Project 2: Targeting signaling networks to overcome therapeutic resistance in pancreatic cancer

项目 2：靶向信号网络克服胰腺癌的治疗耐药性

• 批准号: 10629064
• 负责人: TONY R. HUNTER
• 金额: $ 49.76万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-06 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629064
• 关键词: Acceleration; Affect; Antibodies; Automobile Driving; Autophagocytosis; Blocking Antibodies; CA-19-9 Antigen; Cell Compartmentation; Cell Proliferation; Cell membrane; Cells; Chemoresistance; Clinical; Coculture Techniques; Combined Modality Therapy; Common Carcinoma; Complex; Cytokine Signaling; Data; Diagnosis; Disease; Disease Resistance; Epidermal Growth Factor Receptor; Epithelial Cells; Epithelium; Epitopes; Family; Fibroblasts; Genes; Genetic; Genetically Engineered Mouse; Growth; Heterogeneity; Human; IL6 gene; Immune; Immune checkpoint inhibitor; Intercept; KRAS2 gene; Kinetics; LIF gene; Malignant Neoplasms; Malignant neoplasm of pancreas; Maps; Mediating; Mediator; Methodology; Methods; Modeling; Molecular; Mus; Mutation; Organoids; Pancreas; Pancreatic Ductal Adenocarcinoma; Paracrine Communication; Pathway interactions; Patients; Phenotype; Play; Polysaccharides; Post-Translational Protein Processing; Production; Proteins; Receptor Protein-Tyrosine Kinases; Refractory; Residual state; Resistance; Resistance development; Resolution; Role; STAT3 gene; Signal Pathway; Signal Transduction; Specimen; Stromal Cells; TP53 gene; Therapeutic; Time; Tissues; Tumor Promotion; Work; biobank; cancer cell; cell type; checkpoint inhibition; chemotherapy; conventional therapy; cytokine; experimental study; glycosylation; in vivo; leukemia inhibitory factor receptor; loss of function; mouse model; neoplastic cell; novel; organoid transplantation; pancreatic ductal adenocarcinoma model; pancreatic tumorigenesis; paracrine; pharmacologic; preempt; response; success; targeted treatment; therapy resistant; treatment response; treatment strategy; tumor; tumor growth; tumor microenvironment; tumor-immune system interactions; tumorigenesis

PROJECT SUMMARY – Project 2: Signaling Pancreatic ductal adenocarcinoma (PDA) is most commonly diagnosed at late stages, at which time the disease is refractory to most conventional treatment options. Identifying therapeutic strategies that pre-empt the development of resistance and achieve durable tumor control has been hampered by the enormous complexity of the paracrine signaling network in PDA. STAT3 is a major node in this resistance network. The Hunter lab has shown that production of leukemia inhibitory factor (LIF) by cancer associated fibroblasts (CAFs) is a major driver of STAT3 signaling in tumor cells and promotes chemoresistance. Further, they have shown that LIF and the related cytokines IL6 and OSM may also play a role in STAT3 signaling in the immune microenvironment (IME). In Aim 1, the team will investigate the extent to which induction of STAT3 signaling by these cytokines contributes to tumor growth and therapeutic resistance. High-resolution spatial profiling and single-cell approaches will be used to delineate the cell type-specific alterations in STAT3 activation using syngeneic orthotopic models of PDA following treatment with chemotherapy and blockade of LIF, IL6, and/or OSM, which will be corroborated with the colocalization of cytokines and STAT3 signaling in human patient specimens. As LIF can activate several pro-survival pathways, the importance of STAT3-dependent and -independent mechanisms to PDA growth and chemoresistance will be determined. In addition to chemotherapy, the ability of cytokine blockade to potentiate immune checkpoint inhibition (ICI) will be investigated. Further, the team has discovered that aberrant glycosylation can also modulate the STAT3 pathway. Elevation of the glycan CA19-9 in organoids and in vivo resulted in increased STAT3 signaling in both the epithelial and stromal compartments. In Aim 2, the mechanism by which CA19-9 elevation activates STAT3 will be determined and the role of this signaling pathway in chemoresistance will be dissected in a cell type-specific manner using a novel organoid co-culture platform and a first of its kind genetically engineered mouse model with pancreas-specific KRAS and TRP53 mutations and inducible CA19-9 expression. In addition to STAT3, CA19-9 elevation promotes receptor tyrosine kinase signaling, including activation of the EGFR pathway, which also drives chemoresistance. However, inhibition of either pathway has yielded limited clinical success. The team’s preliminary studies have revealed correlations between elevated CA19-9, resistance to chemotherapy, and induction of autophagy, a pathway known to mediate resistance to both chemotherapeutics and targeted therapies. Therefore, experiments proposed in Aim 3 will systematically dissect how CA19-9 modulation of STAT3, EGFR, and autophagy contribute to therapeutic resistance. It is important to note that most previous mouse studies on these resistance pathways have been performed in the absence of CA19-9 and that >90% of PDA patients are CA19-9 positive. Advanced organoid methodologies will be used to investigate both genetic and pharmacologic perturbation of these pathways, culminating in in vivo explorations of the efficacies of combination treatment strategies in CA19-9-positive PDA.

项目概要-项目2：信号 胰腺导管腺癌（PDA）最常在晚期诊断，此时疾病 对大多数常规治疗方案是难治的。确定预先预防的治疗策略 发展耐药性和实现持久的肿瘤控制受到巨大复杂性的阻碍， 旁分泌信号网络。STAT 3是该电阻网络中的主要节点。亨特实验室 显示由癌症相关成纤维细胞（CAF）产生的白血病抑制因子（LIF）是一个主要驱动因素， STAT 3信号在肿瘤细胞和促进化疗耐药性。此外，他们还表明，LIF和 相关的细胞因子IL 6和OSM也可能在免疫微环境（IME）中的STAT 3信号传导中起作用。 在目标1中，研究小组将研究这些细胞因子诱导STAT 3信号传导的程度， 与肿瘤生长和治疗耐药性有关。高分辨率空间剖面和单细胞方法将是 用于使用PDA的同基因原位模型描绘STAT 3活化中的细胞类型特异性改变 在用化疗和LIF、IL 6和/或OSM的阻断治疗后，这将用 细胞因子和STAT 3信号在人类患者标本中的共定位。由于LIF可以激活多个 促生存途径，STAT 3依赖和非依赖机制对PDA生长的重要性， 将测定化学抗性。除化疗外，细胞因子阻断增强 将研究免疫检查点抑制（ICI）。此外，研究小组还发现， 糖基化也可以调节STAT 3途径。类器官和体内聚糖CA 19 -9的升高 导致上皮和基质区室中STAT 3信号传导增加。在目标2中， CA 19 -9升高激活STAT 3的信号通路，以及该信号通路在 将使用新型类器官共培养平台以细胞类型特异性方式剖析化学抗性， 第一个具有胰腺特异性KRAS和TRP 53突变的基因工程小鼠模型， 可诱导CA 19 -9表达。除了STAT 3，CA 19 -9升高促进受体酪氨酸激酶 信号传导，包括EGFR途径的激活，其也驱动化学抗性。然而，抑制 两种途径都产生了有限的临床成功。该团队的初步研究揭示了 CA 19 -9升高、化疗耐药和诱导自噬之间的关系，自噬是一种已知的 介导对化疗药物和靶向治疗的抗性。因此，Aim中提出的实验 3将系统地剖析CA 19 -9对STAT 3，EGFR和自噬的调节如何有助于治疗 阻力值得注意的是，大多数以前对这些耐药途径的小鼠研究都是 在不存在CA 19 -9的情况下进行，并且>90%的PDA患者是CA 19 -9阳性。高级类器官 方法学将用于研究这些途径的遗传和药理学扰动， 最终在体内探索联合治疗策略在CA 19 -9阳性PDA中的功效。