Targeting HNF1A-mediated therapeutic resistance in pancreatic ductal adenocarcinoma

靶向 HNF1A 介导的胰腺导管腺癌治疗耐药

• 批准号: 10561370
• 负责人: Ethan Vincent Abel
• 金额: $ 47.33万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-03 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10561370
• 关键词: Ablation; Affect; Apoptosis; BCL2 gene; Biological Assay; Biology; Bromodomain; Bromodomains and extra-terminal domain inhibitor; Cancer Etiology; Cell Cycle Progression; Cell Death; Cell Line; Cell Proliferation; Cell Survival; Cells; Cessation of life; ChIP-seq; Clinic; Clinical; Data; Drug Targeting; Drug resistance; Epigenetic Process; Foundations; Genes; Genetic Transcription; Goals; Growth; Human; In Vitro; KRAS2 gene; MAP Kinase Gene; MEKs; Maintenance; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Mutation; Nature; Oncogenes; Oncogenic; Organoids; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Process; Proliferating; Promoter Regions; Property; Proteins; Reader; Refractory; Regulation; Reporter; Resistance; Role; Signal Transduction; Survival Rate; Testing; Therapeutic; Treatment Protocols; Work; cancer cell; cancer stem cell; cancer type; cell growth; combinatorial; conventional therapy; effective therapy; efficacy evaluation; gastrointestinal; genetic approach; genetic manipulation; improved; improved outcome; in vivo; inhibitor; innovation; insight; knock-down; mutant; new therapeutic target; next generation; novel; outcome prediction; overexpression; pancreatic ductal adenocarcinoma cell; patient derived xenograft model; patient stratification; preclinical efficacy; programs; response; stem cell function; targeted treatment; therapy resistant; transcription factor; transcriptome; transcriptome sequencing; treatment response; tumor; tumorigenesis; tumorigenic

Pancreatic ductal adenocarcinoma (PDA) is one of the deadliest cancers with a survival rate of 11% due to its aggressive nature and resistance to therapies. While nearly all PDA cases are driven by mutations in the KRAS gene, efforts to target KRAS or its effectors (e.g., MEK and ERK) are met with adaptive resistance. Pancreatic cancer stem cells (PCSCs), a subpopulation of transcriptionally-plastic cancer cells that are especially drug resistant and particularly tumorigenic, are a critical component of the aggressive and therapy- resistant nature of PDA. There are currently no strategies to target PCSCs, as well as a lack of information regarding their drivers. We previously identified HNF1A, a gastrointestinal-lineage transcription factor, as a novel master regulator of the PCSC state. Our preliminary data show that HNF1A expression can be potently blocked by BET-inhibitors (BETi), a class of drugs which inhibit the epigenetic reader protein BRD4. Interestingly, re-expression of HNF1A rescues cell cycle progression and PCSC-properties in BETi-treated PDA cells, suggesting that HNF1A is a novel and critical target for these drugs. We have also found that HNF1A is a novel driver of resistance to targeting KRAS and downstream MEK/ERK-signaling. Importantly, the use of BETi in combination with MEK- and ERK-inhibitors (MEKi/ERKi) increases growth arrest and cell death in an HNF1A-dependent manner. We hypothesize that HNF1A is directly regulated by BRD4, and is therefore targetable with BETi, and that the inhibition of HNF1A expression with BETi will nullify HNF1A-dependent PCSCs and adaptive resistance to KRAS-ablation. In this proposal, we aim to characterize the regulation of HNF1A and its role in therapeutic response and resistance. In Specific Aim 1, we will characterize the regulation of HNF1A by BETi-target BRD4. Specific Aim 1 will combine genetic manipulation of BRD4, ChIP- PCR, and reporter assays to demonstrate regulation of HNF1A by BRD4. Re-expression of HNF1A in BRD4- depleted cells will 1) define the role of HNF1A in BRD4-mediated cell growth and survival in vitro and in vivo, as well as 2) determine the contribution of HNF1A to the BRD4 transcriptome using RNA-seq/ChIP-seq to identify how HNF1A determines response to BETi. In Specific Aim 2, we will establish BETi as a means to overcome HNF1A-mediated resistance to KRAS-ablation. Specific Aim 2 will use PDA cells with and without ectopic HNF1A expression to examine the contribution of HNF1A to BETi and KRAS-pathway inhibitor activity; we will utilize in vitro and in vivo assays to examine how these drugs affect PCSCs and use RNA-seq/ChIP- seq to determine how HNF1A promotes resistance to targeting KRAS-signaling. Both aims will utilize next- generation bromodomain-selective inhibitors that have not been explored in PDA or combinatorial therapies. The completion of the above studies will dramatically improve our understanding of PDA biology and uncover novel therapeutic targets. By improving our understanding of resistance to KRAS suppression and expanding the therapeutic spectrum of PDA to include PCSCs, more effective treatment of PDA can be achieved.

胰腺导管腺癌（PDA）是最致命的癌症之一，由于其具有高的生存率， 攻击性和对治疗的抵抗力。虽然几乎所有的PDA病例都是由 KRAS基因，靶向KRAS或其效应子的努力（例如，MEK和ERK）遇到适应性抗性。 胰腺癌干细胞（PCSC）是一种转录可塑性癌细胞亚群， 特别是耐药性和特别是致瘤性，是侵袭性和治疗的关键组成部分， PDA的抵抗力。目前没有针对PCSC的策略，也缺乏信息 关于他们的司机。我们先前鉴定了HNF 1A，一种胃肠道谱系转录因子， 新型PCSC状态主调节器。我们的初步数据表明，HNF 1A表达可以有效地 被BET抑制剂（BETi）阻断，BET抑制剂是一类抑制表观遗传阅读蛋白BRD 4的药物。 有趣的是，HNF 1A的重新表达挽救了BETi处理的细胞中的细胞周期进展和PCSC性质。 PDA细胞，这表明HNF 1A是这些药物的一个新的关键靶点。我们还发现 HNF 1A是对靶向KRAS和下游MEK/ERK信号传导的抗性的新驱动因素。重要的是 BETi与MEK和ERK抑制剂（MEKi/ERKi）组合使用增加生长停滞和细胞死亡 HNF 1A依赖性。我们假设HNF 1A直接受BRD 4调节，因此， 用BETi靶向，并且用BETi抑制HNF 1A表达将使HNF 1A依赖性 PCSC和对KRAS消融的适应性抵抗。在本提案中，我们的目标是描述 HNF 1A及其在治疗反应和抗性中的作用。在具体目标1中，我们将描述 通过BETi靶向BRD 4调节HNF 1A。特异性目标1将联合收割机结合BRD 4、ChIP- PCR和报告基因测定以证明BRD 4对HNF 1A的调节。HNF 1A在BRD 4中的再表达- 耗尽的细胞将1）确定HNF 1A在体外和体内BRD 4介导的细胞生长和存活中的作用， 以及2）使用RNA-seq/ChIP-seq确定HNF 1A对BRD 4转录组的贡献， 确定HNF 1A如何确定对BETi的响应。在具体目标2中，我们将建立BETi作为一种手段， 克服HNF 1A介导的对KRAS消融的抗性。具体目标2将使用PDA细胞， 异位HNF 1A表达以检查HNF 1A对BETi和KRAS途径抑制剂活性的贡献; 我们将利用体外和体内试验来检查这些药物如何影响PCSC，并使用RNA-seq/ChIP- seq以确定HNF 1A如何促进对靶向KRAS信号传导的抗性。两个目标都将利用下一个- 因此，本发明提供了新一代溴结构域选择性抑制剂，其尚未在PDA或组合疗法中探索。 上述研究的完成将大大提高我们对PDA生物学的理解，并揭示 新的治疗靶点。通过提高我们对KRAS抑制的抗性的理解， PDA的治疗谱包括PCSC，可以实现更有效的PDA治疗。