Validating the mode of action of ergosterol peroxide as a selective breast cancer inhibitor

验证过氧化麦角甾醇作为选择性乳腺癌抑制剂的作用方式

• 批准号: 10411072
• 负责人: Michelle M Martínez
• 金额: $ 11.7万
• 依托单位: UNIVERSIDAD CENTRAL DEL CARIBE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10411072
• 关键词: 4T1; Address; Affect; African American; Aminoacyl-tRNA hydrolase; Ankyrin Repeat; Apoptosis; Biological; Biological Assay; Biological Availability; Biomedical Research; Biophysics; Breast Cancer Cell; Breast Cancer Model; Breast Cancer Patient; Breast Cancer Treatment; Cell Death; Cell Line; Cell model; Cells; Cellular Assay; Cessation of life; Chemicals; Cholesterol; Cholesterol Homeostasis; Clinic; Clinical Research; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Cytosol; Data; Dependence; Development; Diagnosis; Disease; Dose; Doxycycline; Drug toxicity; ERBB2 gene; Early Diagnosis; Environment; Enzymes; Ergosterol; Excision; Extravasation; Female; Future; Genes; Goals; Hispanic Americans; Homeostasis; Impairment; In complete remission; Knowledge; Lead; Ligation; MCF10A cells; Malignant Neoplasms; Membrane; Mitochondria; Mitochondrial Proteins; Modality; Modeling; Mus; Natural Products; Normal Cell; Normal tissue morphology; Oxidation-Reduction; Oxidative Stress; Pathway interactions; Patient-derived xenograft models of breast cancer; Patients; Peroxides; Pharmacology; Prognosis; Property; Protac; Proteins; Puerto Rico; Recurrence; Research; Resistance; Safety; Scientist; Steroids; Sterols; Structure; Students; Substrate Specificity; Survival Rate; System; Testing; Therapeutic; Therapeutic Agents; Toxic effect; Tumor Volume; Ubiquitin; Ubiquitination; United States; Woman; Xenograft procedure; Zinc Fingers; aggressive breast cancer; anti-cancer therapeutic; antiproliferative agents; base; cancer cell; cancer subtypes; chemotherapy; clinically relevant; efficacy testing; fungus; improved; in vivo; in vivo Model; inhibitor; malignant breast neoplasm; misfolded protein; mitochondrial dysfunction; mortality; mouse model; multicatalytic endopeptidase complex; neoplastic cell; novel; pharmacokinetics and pharmacodynamics; pre-clinical; protein aggregation; protein complex; public health relevance; stem; student participation; targeted treatment; triple-negative invasive breast carcinoma; tumor progression; underrepresented minority student; uptake; valosin-containing protein; women of color

PROJECT SUMMARY Triple Negative Breast Cancer (TNBC) is an aggressive and lethal breast cancer subtype more commonly diagnosed on younger (<40y) women of color (Hispanics and African American), who are more likely to have disease recurrence and who have an overall shorter survival. Furthermore, additional limitations (i.e. drug toxicity, and therapy resistance) persist in the clinic for TNBC patients. Thus, a critical need exists to discover more desirable targeted therapeutic modalities that selectively target TNBC tumor cells while leaving normal cells unaffected, enhancing a complete response and reducing TNBC-associated mortality rates. Accordingly, our goal is to reduce BC mortality by identifying unique vulnerabilities of TNBC that can be exploited through the development of selective therapeutic agents. We recently identified the natural product ergosterol peroxide a steroidal compound found in fungi, as a potent antiproliferative agent against TNBC cell models with little to no effect on noncancerous cells. We demonstrated that EP is more potent towards TNBC than HER2-positive models. EP induces ROS and apoptosis in TNBC cells and reduces tumor volume in TNBC models in vivo. However, EP’s mode of action (MOA) remains to be defined. Extended periods of oxidative stress in the mitochondria lead to accumulation of damaged proteins, which are normally removed by ubiquitination via the VCP/ANKZF1 pathway. We have synthesized EP in gram scale and show that EP accumulates in the cytosol and affects VCP and ANKZF1 interactions, raising the possibility that the VCP/ANKZF1 complex is targeted during its assembly prior to arriving to the mitochondria. Thus, taking advantage of TNBC’s dependency on sterol uptake, removal of misfolded proteins and altered redox homeostasis, we propose that EP serves as a VCP/ANKZF1 complex protein-protein inhibitor (PPI). We hypothesize that EP targets the VCP/ANKZF1 complex, impairing the ability of cancer cells to clear damaged proteins and causing mitochondria dysfunction to induce TNBC cell death. To test the hypothesis we will identify the mechanism by which EP targets the VCP/ANKZF1 complex in TNBC models using biophysical assays and synthesized EP-probes (Aim 1). The results from this Aim will validate that targeting VCP/ANKZF1 complex induces increases misfolded and aggregated proteins, which lead to cancer cell death. We will determine the efficacy of EP using syngeneic models injected with doxycycline inducible ANKZF1 silenced cells, and in TNBC PDXs. EP safety and bio- availability will also be determined and studied by MTD and PK/PD (Aim 2). The result of this aim will provide data needed for the next step towards the translational development of EP. Student participation in this hypothesis-driven research will improve the pipeline for URM students in biomedical research and lead to improved BC treatment.

项目概要 三阴性乳腺癌 (TNBC) 是一种更常见的侵袭性和致命性乳腺癌亚型 诊断为年轻（<40 岁）有色人种女性（西班牙裔和非裔美国人），她们更有可能患有 疾病复发和总体生存期较短的人。此外，还有其他限制（即药物 对于 TNBC 患者来说，临床上仍然存在这种情况。因此，迫切需要发现 更理想的靶向治疗方式，选择性靶向 TNBC 肿瘤细胞，同时保留正常细胞 细胞不受影响，增强完全反应并降低 TNBC 相关死亡率。因此， 我们的目标是通过识别 TNBC 的独特漏洞来降低 BC 死亡率，这些漏洞可以通过 开发选择性治疗剂。我们最近发现了天然产物过氧化麦角甾醇a 在真菌中发现的类固醇化合物，作为针对 TNBC 细胞模型的有效抗增殖剂，几乎没有作用 对非癌细胞的影响。我们证明 EP 对 TNBC 比 HER2 阳性更有效 模型。 EP 可诱导 TNBC 细胞中的 ROS 和细胞凋亡，并减少体内 TNBC 模型中的肿瘤体积。 然而，EP 的作用模式 (MOA) 仍有待定义。长时间的氧化应激 线粒体导致受损蛋白质的积累，这些蛋白质通常通过泛素化通过 VCP/ANKZF1 通路。我们合成了克级的 EP，并表明 EP 在细胞质中积累 并影响 VCP 和 ANKZF1 相互作用，增加了 VCP/ANKZF1 复合物被靶向的可能性 在到达线粒体之前的组装过程中。因此，利用 TNBC 对甾醇的依赖 摄取、去除错误折叠的蛋白质和改变氧化还原稳态，我们建议 EP 作为 VCP/ANKZF1 复合蛋白-蛋白抑制剂 (PPI)。我们假设 EP 针对 VCP/ANKZF1 复杂，损害癌细胞清除受损蛋白质的能力并导致线粒体功能障碍 诱导TNBC细胞死亡。为了检验这个假设，我们将确定 EP 靶向的机制 使用生物物理测定和合成的 EP 探针在 TNBC 模型中形成 VCP/ANKZF1 复合物（目标 1）。这 该目的的结果将验证靶向 VCP/ANKZF1 复合物会诱导错误折叠和 聚集的蛋白质，导致癌细胞死亡。我们将使用同基因确定 EP 的功效 注射强力霉素诱导的 ANKZF1 沉默细胞的模型以及 TNBC PDX。 EP安全和生物 可用性也将通过 MTD 和 PK/PD 来确定和研究（目标 2）。这一目标的结果将提供 下一步 EP 转化开发所需的数据。学生参与本次活动 假设驱动的研究将改善 URM 学生在生物医学研究方面的渠道，并导致 改善 BC 治疗。