Abstract
摘要:藥物協同治療於近年來已成為處置癌症、代謝失調或抑制發炎首要手段。相對於使 用單一藥物,藥物協同作用不僅可增加治療成功率,並可大幅減少藥物劑量使用以 及副作用。然而當前的藥物篩選平台存在常見的缺陷,比如額外的樣本需求(每個 測試需要至少1000顆細胞)或是較長的評估時間(3天至3禮拜),因此大部分的藥物 協同策略未必有理想的成效,特別是針對個人化醫療以及與時間賽跑的病患。 本計畫為了突破以上所提及的研究困境,因此開發一仿生奈米液滴處理平台,該技 術係模仿沙漠甲蟲的背部微結構,此微結構係由石蠟疏水與親水表面所組成。藉由 整合雷射直雕技術,我們製作出新型的仿生微結構介面,該介面可應用於細胞排列 與高通量藥物篩選。初步實驗結果得知,經由平台篩選的最佳化藥物組合可有效降 低單一藥物使用量至28倍。此外,相對於使用標準化96多孔盤來預測腫瘤抑制 ,我們平台的預測結果與小鼠腫瘤動物模型結果呈現高度一致性。整體藥物評估時 間僅只需短短2天。我們亦成功整合血球血癌細胞模型來篩選新型的紅血球分化增 效劑或抗貧血藥物。因此,本兩年計畫所開發的平台不但具有晶片可拋棄式、低耗 費、自動化以及深度類神經網路來加速評估藥物協同效能,於未來可直接被應用於 研究抗癌藥物開發以及血球分化發展過程。 本計畫將會開發並雛型化仿生奈米液滴處理平台,該平台將具備自動化、高效率、 高通量智能分析以及低樣本需求與低耗費特色,並將鎖定抗癌藥物快速篩選、開發 以及開闢貧血治療新契機為首要應用標的。國人因癌致死率逐年增加,而因癌症、 慢性疾病或其他環境因素所衍生的貧血相關疾病也有增加趨勢。有鑑於此,本計畫 提出的平台預期將可有效舒緩此問題,藉由提升藥物協同作用並降低藥物對病患的 副作用,以及大幅度減輕病患等待藥物評估時間與金錢的負擔,達到雙贏的局面。
Abstract: Therapeutic drug synergism intervened in but not limited to the treatment of cancer, metabolic or inflammation disorders has been the crucial avenue rather than using a single effector. This regimen not only improves the therapeutic efficacy by triggering synthetic lethality in target cells, but it also minimizes the side effects by reducing doses of each drug. Nonetheless, it remains inherently challenging. For example, a limited cell count from cancer patient’s samples is typically processed with only 105 cells from one biopsy. Such limitation would be interrogated to achieve potent personalized drug cocktails while adopting conventional multi-well plate assays (at least 1000 cells needed per test). Moreover, the regular evaluation period of 3 days to 3 weeks required for screening drugs is a major concern as well, since long wait time is not applicable for some patients with severe disease.
Our lab has developed, in the last few years, a wax-imprinted nanodroplet processing technique that is inspired from Stenocara gracilipes beetle’s bumpy back surfaces. By incorporating laser-direct writing with such technique, we can control the hydrophobic-hydrophilic interfaces with configurable features of both patterning cells and performing high-throughput drug synergy screening. Leveraging on advantages of utilizing customized liquid dispensing, moreover, cell counts and reagents needed can be retained down to 50 cells and 200 nl for each test, respectively. Results thus far are promising. Both prostate (PC-3) and breast (MCF7 and MDA-MB-231) cancer cell lines were tested and compared in vitro and in vivo. The optimal drug combination, evaluated from the platform, for inhibiting cell proliferation was of average 28-folds in dose reduction compared with single drugs conducted. Furthermore, the prediction of tumor inhibition showed a surprising correspondence with which from tumor-bearing mouse models, in contrast to standard 96-well plate assays. Remarkably, only 2 days were required to achieve actionable evaluations (1 day for experiment and the rest for imaging/computing manually). By this platform, likewise, the feasibility for screening antianemia drugs was also successfully demonstrated in leukemia K562 cell models. Consequently, these findings imply that the platform can serve and boost the high-throughput requirements targeting anticancer or antianemia drug synergy screening.
This two-year proposal aims to develop the technology with the primary goals of (1) producing the disposable bioinspired nanodroplet processing (BioNDP) slide in standard microscope slide dimension and (2) prototyping the BioNDP platform with deep learning analyzer targeting cancer or anemia-associated diseases. The yearly tasks of the entire proposal are as follow:
Year 1: Design of the BioNDP platform and the BioNDP analyzer. Development of tumor-bearing zebrafish models to boost the preclinical drug screening. Development of glycophorin A (GlyA)-inducible cell models to improve the screening of antianemia drugs. Prototyping of the BioNDP slide. Initial testing of the BioNDP platform with cell lines and chemotherapeutic drugs.
Year 2: Finish testing of the BioNDP platform. Comparison of the evaluated results from BioNDP, 96-well plate and zebrafish xenograft model. Finish screening of the antianemia drugs. Finish prototyping of the BioNDP platform.
Keyword(s)
藥物協同作用
個人化醫療
仿生
奈米液滴
高通量
癌症
異種移植
貧血
深度學習
drug synergism
personalized
bioinspired
nanodroplet
high-throughput
cancer
xenograft
anemia
deep learning