High dose acetaminophen with n-acetylcysteine rescue as a novel STAT3 inhibitor with anti-cancer stem cell properties

高剂量对乙酰氨基酚与 n-乙酰半胱氨酸救援作为具有抗癌干细胞特性的新型 STAT3 抑制剂

• 批准号: 10368472
• 负责人: Alexander Neuwelt
• 金额: --
• 依托单位: VA VETERANS ADMINISTRATION HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-10-01 至 2026-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10368472
• 关键词: Acetaminophen; Acetylcysteine; Address; Americas; Analgesics; Antidotes; Award; Binding; Biological Assay; Cancer Model; Cancer Patient; Cancer Relapse; Caring; Cells; Chemoresistance; Cisplatin; Clinical; Clinical Research; Collaborations; Computer software; Data; Disease; Doctor of Philosophy; Dominant-Negative Mutation; Dose; Drug Targeting; Drug usage; Enrollment; Epidermal Growth Factor Receptor; Exhibits; Foundations; Free Radicals; Funding; Gene Proteins; Genes; Genetic Transcription; Glutathione; Goals; Head; Hepatoblastoma; Hepatotoxicity; Human; Hydrogen Bonding; In Vitro; In complete remission; Injury; K-Series Research Career Programs; Knock-out; Knowledge; Laboratory Research; Lead; Length; Liver; Maintenance; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Mentors; Methods; Mitochondria; Modeling; Molecular; Mus; Natural regeneration; Non-Small-Cell Lung Carcinoma; Overdose; Patient Selection; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phenotype; Phosphorylation; Physicians; Physiology; Play; Population; Process; Prognosis; Property; Proteins; Regulation; Research; Research Personnel; Resistance; Resources; Role; STAT3 gene; Scientist; Series; Signal Transduction; Site-Directed Mutagenesis; Stat3 protein; Testing; Time; Toxic effect; Tumorigenicity; Universities; Virginia; Work; Xenograft procedure; anti-cancer; anticancer activity; antitumor effect; base; cancer cell; cancer stem cell; career; chemotherapy; driver mutation; early phase clinical trial; experience; experimental study; in silico; in vivo; inhibitor; insight; lung cancer cell; neoplastic cell; novel; patient derived xenograft model; pre-clinical; predictive marker; response; self-renewal; skills; src Homology Region 2 Domain; stem cell biology; stem cell biomarkers; stem cell growth; stem cell population; symposium; synergism; targeted treatment; tumor; tumor growth; tumor microenvironment; vector

High dose acetaminophen (AAP) with delayed n-acetylcysteine (NAC) rescue has shown promise in early phase clinical trials, inducing disease shrinkage in 8/14 and an objective response in 3/14 assessable patients with diverse tumor types. However, the mechanism of anti-tumor activity of high dose AAP is not well understood. It was previously felt that high dose AAP has anti-tumor activity via similar processes as its mechanism of hepatotoxicity, namely free radical injury and glutathione depletion. However, pre-clinical and clinical evidence suggests that glutathione is not depleted within the tumor in response to high-dose AAP treatment. This suggests an alternate mechanism of anti-tumor activity of high dose AAP. In this proposal, we outline preliminary data suggesting that: 1.) High dose AAP has anti-cancer stem cell (CSC) activity and 2.) High dose AAP functions as a novel inhibitor of STAT3 phosphorylation. STAT3 is a protein that has previously been shown to play an important role in the maintenance and proliferation of a viable CSC population. We hypothesize that the anti-CSC activity in non-small cell lung cancer (NSCLC) of high dose AAP is via a STAT3 dependent mechanism. In specific aim #1, sub-aim A, we propose to evaluate the mechanism of STAT3 inhibition by AAP. We have obtained preliminary data suggesting that AAP binds STAT3 in the SH2 domain via a strong hydrogen bond formed at Lys626. We aim to validate these preliminary findings using orthogonal approaches such as microscale thermopheresis (MST). These studies will be performed in collaboration with co-mentor Umesh Desai, PhD, head of Medicinal Chemistry at Virginia Commonwealth University (VCU). In sub-aim B, we evaluate the mechanism of the anti-CSC activity of high dose AAP. We will generate constructs of human NSCLC with altered STAT3 proteins: empty vector (STAT3 KO), STAT3 K626K (WT) and STAT3 K626A (predicted to have decreased AAP-binding). We will determine the sensitivity of STAT3-altered constructs to the anti-CSC effects of AAP or AAP + NAC. If human NSCLC cells with STAT3 K626A have decreased responsiveness to the anti-CSC effects of high dose AAP, this would suggest that the anti-CSC effects of AAP are mediated by direct binding to STAT3. In specific aim #2, we will evaluate the mechanisms of synergy between chemotherapy (in NSCLC without targetable driver mutation) and targeted therapy (in NSCLC with EGFR or ALK driver alteration) with AAP, both in vitro and in vivo. We will investigate whether AAP-enhances chemotherapy/targeted therapy anti-tumor activity via a a STAT3 dependent mechanism. In this CDA application, Dr. Neuwelt proposes to work with an outstanding group of established scientists whom have the experience and expertise necessary to mentor Dr. Neuwelt on the proposed project. Dr. Neuwelt’s primary mentor will be Dr. Patel, a senior VA-merit funded researcher with expertise in cancer stem cell biology. Dr. Larner, a co-mentor on the project, discovered the role of STAT3 in mitochondrial function and has dedicated much of his career to understanding STAT3 physiology. If awarded a CDA, Dr. Neuwelt will enroll in relevant classes at local Universities, attend scientific conferences, take care of VA-based patients, and grow his research lab, all with the goal of becoming an independent VA-based physician scientist.

具有延迟的N-乙酰半胱氨酸（NAC）救援的高剂量对乙酰氨基酚（AAP）在早期表现出希望 阶段临床试验，诱导8/14的疾病收缩，3/14可评估患者的客观反应 具有潜水员肿瘤类型。但是，高剂量AAP的抗肿瘤活性的机制不好 理解。以前觉得高剂量AAP通过与其类似的过程具有抗肿瘤活性 肝毒性的机制，即自由基损伤和谷胱甘肽耗竭。但是，临床前和 临床证据表明，谷胱甘肽在肿瘤中没有耗尽大剂量AAP 治疗。这表明高剂量AAP抗肿瘤活性的另一种机制。在这个建议中，我们 概述初步数据表明：1。1.）高剂量AAP具有抗癌干细胞（CSC）活性和2。） 高剂量AAP充当STAT3磷酸化的新型抑制剂。 STAT3是一种蛋白质 以前被证明在可行的CSC的维持和增殖中起着重要作用 人口。我们假设高剂量AAP非小细胞肺癌（NSCLC）的抗CSC活性 是通过STAT3依赖机制。在特定的目标＃1（sub-aim a）中，我们建议评估机制 AAP抑制STAT3。我们获得了初步数据，表明AAP在SH2中结合了STAT3 通过在Lys626形成的强氢键通过域。我们旨在使用 诸如微观热过渡（MST）之类的正交方法。这些研究将在 与弗吉尼亚联邦药物化学负责人的Co-Imesh Desai合作 大学（VCU）。在Sub-aim B中，我们评估了高剂量AAP的抗CSC活性的机制。我们 将使用改变的STAT3蛋白质生成人NSCLC的构建体：空矢量（STAT3 KO），STAT3 K626K（WT）和STAT3 K626a（预计已减少了AAP结合）。我们将确定灵敏度 AAP或AAP + NAC的抗CSC效应的STAT3改变构建体的构造。如果具有STAT3的人NSCLC细胞 K626A对高剂量AAP的抗CSC效应的反应性降低了，这表明 AAP的抗CSC效应是通过直接结合与STAT3介导的。在特定的目标＃2中，我们将评估 化学疗法之间的协同作用机制（在没有靶向驱动器突变的NSCLC中）并针对性 在体外和体内的AAP中，治疗（在具有EGFR或ALK驱动器改变的NSCLC中）。我们将调查 AAP-Enhances化学疗法/靶向治疗是否通过STAT3依赖 机制。在此CDA应用程序中，Neuwelt博士的提议与一群未成年人合作 科学家有经验和专业知识所需的经验和专业知识，可以指导Neuwelt博士对拟议项目进行指导。 Neuwelt博士的主要导师将是VA-Merit资助的癌症专业知识的高级资助研究人员Patel博士 干细胞生物学。该项目的院长Larner博士发现了STAT3在线粒体中的作用 功能，并将他的大部分时间都致力于理解STAT3生理学。如果被授予CDA，博士 Neuwelt将参加当地大学的相关课程，参加科学会议，照顾基于VA 患者，并成长他的研究实验室，目的是成为独立的基于VA的物理学 科学家。