Repurposing Glucosylceramide Synthase to Promote Mitochondrial Lethality and Potentiate an Anti-Tumor Immune Response in Triple-Negative Breast Cancer

重新利用葡萄糖神经酰胺合酶促进线粒体致死性并增强三阴性乳腺癌的抗肿瘤免疫反应

• 批准号: 10656615
• 负责人: Johannes F Fahrmann
• 金额: $ 47.78万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-31 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656615
• 关键词: 4T1; Adjuvant Therapy; Alternative Therapies; Binding; Biochemical; Biological Assay; Blood Chemical Analysis; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer cell line; CD8-Positive T-Lymphocytes; Cell Death; Cells; Cellular Assay; Ceramide glucosyltransferase; Ceramides; Chemicals; Clinical; Coupled; Cytotoxic T-Lymphocytes; Dose; Enzymes; Evaluation; Expression Profiling; FDA approved; Family; Gaucher Disease; Glycosphingolipids; Histologic; Immune; Immune response; Immunocompetent; Immunofluorescence Immunologic; Immunohistochemistry; Immunophenotyping; Immunoprecipitation; Immunotherapy; Infiltration; Interferons; Intervention; Lipids; MHC Class I Genes; Mass Spectrum Analysis; Measures; Mediating; Metabolic; Methods; Mitochondria; Mouse Mammary Tumor Virus; Mus; Neoplasm Metastasis; Pathologic; Pathway interactions; Patients; Peptides; Pharmacotherapy; Phenotype; Plasma; Prognosis; Proteomics; Recurrence; Resistance; Role; Signaling Protein; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Sphingomyelins; Stimulator of Interferon Genes; Surface; T cell response; T-Cell Activation; T-Lymphocyte Epitopes; TP53 gene; Technology; Testing; Tissues; Toxic effect; Translating; Treatment-related toxicity; Tumor-Derived; Western Blotting; anti-PD-L1; anti-cancer; anti-tumor immune response; cancer cell; cancer immunotherapy; chemokine; chemotherapy; cytokine; early phase clinical trial; extracellular vesicles; glycation; histopathological examination; improved; in vivo; inhibitor; interest; lipidomics; malignant breast neoplasm; mammary; mass spectrometric imaging; mouse model; novel; novel therapeutic intervention; patient derived xenograft model; pre-clinical; primary endpoint; programs; response; small molecule; success; tandem mass spectrometry; transcriptome sequencing; transcriptomics; triple-negative invasive breast carcinoma; tumor; tumor growth

Project Summary Current approaches to treating triple-negative breast cancer (TNBC) remain unsatisfactory. There remains an unmet need to establish novel alternative therapies that can both exert targeted effects on cancer cells as well as stimulate an anti-cancer immune response that may render additional opportunity for combination with emerging immunotherapy. We have uncovered a novel onco-metabolic feature in TNBC wherein cancer cells exploit a sphingomyelin lipid scavenging phenotype that is targetable through repurposing of the selective glucosylceramide synthase inhibitor eliglustat, an FDA approved drug for treatment of Gaucher Disease. Our preliminary studies demonstrate that eliglustat promotes accumulation of mitotoxic ceramides with resultant shift from survival mitophagy to lethal mitophagy and subsequent cancer cell death. Moreover, we show that eliglustat suppresses tumor growth at clinically achievable doses in TNBC tumor-bearing mice and that the anti-cancer effects of eliglustat are associated with pronounced increases in tumor infiltrating CD4+ and CD8+ T-cells, suggesting an additional role of eliglustat in potentiating an anti-tumor immune response. The primary objectives of this proposal are to establish novel utility for eliglustat as an ‘immunometabolic adjuvant’ for the treatment of TNBC and to also define the mechanism(s) by which eliglustat potentiates an anti-cancer immune response. To test this, we will assess the anti-cancer efficacy of eliglustat in patient-derived xenograft (PDX) models generated from patients with primary treatment-naïve TNBC that did or did not go on to respond to chemotherapy (Specific Aim 1a). To test whether the combination of eliglustat plus anti-PD-L1 yields improved anti-cancer effects compared to either treatment alone, we will use the BRCA1co/co; MMTV-Cre; p53+/- and 4T1 orthotopic syngeneic mouse models of TNBC (Specific Aim 1b). Primary endpoints of interest for in vivo studies will be overall survival and tumor growth; effects on the tumor immunophenotype following intervention will be assess using multiplex immunofluorescence panels and single cell transcriptomics for single cell-level expression profiling of tissues. We also aim to define the mechanisms by which eliglustat induces an anti-cancer immune response. First, we will evaluate the effect of eliglustat on cGAS-STING signaling proteins and downstream pathway activities in TNBC cells (Specific Aim 2a). Next, we will use advanced mass spectrometry technologies coupled with novel isolation methods for extracellular vesicles (EVs) to define the MHC-I bound peptidome on surfaces of TNBC cells and TNBC-derived circulating and intra-tumor EVs following eliglustat treatment. ELIspot and Cytotoxic T Cell-based tumor killing live-cell assays will be used to test the functionality of EVs on activating T-cells ex vivo (Specific Aim 2b). If successful, our potential findings will provide key pre- clinical evidence for the use of eliglustat and justification for moving eliglustat into early phase clinical trials. Given that eliglustat is already FDA-approved for another indication, it has high potential to be readily translated into clinical use, and potentially providing a new paradigm for cancer immunotherapy.

项目总结