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: 10041399
• 负责人: Christina Ferrer
• 金额: $ 15.63万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2022-07-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10041399
• 关键词: Address; Affect; Agar; Anchorage-Independent Growth; Automobile Driving; Biological Assay; Blood Circulation; Breast; Breast cancer metastasis; Cancer Etiology; Cell Line; Cells; Cessation of life; Characteristics; DNA Sequence Alteration; Data; Development; Diagnosis; Disease; Disease Progression; Distal; Doxycycline; Future; Gene Expression Profile; Genes; Genetically Engineered Mouse; Glutathione S-Transferase; Growth; Human; Hydrophobicity; In Vitro; Incidence; Lesion; Liver; Maintenance; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Metastatic Adenocarcinoma; Metastatic to; Modality; Modeling; Molecular; Neoplasm Metastasis; Outcome; Pancreas; Pancreatic Adenocarcinoma; Pancreatic Ductal Adenocarcinoma; Patient-Focused Outcomes; Patients; Pattern; Play; Population; 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; base; cancer cell; cell growth; differential expression; efficacy testing; experimental study; human tissue; in vivo; inhibitor/antagonist; insight; knock-down; lung metastatic; malignant breast neoplasm; mortality; mouse model; non-genetic; novel; pancreatic cancer patients; 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.

项目概要 只有 0.01% 的癌细胞进入循环、存活并产生转移，然而，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 存在胰腺外侵袭和转移性疾病，其五年生存率很低 率，并且没有针对转移性疾病治疗的具体治疗策略。值得注意的是，大部分 对转移机制的研究重点是确定原发肿瘤内的早期驱动因素， 然而，对已形成病变内转移生长的主要驱动因素的识别在很大程度上仍然存在 未经探索。最近，我们证明 SIRT6 失活可显着加速 PDA 发育，导致 Kras-p53 GEM 模型中高度侵袭性的转移性疾病。使用这个 高度侵袭性的PDA转移模型，除了已建立的乳腺癌转移模型外，这 研究旨在确定转移性病变的潜在脆弱性，这些脆弱性可用于治疗患有癌症的患者 晚期转移性疾病。对匹配的原发组织和转移组织进行无偏 RNA 测序 随后通过新开发的 96 孔软琼脂筛选，我们确定了独特所需的因素 已建立的转移细胞的贴壁独立生长。利用此功能屏幕和验证，我们 已确定 Gstt1（谷胱甘肽 S 转移酶 theta 1）是转移性生长的首要候选驱动因素 多种小鼠转移模型。初步数据表明 Gstt1 在 与匹配的原代细胞系相比，转移细胞系和 Gstt1 的抑制显着降低 转移衍生细胞的转移潜力，而不影响原发肿瘤的生长，这表明重要的 Gstt1 在已形成的转移性病变的生长中的作用。此外，在转移性病灶内，Gstt1 显示异质表达模式，其中 Gstt1high 细胞代表一种侵袭性、非增殖性亚细胞 人口。此外，我们还证明 Gstt1 是乳腺肿瘤球形成所必需的 和体外 PDA 转移衍生细胞系，表明 Gstt1 在转移性肿瘤中作为自我更新驱动因素的作用 细胞。在本提案中，我们试图确定 Gstt1high PDA 衍生的转移性病变的独特特征。这 拟议的研究将为 Gstt1 及其下游靶标是否存在提供重要的临床前证明 转移性肿瘤持续生长所必需的，从而确定了可能的治疗窗口 转移性 PDA 的子集。