Development of TGF-beta antagonists for cancer therapy

开发用于癌症治疗的 TGF-β 拮抗剂

• 批准号: 9153704
• 负责人: Lalage Wakefield
• 金额: $ 87.44万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9153704
• 关键词: 4T1; Address; Adult; Advanced Malignant Neoplasm; Adverse effects; Antibodies; Biological; Biological Assay; Biological Markers; Biology; Cell Proliferation; Cell Survival; Cells; Cessation of life; Clinical; Clinical Data; Clinical Treatment; Clinical Trials; Complex; DNA Sequence; Data; Detection; Development; Ecosystem; Embryonic Development; Epithelial; Gene Expression Profile; Generations; Goals; Growth Factor; Heterogeneity; Homeostasis; Human; Immune response; Immunologic Surveillance; Knowledge; Ligation; Literature; Maintenance; Malignant Neoplasms; Messenger RNA; Metastatic breast cancer; Methods; Modeling; Molecular; Monitor; Mus; Mutation; Neoplasm Metastasis; Outcome; Pathway interactions; Patients; Pharmacodynamics; Phase; Play; Pre-Clinical Model; Primary Neoplasm; Protein Isoforms; Proteins; Role; Sampling; Signal Transduction; Staging; Technology; Testing; Therapeutic; Therapeutic Uses; Transforming Growth Factor Beta 2; Transforming Growth Factor beta; Transforming Growth Factor-Beta Overexpression; Transgenic Organisms; Tumor Cell Line; Tumor Promoters; Tumor Suppression; Tumor Suppressor Proteins; Work; angiogenesis; base; cancer therapy; carcinogenesis; cell motility; comparative efficacy; exome; genetic regulatory protein; human TGFB1 protein; immune function; improved; in vivo; malignant breast neoplasm; mouse model; neoplastic cell; neutralizing antibody; novel strategies; outcome forecast; pre-clinical; response; therapeutic target; transcriptomics; transforming growth factor beta3; treatment strategy; tumor; tumor microenvironment; tumor progression; tumorigenesis

Despite the dual role for TGF-beta as both tumor suppressor and tumor promoter in carcinogenesis, preclinical data from our lab and others has previously suggested that strategies to antagonize TGF-beta may selectively reduce the undesirable tumor promoting effects of this growth factor while sparing the desirable effects on tumor suppression and normal homeostasis. Based on these promising preclinical results, several different TGF-beta pathway antagonists are in early phase clinical trials for the treatment of advanced cancer. However, given the complex biology of TGF-beta, the successful development of TGF-beta antagonists for cancer therapy will depend on a clear understanding of how these agents work, and the related question of how to select patients who will benefit from this type of treatment. In our previous work, we performed detailed mechanistic analysis of the mode of action of anti-TGF-beta neutralizing antibodies in the widely used 4T1 transplantable mouse model of metastatic breast cancer. Using a panel of 12 transplantable syngeneic mouse models of metastatic breast cancer, with metastatic burden as the primary therapeutic endpoint, we had previously uncovered heterogeneous responses to TGF-beta antagonism, with inhibition of metastasis in some models, and no effect on or stimulation of metastasis in other models. We have applied discovery-based approaches to address molecular and biological mechanisms underlying the heterogeneity of therapeutic response and to generate useful predictive biomarkers. Whole exome DNA sequencing of the tumor cell lines has shown that none of the commonly occurring mutations in breast cancer are correlated with response to therapy. Transcriptomic analysis of untreated primary tumors in the panel segregates tumors by response to anti-TGF-beta therapy, suggesting that the therapeutic response is dictated by prominent, readily identifiable molecular and biological features of the tumor. Tumors from models showing a desirable response to TGF-beta antagonism are characterized by reduced immune function, enhanced angiogenesis, higher tumor cell proliferation and survival, and transcriptomic evidence of TGF-beta pathway activation in the untreated state. Gene expression signatures that are predictive of response to anti-TGF-beta therapy have been generated and are being tested in additional preclinical models. In our search for useful predictive and pharmacodynamic biomarkers of TGF-beta antagonism, we have developed approaches for quantitative and more sophisticated monitoring of TGF-beta pathway activation in tumors, adapting the ProteinSimple SimpleWestern technology for accurate quantitation of Smad activation in tumor samples, and developing a quantitative brightfield proximity ligation assay for detection of the non-canonical "mixed Smad" signaling complexes that are associated with pathological TGF-beta signaling. We have also begun to address whether it might be possible to improve the therapeutic response to TGF-beta antagonism by selective neutralization of different TGF-beta isoforms. Correlative clinical data and literature evidence suggests that whereas TGF-beta1 is primarily associated with poor outcome, TGF-beta3 may actually oppose TGF-beta1 and be associated with good outcome, providing a rationale for selective neutralization of TGF-beta1 and TGF-beta2, while sparing TGF-beta3. As part of this initiative, we have developed methods for accurate quantitation of TGF-betas in tumor extracts since we have shown that TGF-beta mRNA levels do not correlate well with protein levels for TGF-beta1 and TGF-beta3. We have assessed TGF-beta isoform protein levels across the metastatic tumor panel and find that TGF-beta1:TGF-beta3 ratios vary over a 20-fold range. Therapeutic anti-TGF-beta antibodies with different isoform selectivity will be compared for efficacy in representative models. Through this approach, we hope to generate improved TGF-beta-targeted therapeutics.

尽管TGF-β在癌变中既是肿瘤抑制剂又是肿瘤启动子的双重作用，但我们实验室和其他人的临床前数据以前提出，与TGF-β拮抗TGF-β的策略可能有选择地减少这种生长对这种生长的不良肿瘤促进对肿瘤抑制和正常施加的影响。基于这些有希望的临床前结果，几种不同的TGF-β途径拮抗剂正在早期临床试验中治疗晚期癌症。但是，鉴于TGF-β的复杂生物学，TGF-beta拮抗剂用于癌症治疗的成功发展将取决于对这些药物的工作原理的清晰了解，以及如何选择将从这种类型治疗中受益的患者的相关问题。在我们以前的工作中，我们对广泛使用的4T1转移性乳腺癌小鼠模型中和抗TGF-beta中和抗体的作用方式进行了详细的机械分析。我们使用一组转移性乳腺癌的12个可移植的合成小鼠模型，并具有转移性负担为主要的治疗终点，我们以前已经发现了对TGF-β拮抗作用的异质反应，并且在某些模型中没有抑制转移的反应，并且对其他模型中的转移或刺激没有影响。我们采用了基于发现的方法来解决治疗反应异质性的基础的分子和生物学机制，并产生有用的预测生物标志物。肿瘤细胞系的整个外显子体DNA测序表明，乳腺癌中通常发生的突变均与对治疗的反应相关。小组中未经处理的原发性肿瘤的转录组分析通过对抗TGF-β疗法的反应分离肿瘤，这表明治疗反应取决于肿瘤的突出，易于识别的分子和生物学特征。来自显示对TGF-β拮抗作用的理想反应的模型的肿瘤的特征是免疫功能降低，血管生成增强，较高的肿瘤细胞增殖和存活率以及未经处理状态的TGF-β途径激活的转录组证据。已经产生了预测对抗TGF-β疗法反应的基因表达特征，并正在其他临床前模型中进行了测试。在寻找TGF-β拮抗作用的有用的预测和药效动力学生物标志物中，我们开发了对肿瘤中TGF-β途径激活TGF-beta途径激活的定量和更复杂监测的方法与病理TGF-β信号传导相关的SMAD“信号复合物。我们还开始解决是否有可能通过选择性中和不同的TGF-β同工型来改善对TGF-β拮抗作用的治疗反应。相关临床数据和文献证据表明，尽管TGF-BETA1主要与不良结果有关，但TGF-BETA3实际上可能反对TGF-BETA1，并且与良好的结果相关，提供了选择性中和TGF-BETA1和TGF-BETA1和TGF-BETA2的基本原理，而TGF-BETA2则在tgf-beta2的同时进行了比较。作为该计划的一部分，我们开发了用于准确定量肿瘤提取物TGF-betas的方法，因为我们已经表明TGF-βmRNA水平与TGF-BETA1和TGF-BETA3的蛋白质水平不太相关。我们已经评估了整个转移性肿瘤面板的TGF-β同工型蛋白水平，发现TGF-BETA1：TGF-BETA3比在20倍范围内变化。在代表性模型中，将比较具有不同同工型选择性的治疗性抗TGF-β抗体。通过这种方法，我们希望产生改进的TGF-beta靶向治疗剂。