT 组（6 名参与者中有 3 名）和抗 LAG3 组（6 名参与者中有 2 名）中均观察到持久的临床反应。抗 LAG3 应答者表现出较高比例的初始CD4阳性 T 细胞和较低的程序性细胞死亡蛋白 1（PD-1）阳性效应 T 细胞，而抗 TIGIT 反应者表现出较高的 CD226 表达、自然杀伤细胞（NK）活化以及较低的 CD112 表达。这些数据证明了 TIGIT-LAG3 阻断的临床活性，并揭示了多发性骨髓瘤中通路特异性反应的相关性。

2. Neoadjuvant gemcitabine–cisplatin plus tislelizumab in persons with resectable muscle-invasive bladder cancer: a multicenter, single-arm, phase 2 trial

新辅助吉西他滨-顺铂-替雷利珠单抗治疗可切除的肌肉浸润性膀胱癌症患者：一项多中心、单臂、2期试验

中山大学孙逸仙纪念医院林天歆教授和黄健教授领衔的国内多中心临床研究

Programmed death 1 blockade (tislelizumab) has been approved for metastatic urothelial carcinoma but not as part of neoadjuvant therapy for muscle-invasive bladder cancer (MIBC). In this multicenter single-arm trial (ChiCTR2000037670), 65 participants with cT2-4aN0M0 MIBC received neoadjuvant gemcitabine–cisplatin plus tislelizumab; 57 of them underwent radical cystectomy (RC). The primary endpoint of pathologic complete response (pCR) rate was 50.9% (29/57, 95% confidence interval (CI) 37.3–64.4%) and the pathologic downstaging (secondary endpoint) rate was 75.4% (43/57, 95% CI 62.2–85.9%) in participants undergoing RC. Genomic and transcriptomic analyses revealed three MIBC molecular subtypes (S): S1 (immune-desert) with activated cell-cycle pathway, S2 (immune-excluded) with activated transforming growth factor-β pathway and S3 (immune-inflamed) with upregulated interferon-α and interferon-γ response. Post hoc analysis showed pCR rates of 16% (3/19, S1), 77% (10/13, S2) and 80% (12/15, S3) (P = 0.006). In conclusion, neoadjuvant gemcitabine–cisplatin plus tislelizumab for MIBC was compatible with an enhanced pCR rate.

程序性死亡受体1阻断剂（tislelizumab）已被批准用于转移性尿路上皮癌，但尚未作为肌肉浸润性膀胱癌症（MIBC）新辅助治疗的一部分。在这项多中心单臂试验（ChiCTR2000037670）中，65名cT2-4aN0M0-MIBC患者接受了新辅助吉西他滨-顺铂联合替雷利珠单抗治疗；其中57例行根治性膀胱切除术（RC）。主要终点病理完全缓解（pCR）率为50.9%（29/57，95%置信区间[CI] 37.3–64.4%），接受RC患者的病理降期（次要终点）率为75.4%（43/57，95% CI 62.2–85.9%）。基因组和转录组分析揭示了三种MIBC分子亚型（S）：S1（免疫缺失）具有活化的细胞周期途径，S2（免疫排斥）具有活化转化生长因子-β途径，S3（免疫炎症）具有上调的干扰素-α和干扰素-γ反应。分析显示，pCR率分别为16%（3/19，S1）、77%（10/13，S2）和80%（12/15，S3）（P=0.006）。总之，新辅助吉西他滨-顺铂联合替雷利珠单抗新辅助治疗MIBC可显著提高pCR率。

3. BRD-810 is a highly selective MCL1 inhibitor with optimized in vivo clearance and robust efficacy in solid and hematological tumor models

BRD-810 是一种高选择性 MCL1 抑制剂，具有优化的体内清除率，在实体肿瘤和血液肿瘤模型中具有强大的疗效

麻省理工学院和哈佛大学布罗德研究所Todd R. Golub和Trueline Therapeutics Inc.公司的Ulrike Rauh及其团队

The MCL1 gene is frequently amplified in cancer and codes for the antiapoptotic protein myeloid cell leukemia 1 (MCL1), which confers resistance to the current standard of care. Therefore, MCL1 is an attractive anticancer target. Here we describe BRD-810 as a potent and selective MCL1 inhibitor and its key design principle of rapid systemic clearance to potentially minimize area under the curve-driven toxicities associated with MCL1 inhibition. BRD-810 induced rapid cell killing within 4 h in vitro but, in the same 4-h window, had no impact on cell viability or troponin I release in human induced pluripotent stem cell-derived cardiomyocytes, even at suprapharmacologic concentrations. In vivo BRD-810 induced efficacy in xenograft hematological and solid tumor models despite the short residence time of BRD-810 in plasma. In totality, our data support the hypothesis that short-term inhibition of MCL1 with BRD-810 can induce apoptosis in tumor cells while maintaining an acceptable safety profile. We, therefore, intend to advance BRD-810 to clinical trials.

MCL1 基因在癌症中频繁扩增，其编码的抗凋亡蛋白髓样细胞白血病 1 (MCL1)可导致对当前标准治疗的耐药性。因此，MCL1 是一个有吸引力的抗癌靶点。本研究报道了一种强效且选择性的 MCL1 抑制剂BRD-810，其关键设计原理是快速系统清除以最小化曲线下面积（AUC）驱动的MCL1抑制相关毒性。BRD-810 在体外 4 小时内诱导快速细胞杀伤，但在相同的时间窗口内，即使在超药理浓度下，也不会影响人诱导多能干细胞衍生的心肌细胞的细胞活力或肌钙蛋白 I 释放。在体内实验中，尽管 BRD-810 在血浆中的停留时间很短，但在异种移植血液肿瘤和实体瘤模型中均表现出疗效。总体而言，该研究数据支持以下假设：使用 BRD-810 短期抑制 MCL1 可诱导肿瘤细胞凋亡，同时保持可接受的安全性。因此，研究团队打算将 BRD-810 推进至临床试验阶段。

4. ISB 2001 trispecific T cell engager shows strong tumor cytotoxicity and overcomes immune escape mechanisms of multiple myeloma cells

ISB 2001三特异性T细胞衔接剂显示出强大的肿瘤细胞毒性并克服多发性骨髓瘤细胞的免疫逃逸机制

Certara 公司（纳斯达克代码：CERT），与 Ichnos Glenmark Innovation（IGI）公司携手

Despite recent advances in immunotherapies targeting single tumor-associated antigens, patients with multiple myeloma eventually relapse. ISB 2001 is a CD3+ T cell engager (TCE) co-targeting BCMA and CD38 designed to improve cytotoxicity against multiple myeloma. Targeting of two tumor-associated antigens by a single TCE resulted in superior cytotoxic potency across a variable range of BCMA and CD38 tumor expression profiles mimicking natural tumor heterogeneity, improved resistance to competing soluble factors and exhibited superior cytotoxic potency on patient-derived samples and in mouse models. Despite the broad expression of CD38 across human tissues, ISB 2001 demonstrated a reduced T cell activation profile in the absence of tumor cells when compared to TCEs targeting CD38 only. To determine an optimal first-in-human dose for the ongoing clinical trial (NCT05862012), we developed an innovative quantitative systems pharmacology model leveraging preclinical data, using a minimum pharmacologically active dose approach, therefore reducing patient exposure to subefficacious doses of therapies.

尽管近期靶向单一肿瘤相关抗原的免疫疗法取得了进展，多发性骨髓瘤患者最终仍会复发。ISB 2001是一种同时靶向BCMA和CD38的CD3+ T细胞衔接剂（TCE），旨在提高对多发性骨髓瘤的细胞毒性。单一TCE靶向两种肿瘤相关抗原，在模拟自然肿瘤异质性的BCMA和CD38肿瘤表达谱变化范围内表现出更强的细胞毒性效力，提高了对竞争性可溶性因子的抵抗力，并在患者来源样本和小鼠模型中表现出更强的细胞毒性效力。尽管CD38在人体组织中广泛表达，与仅靶向CD38的TCE相比，ISB 2001在没有肿瘤细胞存在时表现出较低的T细胞激活水平。为了确定正在进行的临床试验（NCT05862012）的最佳首次人体剂量，研究团队开发了一个创新的定量系统药理学模型，利用临床前数据，采用最小药理活性剂量方法，从而减少患者暴露于无效治疗剂量的风险。

5. Glutaminase inhibition in combination with azacytidine in myelodysplastic syndromes: a phase 1b/2 clinical trial and correlative analyses

谷氨酰胺酶抑制联合阿扎胞苷治疗骨髓增生异常综合征：I b/II 期临床试验及相关性分析

MD安德森癌症中心

Malignancies are reliant on glutamine as an energy source and a facilitator of aberrant DNA methylation. We demonstrate preclinical synergy of telaglenastat (CB-839), a selective glutaminase inhibitor, combined with azacytidine (AZA), followed by a single-arm, open-label, phase 1b/2 study in persons with advanced myelodysplastic syndrome (MDS). The dual primary endpoints evaluated clinical activity, safety and tolerability; secondary endpoints evaluated pharmacokinetics, pharmacodynamics, overall survival, event-free survival and duration of response. The dose-escalation study included six participants and the dose-expansion study included 24 participants. Therapy was well tolerated and led to an objective response rate of 70% with (marrow) complete remission in 53% of participants and a median overall survival of 11.6 months, with evidence of myeloid differentiation in responders determined by single-cell RNA sequencing. Glutamine transporter solute carrier family 38 member 1 in MDS stem cells was associated with clinical responses and predictive of worse prognosis in a large MDS cohort. These data demonstrate the safety and efficacy of CB-839 and AZA as a combined metabolic and epigenetic approach in MDS. ClinicalTrials.gov identifier: NCT03047993.

恶性肿瘤依赖谷氨酰胺作为能量来源，同时促进异常DNA甲基化。本研究在临床前研究中证明了选择性谷氨酰胺酶抑制剂 telaglenastat（CB-839）与阿扎胞苷（AZA）联合使用的协同作用，并在晚期骨髓增生异常综合征（MDS）患者中开展了单臂开放标签的1b/2期临床试验。双重主要终点评估了临床疗效、安全性和耐受性；次要终点评估了药代动力学、药效学、总生存期（OS）、无事件生存期（EFS）和缓解持续时间（DOR）。剂量递增研究包括6名参与者，剂量扩展研究包括24名参与者。治疗方案耐受性良好，客观缓解率（ORR）为70%，其中53%的参与者达到骨髓完全缓解（CR），中位总生存期为11.6个月。单细胞RNA测序分析显示，应答者存在髓系分化的证据。研究还发现，MDS干细胞中谷氨酰胺转运蛋白溶质载体家族38成员1（SLC38A1）与临床反应相关，并可预测较差预后（基于大型MDS队列）。这些数据表明，CB-839与AZA联合治疗作为一种代谢与表观遗传结合的策略，在MDS中的安全性和有效性。临床试验注册号：NCT03047993。

6. Spatial analysis reveals targetable macrophage-mediated mechanisms of immune evasion in hepatocellular carcinoma minimal residual disease

空间分析揭示肝细胞癌微小残留病中巨噬细胞介导的免疫逃逸机制及其靶向潜力

斯坦福大学的Dean W. Felsher团队和Renumathy Dhanasekaran团队

Hepatocellular carcinoma (HCC) frequently recurs from minimal residual disease (MRD), which persists after therapy. Here, we identified mechanisms of persistence of residual tumor cells using post-chemoembolization human HCC (n = 108 patients, 1.07 million cells) and a transgenic mouse model of MRD. Through single-cell high-plex cytometric imaging, we identified a spatial neighborhood within which PD-L1 + M2-like macrophages interact with stem-like tumor cells, correlating with CD8+ T cell exhaustion and poor survival. Further, through spatial transcriptomics of residual HCC, we showed that macrophage-derived TGFβ1 mediates the persistence of stem-like tumor cells. Last, we demonstrate that combined blockade of Pdl1 and Tgfβ excluded immunosuppressive macrophages, recruited activated CD8+ T cells and eliminated residual stem-like tumor cells in two mouse models: a transgenic model of MRD and a syngeneic orthotopic model of doxorubicin-resistant HCC. Thus, our spatial analyses reveal that PD-L1+ macrophages sustain MRD by activating the TGFβ pathway in stem-like cancer cells and targeting this interaction may prevent HCC recurrence from MRD.

肝细胞癌（HCC）常因微小残留病灶（MRD）而复发，MRD在治疗后持续存在。本研究通过分析化疗栓塞后的人HCC样本（n = 108名患者，107万个细胞）及MRD的转基因小鼠模型，探讨了残留肿瘤细胞持续存在的机制。通过单细胞高维度成像技术，该研究确定了PD-L1+ M2样巨噬细胞与干细胞样肿瘤细胞之间的空间相互作用关系，并发现这种相互作用与CD8+ T细胞耗竭及不良生存率密切相关。此外，通过残留HCC的空间转录组学分析，该研究揭示了巨噬细胞来源的TGFβ1在介导干细胞样肿瘤细胞持续存在中的关键作用。最后，研究证明，在两种小鼠模型中（包括MRD的转基因模型和阿霉素耐药HCC的同源原位模型），联合阻断Pdl1和Tgfβ能够有效清除免疫抑制性巨噬细胞，招募活化的CD8+ T细胞，并消除残留的干细胞样肿瘤细胞。因此，本研究的空间分析表明，PD-L1+巨噬细胞通过激活干细胞样癌细胞中的TGFβ通路维持MRD的存在，靶向这一相互作用可能预防MRD引起的HCC复发。

7. Spatial single-cell protein landscape reveals vimentinhigh macrophages as immune-suppressive in the microenvironment of hepatocellular carcinoma

空间单细胞蛋白图谱揭示高弧度巨噬细胞在肝癌微环境中的免疫抑制作用

复旦大学代谢与整合生物学研究院王红阳院士及其合作陈磊团队

Tumor microenvironment heterogeneity in hepatocellular carcinoma (HCC) on a spatial single-cell resolution is unclear. Here, we conducted co-detection by indexing to profile the spatial heterogeneity of 401 HCC samples with 36 biomarkers. By parsing the spatial tumor ecosystem of liver cancer, we identified spatial patterns with distinct prognosis and genomic and molecular features, and unveiled the progressive role of vimentin (VIM)high macrophages. Integration analysis with eight independent cohorts demonstrated that the spatial co-occurrence of VIMhigh macrophages and regulatory T cells promotes tumor progression and favors immunotherapy. Functional studies further demonstrated that VIMhigh macrophages enhance the immune-suppressive activity of regulatory T cells by mechanistically increasing the secretion of interleukin-1β. Our data provide deep insights into the heterogeneity of tumor microenvironment architecture and unveil the critical role of VIMhigh macrophages during HCC progression, which holds potential for personalized cancer prevention and drug discovery and reinforces the need to resolve spatial-informed features for cancer treatment.

肝细胞癌(HCC)的肿瘤微环境异质性在空间单细胞分辨率上尚不清楚。本研究通过索引共检测技术，利用36种生物标志物对401例HCC样本的空间异质性进行了分析。通过分析肝癌的空间肿瘤生态系统，发现了具有不同预后、基因组和分子特征的空间模式，并揭示了波形蛋白（VIM）高表达巨噬细胞的渐进作用。与8个独立队列的整合分析表明，VIM高表达巨噬细胞和调节性T细胞的空间共定位促进了肿瘤进展，并有利于免疫治疗。功能研究进一步表明，VIM高表达巨噬细胞通过增加白细胞介素-1β的分泌来增强调节性T细胞的免疫抑制活性。该研究为肿瘤微环境结构的异质性提供了深入的见解，揭示了VIM高表达巨噬细胞在HCC进展中的关键作用，这具有个性化癌症预防和药物发现的潜力，并强调了解决空间信息特征对癌症治疗的重要性。

8. A protein expression atlas on tissue samples and cell lines from cancer patients provides insights into tumor heterogeneity and dependencies

基于癌症患者组织样本和细胞系的蛋白质表达图谱揭示肿瘤异质性和依赖性

美国MD安德森癌症中心梁晗教授研究组、Rehan Akbani研究组、和俄勒冈健康和科学大学Knight癌症研究中心的Gordon Mills教授研究组合作

The Cancer Genome Atlas (TCGA) and the Cancer Cell Line Encyclopedia (CCLE) are foundational resources in cancer research, providing extensive molecular and phenotypic data. However, large-scale proteomic data across various cancer types for these cohorts remain limited. Here, we expand upon our previous work to generate high-quality protein expression data for approximately 8,000 TCGA patient samples and around 900 CCLE cell line samples, covering 447 clinically relevant proteins, using reverse-phase protein arrays. These protein expression profiles offer profound insights into intertumor heterogeneity and cancer dependency and serve as sensitive functional readouts for somatic alterations. We develop a systematic protein-centered strategy for identifying synthetic lethality pairs and experimentally validate an interaction between protein kinase A subunit α and epidermal growth factor receptor. We also identify metastasis-related protein markers with clinical relevance. This dataset represents a valuable resource for advancing our understanding of cancer mechanisms, discovering protein biomarkers and developing innovative therapeutic strategies.

癌症基因组图谱（TCGA）和癌细胞系百科全书（CCLE）是癌症研究中的重要基础资源，提供了广泛的分子和表型数据。然而，这些队列中涵盖多种癌症类型的大规模蛋白质组学数据仍然匮乏。本研究在之前工作的基础上进一步扩展，使用反相蛋白阵列技术生成了约8,000个TCGA患者样本和约900个CCLE细胞系样本的高质量蛋白质表达数据，涵盖447种与临床密切相关的蛋白质。这些蛋白质表达谱为肿瘤间异质性和癌症依赖性提供了深入见解，并作为体细胞突变的功能敏感读数。研究开发了一种以蛋白质为中心的系统化策略来识别合成致死配对，并通过实验验证了蛋白激酶A亚基α与表皮生长因子受体之间的相互作用。此外，本研究还鉴定了与临床相关的转移相关蛋白质标志物。该数据集为深入理解癌症机制、发现蛋白质生物标志物以及开发创新治疗策略提供了宝贵的资源。

2024 年 11 月，第 5 卷第 11 期，共发表13篇文章，其中1篇Comment & Opinion；2篇News & Views；1篇Research Briefings；8篇Research；1篇Amendments & Corrections.

1. CAR-redirected natural killer T cells demonstrate superior antitumor activity to CAR-T cells through multimodal CD1d-dependent mechanisms

CAR 重定向的自然杀伤 T 细胞通过多模式 CD1d 依赖机制表现出优于 CAR-T 细胞的抗肿瘤活性

美国北卡罗来纳大学的Gianpietro Dotti研究团队

Human natural killer T (NKT) cells have been proposed as a promising cell platform for chimeric antigen receptor (CAR) therapy in solid tumors. Here we generated murine CAR-NKT cells and compared them with CAR-T cells in immune-competent mice. Both CAR-NKT cells and CAR-T cells showed similar antitumor effects in vitro, but CAR-NKT cells showed superior antitumor activity in vivo via CD1d-dependent immune responses in the tumor microenvironment. Specifically, we show that CAR-NKT cells eliminate CD1d-expressing M2-like macrophages. In addition, CAR-NKT cells promote epitope spreading and activation of endogenous T cell responses against tumor-associated neoantigens. Finally, we observed that CAR-NKT cells can co-express PD1 and TIM3 and show an exhaustion phenotype in a model of high tumor burden. PD1 blockade as well as vaccination augmented the antitumor activity of CAR-NKT cells. In summary, our results demonstrate the multimodal function of CAR-NKT cells in solid tumors, further supporting the rationale for developing CAR-NKT therapies in the clinic.

人源自然杀伤T（NKT）细胞被认为是实体瘤嵌合抗原受体（CAR）治疗中具有前景的细胞平台。本研究构建了小鼠CAR-NKT细胞，并与免疫健全的小鼠中与CAR-T细胞进行了比较。结果显示，尽管CAR-NKT细胞与CAR-T细胞在体外显示出相似的抗肿瘤效果，但CAR-NKT细胞在体内通过肿瘤微环境中的CD1d依赖性免疫反应表现出更优的抗肿瘤活性。具体而言，该研究发现CAR-NKT细胞能够消除表达CD1d的M2样巨噬细胞。此外，CAR-NKT细胞还促进了表位扩散并激活内源性T细胞对肿瘤相关新抗原的免疫反应。最后，在高肿瘤负荷模型中，研究团队观察到CAR-NKT细胞可以共表达PD1和TIM3，并呈现出耗竭表型。通过阻断PD1和疫苗接种，可以显著增强CAR-NKT细胞的抗肿瘤活性。总之，本研究结果揭示了CAR-NKT细胞在实体瘤中的多模式抗肿瘤功能，为临床开发CAR-NKT疗法提供了进一步支持。

2. Phase separation of phospho-HDAC6 drives aberrant chromatin architecture in triple-negative breast cancer

磷酸化的 HDAC6的相分离驱动三阴性乳腺癌中染色质结构异常

广东省人民医院、中山大学、广州市妇女儿童中心、桂林医科大学附属医院及顺德医院的联合科研团队

How dysregulated liquid–liquid phase separation (LLPS) contributes to the oncogenesis of female triple-negative breast cancer (TNBC) remains unknown. Here we demonstrate that phosphorylated histone deacetylase 6 (phospho-HDAC6) forms LLPS condensates in the nuclei of TNBC cells that are essential for establishing aberrant chromatin architecture. The disordered N-terminal domain and phosphorylated residue of HDAC6 facilitate effective LLPS, whereas nuclear export regions exert a negative dominant effect. Through phase-separation-based screening, we identified Nexturastat A as a specific disruptor of phospho-HDAC6 condensates, which effectively suppresses tumor growth. Mechanistically, importin-β interacts with phospho-HDAC6, promoting its translocation to the nucleus, where 14-3-3θ mediates the condensate formation. Disruption of phospho-HDAC6 LLPS re-established chromatin compartments and topologically associating domain boundaries, leading to disturbed chromatin loops. The phospho-HDAC6-induced aberrant chromatin architecture affects chromatin accessibility, histone acetylation, RNA polymerase II elongation and transcriptional profiles in TNBC. This study demonstrates phospho-HDAC6 LLPS as an emerging mechanism underlying the dysregulation of chromatin architecture in TNBC.

失调的液-液相分离(LLPS)如何促进女性三阴性乳腺癌(TNBC)的发生仍不清楚。在这里，研究人员证明磷酸化的组蛋白去乙酰化酶6 (phospho-HDAC6)在TNBC细胞的细胞核中形成LLPS凝聚物，这是建立异常染色质结构所必需的。HDAC6的无序N端结构域和磷酸化残基促进了有效的LLPS，而核输出区则起负主导作用。通过相分离筛选发现，Nexturastat A是磷酸化-HDAC6凝聚物的特异性破坏剂，可有效抑制肿瘤生长。在机制上，importin-β与磷酸HDAC6相互作用，促进其转运到细胞核，其中14-3-3θ介导凝析物的形成。磷酸化-HDAC6 LLPS的破坏重建了染色质区室和拓扑相关结构域边界，导致染色质环受到干扰。磷酸化-HDAC6诱导的异常染色质结构影响TNBC中染色质的可及性、组蛋白乙酰化、RNA聚合酶II延长和转录谱。该研究表明，磷酸化-HDAC6 LLPS是TNBC中染色质结构失调的一种新兴机制。

3. The neuroendocrine transition in prostate cancer is dynamic and dependent on ASCL1

前列腺癌的神经内分泌转变是动态的并且依赖ASCL1

纪念斯隆-凯特琳癌症中心Charles L. Sawyers教授团队

Lineage plasticity is a hallmark of cancer progression that impacts therapy outcomes, yet the mechanisms mediating this process remain unclear. Here, we introduce a versatile in vivo platform to interrogate neuroendocrine lineage transformation throughout prostate cancer progression. Transplanted mouse prostate organoids with human-relevant driver mutations (Rb1−/−; Trp53−/−; cMyc+ or Pten−/−; Trp53−/−; cMyc+) develop adenocarcinomas, but only those with Rb1 deletion advance to aggressive, ASCL1+ neuroendocrine prostate cancer (NEPC) resistant to androgen receptor signaling inhibitors. Notably, this transition requires an in vivo microenvironment not replicated by conventional organoid culture. Using multiplexed immunofluorescence and spatial transcriptomics, we reveal that ASCL1+ cells arise from KRT8+ luminal cells, progressing into transcriptionally heterogeneous ASCL1+;KRT8− NEPC. Ascl1 loss in established NEPC causes transient regression followed by recurrence, but its deletion before transplantation abrogates lineage plasticity, resulting in castration-sensitive adenocarcinomas. This dynamic model highlights the importance of therapy timing and offers a platform to identify additional lineage plasticity drivers.

谱系可塑性是影响治疗结果的癌症进展的标志，但介导这一过程的机制仍不清楚。在这里，本研究介绍了一个多功能的体内平台，用于探究前列腺癌进展中的神经内分泌谱系转化。具有人相关驱动基因突变（Rb1-/-；Trp53-/-；cMyc+或Pten-/-；Trp55-/-；c Myc+）的移植小鼠前列腺类器官发展为腺癌，但只有Rb1缺失的小鼠前列腺类肿瘤发展为对雄激素受体信号抑制剂具有耐药性的侵袭性ASCL1+神经内分泌癌症癌（NEPC）。值得注意的是，这种转变需要一个传统类器官培养无法复制的体内微环境。使用多重免疫荧光和空间转录组学，团队发现ASCL1+细胞起源于KRT8+管腔细胞，进展为转录异质性ASCL1+；KRT8-NEPC。在已建立的NEPC中，Ascl1的缺失会导致短暂的消退，随后复发，但在移植前删除它会消除谱系可塑性，产生去势敏感性腺癌。该动态模型强调了治疗时机的重要性，并为识别其他谱系可塑性驱动因素提供了平台。

4. Two distinct epithelial-to-mesenchymal transition programs control invasion and inflammation in segregated tumor cell populations

两种不同的上皮-间充质转化程序调控分离肿瘤细胞群体的侵袭与炎症反应

西班牙神经科学研究所等机构团队

Epithelial-to-mesenchymal transition (EMT) triggers cell plasticity in embryonic development, adult injured tissues and cancer. Combining the analysis of EMT in cell lines, embryonic neural crest and mouse models of renal fibrosis and breast cancer, we find that there is not a cancer-specific EMT program. Instead, cancer cells dedifferentiate and bifurcate into two distinct and segregated cellular trajectories after activating either embryonic-like or adult-like EMTs to drive dissemination or inflammation, respectively. We show that SNAIL1 acts as a pioneer factor in both EMT trajectories, and PRRX1 drives the progression of the embryonic-like invasive trajectory. We also find that the two trajectories are plastic and interdependent, as the abrogation of the EMT invasive trajectory by deleting Prrx1 not only prevents metastasis but also enhances inflammation, increasing the recruitment of antitumor macrophages. Our data unveil an additional role for EMT in orchestrating intratumor heterogeneity, driving the distribution of functions associated with either inflammation or metastatic dissemination.

上皮-间充质转化（EMT）在胚胎发育、成人损伤组织修复以及癌症中触发细胞可塑性。通过结合细胞系、胚胎神经嵴以及肾纤维化和乳腺癌小鼠模型中EMT的分析，该研究发现并不存在癌症特异性的EMT程序。相反，癌细胞在激活胚胎样或成体样EMT后，分别驱动播散或炎症反应，并去分化为两个不同且分离的细胞轨迹。研究证明SNAIL1在这两种EMT轨迹中均作为关键的先锋因子，而PRRX1驱动胚胎样侵袭轨迹的进程。该研究还发现，这两种轨迹具有可塑性并相互依赖：通过删除Prrx1阻断EMT侵袭轨迹不仅可以抑制转移，还能增强炎症反应，从而增加抗肿瘤巨噬细胞的募集。该研究揭示了EMT在协调肿瘤内异质性方面的新作用，驱动与炎症或转移扩散相关的功能分布。

5. Developmental mosaicism underlying EGFR-mutant lung cancer presenting with multiple primary tumors

以多发原发性肿瘤为表现的EGFR突变型肺癌的发育嵌合机制

Gad Getz 和 Daniel A. Haber 教授领导的哈佛大学-麻省总医院癌症研究团队

Although the development of multiple primary tumors in smokers with lung cancer can be attributed to carcinogen-induced field cancerization, the occurrence of multiple tumors at presentation in individuals with EGFR-mutant lung cancer who lack known environmental exposures remains unexplained. In the present study, we identified ten patients with early stage, resectable, non-small cell lung cancer who presented with multiple, anatomically distinct, EGFR-mutant tumors. We analyzed the phylogenetic relationships among multiple tumors from each patient using whole-exome sequencing (WES) and hypermutable poly(guanine) (poly(G)) repeat genotyping as orthogonal methods for lineage tracing. In four patients, developmental mosaicism, assessed by WES and poly(G) lineage tracing, indicates a common non-germline cell of origin. In two other patients, we identified germline EGFR variants, which confer moderately enhanced signaling when modeled in vitro. Thus, in addition to germline variants, developmental mosaicism defines a distinct mechanism of genetic predisposition to multiple EGFR-mutant primary tumors, with implications for their etiology and clinical management.

尽管吸烟者的肺癌多原发灶形成可归因于致癌物诱导的区域癌化，但在缺乏已知环境暴露的EGFR突变肺癌个体中出现多发性肿瘤的机制仍未明确。本研究分析了10例早期、可切除的非小细胞肺癌患者，这些患者表现为多个解剖学位置不同的EGFR突变肿瘤。本研究利用全外显子组测序（WES）和高突变性poly嘌呤（鸟嘌呤 poly(G)）重复序列分型作为正交谱系追踪的互补策略，分析了每位患者多个肿瘤之间的系统发育关系。在4例患者中，通过WES和poly(G)谱系追踪评估的发育嵌合现象表明这些肿瘤来源于一个共同的非种系起源细胞。而在另外两例患者中，鉴定出种系EGFR变异，这些变异在体外建模中显示中度增强的信号传导能力。因此，除了种系变异外，发育嵌合性还揭示了一种驱动多发EGFR突变原发性肿瘤的独特遗传易感机制，这对其病因研究和临床管理具有重要意义。

6. Global loss of promoter–enhancer connectivity and rebalancing of gene expression during early colorectal cancer carcinogenesis

结直肠癌早期致癌过程中启动子-增强子连接的整体缺失和基因表达的再平衡

斯坦福大学Michael P. Snyder遗传学研究团队

Although three-dimensional (3D) genome architecture is crucial for gene regulation, its role in disease remains elusive. We traced the evolution and malignant transformation of colorectal cancer (CRC) by generating high-resolution chromatin conformation maps of 33 colon samples spanning different stages of early neoplastic growth in persons with familial adenomatous polyposis (FAP). Our analysis revealed a substantial progressive loss of genome-wide cis-regulatory connectivity at early malignancy stages, correlating with nonlinear gene regulation effects. Genes with high promoter–enhancer (P–E) connectivity in unaffected mucosa were not linked to elevated baseline expression but tended to be upregulated in advanced stages. Inhibiting highly connected promoters preferentially represses gene expression in CRC cells compared to normal colonic epithelial cells. Our results suggest a two-phase model whereby neoplastic transformation reduces P–E connectivity from a redundant state to a rate-limiting one for transcriptional levels, highlighting the intricate interplay between 3D genome architecture and gene regulation during early CRC progression.

尽管三维 （3D） 基因组结构对基因调控至关重要，但其在疾病中的作用尚未完全阐明。该研究通过构建 33 例家族性腺瘤性息肉病（FAP）患者不同早期瘤变阶段的结肠样本高分辨率染色质构象图谱，系统解析了结直肠癌（CRC）演进与恶性转化的分子特征。分析显示，在早期恶性转化阶段即存在全基因组顺式调控连接性的大幅度渐进性缺失，这一现象与非线性基因调控效应相关。在未受影响的粘膜中具有高启动子-增强子 （P-E） 连接的基因与基线表达升高无关，但在晚期往往上调。与正常结肠上皮细胞相比，抑制高度连接的启动子优先抑制 CRC 细胞中的基因表达。该研究结果表明了一个两阶段模型，其中肿瘤转化将 P-E 连接从冗余状态降低到转录水平的限速状态，突出了 CRC 早期进展期间 3D 基因组结构和基因调控之间错综复杂的相互作用。

7. Differential chromatin accessibility and transcriptional dynamics define breast cancer subtypes and their lineages

不同的染色质可及性和转录动态定义了乳腺癌的亚型及其谱系

美国圣路易斯市华盛顿大学Li Ding团队

Breast cancer (BC) is defined by distinct molecular subtypes with different cells of origin. The transcriptional networks that characterize the subtype-specific tumor-normal lineages are not established. In this work, we applied bulk, single-cell and single-nucleus multi-omic techniques as well as spatial transcriptomics and multiplex imaging on 61 samples from 37 patients with BC to show characteristic links in gene expression and chromatin accessibility between BC subtypes and their putative cells of origin. Regulatory network analysis of transcription factors underscored the importance of BHLHE40 in luminal BC and luminal mature cells and KLF5 in basal-like tumors and luminal progenitor cells. Furthermore, we identify key genes defining the basal-like (SOX6 and KCNQ3) and luminal A/B (FAM155A and LRP1B) lineages. Exhausted CTLA4-expressing CD8+ T cells were enriched in basal-like BC, suggesting an altered means of immune dysfunction. These findings demonstrate analysis of paired transcription and chromatin accessibility at the single-cell level is a powerful tool for investigating cancer lineage and highlight transcriptional networks that define basal and luminal BC lineages.

乳腺癌（BC）根据分子特征可分为不同的亚型，每种亚型可能来源于不同的细胞类型，但目前尚未建立表征亚型特异性肿瘤-正常谱系的转录网络，其具体起源和调控机制尚未完全明确。在本项工作中应用批量、单细胞和单核多组学技术，以及空间转录组学和多重成像技术，对来自37名乳腺癌患者的61份样本进行了深入研究，以展示乳腺癌亚型与其假定的起源细胞之间的特征性联系。结果表明，不同亚型乳腺癌的基因表达和染色质可及性与其假定的起源细胞存在特异性关联。研究发现BHLHE40在管腔乳腺癌和管腔成熟细胞中的重要作用，而KLF5在基底样乳腺癌和管腔祖细胞中发挥关键功能。此外，基因SOX6和KCNQ3定义了基底样乳腺癌谱系，而FAM155A和LRP1B是管腔A/B乳腺癌谱系的重要标志。同时，基底样乳腺癌中富集表达CTLA4的耗竭型CD8+ T细胞，提示其免疫功能存在异常。本研究首次结合单细胞水平的转录与染色质可及性分析，揭示了乳腺癌亚型的转录网络和谱系特征，为理解癌症发生和精准治疗提供了新的思路。

8. Multiomic analysis of familial adenomatous polyposis reveals molecular pathways associated with early tumorigenesis

家族性腺瘤性息肉病的多组学分析揭示了与早期肿瘤发生相关的分子通路

斯坦福大学医学院遗传学系James M. Ford & Michael P. Snyder癌症多组学研究团队

Familial adenomatous polyposis (FAP) is a genetic disease causing hundreds of premalignant polyps in affected persons and is an ideal model to study transitions of early precancer states to colorectal cancer (CRC). We performed deep multiomic profiling of 93 samples, including normal mucosa, benign polyps and dysplastic polyps, from six persons with FAP. Transcriptomic, proteomic, metabolomic and lipidomic analyses revealed a dynamic choreography of thousands of molecular and cellular events that occur during precancerous transitions toward cancer formation. These involve processes such as cell proliferation, immune response, metabolic alterations (including amino acids and lipids), hormones and extracellular matrix proteins. Interestingly, activation of the arachidonic acid pathway was found to occur early in hyperplasia; this pathway is targeted by aspirin and other nonsteroidal anti-inflammatory drugs, a preventative treatment under investigation in persons with FAP. Overall, our results reveal key genomic, cellular and molecular events during the earliest steps in CRC formation and potential mechanisms of pharmaceutical prophylaxis.

家族性腺瘤性息肉病（FAP）是一种遗传性疾病，可导致患者患上数百个癌前息肉，是研究早期癌前状态向结直肠癌（CRC）转变的理想模型。作者对6名FAP患者的93个样本进行了深度多组学分析，包括正常粘膜、良性息肉和发育不良息肉。通过转录组学、蛋白质组学、代谢组学和脂质组学分析揭示了癌前向癌症形成转变期间发生的数千个分子和细胞事件的动态变化。这些过程包括细胞增殖、免疫应答、代谢改变（包括氨基酸和脂质）、激素和细胞外基质蛋白。有趣的是，作者发现花生四烯酸途径的激活发生在增生的早期，而阿司匹林和其他非甾体抗炎药（正在研究中的FAP患者的预防性治疗药物）能够靶向作用于这一途径。总的来说，研究结果揭示了CRC形成早期阶段的关键基因组、细胞和分子特征以及药物预防的潜在机制。

2024 年 12 月，第 5 卷第 12 期，共发表33篇文章，其中1篇Editorial；2篇News；5篇Comment & Opinion；12篇Research Highlights；2篇News & Views；2篇Research Briefings；1篇Reviews；8篇Research

1. Unique structural configuration of EV-DNA primes Kupffer cell-mediated antitumor immunity to prevent metastatic progression

EV-DNA独特的结构启动了Kupffer细胞介导的抗肿瘤免疫反应，以防止癌症转移进展

美国威尔康奈尔医学院、纪念斯隆-凯特琳癌症中心和韩国延世大学的研究人员

Extracellular vesicles (EVs) transport biomolecules that mediate intercellular communication. We previously showed that EVs contain DNA (EV-DNA) representing the entire genome. However, the mechanism of genomic EV-DNA packaging and its role in cancer remain elusive. We now demonstrate that EV-DNA is predominantly localized on the vesicle surface and associated with uniquely modified and cleaved histones. Moreover, a genome-wide clustered regularly interspaced short palindromic repeats knockout screen revealed that immune developmental pathways and genes, including apoptotic peptidase activating factor 1 (APAF1) and neutrophil cytosolic factor 1 (NCF1), regulate EV-DNA packaging. Furthermore, in colorectal cancer models, uptake of EV-DNA by pre-metastatic liver Kupffer cells (KCs) activated DNA damage responses. This activation rewired KC cytokine production and promoted the formation of tertiary lymphoid structures, thereby suppressing liver metastasis. Conversely, loss of APAF1 decreased EV-DNA packaging and promoted liver metastasis. Importantly, colorectal cancer biopsy EV-DNA secretion could serve as a predictive biomarker for postoperative metastasis. Taken together, our findings indicate that uniquely chromatinized EV-DNA induces antitumor immunity.

细胞外囊泡（EVs）运输介导细胞间通信的生物分子。之前的研究表明，EVs包含代表整个基因组的DNA（EV-DNA）。然而，基因组EV-DNA的包装机制及其在癌症中的作用仍然不明确。本研究现表明，揭示EV-DNA主要定位于囊泡表面，并与独特修饰及剪切形式的组蛋白结合。此外，通过全基因组CRISPR敲除筛选发现，免疫发育通路及基因（包括凋亡肽酶激活因子1（APAF1）和中性粒细胞胞质因子1（NCF1））调控EV-DNA的包装。进一步的研究表明，在结直肠癌模型中，转移前肝脏Kupffer细胞（KCs）摄取EV-DNA后激活了DNA损伤反应。该反应重塑了KCs的细胞因子产生，并促进了三级淋巴结构的形成，从而抑制了肝脏转移。相反，APAF1的缺失减少了EV-DNA的包装，并促进了肝脏转移。重要的是，结直肠癌活检中的EV-DNA分泌可以作为术后转移的预测生物标志物。综上所述，研究发现，独特的染色质化EV-DNA能够诱导抗肿瘤免疫反应。

2. TIGIT and PD-L1 co-blockade promotes clonal expansion of multipotent, non-exhausted antitumor T cells by facilitating co-stimulation

TIGIT 和 PD-L1 共阻断通过促进共刺激促进多能、非耗竭抗肿瘤 T 细胞的克隆扩增

Genentech公司Ira Mellman团队

Blockade of immune checkpoints PD-1 and TIGIT has demonstrated activity in mouse tumor models and human patients with cancer. Although these coinhibitory receptors can restrict signaling in CD8+ T cells by regulating their associated co-stimulatory receptors CD28 and CD226, the functional consequences of combining PD-1 and TIGIT blockade remain poorly characterized. In mouse tumor models, we show that combination blockade elicited CD226-driven clonal expansion of tumor antigen-specific CD8+ T cells. The expanded clones emerged from a population of stem-like cells in draining lymph nodes, entering the blood as a previously unidentified single-phenotype, multiclonal population. Upon reaching the tumor, these transiting cells expanded further and differentiated into effector or exhausted T cells, with combination blockade restricting entry into the exhaustion pathway by favoring co-stimulation. Thus, PD-1 and TIGIT inhibition helps shape the repertoire of tumor-reactive CD8+ T cells in draining lymph nodes and determines their immunological fate in the tumor to enhance therapeutic benefit. Analysis of clinical trial samples suggests a similar mechanism may also occur in patients with cancer.

在肿瘤免疫治疗领域，PD-1与TIGIT联合阻断策略展现出协同抗肿瘤效应。尽管这些共抑制受体可以通过调节其相关的共刺激受体 CD28 和 CD226 来限制 CD8+ T 细胞中的信号传导，但联合 PD-1 和 TIGIT 阻断的功能后果仍然难以表征。在小鼠肿瘤模型中，该研究表明联合阻断引起了 CD226 驱动的肿瘤抗原特异性 CD8+ T 细胞的克隆扩增。扩增的克隆来自引流淋巴结中的干细胞样细胞群，作为以前未识别的单表型、多克隆细胞群进入血液。到达肿瘤后，这些过境细胞进一步扩增并分化为效应或耗竭 T 细胞，联合阻断通过有利于共刺激来限制进入耗竭途径。因此，PD-1 和 TIGIT 抑制有助于塑造引流淋巴结中肿瘤反应性 CD8+ T 细胞的库，并确定它们在肿瘤中的免疫命运以增强治疗效果。对临床试验样本的分析表明，类似的机制也可能发生在癌症患者中。

3. Recapitulating the adenoma–carcinoma sequence by selection of four spontaneous oncogenic mutations in mismatch-repair-deficient human colon organoids

通过选择错配修复缺陷型人类结肠类器官中的四种自发致癌突变来重建腺瘤-癌序列

荷兰乌特勒支大学Hans Clevers教授团队

Carcinogenesis results from the sequential acquisition of oncogenic mutations that convert normal cells into invasive, metastasizing cancer cells. Colorectal cancer exemplifies this process through its well-described adenoma–carcinoma sequence, modeled previously using clustered regularly interspaced short palindromic repeats (CRISPR) to induce four consecutive mutations in wild-type human gut organoids. Here, we demonstrate that long-term culture of mismatch-repair-deficient organoids allows the selection of spontaneous oncogenic mutations through the sequential withdrawal of Wnt agonists, epidermal growth factor (EGF) agonists and the bone morphogenetic protein (BMP) antagonist Noggin, while TP53 mutations were selected through the addition of Nutlin-3. Thus, organoids sequentially acquired mutations in AXIN1 and AXIN2 (Wnt pathway), TP53, ACVR2A and BMPR2 (BMP pathway) and NRAS (EGF pathway), gaining complete independence from stem cell niche factors. Quadruple-pathway (Wnt, EGF receptor, p53 and BMP) mutant organoids formed solid tumors upon xenotransplantation. This demonstrates that carcinogenesis can be recapitulated in a DNA repair-mutant background through in vitro selection that targets four consecutive cancer pathways.

结直肠癌的发生遵循经典的腺瘤-癌变序列，这一过程可在体外通过基因编辑与微环境调控进行模拟。本研究利用错配修复缺陷的肠道类器官模型，通过逐步撤除Wnt激动剂、表皮生长因子（EGF）激动剂及骨形态发生蛋白（BMP）拮抗剂Noggin，选择性地诱导AXIN1/AXIN2（Wnt通路）、TP53（p53通路）、ACVR2A/BMPR2（BMP通路）及NRAS（EGF通路）等关键基因的连续突变。获得四重通路（Wnt/EGFR/p53/BMP）联合突变的类器官在异种移植中形成实体肿瘤，证实DNA修复缺陷背景下可通过体外选择压力重现体内癌变过程。

4. Selenocysteine tRNA methylation promotes oxidative stress resistance in melanoma metastasis

硒半胱氨酸tRNA甲基化促进黑色素瘤转移中的氧化应激抵抗

威尔康奈尔医学研究院Elena Piskounova团队

Selenocysteine-containing proteins play a central role in redox homeostasis. Their translation is a highly regulated process and is dependent on two tRNASec isodecoders differing by a single 2′-O-ribose methylation called Um34. Here we characterized FTSJ1 as the Um34 methyltransferase and show that its activity is required for efficient selenocysteine insertion at the UGA stop codon during translation. Specifically, loss of Um34 leads to ribosomal stalling and decreased UGA recoding. FTSJ1-deficient cells are more sensitive to oxidative stress and show decreased metastatic colonization in xenograft models of melanoma metastasis. We found that FTSJ1 mediates efficient translation of selenoproteins essential for the cellular antioxidant response. Our findings uncover a role for tRNASec Um34 modification in oxidative stress resistance and highlight FTSJ1 as a potential therapeutic target specific for metastatic disease.

含硒半胱氨酸（Sec）蛋白在氧化还原稳态中起核心作用在维持氧化还原和内质网的稳态起着重要作用，已知有25种硒蛋白，其中11种参与细胞抗氧化反应。根据报道，硒代半胱氨酸特异性tRNA（tRNASec）仅在四个位置发生修饰，其中在反密码子环中的修饰N6-异戊烯腺苷37（i6A37）和5-甲氧基羰基甲基Um34（mcm5 Um34），其中硒蛋白翻译受到硒代半胱氨酸tRNA（tRNASec）上唯一的摆动位2'-O-核糖甲基化修饰（Um34）的调控。本研究鉴定了FTSJ1是Um34修饰的甲基转移酶，并展示了其活性对于高效硒半胱氨酸插入UGA终止密码子至关重要。特别是，Um34的缺失会导致核糖体停滞和UGA重新编码水平下降。FTSJ1缺陷细胞对氧化应激更敏感，并在黑色素瘤转移的移植模型中表现为降低转移定植。研究还发现FTSJ1介导了对细胞抗氧化反应至关重要的硒蛋白的有效翻译。这些发现揭示了tRNASec Um34修饰在氧化应激抵抗中的作用，并突出了FTSJ1作为针对转移性疾病的潜在治疗靶点。

5. Transcription and DNA replication collisions lead to large tandem duplications and expose targetable therapeutic vulnerabilities in cancer

转录与DNA复制冲突导致大规模串联重复，并揭示癌症中可靶向的治疗弱点

芝加哥大学和加州大学旧金山分校的癌症研究团队

Despite the abundance of somatic structural variations (SVs) in cancer, the underlying molecular mechanisms of their formation remain unclear. In the present study, we used 6,193 whole-genome sequenced tumors to study the contributions of transcription and DNA replication collisions to genome instability. After deconvoluting robust SV signatures in three independent pan-cancer cohorts, we detected transcription-dependent, replicated-strand bias, the expected footprint of transcription–replication collision (TRC), in large tandem duplications (TDs). Large TDs are abundant in female-enriched, upper gastrointestinal tract and prostate cancers. They are associated with poor patient survival and mutations in TP53, CDK12 and SPOP. Upon inactivating CDK12, cells display significantly more TRCs, R-loops and large TDs. Inhibition of WEE1, CHK1 and ATR selectively inhibits the growth of cells deficient in CDK12. Our data suggest that large TDs in cancer form as a result of TRCs and their presence can be used as a biomarker for prognosis and treatment.

尽管癌症中存在大量体细胞结构变异（SV），其形成的潜在分子机制仍未完全明了。研究对6,193个全基因组测序的肿瘤样本进行了分析，探讨了转录与DNA复制碰撞（TRC）对基因组不稳定性的贡献。通过对三个独立的泛癌队列中的SV特征进行去卷积分析，该研究在大片段串联重复（TD）中检测到了转录依赖性和复制链偏差的特征，这符合TRC的预期分子足迹。研究发现，大型TD在女性高发的上消化道癌症和前列腺癌中尤为丰富，并与较差的患者生存率以及TP53、CDK12和SPOP基因突变密切相关。在灭活CDK12后，细胞中观察到显著增多的TRC、R环和大型TD。此外，抑制WEE1、CHK1和ATR可选择性抑制CDK12缺陷细胞的生长。本研究表明，大型TD在癌症中由TRC引发，其存在可作为预后和治疗的潜在生物标志物。

6. The pro-oncogenic noncanonical activity of a RAS•GTP:RanGAP1 complex facilitates nuclear protein export

RAS•GTP:RanGAP1 复合物的促癌非典型活性促进核蛋白转出

美国国家癌症研究所的Douglas R. Lowy和Brajendra K. Tripathi团队

Canonical RAS signaling, including PI3K/AKT- and RAF/MEK-dependent activities, results mainly from RAS•GTP interaction with its effectors at the plasma membrane. Here, we identified a fundamental, oncogenic, noncanonical RAS•GTP activity that increases XPO1-dependent export of nuclear protein cargo into the cytoplasm and is independent of PI3K/AKT and RAF/MEK signaling. This RAS-dependent step acts downstream from XPO1 binding to nuclear protein cargo and is mediated by a perinuclear protein complex between RAS•GTP and RanGAP1 that facilitates hydrolysis of Ran•GTP to Ran•GDP, which promotes release of nuclear protein cargo into the cytoplasm. The export of nuclear EZH2, which promotes cytoplasmic degradation of the DLC1 tumor suppressor protein, is a biologically important component of this pro-oncogenic activity. Conversely, preventing nuclear protein export contributes to the antitumor activity of KRAS inhibition, which can be further augmented by reactivating the tumor suppressor activity of DLC1 or potentially combining RAS inhibitors with other cancer treatments.

典型的 RAS 信号传导，包括 PI3K/AKT 和 RAF/MEK 依赖性活性，主要由 RAS•GTP 与其在质膜上的效应因子相互作用产生。在这篇文章中，研究人员确定了一种基本的、致癌的、非经典的 RAS•GTP 活性，该活性可增加 XPO1 依赖性核蛋白货物向细胞质的输出，并且独立于 PI3K/AKT 和 RAF/MEK 信号传导。这个依赖于 RAS 的步骤在 XPO1 与核蛋白货物结合的下游起作用，并由 RAS•GTP 和 RanGAP1 之间的核周蛋白复合物介导，该复合物促进 Ran•GTP 水解为 Ran•GDP 从而促进核蛋白货物释放到细胞质中。核 EZH2 的输出促进 DLC1 肿瘤抑制蛋白的细胞质降解，是这种促癌活性的重要生物学组成部分。反过来，阻止核蛋白输出有助于 KRAS 抑制的这种抗肿瘤活性，通过重新激活 DLC1 的肿瘤抑制活性或可能将 RAS 抑制剂与其他癌症治疗相结合，可以进一步增强 KRAS 抑制的抗肿瘤活性。

7. Single-cell transcriptomic landscape deciphers olfactory neuroblastoma subtypes and intra-tumoral heterogeneity

单细胞转录组学景观破译嗅觉神经母细胞瘤亚型和肿瘤内异质性

复旦大学附属眼耳鼻喉科医院余洪猛团队

Olfactory neuroblastoma (ONB) is a rare malignancy known to originate from the olfactory epithelium. The complex tumor ecosystem of this pathology remains unclear. Here, we explored the cellular components within ten ONB tumors and one olfactory mucosa sample based on single-cell RNA profiles. We showed the intra-tumoral heterogeneity by identifying five unique expression programs among malignant epithelial cells. A distinct three-classification system (neural, basal, mesenchymal) for ONB was established according to the distinguished gene expression patterns. Biomarkers for categorizing bulk tumors into uncharacterized subtypes were elucidated. Different responses towards certain chemotherapy regimens could be cautiously inferred according to the molecular features representing the three tumor types, thus helping with precision chemotherapy. We also analyzed subclusters of the tumor microenvironment (TME) and the interactions among different cell types within the TME. The relative abundance of immunosuppressive tumor-associated macrophages suggests potential benefits of immunotherapies targeting macrophages.

嗅觉神经母细胞瘤（ONB）是一种罕见的恶性肿瘤，已知起源于嗅觉上皮。这种病理学的复杂肿瘤生态系统仍不清楚。该研究基于单细胞RNA谱探索了10个ONB肿瘤和1个嗅粘膜样本中的细胞成分。该研究通过在恶性上皮细胞中鉴定5个独特的表达程序来显示肿瘤内异质性。根据不同的基因表达模式，建立了ONB的不同三分类系统（神经、基底、间质型）。阐明了将大块肿瘤分类为未表征亚型的生物标志物。根据代表三种肿瘤类型的分子特征，可以谨慎推断对某些化疗方案的不同反应，从而有助于精准化疗。该研究还分析了肿瘤微环境（TME）的子簇以及TME内不同细胞类型之间的相互作用。免疫抑制性肿瘤相关巨噬细胞的相对丰度表明靶向巨噬细胞的免疫疗法具有潜在益处。

8. Dura immunity configures leptomeningeal metastasis immunosuppression for cerebrospinal fluid barrier invasion

硬脑膜免疫调控软脑膜转移的免疫抑制以促进脑脊液屏障侵袭

复旦大学脑科学转化研究院/附属华山医院的迟喻丹课题组及浙江清华长三角研究院的张强强研究员

The cerebrospinal fluid (CSF) border accommodates diverse immune cells that permit peripheral cell immunosurveillance. However, the intricate interactions between CSF immune cells and infiltrating cancer cells remain poorly understood. Here we use fate mapping, longitudinal time-lapse imaging and multiomics technologies to investigate the precise origin, cellular crosstalk and molecular landscape of macrophages that contribute to leptomeningeal metastasis (LM) progression. Mechanically, we find that dura-derived LM-associated macrophages (dLAMs) migrate into the CSF in a matrix metalloproteinase 14 (MMP14)-dependent manner. Furthermore, we identify that dLAMs critically require the presence of secreted phosphoprotein 1 (SPP1) in cancer cells for their recruitment, fostering an immunosuppressed microenvironment characterized by T cell exhaustion and inactivation. Conversely, inhibition of the SPP1–MMP14 axis can impede macrophages from bypassing the border barrier, prevent cancer cell growth and improve survival in LM mouse models. Our findings reveal an unexpectedly private source of innate immunity within the meningeal space, shed light on CSF barrier dysfunction dynamics and supply potential targets of clinical immunotherapy.

脑脊液（CSF）边界容纳多种免疫细胞，允许外周细胞免疫监视。然而，脑脊液免疫细胞与浸润性癌细胞之间复杂的相互作用仍然知之甚少。本研究利用命运追踪、纵向时间推移成像和多组学技术，探讨了促进软脑膜转移（LM）进展的巨噬细胞的具体起源、细胞间相互作用及分子特征。研究发现硬脑膜衍生的LM相关巨噬细胞（dLAMs）以基质金属蛋白酶14 （MMP14）依赖的方式迁移到CSF中。此外，dLAMs需要癌细胞中分泌磷酸化蛋白1 （SPP1）的存在才能募集，从而形成以T细胞衰竭和失活为特征的免疫抑制微环境。相反，抑制SPP1-MMP14轴可以阻止巨噬细胞绕过边界屏障，阻止癌细胞生长，提高LM小鼠模型的存活率。该发现揭示了脑膜空间内先天免疫的一个意想不到的来源，揭示了脑脊液屏障功能障碍动力学，并提供了临床免疫治疗的潜在靶点。

汇报人：李娅妮

导师：郑芸

审核：饶郁芳、任建君