Molecular Mechanisms and Translational studies in Colon Cancer

结肠癌的分子机制和转化研究

• 批准号: 9240497
• 负责人: Lopa Mishra
• 金额: --
• 依托单位: U.S. DEPT/VETS AFFAIRS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-10-01 至 2020-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9240497
• 关键词: Address; Age; Animal Model; Anti-CEA Antibody; Antibodies; Beckwith-Wiedemann Syndrome; Benign; Biological Markers; Biological Models; Cancer Etiology; Carcinoma; Caring; Case Study; Cell physiology; Colon; Colon Carcinoma; Colonic Adenoma; Colorectal; Colorectal Adenoma; Colorectal Cancer; Colorectal Neoplasms; Comorbidity; DNA; Data Set; Development; Diagnosis; Disease; Evaluation; Exposure to; Family; Future; Genes; Genome; Genomics; Goals; Healthcare Systems; Homeostasis; Human; Human Cell Line; Incidence; Inherited; Interleukin-6; Label; Lead; MAP Kinase Gene; Malignant Neoplasms; Malignant neoplasm of gastrointestinal tract; Measures; Mediating; Military Personnel; Mismatch Repair; Molecular; Mucous Membrane; Mutant Strains Mice; Mutation; Oncogenic; PIK3CG gene; Pathway interactions; Phenocopy; Polyps; Population; Prevention program; Prevention strategy; Preventive screening; Process; Prognostic Factor; Property; Quality of life; Radiation; Regulation; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; Role; STAT3 gene; Sampling; Screening for cancer; Sequence Analysis; Side; Signal Transduction; Small Interfering RNA; Stem cells; Subgroup; Survival Rate; Syndrome; TGFBR1 gene; TP53 gene; Testing; The Cancer Genome Atlas; Tissue Sample; Toxin; Transforming Growth Factor beta; Transforming Growth Factor beta Receptors; Translational Research; Treatment-related toxicity; Tumor Suppressor Proteins; Tumorigenicity; United States; Vaccines; Veterans; actionable mutation; adenoma; base; cancer invasiveness; cancer stem cell; colorectal cancer prevention; exome; genome sequencing; genomic profiles; high risk; improved; in vivo; inhibitor/antagonist; insight; member; mortality; mouse model; neoplastic; novel; programs; therapeutic development; transcriptome; transcriptome sequencing; transcriptomics; translational study; tumor; tumorigenesis; tumorigenic; whole genome

Colorectal cancer (CRC) remains the second most common cause of cancer mortality worldwide, and among Veterans in the United States, the third despite the successful implementation of successful preventive screening measures. One contributing factor that has newly emerged is the rapid development of cancers in small, sessile adenomas (polyps), with mutational processes and signatures that drive early CRC tumorigenesis. Yet, to date, biomarkers and risk factors for polyps (adenomas) that inordinately and rapidly develop into colon cancer remain poorly defined. We recently observed surprisingly high mutational profiles through whole-genome sequence (WGS) analysis in 2 of 4 pairs of benign colorectal adenoma tissue samples. Extending this to unsupervised hierarchical clustered transcriptomic analysis of a further 7 pairs of adenomas reveals distinct mutational signatures regardless of adenoma size. Strikingly, we observe mutations in the TGF-β pathway and CEA-associated genes in 4 out of 11 adenomas, overlapping with the Wnt pathway. Immunohistochemical labeling reveals a nearly 5-fold increase in CEA levels in 23% of adenoma samples tested, with a concomitant loss of TGF-β signaling. We have defined a functional role by which the CEA B3 domain interacts with TGFBR1, potentially inactivating the tumor suppressor function of TGF-β signaling. Raised CEA levels occur with concomitant loss of TGF-β signaling and activation of oncogenic molecules that include STAT3. Furthermore, deletion of Smad3/4 adaptor β2SP results in loss of TGF-β signaling, with a spontaneous formation of adenomas as well as overt CRC, and presents a strong mouse model of CRC. The overall hypothesis of this application is that disruption of the TGF-β tumor suppressor pathway, and concomitantly raised levels of oncogenic molecules such as CEA, STAT3, disrupt normal colonic mucosa and promote oncogenesis in CRC. Our aims are to: AIM 1a) Expand and validate the significance of loss of the TGF-β pathway associated with high CEA and/or CEACAM6 levels as prognostic factors in advanced adenomas that rapidly progress to CRC through i) First, whole transcriptomic sequence, immunohistochemical and RT-PCR analyses of 40 additional adenomas ii) Secondly, by examining The Cancer Genome Atlas (TCGA) Pan gastrointestinal cancer (GI) datasets to obtain a complete evaluation of our findings. 1b) Determine whether CEA/CEACAM6 and STAT3 with loss of TGF-β signaling are drivers of early CRC in vivo. 2 a) Determine the molecular mechanisms by which CEA inhibits the TGF-β tumor suppressor pathway in early and advanced CRC. We plan to accomplish this through i) Analyzing the role of CEA and CEACAM6 in the disruption of the TGF-β pathway and concomitant IL-6/STAT3 activation in CRC. ii) Determine antitumor activity of CEA inhibition, with and without STAT3 inhibitors and characterize their mechanisms of action. Through translational studies- interrogating human cell lines/genomics, and animal model systems of colon cancer, this application proposes an integrated approach for human CRC, providing novel insights into early and advanced CRC. With vaccines available, as well as new low toxicity therapeutics, identifying specific populations particularly among Veterans with multiple co-morbidities, that could respond to treatment, these goals are centrally important for improving the quality of life under treatment, as well as the mortality from this common cancer.

结直肠癌 (CRC) 仍然是全球第二大癌症死亡原因， 美国退伍军人第三次尽管成功实施了成功的预防措施 筛查措施。新出现的一个促成因素是癌症的快速发展 小型无蒂腺瘤（息肉），具有驱动早期 CRC 的突变过程和特征 肿瘤发生。然而，迄今为止，息肉（腺瘤）的生物标志物和危险因素异常且迅速地 发展成结肠癌仍不明确。我们最近观察到令人惊讶的高突变谱 通过对 4 对良性结直肠腺瘤组织样本中的 2 对进行全基因组序列 (WGS) 分析。 将其扩展到另外 7 对腺瘤的无监督分层聚类转录组分析 无论腺瘤大小如何，都显示出不同的突变特征。引人注目的是，我们观察到突变 11 个腺瘤中有 4 个存在 TGF-β 通路和 CEA 相关基因，与 Wnt 通路重叠。 免疫组织化学标记显示 23% 的腺瘤样本中 CEA 水平增加了近 5 倍 经测试，伴随着 TGF-β 信号传导的丧失。我们定义了 CEA B3 的职能角色 结构域与 TGFBR1 相互作用，可能使 TGF-β 信号传导的肿瘤抑制功能失活。 CEA 水平升高伴随着 TGF-β 信号传导的丧失和致癌分子的激活， 包括 STAT3。此外，删除 Smad3/4 接头 β2SP 会导致 TGF-β 信号传导丧失， 腺瘤的自发形成以及明显的结直肠癌，并提出了强大的结直肠癌小鼠模型。这 本申请的总体假设是 TGF-β 肿瘤抑制途径的破坏，以及 同时升高致癌分子（如 CEA、STAT3）水平，破坏正常结肠 粘膜并促进结直肠癌的肿瘤发生。我们的目标是： 目标 1a) 扩展并验证与高 CEA 和/或相关的 TGF-β 通路缺失的重要性 CEACAM6 水平作为晚期腺瘤的预后因素，通过 i) 首先， 40 个其他腺瘤的全转录组序列、免疫组织化学和 RT-PCR 分析 ii) 其次，通过检查癌症基因组图谱（TCGA）泛胃肠道癌症（GI）数据集来获得 对我们的发现的完整评估。 1b) 确定 CEA/CEACAM6 和 STAT3 是否丢失 TGF-β 信号传导是体内早期 CRC 的驱动因素。 2 a) 确定 CEA 抑制 TGF-β 肿瘤抑制途径的分子机制 早期和晚期 CRC。我们计划通过 i) 分析 CEA 和 CEACAM6 在 CRC 中 TGF-β 通路的破坏以及伴随的 IL-6/STAT3 激活。 ii) 确定抗肿瘤作用 使用和不使用 STAT3 抑制剂时 CEA 抑制活性并表征其作用机制。 通过转化研究——探究人类细胞系/基因组学和结肠动物模型系统 癌症，该应用提出了一种针对人类 CRC 的综合方法，为早期癌症提供了新的见解 和高级 CRC。借助可用的疫苗以及新的低毒性疗法，确定特定的 人群，特别是患有多种合并症的退伍军人，可能对治疗有反应，这些 目标对于改善治疗下的生活质量以及由此导致的死亡率至关重要 常见癌症。