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.

项目概要 胰腺神经内分泌肿瘤（pNET）是胰腺第二常见的恶性肿瘤， 总生存期为 3.6 年，成功的手术是唯一有可能治愈的治疗方法。然而， 大约 40-95% 的 pNET 在初次诊断时已发生转移，且局部复发 切除床为常态。此外，pNET 肝转移患者通常会出现衰弱症状。 症状如无法控制的腹泻、潮红、皮疹和心力衰竭。系统性的还很少 已被证明在临床上有用的疗法，以及那些仍然具有广泛变化的反应率的疗法 并且具有较差的副作用。该提案的目标是确定 Notch2 在 NET 的增殖、转移和激素分泌表型，并将 Notch2 评估为 患者结果的预测因子。我们实验室的初步数据表明，Notch2 的过度表达导致 增加 NET 细胞的增殖率，同时减少其激素分泌。我们的数据还 表明与原发性肿瘤相比，Notch2 在转移性 pNET 中表达上调。来自一位的开创性数据 迄今为止关于 pNET 的最大基因组研究也确定了 Notch2 是 pNET 的关键主调节因子 转移，代表疾病进展所需的依赖关系的关键融合 转移的建立。因此，我的假设是，Notch2 具有致癌作用 NET 并驱动肿瘤进展，导致更具侵袭性的表型并预示着更糟的情况 患者预后。为了评估这个假设，我们将进行表型表征（增殖、 Notch2 过表达 pNET 细胞系上的激素产生、迁移） 稳定转染。我们将类似地通过 siRNA 评估具有 Notch2 敲低的 pNET 细胞系，以及 使用 CRISPR-Cas9 生成稳定的敲除 pNET 细胞系。然后我们将在体内进行类似的研究 使用肝转移小鼠模型，其中注射Notch2过表达和Notch2 pNET细胞系 进入无胸腺小鼠。使用该模型，我们将通过 microCT 评估肿瘤生长和转移，此外 16 周期间的激素分泌。然后我们将对肿瘤和小鼠进行终点分析 器官。最后，我们将使用 pNET 组织对 Notch2 的表达进行免疫组织化学分析 来自阿拉巴马大学接受手术切除的人类患者的微阵列 在伯明翰评估Notch2表达、Notch2通路之间的关系 成分和各种患者结果测量。这项研究将产生有关影响的有价值的信息 pNET 中的 Notch2 有助于指导未来的靶向治疗工作并有助于了解 pNET 生物学。