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.

项目摘要 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，并且增加 谷胱甘肽发生在KRAS突变的肺腺癌中。谷胱甘肽也可能导致s-谷胱甘肽化 （PSSG），GSH与蛋白质中的反应性半胱氨酸的结合，可以通过de-逆转 谷胱甘肽酶，谷胱甘肽（GLRX）。而谷胱甘肽/PSSG系统已成为关键调节器 氧化还原信号传导对其在肺腺癌（LUAD）中的破坏知之甚少。我们使令人兴奋的 发现GLRX表达在人luAD中降低，而KRASG12D诱导的肺肿瘤发生是 缺乏GLRX的小鼠增强。在氧化还原蛋白质组学屏幕中，我们揭示了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-的调节剂。系统XC- IS 出口谷氨酸并进口胱氨酸的半胱氨酸/谷氨酸抗植物 对于半胱氨酸，谷胱甘肽合成中氨基酸的速率限制。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。我们做了 令人兴奋的发现，即OTUB1的S-谷胱甘肽化在SLC7A11的稳定中很重要，从而导致 GSH的增加。与相邻的对照组织相比，OTUB1和SLC7A11的表达在 卢德。这些令人兴奋的发现导致我们假设突变体中谷氨酸蛋白表达的降低 KRAS驱动的肺腺癌导致系统XC活性通过S-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，我们假设增强谷氨酸毒素的途径会减少 GSH水平导致顺铂诱导的肺癌细胞杀死的效力提高。在特定的目标1中我们 将确定GLRX状态是否控制KRASG12D诱导的肿瘤发生。特定目标2提议 探讨了otub1 s-谷胱甘肽化和系统xc-在krasg12d-的GSH增强中的重要性 在特定目标3中诱导肿瘤，我们旨在解决AAV介导的GLRX转导的影响 在小鼠原发性LUAD肿瘤器官和KRASG12D驱动的肿瘤中，顺铂诱导的肿瘤杀死。完成 该提案将提供有关GLRX-蛋白S-谷氨酸氧化氧化还原系统作用的新见解 LUAD的发病机理，并提供了增加肺部肿瘤中GLRX的理由，以促进杀戮。 GLRX的药物开发有可能提供新的治疗策略来打击LUAD， 结合护理疗法的结合。