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，五年生存率为惨淡 即使目前的治疗也9％。 PDAC有两个分子肿瘤亚型；基底pDAC更多 积极进取，与较短的生存时间有关。最近的临床试验发现基底样PDAC 患者无法对包括Folfirinox在内的一线化学疗法做出反应，因此至关重要 该亚型的治疗选择。为了研究PDAC亚型，使用了类器官，因为它们已显示为 与常规的2D细胞系不同，代表这两种分子亚型。首先，本研究旨在识别基因 使用CRISPR/CAS9的功能丧失 筛网靶向基底样PDAC类器官中的395个基础标记基因。合成的致命命中可能是 将来被研究为与Folfirinox结合使用的药物靶标作为一种更有效的治疗 基底PDAC的策略。为了研究基底样PDAC的生物学，我将研究 使用器官模型的PDAC亚型中细胞极性的失调。初步数据表明 基底样PDAC的特征是细胞极性丧失，而经典PDAC则保持它。基础状 类器官不会组织形成像经典类正弦和最差异的空心流明 亚型之间的磷酸化蛋白质和途径涉及细胞粘附和细胞极性。失调 细胞极性可能会增加癌细胞侵袭，这可能导致基底样PDAC的结果不佳。 使用荧光显微镜，这项研究将确定细胞极性标记的定位是否为 与经典相比，基底样PDAC的改变与细胞极性丧失相比。它也会 可以测试细胞极性基因的遗传耗竭是否导致从经典到基础样的转变 表型，通过损失器官流明和增加的浸润来衡量。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 一个更古典的表型，因为经典PDAC对当前治疗的反应更快。总体而言，这个项目 将促进我们对PDAC子类型生物学的理解，并朝着发展 基底样PDAC的治疗策略。该项目将得到该项目的支持 UNC Chapel Hill和药理学系以及博士的授权。 Jen Jen Yeh和 Gaorav Gupta。