Tolerability and safety of EUS-injected adenovirus-mediated double-suicide gene therapy with chemotherapy in locally advanced pancreatic cancer: Phase I Trial.
EUS 注射腺病毒介导的双自杀基因治疗联合化疗治疗局部晚期胰腺癌的耐受性和安全性: I 期试验。
- 作者列表："Lee JC","Shin DW","Park H","Kim J","Youn Y","Kim JH","Kim J","Hwang JH
BACKGROUND AND AIMS:Locally advanced pancreatic cancer (LAPC) is challenging. Here we aimed to evaluate the tolerability and safety of Ad5-yCD/mutTK(SR39)rep-ADP (Ad5-DS), a replication-competent adenovirus-mediated double-suicide gene therapy in combination with gemcitabine in LAPC patients. METHODS:Newly diagnosed LAPC patients were enrolled in this single-center, open label, 3+3 dose-escalation phase I trial (NCT02894944). Ad5-DS was injected into the pancreatic mass with endoscopic ultrasonography-guided fine needles combined with oral 5-fluorocytosine and valganciclovir, and standard dose of intravenous gemcitabine. The doses of Ad5-DS in cohorts 1 to 3 were 1 × 1011, 3 × 1011, and 1 × 1012 vp/mL, respectively. Patients were observed for dose-limiting toxicity (DLT) for 8 weeks after Ad5-DS injection. Toxicity within 12 weeks, tumor response in 12 weeks, disease progression in 6.5 months, and detection of adenoviral DNA particle in 8 weeks were also assessed. RESULTS:Among the 11 enrolled patients, 9 completed the evaluation period and 2 withdrew their consent. No DLT was reported; thus, the maximum tolerated dose (MTD) was not reached. No additional toxicity was reported in 9 to 12 weeks. One patient showed a partial response and 8 showed stable disease at 12 weeks. Two patients showed disease progression at 6.5 months (median progression-free survival, 11.4 months). At 8 weeks, serum adenoviral DNA particles were detected in 4 patients (median, 55 days). CONCLUSION:Combination of intratumoral Ad5-DS and gemcitabine is safe and well-tolerated in patients with locally advanced pancreatic cancer. This warrants further investigation in a larger clinical trial.
背景和目的: 局部晚期胰腺癌 (LAPC) 具有挑战性。在这里，我们旨在评估 Ad5-yCD/mutTK (SR39) rep-ADP (Ad5-DS) 的耐受性和安全性, 一种复制活性腺病毒介导的双自杀基因治疗与吉西他滨联合治疗 LAPC 患者。 方法: 新诊断的 LAPC 患者入选本单中心、开放标签、 3 + 3 剂量递增 I 期试验 (NCT02894944)。采用超声内镜引导下细针联合口服 5-氟胞嘧啶和 lovir 更昔洛韦及标准剂量吉西他滨静脉注射 Ad5-DS。队列 1 ~ 3 中 Ad5-DS 的剂量分别为 1 × 1011 、 3 × 1011 和 1 × 1012 vp/mL。观察患者注射 Ad5-DS 后 8 周的剂量限制性毒性 (DLT)。12 周内毒性，12 周肿瘤反应，6.5 个月疾病进展，8 周检测腺病毒 DNA 颗粒也被评估。 结果: 11 例入组患者中，9 例完成评估期，2 例撤回同意。未报告 DLT; 因此，未达到最大耐受剂量 (MTD)。在 9 至 12 周内未报告额外毒性。1 例患者显示部分缓解，8 例患者在 12 周时疾病稳定。2 例患者在 6.5 个月时出现疾病进展 (中位无进展生存期，11.4 个月)。8 周时，检测 4 例患者 (中位数，55 天) 血清腺病毒 DNA 颗粒。 结论: Ad5-DS 联合吉西他滨治疗局部晚期胰腺癌安全、耐受性好。这值得在更大的临床试验中进一步研究。
METHODS::Aims: Radiotherapy is predominantly used as one of the treatment modalities to treat local tumor in colorectal cancer (CRC). Hindrance in disease treatment can be attributed to radio-tolerance of cancer stem cells (CSCs) subsistence in the tumor. Understanding the radio-resistant property of CSCs might help in the accomplishment of targeted radiotherapy treatment and increased disease-free survival. Telomeric RAP1 contributes in modulation of various transcription factors leading to aberrant cell proliferation and tumor cell migration. Therefore, we investigated the role of RAP1 in maintaining resistance phenotype and acquired stemness in radio-resistant cells.Main Methods: Characterization of HCT116 derived radio-resistant cell (HCT116RR) was performed by cell survival and DNA damage profiling. RAP1 silenced cells were investigated for DNA damage and expression of CSC markers through western blotting and Real-time PCR post-irradiation. Molecular docking and co-immunoprecipitation study were performed to investigate RAP1 and KLF4 interaction followed by RAP1 protein status profiling in CRC patient.Key findings: We established radio-resistant cells, which showed tolerance to radiotherapy and elevated expression of CSC markers along with RAP1. RAP1 silencing showed enhanced DNA damage and reduced expression of CSC markers post-irradiation. We observed strong physical interaction between RAP1 and KLF4 protein. Furthermore, higher RAP1 expression was observed in the tumor of CRC patients. Dataset analysis also revealed that high expression of RAP1 expression is associated with poor prognosis.Significance: We conclude that higher expression ofRAP1 implicates its possible role in promoting radio-resistance in CRC cells by modulating DNA damage and CSC phenotype.
METHODS::Cancer stem-like cells are rare immortal cells within tumor, which are thought to play important roles in ionizing radiation (IR) therapy-resistance. Quercetin is a natural flavonoid with potential anti-cancer properties without significant cytotoxicity in normal tissues. In this study, we demonstrated that quercetin-IR combination treatment exhibited more dramatic anti-cancer effect than either quercetin or IR treatment alone via targeting colon cancer stem cells (CSCs) and inhibiting the Notch-1 signaling. These effects were further verified by in vivo studies which showed remarkable decrease of the CSCs markers and the expression of Notch-1 signaling proteins in human colon cancer xenografts in nude mice. Co-treatment with quercetin and low dose of radiation significantly reduced the expressions of all five proteins of γ-secretase complex in HT-29 and DLD-1 cells. In addition, ectopic expression of the Notch intracellular domain (NICD) partly reversed the inhibition effects by the combination therapy. In conclusion, our results indicated that the combination of quercetin (20 μM) and IR (5Gy) might be a promising therapeutic strategy for colon cancer treatment by targeting colon cancer stem-like cells and inhibiting the Notch-1 signaling. In future studies, we intend to further explore the potential therapeutic efficacy of the quercetin-radiation combination treatment in clinical trials.
METHODS:OBJECTIVES:Long-term prevention of metastatic disease remains a challenge in locally advanced rectal cancer, and robust pretreatment prognostic factors for metastatic progression are lacking. We hypothesized that detecting circulating tumor-specific DNA (ctDNA) based on hypermethylation of the neuropeptide Y gene (meth-ctDNA) could be a prognostic marker in the neoadjuvant setting; we examined this in a secondary, explorative analysis of a prospective trial. MATERIALS AND METHODS:Serum samples were prospectively collected in a phase III trial for locally advanced rectal cancer. Positivity for and fractional abundance of meth-ctDNA in baseline samples were estimated. Overall survival (OS) and the rate of distant metastases were compared between meth-ctDNA positive and negative patients; other prognostic factors were controlled for in multivariate Cox regression. Importance of quantitative load was examined by considering the fractional abundance of meth-ctDNA relative to total circulating DNA. RESULTS:Baseline serum samples were available for 146 patients. In total, 30 patients had presence of meth-ctDNA, with no correlation with cT (P=0.8) or cN (P=0.6) stages. Median follow-up was 10.6 years for OS and 5.1 years for freedom from distant metastases. Patients with meth-ctDNA had significantly worse 5-year OS (47% vs. 69%), even when controlling for other prognostic factors (hazard ratio=2.08; 95% confidence interval, 1.23-1.51). This seemed mainly driven by disparity in the rate of distant metastases (55% vs. 72% at 5 y, P=0.01); hazard ratio=2.20 (95% confidence interval, 1.19-4.07, P=0.01) in multivariate analysis. Increased quantitative load was highly significant for worse outcomes. CONCLUSIONS:Meth-ctDNA could be a potential prognostic marker in the neoadjuvant setting and may, if validated, identify patients at increased risk of distant metastases.