Characterizing the Role of NOTCH2 in Neuroendocrine Tumors

表征 NOTCH2 在神经内分泌肿瘤中的作用

• 批准号: 10760205
• 负责人: Brendon Robert Herring
• 金额: $ 4.13万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10760205
• 关键词: Affect; Alabama; Amino Acids; Automobile Driving; Beds; Biology; CRISPR/Cas technology; Cancerous; Cell Nucleus; Cell Proliferation; Cells; Chromogranin A; Clinical; Complex; Data; Dependence; Development; Diagnosis; Diarrhea; Disease; Disease Progression; Disease remission; Epithelium; Exanthema; Excision; Flushing; Foundations; Future; Gene Targeting; Genes; Genomics; Goals; Growth; Heart failure; Histone Deacetylase Inhibitor; Hormone secretion; Hormones; Human; In Vitro; Incidence; Islet Cell Tumor; Knock-out; Ligand Binding; Maintenance; Malignant neoplasm of pancreas; Malignant neoplasm of urinary bladder; Measurement; Mentors; Mesenchymal; Metastatic Neoplasm to the Liver; Modeling; Mus; NOTCH3 gene; Neoplasm Metastasis; Neoplasms; Neuroendocrine Tumors; Non-Small-Cell Lung Carcinoma; Notch Signaling Pathway; Nude Mice; Oncogenic; Oncology; Operative Surgical Procedures; Organ; Outcome; Outcome Measure; Palliative Care; Pancreatic Ductal Carcinoma; Pathway Analysis; Pathway interactions; Patient-Focused Outcomes; Patients; Pattern; Phenotype; Physicians; Play; Primary Neoplasm; Primary carcinoma of the liver cells; Production; Progression-Free Survivals; Proliferating; Proteins; Recurrence; Research; Role; Sampling; Scanning; Scientist; Seminal; Serum; Signal Pathway; Signal Transduction; Small Interfering RNA; Symptoms; Systemic Therapy; Testing; Time; Tissue Microarray; Training; Transfection; Tumor Biology; Tumor Cell Line; Universities; Widespread Disease; Work; Xenograft Model; career; cell growth; cell motility; clinically relevant; curative treatments; disabling symptom; effective therapy; experience; gastrointestinal; gene network; improved; in vivo; insight; knock-down; lung Carcinoma; medulloblastoma; microCT; migration; mortality; mouse model; neoplastic cell; notch protein; novel; novel therapeutics; overexpression; patient prognosis; public health relevance; reduce symptoms; response; side effect; targeted treatment; treatment strategy; tumor; tumor growth; tumor progression; tumor xenograft; tumorigenesis

PROJECT SUMMARY Pancreatic neuroendocrine tumors (pNETs) are the second most common malignancy of the pancreas, with an overall survival of 3.6 years and successful surgery the only treatment offering potential for cure. However, around 40-95% of pNETs are metastatic at the time of initial diagnosis, with local recurrence within the resection bed as the norm. Furthermore, patients with liver metastases from pNETs often have debilitating symptoms such as uncontrollable diarrhea, flushing, skin rashes, and heart failure. There are few systemic therapies that have proven to be clinically useful, and those that have still bear widely variable response rates and have poor side effect profiles. The goal of this proposal is to determine the role of Notch2 in the proliferation, metastasis, and hormone-secreting phenotype of NETs, and evaluate Notch2 as a predictor of patient outcomes. Preliminary data from our lab suggests that overexpression of Notch2 results in an increase of the proliferative rate of NET cells, while decreasing their hormone secretion. Our data also suggest that Notch2 is upregulated in metastatic pNETs compared to primary tumors. Seminal data from one of the largest genomic studies to date on pNETs has also identified Notch2 as a key master-regulator of pNET metastasis, representing a key convergence of dependencies required for disease progression and the establishment of metastasis. Therefore, it is my hypothesis that Notch2 functions in an oncogenic role in NETs and drives tumor progression, resulting in a more aggressive phenotype and portending worse patient prognosis. To evaluate this hypothesis, we will conduct phenotypic characterization (proliferation, hormone production, migration) on Notch2-overexpressing pNET cell lines that have been transiently and stably transfected. We will similarly evaluate pNET cell lines with Notch2 knockdown via siRNA, as well as stable knockout pNET cell lines generated using CRISPR-Cas9. We will then conduct similar studies in vivo using a liver metastasis mouse model whereby Notch2-overexpressing and Notch2 pNET cell lines are injected into athymic mice. Using this model, we will evaluate tumor growth and metastasis via microCT, in addition to hormone secretion over a 16-week period. We will then conduct endpoint analysis of tumors and mouse organs. Lastly, we will immunohistochemically analyze the expression of Notch2 using pNET tissue microarrays derived from human patients that have undergone surgical resection at the University of Alabama at Birmingham and evaluate the relationships between the expression of Notch2, Notch2 pathway components, and various patient outcome measures. This study will yield valuable information on the effects of Notch2 in pNETs that can help to guide future targeted therapeutic efforts and inform an understanding of pNET biology.

项目摘要 胰腺神经内分泌肿瘤（PNETS）是胰腺的第二常见恶性肿瘤， 总体生存率为3。6年和成功手术，这是唯一提供治愈潜力的治疗方法。然而， 在初次诊断时，大约40-95％的PNET是转移性的，局部复发 切除床作为常态。此外，来自PNET的肝转移的患者通常会使人衰弱 诸如无法控制的腹泻，潮红，皮疹和心力衰竭之类的症状。很少有系统性 被证明在临床上有用的疗法，而那些仍具有较大响应率的疗法 并且副作用曲线差。该提议的目的是确定Notch2在 网络的增殖，转移和激素分泌表型，并评估Notch2 预测患者预后。我们实验室的初步数据表明Notch2结果过表达 在净细胞增殖速率的增加，同时减少其激素分泌。我们的数据也是如此 表明与原发性肿瘤相比，转移性PNET中Notch2被上调。来自一个的精确数据 迄今为止在PNET上最大的基因组研究中，Notch2也确定为PNET的关键主调节剂 转移，代表疾病进展所需的依赖关系的关键融合和 建立转移。因此，我的假设是Notch2在致癌作用中起作用 网和驱动肿瘤的进展，导致更具侵略性的表型，并且定期更糟 患者预后。为了评估这一假设，我们将进行表型表征（增殖， 激素的产生，迁移）在过表达Notch2的PNET细胞系上，该细胞系已瞬时，并且 稳定转染。我们将通过siRNA和 使用CRISPR-CAS9生成的稳定敲除PNET细胞系。然后，我们将在体内进行类似的研究 使用肝脏转移鼠标模型，从而注入Notch2过表达和Notch2 PNET细胞系 进入无胸腺小鼠。使用此模型，我们还将通过Microct评估肿瘤的生长和转移 在16周期间的激素分泌。然后，我们将对肿瘤和小鼠进行端点分析 器官。最后，我们将使用PNET组织进行免疫组织化学分析Notch2的表达 来自阿拉巴马大学接受手术切除的人类患者的微阵列 在伯明翰并评估Notch2的表达之间的关系Notch2途径 组件和各种患者结果指标。这项研究将产生有关影响的宝贵信息 PNET中的Notch2可以帮助指导未来的针对性的治疗努力并告知人们对 PNET生物学。