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.

项目摘要 当前治疗三阴性乳腺癌（TNBC）的方法仍然不令人满意。那里 建立新型替代疗法仍然是未满足的需求，既可以对 癌细胞以及刺激抗癌免疫反应，这可能会导致额外的机会 与新兴的免疫疗法结合使用。我们在TNBC中发现了一种新型的onco-亚代谢特征 其中癌细胞利用鞘脂蛋白脂质清除表型，可通过重新利用而定位 选择性葡萄糖基酰胺合酶抑制剂Eliglustat，一种经FDA批准的药物用于治疗Gaucher 疾病。我们的初步研究表明，Eliglustat促进有丝毒神经酰胺的积累 从生存线粒体到致命的线粒能和随后的癌细胞死亡。而且，我们显示 Eliglustat抑制了TNBC肿瘤小鼠临床上可达到的剂量时肿瘤的生长，并且 Eliglustat的抗癌作用与肿瘤浸润CD4+和CD8+的明显增加有关 T细胞，表明Eliglustat在增强抗肿瘤免疫响应中的其他作用。主要 该提案的目标是为Eliglustat建立新的实用性，以作为“免疫代谢调整” TNBC的处理并定义了Eliglustat增强抗癌免疫的机制 回复。为了测试这一点，我们将评估Eliglustat在患者衍生异种移植物（PDX）中的抗癌效率 从未接受过或没有反应的初级治疗的患者产生的模型 化学疗法（特定目标1A）。测试Eliglustat加抗PD-L1的组合是否有所提高 抗癌作用与仅任何一种治疗相比，我们将使用BRCA1CO/CO； mmtv-cre; p53 +/-和 TNBC的4T1原位合成小鼠模型（特定AIM 1B）。体内感兴趣的主要终点 研究将是总体生存和肿瘤生长；干预后对肿瘤免疫表型的影响 将使用多重免疫荧光面板和单细胞转录组学评估单细胞级 组织的表达分析。我们还旨在定义Eliglustat诱导抗癌的机制 免疫反应。首先，我们将评估Eliglustat对CGAS插入信号蛋白和 TNBC细胞中的下游途径活动（特定AIM 2A）。接下来，我们将使用高级质谱法 技术以及新的细胞外蔬菜（EV）的新型隔离方法定义MHC-I结合 TNBC细胞表面和TNBC衍生的循环和肿瘤内电动汽车的肽组在Eliglustat之后 治疗。 ELISPOT和细胞毒性T细胞基肿瘤杀死活细胞测定法将用于测试功能 激活T细胞的电动汽车（特定目标2B）。如果成功的话，我们的潜在发现将为关键预先提供 使用Eliglustat和理由将Eliglustat转移到早期临床试验中的临床证据。 鉴于Eliglustat已经被FDA批准了另一种迹象，因此很容易翻译它的潜力 进入临床用途，并有可能为癌症免疫疗法提供新的范式。