C16-Ceramide Nano-Liposomes Reverse Multi-Drug Resistance

C16-神经酰胺纳米脂质体逆转多药耐药性

• 批准号: 9106835
• 负责人: Richard N Kolesnick
• 金额: $ 39.21万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9106835
• 关键词: ABCB1 gene; Address; Affect; Anthracyclines; Apoptotic; Biochemical; Biological Assay; Breast; Cell Culture Techniques; Cell Death; Cell Nucleus; Cells; Ceramides; Clinic; Clinical; Collaborations; Colon; Complement; Cytoplasm; Cytoplasmic Vesicles; Data; Daunorubicin; Defect; Development; Diffuse; Disease; Doxorubicin; Doxorubicin Hydrochloride Liposome; Drug Transport; Engineering; Evaluation; Exhibits; Failure; Fatty Acids; Goals; Human; In Vitro; Investigation; Kidney; Lead; Legal patent; Length; Letters; Lipids; Liposomes; Location; Lung; Malignant Neoplasms; Mediating; Membrane; Membrane Lipids; Memorial Sloan-Kettering Cancer Center; Metabolism; Multi-Drug Resistance; N-palmitoylsphingosine; Nuclear; Nuclear Envelope; P-Glycoprotein; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Photography; Positioning Attribute; Preparation; Prevalence; Process; Reporting; Research; Research Design; Resistance; Specificity; Sphingolipids; Sphingomyelins; Sphingosine; System; Time; Universities; Vacuole; Vertebral column; Vesicle; Vesicle Transport Pathway; Work; acid sphingomyelinase; base; cancer therapy; cell killing; clinically relevant; design; drug development; effective therapy; experience; in vivo; inhibitor/antagonist; nano; nanoliposome; neoplastic cell; overexpression; preclinical study; programs; public health relevance; restoration; serine palmitoyltransferase; trafficking; tumor; tumor xenograft

 DESCRIPTION (provided by applicant): Cells with multidrug resistance (MDR) due to aberrant expression of the lipid transporter P-glycoprotein (P-gp) display a wide range of biochemical changes that affect membrane lipid composition. Despite significant clinical effort, no effective therapy currently exists to reverse P-gp mediated MDR in human cancers. Recently we discovered that distinct MDR cells exhibit constitutive activation of the de novo pathway of ceramide synthesis as a result of enhanced serine palmitoyltransferase activity, resulting in increased ceramide content (1.7-1.9 fold) and sphingomyelin levels (2-3 fold). Evaluation of the mechanism of daunorubicin resistance in sphingolipid-disordered ADX cells revealed that drug traffics to the nucleus of daunorubicin-sensitive parental DC-3F cells, whereas it mislocalizes into large cytoplasmic vacuoles in ADX MDR cells. Furthermore application of small amounts of exogenous acid sphingomyelinase to generate endogenous ceramide, or provision of exogenous long-chain natural C16-ceramide, result in rapid translocation of vacuolar daunorubicin into the nucleus (within minutes), and 25-fold enhanced cell kill. We posit that MDR chemoresistance results in part from failure to generate the fusogenic lipid ceramide in a vesicular trafficking system that normally transports daunorubicin into the nucleus, reversible by exogenous ceramide. Natural ceramides are categorized into long chain (C16:0-C20:0) and very long chain (C22:0-C24:1) species relative to the length of the N-acylated fatty acid at the second position of the sphingosine backbone. We recently showed that different ceramide species possess distinct biologic attributes with long chain C16:0 ceramide being pro-apoptotic, while very long chain C24:0, C24:1 ceramides are anti-apoptotic. We now show that inclusion of C16:0 ceramide, but no other natural ceramide species, in a nano-liposomal preparation permits rapid translocation of daunorubicin from cytoplasmic vesicles to the nucleus of ADX MDR cells. Based on these data we initiated a collaboration with Chezy Barenholz, who engineered Doxil, the first successful liposomal drug that delivers doxorubicin systemically to tumors. Patent protection for Doxil has recently expired. The overall purpose of this application is to set the groundwork for the development of C16-ceramide Doxil-like liposomes for cancer therapy through three specific in vitro and in vivo aims. Successful completion of these studies will resul in a direct path of drug development as Chezy Barenholz has agreed in principle to generate C16-ceramide/daunorubicin liposomes for the clinic in collaboration with Memorial Sloan-Kettering Cancer Center.

 描述（由适用提供）：由于脂质转运蛋白P-糖蛋白（P-GP）异常表达，具有多药电阻（MDR）的细胞显示出影响膜脂质组成的广泛的生化变化。尽管临床努力很大，但目前尚无有效的治疗来逆转P-gp介导的人类癌症的MDR。最近，我们发现，由于增强的丝氨酸棕榈酰转移酶活性的增强，不同的MDR细胞暴露了神经酰胺合成的从头途径的组成型激活，从而导致神经酰胺含量增加（1.7-1.9倍）和鞘磷脂水平（2-3倍）。评估鞘脂降低的ADX细胞中多霉素耐药性的机制表明，药物传播到dunorumubicin敏感的父母DC-3F细胞的细胞核，而它将其错误地定位于ADX MDR细胞中的大型细胞质液体。此外，少量的外源酸鞘磷脂酶的应用以产生内源性神经酰胺，或提供外源性长链天然C16-钙酰胺，从而导致真空肺道努比霉素快速易位到核（几分钟之内），并使25倍增强的细胞杀死。我们肯定的是，MDR化学抗性部分是由于未能在囊泡运输系统中产生熔融脂质神经酰胺，该系统通常会通过外源性神经酰胺可逆地将daunorubicin转运到核中。天然神经酰胺分为长链（C16：0-C20：0）和非常长的链（C22：0-C24：1），相对于N-酰基脂肪酸在鞘氨酸骨架的第二位置的长度相对于N-酰基脂肪酸的长度。我们最近表明，不同的神经酰胺物种具有独特的生物学特性，长链C16：0神经酰胺是促凋亡的，而长链C24：0，C24：1神经酰胺是抗凋亡的。现在，我们表明，在纳米脂质体制备中包含C16：0神经酰胺，但没有其他天然神经酰胺种类允许将daunorubicin从细胞质蔬菜迅速转移到ADX MDR细胞的核心。基于这些数据，我们与Chezy Barenholz进行了合作，他设计了Doxil，他设计了Doxil，这是第一种成功的脂质体药物，从系统地将阿霉素递送至肿瘤。 Doxil的专利保护最近已过期。该应用的总体目的是为开发C16-塞他酰胺类似Doxil样脂质体开发癌症治疗，以通过三种特定的体外和体内目标为癌症治疗。这些研究的成功完成将导致药物开发的直接途径，因为Chezy Barenholz原则上同意与纪念Sloan-Kettering Cancer Center合作，为临床机构生成C16-Ceramide/daunorubicin脂质体。