Targeted Therapeutics of LGL Leukemia utilizing Ceramide Nanoliposomes

项目摘要 这项提议的广泛长期目标是设计、开发和优化下一代 聚乙二醇化C6-神经酰胺阳离子脂质体制剂治疗大颗粒淋巴细胞的研究 (LGL)白血病。LGL白血病源于T细胞或自然杀伤细胞(NK)的克隆性增殖 细胞。目前还没有已知的治疗LGL白血病的方法。白血病LGL耐药 Fas诱导的细胞凋亡和高水平激活的STAT3，这是致癌的关键介质 发信号。抑制白血病LGL中的STAT3导致生存蛋白Mcl-1和AN的减少 对Fas介导的细胞凋亡的敏感性增加。 这一提议的基础是第一代C6-神经酰胺纳米脂质体，它已经被 作为平台技术被国家纳米技术表征实验室采用 癌症研究所。具体目标集中在一个中心假设上，即纳米级、非纳米级 聚集型C6-神经酰胺脂质体具有极高的敏感性、选择性、无毒、隐形、 用于系统靶向疏水性化疗药物的生物可降解和响应平台 和/或siRNA作用于LGL白血病细胞。多功能仪器的潜在临床应用 纳米脂质体，旨在提供神经酰胺(C6神经酰胺)的细胞渗透性类似物以及 靶向特异性siRNA和/或甲氨蝶呤将在LGL白血病的Fischer F344大鼠模型中进行测试。 假设甲氨蝶呤和/或siRNA同时在神经酰胺掺入的 脂质体对LGL白血病有协同治疗作用。 第一个具体目标将检验脂质体短链全身性递送的假设 神经酰胺在已建立的动物模型中抑制LGL白血病。《特定目标2》将检验这一假设 聚乙二醇化的阳离子脂质体可输送抑制性siRNA和化疗药物 在体外和体内转化成LGL白血病细胞，从而增强其抗白血病活性。特定的 目的3将测试一种针对CD8的单抗的结合假设，CD8是人类免疫缺陷病毒的特异性标志物 LGL白血病，将在体内外靶向治疗LGL白血病细胞的脂质体。 特定目标3的实验是免疫脂质体传递的重要原则验证研究 通过靶向癌细胞上表达的表面抗原。这第二代的发展 本提案中概述的癌症纳米技术平台具有改善癌症的潜力 治疗学。 。项目叙事 这项建议的目标是开发治疗大颗粒淋巴细胞(LGL)白血病的方法 新型癌症纳米技术平台。我们建议给药系统靶向的C6-神经酰胺 阳离子纳米免疫脂质体制剂将甲氨蝶呤和siRNAs包裹到已建立的 LGL白血病动物模型的建立。这些研究将为利用这种治疗方法提供基础 在患有这种不治之症的人类受试者中的方法。

Project Summary The broad long term goal of this proposal is to design, develop and optimize the next-generation of pegylated C6-ceramide cationic liposomal formulations for treatment of large granular lymphocyte (LGL) leukemia. LGL leukemia arises from clonal proliferation of either T cells or natural killer (NK) cells. There is no known curative therapy for patients with LGL leukemia. Leukemic LGL are resistant to Fas-induced apoptosis and display high levels of activated STAT3, a critical mediator of oncogenic signaling. Inhibition of STAT3 in leukemic LGL causes a decrease in survival protein Mcl-1 and an increased sensitivity to Fas-mediated apoptosis. The foundation of this proposal is a first generation C6-ceramide nanoliposome which has been adopted as platform technology by the Nanotechnology Characterization Laboratory of the National Cancer Institute. The specific aims are focused around the central hypothesis that nano-scale, non- aggregating C6-ceramide liposomes offer exquisitely sensitive, selective, non-toxic, stealthy, biodegradable and responsive platforms for systemically targeting hydrophobic chemotherapeutic drugs and/or siRNA to LGL leukemic cells. The potential clinical usefulness of multi-functional nanoliposomes, designed to deliver cell-permeable analogs of ceramide (C6 ceramide) as well as target-specific siRNA and/or Methotrexate, will be tested in a Fischer F344 rat model of LGL leukemia. It is hypothesized that simultaneous delivery of Methotrexate and/or siRNA in a ceramide-incorporated liposome will have synergistic efficacy in LGL leukemia. The first specific Aim will test the hypothesis that systemic delivery of liposomal short-chain ceramide inhibits LGL leukemia in an established animal model. Specific Aim 2 will test the hypothesis that the pegylated cationic liposomes allow delivery of inhibitory siRNA and chemotherapeutic drugs into LGL leukemic cells in vitro and in vivo, such that their anti-leukemic activity is enhanced. Specific Aim 3 will test the hypothesis that conjugation of a monoclonal antibody to CD8, the specific marker for LGL leukemia, will ¿target¿ the therapeutic liposomes to LGL leukemic cells in vitro and in vivo. Experiments in specific aim 3 are important proof-of-principle studies for delivery of immunoliposomes by targeting surface antigens expressed on cancer cells. Development of this second generation cancer nanotechnology platform as outlined in this proposal has potential for improved cancer therapeutics. . Project Narrative The goal of this proposal is to develop therapy for large granular lymphocyte (LGL) leukemia using a novel cancer nanotechnology platform. We propose to administer systemically targeted C6-ceramide cationic nanoimmunoliposome formulations encapsulating methotrexate and siRNAs to an established animal model of LGL leukemia. These studies will provide the foundation for utilizing this therapeutic approach in human subjects with this incurable disease.