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 只有0.01%的癌细胞进入循环、存活并产生转移，但转移性疾病 占癌症死亡人数的90%在胰腺导管腺癌（PDA）中，大多数患者 存在胰腺外浸润和转移性疾病，其5年生存率很低 率和针对转移性疾病的治疗没有特定的治疗策略。值得注意的是， 对转移机制的研究集中在鉴别原发性肿瘤内的早期驱动因素， 然而，在已建立的病灶内转移性生长的中心驱动因素的识别在很大程度上仍然存在， 未开发的最近，我们已经证明SIRT 6失活显著加速了PDA， 发展，导致Kras-p53 GEM模型中的高度侵袭性转移性疾病。采用这种 高度侵袭性PDA转移模型，除了已建立的乳腺癌转移模型之外， 一项旨在确定转移性病变的潜在脆弱性的研究，可用于治疗 晚期转移性疾病对匹配的原发性和转移性组织进行无偏倚的RNA-Seq 随后通过新开发的96孔软琼脂筛选，我们确定了 已建立的转移性细胞的贴壁非依赖性生长。利用此功能筛选和验证，我们 已经确定Gstt 1（谷胱甘肽S-转移酶θ 1）是转移性生长的最佳候选驱动因素， 转移的多种小鼠模型。初步数据表明，Gstt 1的差异表达， 与匹配的原代衍生细胞系相比，转移细胞系和Gstt 1抑制显著降低 转移潜能的肿瘤细胞，而不影响原发性肿瘤的生长，这表明一个重要的 Gstt 1在已建立的转移性病变的生长中的作用。此外，在转移性病变中，Gstt 1 显示异质性表达模式，其中Gstt 1high细胞代表侵袭性、非增殖性亚细胞， 人口此外，我们已经证明Gstt 1是乳腺癌肿瘤球形成所必需的。 和PDA转移衍生的细胞系，表明Gstt 1在转移性肿瘤中作为自我更新的驱动因素的作用。 细胞在这个建议中，我们试图确定独特的Gstt 1high PDA衍生的转移性病变的特点。的 拟议的研究将提供重要的临床前证明Gstt 1及其下游靶点是否 是转移性肿瘤持续生长所必需的，因此确定了一个可能的治疗窗口。 转移性PDA的子集。