Role of Gstt1 in metastatic maintenance and self-renewal in PDA

Gstt1 在 PDA 转移维持和自我更新中的作用

• 批准号: 10704159
• 负责人: Christina Ferrer
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 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 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10704159
• 关键词: Acceleration; Address; Affect; Agar; Anchorage-Independent Growth; Automobile Driving; Biological Assay; Breast; Breast cancer metastasis; Cancer Etiology; Cell Line; Cells; Cessation of life; Characteristics; Circulation; DNA Sequence Alteration; Data; Development; Diagnosis; Disease; Disease Progression; Distal; Doxycycline; Early identification; Future; Gene Expression Profile; Genes; Genetically Engineered Mouse; Glutathione S Transferase A; Glutathione S-Transferase; Glutathione Transferase Inhibitor; Growth; Human; Hydrophobicity; In Vitro; Incidence; Invaded; Lesion; Liver; Maintenance; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Metastatic Adenocarcinoma; Modality; Modeling; Molecular; Neoplasm Metastasis; Outcome; Pancreas; Pancreatic Adenocarcinoma; Pancreatic Ductal Adenocarcinoma; Patient-Focused Outcomes; Patients; Pattern; Play; Primary Neoplasm; Proteins; Reduced Glutathione; Research; Role; Solid Neoplasm; Specimen; Stains; Survival Rate; System; TP53 gene; Testing; Therapeutic; Tissue-Specific Gene Expression; Tissues; Validation; Work; cancer cell; cell growth; differential expression; efficacy testing; experimental study; human tissue; in vivo; inhibitor; insight; knock-down; lung metastatic; malignant breast neoplasm; mortality; mouse model; non-genetic; novel; pancreatic cancer patients; pancreatic ductal adenocarcinoma model; pre-clinical; self-renewal; small hairpin RNA; small molecule inhibitor; therapeutic target; transcriptome sequencing; tumor; tumor growth

PROJECT SUMMARY Only 0.01% of cancer cells enter circulation, survive & produce metastasis, however, metastatic disease accounts for 90% of cancer-related deaths. In pancreatic ductal adenocarcinoma (PDA), the majority of patients present with extra-pancreatic invasion and metastatic disease for which the there is a dismal five-year survival rate and no specific therapeutic strategies directed at the treatment of metastatic disease. Notably, much of the research into the mechanisms of metastasis has been focused on identifying early drivers within primary tumors, however, the identification of central drivers of metastatic outgrowth within established lesions largely remain unexplored. Recently, we have demonstrated that SIRT6 inactivation dramatically accelerates PDA development, resulting in highly aggressive metastatic disease in the Kras-p53 GEM model. Employing this highly aggressive PDA metastasis model, in addition to an established breast cancer metastasis model, this study aims to identify potential vulnerabilities of metastatic lesions that could be exploited to treat patients with advanced metastatic disease. Performing unbiased RNA-Seq on matched primary and metastatic tissues followed by a newly developed 96-well soft agar screen, we identified factors that are uniquely required for anchorage-independent growth of established metastatic cells. Utilizing this functional screen and validation, we have identified Gstt1 (glutathione S-transferase theta 1) as a top candidate driver of metastatic outgrowth in multiple mouse models of metastasis. Preliminary data demonstrates that Gstt1 is differentially expressed in metastatic cell lines compared to matched primary-derived cell lines and inhibition of Gstt1 significantly reduced metastatic potential of metastases-derived cells, without affecting primary tumor growth, suggesting an important role for Gstt1 in the outgrowth of established metastatic lesions. Additionally, within metastatic lesions, Gstt1 shows a heterogenous expression pattern, where Gstt1high cells represent an aggressive, non-proliferative sub- population. Additionally, we have demonstrated that Gstt1 is required for the formation of tumor spheres in breast and PDA metastatic-derived cell lines in vitro, suggesting a role for Gstt1 as a driver of self-renewal in metastatic cells. In this proposal we seek to identify characteristics unique to Gstt1high PDA-derived metastatic lesions. The proposed studies will provide important preclinical demonstration of whether Gstt1 and its downstream targets are required for sustained growth of metastatic tumors, thus identifying a possible therapeutic window for this subset of metastatic PDA.

项目摘要 然而，只有0.01％的癌细胞进入循环，生存并产生转移性疾病 占癌症相关死亡的90％。在胰腺导管腺癌（PDA）中，大多数患者 存在胰腺外入侵和转移性疾病，其中有五年的生存期很沮丧 率和没有针对转移性疾病治疗的特定治疗策略。值得注意的是，很多 对转移机制的研究一直集中在确定原发性肿瘤中的早期驱动因素， 但是，确定既定病变内转移性侵蚀的中央驱动因素在很大程度上仍然存在 未探索。最近，我们已经证明了SIRT6灭活急剧加速了PDA 开发，导致KRAS-P53宝石模型中高度侵略性的转移性疾病。采用这个 高度侵略性的PDA转移模型，除了建立的乳腺癌转移模型外， 研究旨在确定可以利用的转移性病变的潜在脆弱性，以治疗患者 晚期转移性疾病。在匹配的原始和转移组织上执行无偏的RNA-Seq 其次是新开发的96孔软琼脂屏幕，我们确定了这些因素 已建立的转移细胞独立于锚固的生长。利用此功能屏幕和验证，我们 已将GSTT1（谷胱甘肽S-转移酶Theta 1）确定为转移性产物的顶级候选驱动力 多种转移的小鼠模型。初步数据表明，GSTT1在 与匹配的原始细胞系相比，转移细胞系和GSTT1的抑制显着降低 转移酶衍生细胞的转移潜力，而不会影响原发性肿瘤的生长，这表明一个重要的 GSTT1在既定转移性病变的生长中的作用。此外，在转移性病变内GSTT1 显示出异质的表达模式，其中GSTT1高细胞代表了侵略性的，无增强的亚 - 人口。此外，我们已经证明了乳房中肿瘤球的形成需要GSTT1 和PDA转移性细胞系的体外，这表明GSTT1是自我更新的驱动因素的作用 细胞。在此提案中，我们试图确定GSTT1高PDA衍生的转移性病变所特有的特征。这 拟议的研究将为GSTT1及其下游目标提供重要的临床前证明 是转移性肿瘤持续生长所必需的，因此为此确定了可能的治疗窗口 转移性PDA的子集。