Microfluidics-Based Platform for Screening Combinatorial Drug Treatments

基于微流控的组合药物治疗筛选平台

• 批准号: 7648208
• 负责人: ARUL JAYARAMAN
• 金额: $ 16.39万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7648208
• 关键词: Ablation; Agonist; Antibodies; Apoptosis; Apoptotic; Attention; Binding; Cell Death; Cells; Cessation of life; Clinic; Combined Modality Therapy; Development; Disease; Dose; Epithelial; Evaluation; Exposure to; FDA approved; Family; Goals; Human; Individual; Investigation; Lead; Libraries; Ligands; Liquid substance; Mediating; Methods; Microfabrication; Microfluidics; Miniaturization; Mutagens; Mutation; Necrosis; Neoplastic Cell Transformation; Normal Cell; Outcome; PC3 cell line; Peptides; Pharmaceutical Preparations; Phenotype; Prostate; Radiation; Radio; Research; Resistance; Sampling; Screening procedure; Small Interfering RNA; Stimulus; Stromal Cells; TNFRSF10A gene; TNFRSF10B gene; Technology; Therapeutic; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Work; base; cancer cell; cancer therapy; chemosensitizing agent; chemotherapy; combinatorial; cytokine; cytotoxicity; death receptor-4; high throughput screening; human TNFRSF10A protein; member; neoplastic cell; novel; public health relevance; receptor; resistance mechanism; response; small molecule; small molecule libraries; treatment strategy; tumor; tumorigenic

DESCRIPTION (provided by applicant): The transformation of neoplastic disease to a highly tumorigenic, metastatic, and radio / chemotherapy resistant phenotype is a result of the accumulation of a number of genetic changes. Combination therapies have emerged as powerful alternatives to `single-agent therapies' since they enhance cell death by overcoming multiple resistance mechanisms in cancer cells. Tumor Necrosis Factor-1 Related Apoptosis Inducing Ligand (TRAIL) and agonistic antibodies to Death Receptor (DR) 4 and 5 have attracted significant attention due to their ability to selectively induce apoptosis in transformed (malignant) cells while demonstrating little cytotoxicity in normal cells. Despite this promise, many tumor cells are resistant to TRAIL and as a result, radiation and chemotherapeutic genotoxins have been employed to sensitize cancer cells to death receptor- mediated apoptosis. While this approach has been moderately successful, traditional `single-chemosensitizer discovery' approaches are inherently low-throughput and do not allow for comparison of different sensitizing agents. We hypothesize that screening small molecule libraries can result in the rapid identification of new combination treatment strategies and candidates and enable comparison of different molecules. In this proposal, we describe a novel cancer cell-based screening paradigm in which we integrate traditional well plate based screening and microfabrication / microfluidics technologies for the identification and mechanistic evaluation of small-molecules that sensitize cancer cells to death receptor-mediated apoptosis. Well-plate screening methods will be employed in the primary screening of a library of ~2,500 FDA approved drugs for identifying lead candidates. Microfluidic cancer cell arrays will then be employed to carry out stringent secondary screening of leads selected from the primary screen; it is hypothesized that the ability to interrogate cells with combinatorial stimuli using microfluidics can lead to the parallelization and miniaturization of the secondary screening step. The higher throughput required in the primary screening and the combinatorial dosing requirements in the secondary screening justify our choices of using conventional 96-well plates for the former and a microfluidcs-based cancer cell array for the latter. It is anticipated that the proposed research will lead to the development of a novel integrated screening paradigm based on well-plates and microfluidic cell arrays for the discovery of small molecules for combination treatments and the identification and mechanistic evaluation of approximately 20-50novel candidates that enhance cancer cell death in combination with death receptor agonists. The proposed screening platform has implications beyond the present work; the platform can be extended to screen/investigate various factors influencing combination therapies including, other therapeutic candidates (e.g. peptides, siRNA, etc.), soluble factors (e.g. cytokines, metabolites, etc.), and tumor microenvironment effects. Project Narrative This proposal aims to identify effective combination treatments for cancer therapy using high-throughput screening. A novel HTS paradigm based on the integration of traditional well-plate screening and a microfluidic cancer cell array will be used. The microfluidic cell array will be developed as a platform for investigating synergistic interactions between candidate molecules. Successful completion of this work can lead to the development of new combination anti-cancer therapies and in other cell-based high- throughput screening applications. PUBLIC HEALTH RELEVANCE: This proposal aims to identify effective combination treatments for cancer therapy using high-throughput screening. A novel HTS paradigm based on the integration of traditional well-plate screening and a microfluidic cancer cell array will be used. The microfluidic cell array will be developed as a platform for investigating synergistic interactions between candidate molecules. Successful completion of this work can lead to the development of new combination anti-cancer therapies and in other cell-based high through put screening applications.

描述（由申请方提供）：肿瘤性疾病转化为高度致瘤性、转移性和放疗/化疗耐药表型是许多遗传变化累积的结果。联合疗法已成为“单一药剂疗法”的有力替代品，因为它们通过克服癌细胞中的多种耐药机制来促进细胞死亡。肿瘤坏死因子-1相关的凋亡诱导配体（TRAIL）和死亡受体（DR）4和5的激动性抗体由于其在转化（恶性）细胞中选择性诱导凋亡的能力而引起了显著的关注，同时在正常细胞中表现出很小的细胞毒性。尽管有这种前景，但许多肿瘤细胞对TRAIL具有抗性，因此，放射和化疗基因毒素已被用于使癌细胞对死亡受体介导的细胞凋亡敏感。虽然这种方法取得了一定的成功，但传统的“单一化学增敏剂发现”方法本身通量低，不允许比较不同的增敏剂。我们假设筛选小分子文库可以快速鉴定新的组合治疗策略和候选物，并能够比较不同的分子。在这个提议中，我们描述了一种新的基于癌细胞的筛选模式，其中我们整合了传统的基于孔板的筛选和微加工/微流体技术，用于识别和机械评价使癌细胞对死亡受体介导的凋亡敏感的小分子。将采用孔板筛选方法对约2，500种FDA批准的药物库进行初步筛选，以确定主要候选药物。然后将采用微流体癌细胞阵列对从初级筛选中选择的先导物进行严格的次级筛选;假设使用微流体用组合刺激询问细胞的能力可以导致次级筛选步骤的并行化和小型化。初级筛选中所需的较高通量和次级筛选中的组合给药要求证明了我们选择使用常规96孔板用于前者和基于微流体的癌细胞阵列用于后者的合理性。预计拟议的研究将导致开发一种新的综合筛选模式，基于孔板和微流体细胞阵列，用于发现用于联合治疗的小分子，以及鉴定和机制评估约20- 50种与死亡受体激动剂联合增强癌细胞死亡的新候选物。所提出的筛选平台具有超出本工作的意义;该平台可以扩展到筛选/研究影响组合疗法的各种因素，包括其他治疗候选物（例如肽、siRNA等），可溶性因子（例如细胞因子、代谢物等），和肿瘤微环境的影响。项目叙述本提案旨在使用高通量筛选确定有效的癌症治疗联合治疗。将使用基于传统孔板筛选和微流体癌细胞阵列的集成的新HTS范例。微流控细胞阵列将被开发为研究候选分子之间协同相互作用的平台。这项工作的成功完成可以导致开发新的组合抗癌疗法和其他基于细胞的高通量筛选应用。 公共卫生相关性：该提案旨在通过高通量筛查确定有效的癌症联合治疗。将使用基于传统孔板筛选和微流体癌细胞阵列的集成的新HTS范例。微流控细胞阵列将被开发为研究候选分子之间协同相互作用的平台。这项工作的成功完成可以导致新的组合抗癌疗法的开发和其他基于细胞的高通量筛选应用。

项目成果

Parallel screening of FDA-approved antineoplastic drugs for identifying sensitizers of TRAIL-induced apoptosis in cancer cells.

• DOI: 10.1186/1471-2407-11-470
• 发表时间: 2011-11-01
• 期刊: BMC cancer
• 影响因子: 3.8
• 作者: Taylor DJ;Parsons CE;Han H;Jayaraman A;Rege K
• 通讯作者: Rege K

Slow Release Kinetics of Mitoxantrone from Ordered Mesoporous Carbon Films.

有序介孔碳膜中米托蒽醌的缓释动力学。

• DOI: 10.1016/j.micromeso.2012.05.003
• 发表时间: 2012
• 期刊: Microporous and mesoporous materials : the official journal of the International Zeolite Association
• 作者: Labiano,Alpha;Dai,Mingzhi;Taylor,David;Young,Wen-Shiue;Epps3rd,ThomasH;Rege,Kaushal;Vogt,BryanD
• 通讯作者: Vogt,BryanD