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.

项目总结