Identifying targets for combination therapy with FOLFIRINOX and investigating cell polarity loss as a potential driver of invasion in basal-like PDAC

确定 FOLFIRINOX 联合治疗的靶标并研究细胞极性丧失作为基底样 PDAC 侵袭的潜在驱动因素

• 批准号: 10347174
• 负责人: Sandra Zarmer
• 金额: $ 3.52万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10347174
• 关键词: 3-Dimensional; Apical; Basal Cell; Biological; Biological Assay; Biology; CRISPR screen; CRISPR/Cas technology; Cancer Etiology; Cell Adhesion; Cell Line; Cell Polarity; Cessation of life; Clinical Trials; Combination Drug Therapy; Combined Modality Therapy; Data; Disease; Dose; Drug Targeting; Epithelial Cells; Exhibits; Fluorescence Microscopy; Future; Genes; Genetic; Image Analysis; Immunocompromised Host; Immunofluorescence Immunologic; Implant; Individual; Knock-out; Knowledge; Lead; Lethal Genes; Malignant neoplasm of pancreas; Measures; Mediating; Mentorship; Modeling; Molecular; Mus; Organoids; Outcome; Paclitaxel; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Pharmacology; Phenotype; Proteins; Resources; Role; Stains; Structure; Survival Rate; Testing; Therapeutic; Tight Junctions; Time; Tissues; Tumor Subtype; arm; cancer cell; chemotherapy; combinatorial; gemcitabine; improved outcome; infiltrating duct carcinoma; loss of function; molecular subtypes; novel; patient derived xenograft model; phosphoproteomics; rational design; response; screening; subcutaneous; targeted agent; targeted treatment; therapeutic development; transcriptomics; treatment arm; treatment comparison; treatment strategy; tumor; tumor growth

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with a dismal five-year survival rate of 9% even with current treatments. There are two molecular tumor subtypes of PDAC; basal-like PDAC is more aggressive and associated with shorter survival times. Recent clinical trials have found that basal-like PDAC patients fail to respond to first-line chemotherapies including FOLFIRINOX, so it is critical to find better treatment options for this subtype. To study PDAC subtypes, organoids are utilized as they have been shown to represent both molecular subtypes, unlike conventional 2D cell lines. First, this study aims to identify genes whose depletion increases basal-like PDAC sensitivity to FOLFIRINOX using a CRISPR/Cas9 loss-of-function screen targeting 395 basal-like marker genes in basal-like PDAC organoids. The synthetic lethality hits may be studied in the future as drug targets for use in combination with FOLFIRINOX as a more efficacious therapeutic strategy for basal-like PDAC. To study the biology of basal-like PDAC, I will investigate the role of dysregulation of cell polarity in the PDAC subtypes using organoid models. Preliminary data suggest that basal-like PDAC is characterized by a loss of cell polarity while classical PDAC maintains it. Basal-like organoids do not organize to form hollow lumens like classical organoids and the most differentially phosphorylated proteins and pathways between subtypes involve cell adhesion and cell polarity. Dysregulation of cell polarity may increase cancer cell invasion, potentially contributing to poor outcomes in basal-like PDAC. Using fluorescence microscopy, this study will determine whether the localization of cell polarity markers is altered in basal-like PDAC compared to classical in a manner consistent with a loss of cell polarity. It will also be tested whether genetic depletion of cell polarity genes results in a shift from classical to basal-like phenotype, as measured by loss of organoid lumens and increased invasion. Understanding this facet of subtype biology could lead to future therapeutic strategies leveraging cell polarity to convert basal-like PDAC to a more classical phenotype, as classical PDAC is more responsive to current treatments. Overall, this project will advance our understanding of the biology underlying PDAC subtypes and move toward the development of therapeutic strategies for basal-like PDAC. This project will be supported by the extensive resources of the UNC Chapel Hill and the Department of Pharmacology as well as by co-mentorship by Drs. Jen Jen Yeh and Gaorav Gupta.

抽象的 胰腺导管腺癌 (PDAC) 是一种致命疾病，五年生存率很低 即使采用目前的治疗方法，仍为 9%。 PDAC 有两种分子肿瘤亚型；类似基底的 PDAC 更 具有攻击性并与较短的生存时间相关。最近的临床试验发现，basal-like PDAC 患者对包括 FOLFIRINOX 在内的一线化疗没有反应，因此寻找更好的药物至关重要 该亚型的治疗方案。为了研究 PDAC 亚型，使用了已证实的类器官 与传统的二维细胞系不同，代表两种分子亚型。首先，本研究旨在鉴定基因 使用 CRISPR/Cas9 功能丧失，其消耗增加了对 FOLFIRINOX 的类基础 PDAC 敏感性 针对 basal 样 PDAC 类器官中 395 个 basal 样标记基因的筛选。合成杀伤力可能是 未来将作为药物靶点进行研究，与 FOLFIRINOX 联合使用作为更有效的治疗方法 类似 basal PDAC 的策略。为了研究 basal-like PDAC 的生物学，我将研究 使用类器官模型研究 PDAC 亚型中细胞极性的失调。初步数据表明 基底样 PDAC 的特点是细胞极性丧失，而经典 PDAC 则保持细胞极性。基底样 类器官不会像经典类器官那样组织形成中空管腔，并且差异最大 磷酸化蛋白质和亚型之间的通路涉及细胞粘附和细胞极性。失调 细胞极性的改变可能会增加癌细胞的侵袭，可能导致基底样 PDAC 的不良结果。 使用荧光显微镜，本研究将确定细胞极性标记的定位是否是 与经典 PDAC 相比，基底样 PDAC 发生了改变，其方式与细胞极性的丧失一致。它还将 测试细胞极性基因的遗传耗竭是否会导致从经典型向基底样型的转变 表型，通过类器官管腔损失和侵袭增加来衡量。了解这个方面 亚型生物学可能会导致未来的治疗策略利用细胞极性将基底样 PDAC 转化为 更经典的表型，因为经典 PDAC 对当前治疗更敏感。总体来说，这个项目 将增进我们对 PDAC 亚型生物学基础的理解，并朝着开发 基础样 PDAC 的治疗策略。该项目将得到广泛资源的支持 北卡罗来纳大学教堂山分校和药理学系以及博士的共同指导。叶珍珍和 高拉夫·古普塔.