Khalafi Shayan, Zhu Shoumin, Khurana Rimpi, Lohse Ines, Giordano Silvia, Corso Simona, Al-Ali Hassan, Brothers Shaun P, Wahlestedt Claes, Schürer Stephan, El-Rifai Wael
Department of Surgery, Miller School of Medicine, University of Miami, Rosenstiel Medical Science Bldg, 1600 NW 10th Ave, Room 4007, Miami, FL 33136-1015, United States.
Department of Surgery, Miller School of Medicine, University of Miami, Rosenstiel Medical Science Bldg, 1600 NW 10th Ave, Room 4007, Miami, FL 33136-1015, United States; Sylvester Comprehensive Cancer Center, Miller School of Medicine, University of Miami, Miami, FL 33136, United States.
Transl Oncol. 2022 Jan;15(1):101260. doi: 10.1016/j.tranon.2021.101260. Epub 2021 Nov 1.
Gastric cancer (GC) is frequently characterized by resistance to standard chemotherapeutic regimens and poor clinical outcomes. We aimed to identify a novel therapeutic approach using drug sensitivity testing (DST) and our computational SynerySeq pipeline. DST of GC cell lines was performed with a library of 215 Federal Drug Administration (FDA) approved compounds and identified clofarabine as a potential therapeutic agent. RNA-sequencing (RNAseq) of clofarabine treated GC cells was analyzed according to our SynergySeq pipeline and identified pictilisib as a potential synergistic agent. Clonogenic survival and Annexin V assays demonstrated increased cell death with clofarabine and pictilisib combination treatment (P<0.01). The combination induced double strand breaks (DSB) as indicated by phosphorylated H2A histone family member X (γH2AX) immunofluorescence and western blot analysis (P<0.01). Pictilisib treatment inhibited the protein kinase B (AKT) cell survival pathway and promoted a pro-apoptotic phenotype as evidenced by quantitative real time polymerase chain reaction (qRT-PCR) analysis of the B-cell lymphoma 2 (BCL2) protein family members (P<0.01). Patient derived xenograft (PDX) data confirmed that the combination is more effective in abrogating tumor growth with prolonged survival than single-agent treatment (P<0.01). The novel combination of clofarabine and pictilisib in GC promotes DNA damage and inhibits key cell survival pathways to induce cell death beyond single-agent treatment.
胃癌（GC）的常见特征是对标准化疗方案耐药且临床预后较差。我们旨在通过药物敏感性测试（DST）和我们的计算SynerySeq流程来确定一种新的治疗方法。使用包含215种美国食品药品监督管理局（FDA）批准化合物的文库对GC细胞系进行DST，并确定氯法拉滨为潜在治疗药物。根据我们的SynergySeq流程分析氯法拉滨处理的GC细胞的RNA测序（RNAseq），并确定匹地昔布为潜在的协同药物。克隆形成存活和膜联蛋白V检测表明，氯法拉滨和匹地昔布联合治疗可增加细胞死亡（P<0.01）。如磷酸化H2A组蛋白家族成员X（γH2AX）免疫荧光和蛋白质印迹分析所示，联合治疗诱导双链断裂（DSB）（P<0.01）。匹地昔布治疗抑制蛋白激酶B（AKT）细胞存活途径，并促进促凋亡表型，B细胞淋巴瘤2（BCL2）蛋白家族成员的定量实时聚合酶链反应（qRT-PCR）分析证明了这一点（P<0.01）。患者来源的异种移植（PDX）数据证实，与单药治疗相比，联合治疗在消除肿瘤生长和延长生存期方面更有效（P<0.01）。氯法拉滨和匹地昔布在GC中的新联合治疗促进DNA损伤并抑制关键细胞存活途径，从而诱导细胞死亡，其效果优于单药治疗。
