Glutaredoxin, Glutathione Metabolism and Lung Cancer

谷氧还蛋白、谷胱甘肽代谢与肺癌

• 批准号: 10657945
• 负责人: Yvonne M. W. Janssen-Heininger
• 金额: $ 53.51万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-10 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657945
• 关键词: Ablation; Address; Adenoviruses; Amino Acids; Antioxidants; Attenuated; Binding; Biochemistry; Breeding; Cessation of life; Cisplatin; Clinical; Cysteine; Cystine; Dependovirus; Development; Enterobacteria phage P1 Cre recombinase; Environment; Glutamates; Glutathione; Glutathione Metabolism Pathway; Growth; Homeostasis; Human; Immune checkpoint inhibitor; In Vitro; KRAS2 gene; KRASG12D; Link; Lung Adenocarcinoma; Lung Neoplasms; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Metabolic; Metabolism; Mus; NADP; Oncogenic; Organoids; Oxidants; Oxidation-Reduction; Oxidative Stress; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Property; Protein S; Proteins; Proteomics; Refractory; Regimen; Resistance; Role; Signal Transduction; Structure; Sulfhydryl Compounds; System; Testing; Therapeutic; Time; Tissues; Treatment Efficacy; Tumor Burden; Ubiquitination; Xenograft procedure; antiporter; combat; glutaredoxin; glutaredoxin 2; human subject; insight; lung cancer cell; lung tumorigenesis; mutant; neoplastic cell; novel therapeutic intervention; ovarian neoplasm; penicillamine-glutathione mixed disulfide; preclinical development; prevent; side effect; standard of care; tumor; tumor growth; tumorigenesis; tumorigenic; ubiquitin isopeptidase

PROJECT SUMMARY Glutathione (GSH) is a small thiol antioxidant that is critical in maintaining redox homeostasis, and increases in glutathione occur in KRAS-mutant lung adenocarcinoma. Glutathione can also lead to S-glutathionylation (PSSG), the conjugation of GSH to reactive cysteines in proteins, which can be reversed by the de- glutathionylase, glutaredoxin (GLRX). While the glutaredoxin/PSSG system has emerged as a key regulator of redox signaling, little is known about its disruption in lung adenocarcinoma (LUAD). We have made the exciting discovery that GLRX expression is decreased in human LUAD and that KrasG12D-induced lung tumorigenesis is enhanced in mice lacking Glrx. In a redox proteomics screen we revealed increases in S-glutathionylation of ovarian tumor deubiquitinase, OTUB1, which was recently identified as a regulator of System XC-. System XC- is a cystine/glutamate antiporter that exports glutamate and imports cystine, which is in turn reduced intracellularly to cysteine, the rate limiting amino acid in glutathione synthesis. System XC- is composed of SLC7A11 and SLC3A2. OTUB1 binds to SLC7A11 to prevent its ubiquitination and proteasomal degradation. We have made the exciting discovery that S-glutathionylation of OTUB1 is important in the stabilization of SLC7A11, leading to increases in GSH. Compared to adjacent control tissue, expression of OTUB1 and SLC7A11 are increased in LUAD. These exciting findings led us to hypothesize that decreases in glutaredoxin expression in mutant KRAS-driven lung adenocarcinoma cause increases in system XC- activity through the S- glutathionylation of ovarian tumor deubiquitinase 1 (OTUB1), augmenting glutathione and promoting survival of lung cancer cells. Furthermore, we hypothesize that avenues to augment glutaredoxin will diminish GSH levels leading to increased potency of cisplatin-induced killing of lung cancer cells. In Specific Aim 1 we will determine whether GLRX status controls KrasG12D-induced tumorigenesis. Specific Aim 2 proposes to address the importance of OTUB1 S-glutathionylation and System XC- in the augmentation of GSH in KrasG12D- induced tumors while in Specific Aim 3 we aim to address the impact of AAV-mediated transduction of GLRX on cisplatin-induced tumor killing in primary LUAD tumor organoids and KrasG12D-driven tumors in mice. Completion of this proposal will provide new insights into the role of the GLRX-protein S-glutathionylation redox system in the pathogenesis of LUAD and offers a rationale for augmenting GLRX in lung tumors in order to promote killing. The pharmaceutical development of GLRX has the potential to offer new therapeutic strategies to combat LUAD, in combination with standard of care therapy.

项目概要 谷胱甘肽 (GSH) 是一种小硫醇抗氧化剂，对于维持氧化还原稳态至关重要，并可增加 谷胱甘肽出现在 KRAS 突变型肺腺癌中。谷胱甘肽还可导致 S-谷胱甘肽化 (PSSG)，GSH 与蛋白质中反应性半胱氨酸的结合，可以通过 de- 谷胱甘肽酶、谷氧还蛋白（GLRX）。虽然谷氧还蛋白/PSSG系统已成为关键的调节因子 尽管氧化还原信号传导在肺腺癌 (LUAD) 中受到破坏，但人们对其的破坏知之甚少。我们创造了令人兴奋的 发现人 LUAD 中 GLRX 表达降低，KrasG12D 诱导的肺肿瘤发生 在缺乏 Glrx 的小鼠中增强。在氧化还原蛋白质组学筛选中，我们发现 S-谷胱甘肽的增加 卵巢肿瘤去泛素酶 OTUB1，最近被确定为系统 XC- 的调节因子。系统 XC- 是 胱氨酸/谷氨酸逆向转运蛋白，输出谷氨酸并输入胱氨酸，从而在细胞内减少 半胱氨酸，谷胱甘肽合成中的限速氨基酸。系统XC-由SLC7A11和 SLC3A2。 OTUB1 与 SLC7A11 结合以防止其泛素化和蛋白酶体降解。我们已经做了 令人兴奋的发现是 OTUB1 的 S-谷胱甘肽化对于 SLC7A11 的稳定非常重要，从而导致 谷胱甘肽 (GSH) 增加。与邻近对照组织相比，OTUB1 和 SLC7A11 的表达在 卢阿德。这些令人兴奋的发现使我们推测突变体中谷氧还蛋白表达减少 KRAS 驱动的肺腺癌通过 S- 导致系统 XC- 活性增加 卵巢肿瘤去泛素酶 1 (OTUB1) 的谷胱甘肽化，增强谷胱甘肽并促进 肺癌细胞的存活。此外，我们假设增加谷氧还蛋白的途径将会减少 GSH 水平导致顺铂诱导的肺癌细胞杀伤效力增强。在具体目标 1 中，我们 将确定 GLRX 状态是否控制 KrasG12D 诱导的肿瘤发生。具体目标 2 建议 阐述 OTUB1 S-谷胱甘肽化和 System XC- 在 KrasG12D- 中 GSH 增强中的重要性 诱导肿瘤，而在具体目标 3 中，我们旨在解决 AAV 介导的 GLRX 转导对肿瘤的影响 顺铂诱导的原发性 LUAD 肿瘤类器官和 KrasG12D 驱动的小鼠肿瘤的杀伤作用。完成 该提案将为 GLRX 蛋白 S-谷胱甘肽氧化还原系统的作用提供新的见解 LUAD 的发病机制，并为增强肺部肿瘤中的 GLRX 以促进杀伤提供了理论依据。 GLRX 的药物开发有潜力提供新的治疗策略来对抗 LUAD， 与标准护理疗法相结合。