GLUCOSYLCERAMIDE SYNTHASE IN A NOVEL TARGET FOR CANCER TREATMENT

葡萄糖神经酰胺合成酶作为癌症治疗的新靶点

• 批准号: 7720009
• 负责人: Yong-Yu Liu
• 金额: $ 9.31万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7720009
• 关键词: Affect; Anthracycline Antibiotics; Anthracyclines; Apoptosis; Bone Marrow; Breast Cancer Cell; Cancer cell line; Cells; Ceramide glucosyltransferase; Ceramides; Computer Retrieval of Information on Scientific Projects Database; Doxorubicin; Drug resistance; Drug-sensitive; Enzymes; Funding; Gene Expression; Genes; Grant; Human; In Vitro; Institution; Malignant Neoplasms; Mixed-Backbone Oligonucleotide; Modeling; Mononuclear; Mus; Neoplasm Metastasis; Protein Overexpression; Research; Research Personnel; Resistance; Resources; Source; Taxane Compound; United States National Institutes of Health; Vinca Alkaloids; Week; cancer therapy; chemotherapy; cytotoxicity; day; design; glycosylation; improved; in vivo; malignant breast neoplasm; novel; novel therapeutics; taxane; therapeutic target; tumor; tumor growth

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Characterizing gene underlying drug resistance, as a novel therapeutic target can significantly improve cancer treatment. Glucosylceramide synthase (GCS), a key enzyme for ceramide glycosylation, is overexpressed in various drug-resistant cancer cell lines and in metastatic tumors. High GCS activity confers cell resistance by disrupting ceramide-induced apoptosis that is tightly associated with the efficiency of anthracycline, vinca alkaloids and taxanes. To determine whether GCS is a target for the reversal of drug resistance, we evaluated the effects of antisense approach against GCS in vitro and in vivo. MBO-asGCS (mixed-backbone oligonucleotide, 20-mer) was designed to silence human GCS gene. MBO-asGCS substantially increased doxorubicin sensitivity in drug-resistant human (MCF-7-AdrR) and murine breast cancer cells (EMT6/AR1) by 83-fold and 43-fold, respectively. In contrast, MBO-asGCS only increased the doxorubicin cytotoxicity by twofold in drug-sensitive cells and did not affect that cytotoxicity in human bone marrow mononuclear cells. In the orthotopic tumor model for drug-resistant breast cancer (MCF-7-AdrR), we found that MBO-asGCS significantly inhibited tumor growth and sensitized tumors to chemotherapy. The administration of MBO-asGCS (1 mg/kg/3-day, 42 days) inhibited tumor growth more than 64% (356 vs 983 mm3, N=10) and increased doxorubicin-sensitivity by 58% (187 vs. 432 mm3, N=10), compared to MBO-scrambled alone and the combination of MBO-scrambled with doxorubicin (2 mg/kg/week), respectively. Further assessments of GCS gene expression levels, ceramide glycosylation, and apoptosis demonstrated that the effects of MBO-asGCS in vivo rely on the suppression of endogenous GCS overexpressed and the enhancement of ceramide-induced apoptosis. These evidences pinpoint that GCS is a novel target for the reversal of drug resistance in cancers.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 表征潜在耐药性的基因，作为一种新型的治疗靶标可以显着改善癌症治疗。葡萄糖基酶合酶（GCS）是神经酰胺糖基化的关键酶，在各种耐药的癌细胞系和转移性肿瘤中过表达。高GCS活性通过破坏神经酰胺诱导的凋亡来赋予细胞耐药性，而神经酰胺诱导的凋亡与蒽环类，VINCA生物碱和紫杉烷的效率紧密相关。为了确定GCS是否是抗药性逆转的靶标，我们评估了反义方法对GC在体外和体内的影响。 MBO-ASGC（混合背骨寡核苷酸，20-mer）旨在使人GCS基因保持沉默。 MBO-ASGC在耐药的人（MCF-7-ADRR）和鼠乳腺癌细胞（EMT6/AR1）中大大提高了阿霉素敏感性，分别为83倍和43倍。相比之下，MBO-ASGC仅在药物敏感细胞中增加了阿霉素细胞毒性，并且不影响人骨髓单核细胞中的细胞毒性。在用于药物耐药性乳腺癌（MCF-7-ADRR）的原位肿瘤模型中，我们发现MBO-ASGC显着抑制肿瘤的生长，并使肿瘤对化学疗法敏感。 MBO-ASGC（1 mg/kg/3天，42天）的施用抑制了肿瘤的增长超过64％（356 vs 983 mm3，n = 10），阿霉素敏感性增加了58％（187 vs.432 mm3，n = 10），与单独的MBO-SCRAMINIB和MBO-SCRAMINIB和CRAMBINIB（MBO）相比mg/kg/kg）分别。对GCS基因表达水平，神经酰胺糖基化和凋亡的进一步评估表明，MBO-ASGC在体内的作用依赖于过表达的内源性GC的抑制以及神经酰胺诱导的凋亡的增强。这些证据表明，GC是癌症中耐药性逆转的新目标。