Regulation of Death Receptor 5 folding and apoptotic signaling by AGR2

AGR2 对死亡受体 5 折叠和细胞凋亡信号的调节

• 批准号: 10042651
• 负责人: Brian K. Law
• 金额: $ 38.88万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-16 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10042651
• 关键词: Active Sites; Animals; Anterior; Antineoplastic Agents; Apoptosis; Apoptotic; Binding; Breast; Breast Epithelial Cells; CASP3 gene; CASP8 gene; Cell Death; Cell surface; Cells; Chemicals; Combined Modality Therapy; Cysteine; Development; Disulfides; Down-Regulation; Drug resistance; ERBB2 gene; ERBB3 gene; Endoplasmic Reticulum; Epidermal Growth Factor Receptor; Epithelial Cell Proliferation; Event; Exhibits; Extracellular Domain; Family; Genetic; In Vitro; Knock-out; Knowledge; Ligands; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of ovary; Malignant neoplasm of pancreas; Mammary Neoplasms; Mass Spectrum Analysis; Mediating; Modeling; Modification; Molecular; Molecular Chaperones; Monoclonal Antibodies; Normal tissue morphology; Oncoproteins; Outcome; Patients; Pharmacology; Play; Protein Disulfide Isomerase; Protein Tyrosine Kinase; Publishing; Refractory; Regimen; Regulation; Reporting; Resistance; Role; Signal Transduction; Site-Directed Mutagenesis; TNFRSF10A gene; TNFRSF10B gene; TNFSF10 gene; Testing; Therapeutic; Toxic effect; Up-Regulation; Weight; Xenopus laevis; aggressive therapy; angiogenesis; anti-cancer; antibody inhibitor; cancer cell; conventional therapy; cytokine; disulfide bond; enzyme activity; improved; in vivo; inhibitor/antagonist; kinase inhibitor; knock-down; lapatinib; malignant breast neoplasm; malignant stomach neoplasm; molecular targeted therapies; novel; novel strategies; overexpression; prevent; prostate cancer cell; receptor; receptor downregulation; response; senescence; small molecule inhibitor; targeted agent; targeted treatment; triple-negative invasive breast carcinoma; tumor; tumor xenograft

Description The Epidermal Growth Factor Receptor-family tyrosine kinases EGFR and HER2 are overexpressed in a significant proportion of aggressive, treatment-refractory breast cancers. This includes HER2+ tumors that have acquired resistance to HER2-specific monoclonal antibodies and kinase inhibitors and Triple-Negative Breast Cancers (TNBCs), of which approximately 50% overexpress EGFR. Agents that selectively kill EGFR+ or HER2+ tumors through a novel mechanism of action could improve the survival of patients with these treatment-refractory cancers. Our team reported a new class of anticancer compounds, termed DDAs, with selective toxicity against EGFR+ and HER2+ cancers. Published and preliminary results demonstrate that DDAs kill patient-derived breast xenograft tumors without detectable toxicity to experimental animals. DDAs kill cancer cells by activating Death Receptors 4 and 5 (DR4/5), the receptors for the anticancer cytokine TRAIL. However, DDAs activate DR4/5 in a novel TRAIL-independent manner. DR4/5 activation by DDAs results from a combination of elevated DR5 expression, disulfide bond-dependent DR4/5 clustering, and localization of DR4/5 to the cell surface. These events occur specifically in EGFR or HER2 overexpressing cancer cells and result in DR4/5 activation of the Caspase 8-Caspase 3 pro-apoptotic cascade. Consistent with this in vitro selectivity, tumor studies demonstrate DDA-induced apoptosis of cancer cells without toxicity to adjacent normal tissues. This project will test the hypothesis that the protein disulfide isomerase AGR2 catalyzes/chaperones native disulfide bonding of EGFR, HER2, DR4, and DR5. We further hypothesize that DDAs inhibit AGR2 enzyme activity and that EGFR and HER2 strongly potentiate DDA actions by competing with DR4/5 for the limited pool of AGR2. Specific Aim 1 is to elucidate the mechanisms by which disulfide bonding within the extracellular domains of DR4/5 restrains their basal pro-apoptotic activity, and demonstrate that genetic or pharmacologic DR5 activation in cancer cells is sufficient to induce tumor regression. Specific Aim 2 is to investigate the mechanisms by which DDAs block AGR2 PDI activity and to demonstrate that DR4/5 activation mediates cancer cell apoptosis in response to ablating AGR2 activity. Specific Aim 3 is to establish the efficacy of DDAs against patient-derived breast tumors and verify that the mechanisms of DDA action in vivo match those identified in vitro. DDA efficacy will be compared with standard therapies in single agent and combination regimens. In summary, this exploratory project will investigate DDAs as a new class of anticancer agents, elucidate a novel AGR2/Death Receptor 4/5 cascade that mediates DDA actions, and establish DDA efficacy against patient-derived models of metastatic and drug-resistant HER2+ breast cancer.

描述 表皮生长因子受体家族酪氨酸激酶 EGFR 和 HER2 在 侵袭性、难治性乳腺癌的比例很高。这包括 HER2+ 肿瘤 已获得对 HER2 特异性单克隆抗体和激酶抑制剂和三阴性的耐药性 乳腺癌 (TNBC)，其中约 50% 过表达 EGFR。选择性杀死 EGFR+ 的药物 或 HER2+ 肿瘤通过一种新的作用机制可以提高这些患者的生存率 难治性癌症。我们的团队报告了一类新的抗癌化合物，称为 DDA， 针对 EGFR+ 和 HER2+ 癌症的选择性毒性。已发表的初步结果表明 DDA 可杀死患者来源的乳腺异种移植肿瘤，且对实验动物没有可检测到的毒性。 DDA 杀死 通过激活死亡受体 4 和 5 (DR4/5)（抗癌细胞因子 TRAIL 的受体）来抑制癌细胞。 然而，DDA 以一种新的不依赖于 TRAIL 的方式激活 DR4/5。 DDA 激活 DR4/5 的结果是 DR5 表达升高、二硫键依赖性 DR4/5 聚类和定位的组合 DR4/5 至细胞表面。这些事件特别发生在 EGFR 或 HER2 过表达的癌细胞中，并且 导致 DR4/5 激活 Caspase 8-Caspase 3 促凋亡级联。与体外实验一致 选择性，肿瘤研究证明 DDA 诱导癌细胞凋亡，且对邻近细胞没有毒性 正常组织。该项目将检验蛋白质二硫键异构酶 AGR2 的假设 催化/陪伴 EGFR、HER2、DR4 和 DR5 的天然二硫键。我们进一步假设 DDA 抑制 AGR2 酶活性，并且 EGFR 和 HER2 通过竞争强烈增强 DDA 作用 DR4/5 用于 AGR2 的有限池。具体目标 1 是阐明二硫化物的作用机制 DR4/5 胞外域内的结合抑制其基础促凋亡活性，并证明 癌细胞中的遗传或药理 DR5 激活足以诱导肿瘤消退。具体的 目标 2 是研究 DDA 阻断 AGR2 PDI 活性的机制并证明 DR4/5 激活介导癌细胞凋亡，以响应 AGR2 活性的消除。具体目标 3 是 确定 DDA 对患者源性乳腺肿瘤的功效，并验证 DDA 的机制 体内作用与体外鉴定的作用相匹配。 DDA 疗效将与单次标准疗法进行比较 剂和组合方案。总之，这个探索性项目将研究 DDA 作为一类新的 抗癌药物，阐明介导 DDA 作用的新型 AGR2/死亡受体 4/5 级联，以及 建立 DDA 对源自患者的转移性和耐药 HER2+ 乳腺癌模型的疗效。

项目成果

Anticancer Agents Derived from Cyclic Thiosulfonates: Structure-Reactivity and Structure-Activity Relationships.

环状硫代磺酸盐衍生的抗癌剂：结构-反应性和结构-活性关系。

• 发表时间: 2022-07-19
• 影响因子: 3.4
• 作者: Ghilardi, Amanda F;Yaaghubi, Elham;Ferreira, Renan B;Law, Mary E;Yang, Yinuo;Davis, Bradley J;Schilson, Christopher M;Ghiviriga, Ion;Roitberg, Adrian E;Law, Brian K;Castellano, Ronald K
• 通讯作者: Castellano, Ronald K

Inhibitors of ERp44, PDIA1, and AGR2 induce disulfide-mediated oligomerization of Death Receptors 4 and 5 and cancer cell death.

ERp44、PDIA1 和 AGR2 抑制剂可诱导二硫键介导的死亡受体 4 和 5 寡聚化以及癌细胞死亡。

• 发表时间: 2022-05-28
• 影响因子: 9.7
• 作者: Law, Mary E;Yaaghubi, Elham;Ghilardi, Amanda F;Davis, Bradley J;Ferreira, Renan B;Koh, Jin;Chen, Sixue;DePeter, Sadie F;Schilson, Christopher M;Chiang, Chi;Heldermon, Coy D;Nørgaard, Peter;Castellano, Ronald K;Law, Brian K
• 通讯作者: Law, Brian K

Disulfide bond-disrupting agents activate the tumor necrosis family-related apoptosis-inducing ligand/death receptor 5 pathway.

二硫键破坏剂激活肿瘤坏死家族相关的凋亡诱导配体/死亡受体 5 通路。

• 发表时间: 2019-12-10
• 影响因子: 7
• 作者: Wang, Mengxiong;Law, Mary E;Davis, Bradley J;Yaaghubi, Elham;Ghilardi, Amanda F;Ferreira, Renan B;Chiang, Chi;Guryanova, Olga A;Kopinke, Daniel;Heldermon, Coy D;Castellano, Ronald K;Law, Brian K
• 通讯作者: Law, Brian K

DNMT3A Harboring Leukemia-Associated Mutations Directs Sensitivity to DNA Damage at Replication Forks.

DNMT3A 含有白血病相关突变，可提高复制叉处 DNA 损伤的敏感性。

• 发表时间: 2022
• 作者: Venugopal, Kartika;Feng, Yang;Nowialis, Pawel;Xu, Huanzhou;Shabashvili, Daniil E;Berntsen, Cassandra M;Kaur, Prabhjot;Krajcik, Kathryn I;Taragjini, Christina;Zaroogian, Zachary;Casellas Román, Heidi L;Posada, Luisa M;Gunaratne, Chamara;Li, Ji
• 通讯作者: Li, Ji