Nanobiotechnology for the Treatment of Mantle Cell Lymphoma

纳米生物技术治疗套细胞淋巴瘤

• 批准号: 7746996
• 负责人: TRUDY M FORTE
• 金额: $ 10.94万
• 依托单位: LYPRO BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7746996
• 关键词: Acute Promyelocytic Leukemia; Aggressive course; Amino Acid Sequence; Animal Model; Antibodies; Antigens; Apolipoprotein A-I; Apolipoproteins; Apoptosis; Autophagocytosis; B lymphoid malignancy; B-Lymphocytes; Binding; Biological; C-terminal; CD20 Antigens; Caspase; Categories; Cell Culture Techniques; Cell Line; Cell Survival; Cells; Chemosensitization; Chimera organism; Chimeric Proteins; Code; Controlled Study; Cultured Cells; Disease; Disease remission; Dose; Drug Delivery Systems; Effectiveness; Escherichia coli; Evaluation; Exhibits; Exposure to; Flow Cytometry; Gene Targeting; Human; Induction of Apoptosis; Investigation; Juvenile Myelomonocytic Leukemia; Kaposi Sarcoma; Lipids; Lymphoma; MS4A1 gene; Malignant Neoplasms; Mantle Cell Lymphoma; Measurement; Mediating; Methods; Modeling; Neuroblastoma; Non-Hodgkin' s Lymphoma; Nuclear Hormone Receptors; Oncogenes; Parents; Plasmid Cloning Vector; Production; Property; Protein Engineering; Proteins; RXR; Reaction Time; Reactive Oxygen Species; Recombinant Fusion Proteins; Research; Research Design; Research Personnel; Resistance; Retinoic Acid Receptor; Retinoids; Role; Surface; Time; Tissues; Treatment Protocols; Tretinoin; Western Blotting; base; cancer therapy; cell growth; cell type; high risk; in vivo; member; nanobiotechnology; nanoscale; novel; novel strategies; outcome forecast; particle; public health relevance; targeted delivery

DESCRIPTION (provided by applicant): Mantle cell lymphoma (MCL) is a B-cell malignancy that is characterized by dysregulation of various oncogenes. Building on evidence that retinoic acid and its derivative, all trans retinoic acid (ATRA), are useful agents that potentiate apoptosis or anti-proliferative effects, it is proposed to pursue a strategy of targeted delivery of ATRA to MCL cells in culture. Using a novel delivery vehicle wherein ATRA is solubilized in nanoscale, protein stabilized lipid particles, termed nanodisks (ND), protein engineering methods will be employed to target ATRA-ND to the CD20 antigen present on the surface of B lymphocytes. This will be achieved by construction of a single chain variable antibody (scFv) apolipoprotein (apo) fusion protein. It is hypothesized that 1-CD20 scFv7apoA-I fusion protein will be capable of forming ND while retaining the antigen recognition properties of the parent scFv. Recombinant fusion protein will be expressed in E. coli, isolated and characterized. The ability of the chimera to associate with lipid and induce formation of ND will be determined while the antigen recognition properties of the 1-CD20 scFv portion of the fusion protein will be evaluated by Western blot and flow cytometry. ND prepared with wild type apoA-I will serve as control for these studies. In a second aim the effect of 1-CD20 scFv7apoA-I ATRA-ND on MCL cells in culture will be assessed. It is hypothesized that retinoid containing, targeted ND, will display enhanced induction of apoptosis in cell culture models of MCL. Different MCL cell lines will be employed in studies designed to evaluate targeting efficiency, concentration effectiveness and cell viability following exposure to CD20 targeted scFv7apoA-I ATRA-ND. Cultured cells will be exposed to control ATRA-ND and 1-CD20 scFv7apoA-I ATRA-ND followed by measurements of cell viability, apoptosis and autophagy. Dose- response and time course studies will be conducted to define optimal conditions. Studies will also be performed with ND harboring related synthetic and natural retinoids. The results of these studies will expand the potential of ND mediated drug delivery by demonstrating cell / tissue specific targeting, providing a framework for in vivo studies of targeted ND in animal models of MCL. PUBLIC HEALTH RELEVANCE: Despite progress made in the broad category of lymphomas, mantle cell lymphoma (MCL) remains a poorly treated disease with median survival time of approximately 3 to 4 years. New therapy regimens have increased the complete remission rate but they have done little to change overall survival. Research proposed herein will evaluate the effectiveness of targeted drug payload delivery to cultured MCL cells. These studies will establish a novel approach with broad applicability, establishing targeted nanodisks as a platform that can be used with other forms of cancer, potentially decreasing the public burden associated with this disease.

描述（由申请人提供）：套细胞淋巴瘤（MCL）是一种B细胞恶性肿瘤，其特征是各种癌基因的失调。基于维甲酸及其衍生物全反式维甲酸（ATRA）是增强细胞凋亡或抗增殖作用的有用药剂的证据，提出了追求将ATRA靶向递送至培养物中的MCL细胞的策略。使用其中ATRA溶解在纳米级蛋白质稳定的脂质颗粒（称为纳米盘（ND））中的新型递送载体，将采用蛋白质工程方法将ATRA-ND靶向存在于B淋巴细胞表面上的CD 20抗原。这将通过构建单链可变抗体（scFv）载脂蛋白（apo）融合蛋白来实现。假设1-CD 20 scFv 7apoA-I融合蛋白将能够形成ND，同时保留亲本scFv的抗原识别特性。重组融合蛋白将在E.大肠杆菌，分离和表征。将测定嵌合体与脂质结合并诱导ND形成的能力，同时通过Western印迹和流式细胞术评价融合蛋白的1-CD 20 scFv部分的抗原识别特性。用野生型apoA-I制备的ND将作为这些研究的对照。在第二个目的中，将评估1-CD 20 scFv 7apoA-I ATRA-ND对培养物中的MCL细胞的影响。据推测，含有类维生素A的靶向ND将在MCL的细胞培养模型中显示增强的细胞凋亡诱导。不同的MCL细胞系将用于设计为评估暴露于靶向CD 20的scFv 7apoA-I ATRA-ND后的靶向效率、浓度有效性和细胞活力的研究中。将培养的细胞暴露于对照ATRA-ND和1-CD 20 scFv 7apoA-I ATRA-ND，然后测量细胞活力、凋亡和自噬。将进行剂量反应和时间过程研究以确定最佳条件。还将对含有相关合成和天然类维生素A的ND进行研究。这些研究的结果将通过证明细胞/组织特异性靶向来扩大ND介导的药物递送的潜力，为在MCL动物模型中靶向ND的体内研究提供框架。 公共卫生相关性：尽管在广泛的淋巴瘤类别中取得了进展，但套细胞淋巴瘤（MCL）仍然是一种治疗效果不佳的疾病，中位生存时间约为3至4年。新的治疗方案提高了完全缓解率，但对总生存率的影响很小。本文提出的研究将评估靶向药物有效载荷递送至培养的MCL细胞的有效性。这些研究将建立一种具有广泛适用性的新方法，建立靶向纳米盘作为可用于其他形式癌症的平台，从而可能减少与这种疾病相关的公共负担。