整合細胞微陣列晶片平台及斑馬魚腫瘤異種移植模型以加速評估對人類乳腺癌之最佳化療藥物組合

Abstract

乳腺癌為女性最嚴重之惡性腫瘤。儘管乳腺癌已有多種治療方法，化療藥物之協同作用依舊具或多或少之療效。協同作用不僅能增強化療功效，並能減輕患者副作用及抗藥性。然而，來自原發性腫瘤之細胞數量有限，且對於高通量篩選所需大量藥物仍具挑戰性。在此，我們提供了一種新型細胞微陣列晶片系統，它由蠟排列孔洞之晶片及一臺自動液體分配裝置所組成，特別適用於高通量藥物協同作用篩選。我們導入四種常用化療藥物，包含順鉑、5-氟尿嘧啶、環磷酰胺及依托泊苷和兩種乳腺癌細胞，MCF7與MDA-MB-231，進行概念性驗證。並利用協同組合指數方程式運算來篩選出最佳化療藥物組合。此外，晶片系統所評估之最佳藥物組合將再利用斑馬魚腫瘤異種移植模型在動物體內進一步驗證。結果顯示，與單一藥物相比，篩選出之最佳藥物協同作用能減少單一藥物約14.3倍之劑量。與標準96孔盤相比，此晶片可降低500倍之藥物體積。此外，透過注射乳癌細胞至斑馬魚模型，我們成功地證明最佳藥物組合可有效抑制21％之腫瘤生長。值得注意的是，細胞微陣列晶片可以比96孔盤更準確地預測動物實驗結果。總之，本研究整合細胞微陣列晶片及斑馬魚腫瘤異種移植模型，能加速評估對人類乳腺癌之最佳化療藥物組合，以提供個人化藥物之早期篩選及未來新藥開發之新平台。

Breast cancer has been recently revealed as the most deadly cancer to females, hitting an astounding 15-year reduction of life. Although numerous planning treatments have been launched, synergistic interactions of current anti-cancer drugs still stun the target. The Synergistic interactions not only improve breast cancer chemotherapy efficacy, but reduce drug resistance and side-effects as well. However, the shortages of less cell amount from primary tumors and massive drugs needed still remain challenging for high throughput evaluation. In this research, we present a cellular microarray ParaStamp (CMP) chip system, which combines of wax-well-arrayed chips and an automatic liquid dispensing machine, particularly for high throughput drug synergy screening. For the proof-of-conceptual demonstration, we conduct four existent chemotherapeutic drugs (e.g. cisplatin, 5-Fluorouracil, cyclophosphamide, and etoposide) and two breast cancer cell lines (e.g. MCF7 and MDA-MB-231 cells) into the system. A screening strategy based on the combination index (CI) equation is then utilized to identify the optimal drug combinations. In addition, the optimizations are further verified by zebrafish (ZF) tumor xenograft models. Results show that the optimal drug combination screened can cause the dose reduction down to approximately 14.3 folds compared with single drugs conducted. In contrast to standard 96-well plate assay, the study conductor demonstrates that the volume of each tested drug can be retained up to 500 folds. Moreover, a significant 21% inhibition of MCF7 breast tumors engrafted in ZF models is successfully presented by the identified drug combination. Remarkably, the CMP chip could predict the in vivo efficacy more accurately than 96-well plate assays. Taken together, our findings demonstrate the integration of the CMP chip platform and zebrafish tumor xenograft model could improve the outcome for breast cancer chemotherapy. It may further offer new opportunities to enhance personalized medicine and drug discovery.