Targeting of the Hedgehog/GLI and mTOR pathways in advanced pancreatic cancer,a phase 1 trial of Vismodegib and Sirolimus combination

Background/ObjectivesPreclinical data indicated a functional and molecular interaction between Hedgehog (HH)/GLI and PI3K-AKT-mTOR pathways promoting pancreatic ductal adenocarcinoma (PDAC). A phase I study was conducted of Vismodegib and Sirolimus combination to evaluate maximum tolerated dose (MTD) and preliminary anti-tumor efficacy.MethodsCohort I included advanced solid tumors patients following a traditional 3 + 3 design. Vismodegib was orally administered at 150 mg daily with Sirolimus starting at 3 mg daily, increasing to 6 mg daily at dose level 2. Cohort II included only metastatic PDAC patients. Anti-tumor efficacy was evaluated every two cycles and target assessment at pre-treatment and after a single cycle.ResultsNine patient were enrolled in cohort I and 22 patients in cohort II. Twenty-eight patients were evaluated for dose-limiting toxicities (DLTs). One DLT was observed in each cohort, consisting of grade 2 mucositis and grade 3 thrombocytopenia. The MTD for Vismodegib and Sirolimus were 150 mg daily and 6 mg daily, respectively. The most common grade 3–4 toxicities were fatigue, thrombocytopenia, dehydration, and infections. A total of 6 patients had stable disease. No partial or complete responses were observed. Paired biopsy analysis before and after the first cycle in cohort II consistently demonstrated reduced GLI1 expression. Conversely, GLI and mTOR downstream targets were not significantly affected.ConclusionsThe combination of Vismodegib and Sirolimus was well tolerated. Clinical benefit was limited to stable disease in a subgroup of patients. Targeting efficacy demonstrated consistent partial decreases in HH/GLI signaling with limited impact on mTOR signaling. These findings conflict with pre-clinical models and warrant further investigations.     

猴头菌提取物颗粒联合美沙拉秦治疗溃疡性结肠炎的临床研究

目的探讨猴头菌提取物颗粒联合美沙拉秦肠溶片治疗溃疡性结肠炎的临床疗效。方法选取2016年4月—2018年4月咸阳市第一人民医院收治的120例溃疡性结肠炎患者作为研究对象,将所有患者根据随机数字表法分为对照组和治疗组,每组各60例。对照组患者口服美沙拉秦肠溶片,1片/次,3次/d;治疗组患者在对照组治疗的基础上口服猴头菌提取物颗粒,3g/次,3次/d。两组患者均持续治疗4周。观察两组的临床疗效,比较两组的血清炎性因子水平、疾病活动指数(DAI)评分、临床症状改善时间。结果治疗后,对照组和治疗组的总有效率分别为81.67%、95.00%,两组比较差异有统计学意义(P0.05)。治疗后,两组患者白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平均显著降低,同组治疗前后比较差异有统计学意义(P0.05);并且治疗组血清炎性因子水平明显低于对照组,两组比较差异有统计学意义(P0.05)。治疗后,两组患者DAI评分显著降低,同组治疗前后比较差异有统计学意义(P0.05);并且治疗组DAI评分明显低于对照组,两组比较差异有统计学意义(P0.05)。治疗后,治疗组患者治愈时间、黏液脓血便消失时间均短于对照组,两组比较差异有统计学意义(P0.05)。结论猴头菌提取物颗粒联合美沙拉秦肠溶片治疗溃疡性结肠炎具有较好的治疗效果,可改善患者临床症状,降低血清炎性因子水平,具有一定临床推广应用价值。     

Indian Hedgehog在牵张力促进成骨细胞增殖中的作用研究

目的观察Hedgehog（Hh）信号在牵张力调控成骨细胞增殖中的作用。方法体外分离和培养新生大鼠颅顶骨成骨细胞,应用Flexcell 4000TM加力系统施加3%和6%的2种牵张应变。用N端Hedgehog重组蛋白（N-Shh）、Hedgehog通道抑制剂环巴胺（cy）和机械通道抑制剂三氯化钆（GdCl3）对成骨细胞进行干预。分别采用细胞计数、甲基噻唑基四唑（MTT）比色法和流式细胞仪检测成骨细胞的增殖。采用SAS 8.0软件包对结果进行统计分析。结果在成骨细胞体外培养过程中,牵张力促进了成骨细胞的增殖,以6%牵张应变更显著;牵张力仍能促进经cy处理后的成骨细胞的增殖,但这种促进作用能被GdCl3抑制。结论牵张力促进成骨细胞增殖的作用有一部分是通过上调Ihh的表达实现的。     

蝎毒多肽干预K562/BALB/c-nu小鼠Hedgehog通路传导的机制研究

目的探究蝎毒多肽干预K562/BALB/c-nu白血病荷瘤小鼠Hedgehog通路信号传导的机制,从基因、蛋白水平解析蝎毒多肽抑制慢性粒细胞白血病发展的分子机制和作用靶点。方法将K562/BALB/c-nu白血病荷瘤小鼠分为对照组、模型组、伊马替尼(50 mg/kg)组和蝎毒多肽高、中、低剂量(0.3、0.6、1.2 mg/kg)组,药物干预14 d后,实时荧光定量PCR(q RT-PCR)法检测小鼠瘤组织Hedgehog信号通路上游因子Shh、Ptch、Smo m RNA表达水平,Western blotting法检测Shh、Ptch、Smo蛋白表达水平,ELISA法检测小鼠瘤组织Hedgehog信号通路下游因子Gli1蛋白表达水平。结果与模型组比较,蝎毒多肽干预组小鼠瘤组织Shh m RNA和蛋白表达水平均升高;蝎毒多肽低、中剂量组小鼠瘤组织Ptch、Smo m RNA及蛋白表达水平均降低;伊马替尼组各上游因子与模型组比较差异不显著;蝎毒多肽低、中剂量组小鼠瘤组织Gli1蛋白表达水平降低,蝎毒多肽高剂量组、伊马替尼组Gli1蛋白表达水平无显著差异。结论蝎毒多肽能够抑制Hedgehog信号通路上游因子Ptch、Smo以及下游因子Gli1的表达,而伊马替尼对Hedgehog信号通路无明显抑制作用。     

刺猬蛋白在宫颈癌组织中的表达及其临床意义

目的检测刺猬蛋白(Sonic Hedgehog,SHH)在正常宫颈、宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)及宫颈癌组织中的表达情况,探讨其与宫颈癌发生发展的关系。方法采用免疫组化SP法,检测正常宫颈组织、CIN及宫颈癌组织中SHH的表达。结果 SHH在宫颈癌组织中高表达。结论 SHH在宫颈癌中的异常表达表明Hedgehog(Hh)信号传导通路可能参与宫颈癌的发生过程。     

A ciliopathy with hydrocephalus,isolated craniosynostosis,hypertelorism, and clefting caused by deletion of Kif3a

Malformations of the facial midline are a consistent feature among individuals with defects in primary cilia. Here, we provide a framework in which to consider how these primary cilia-dependent facial anomalies occur. We generated mice in which the intraflagellar transport protein Kif3a was deleted in cranial neural crest cells. The Kif3a phenotypes included isolated metopic craniosynostosis, delayed closure of the anterior fontanelles, and hydrocephalus, as well as midline facial anomalies including hypertelorism, cleft palate, and bifid nasal septum. Although all cranial neural crest cells had truncated primary cilia as a result of the conditional deletion, only those in the midline showed evidence of hyper-proliferation and ectopic Wnt responsiveness. Thus, cranial neural crest cells do not rely on primary cilia for their migration but once established in the facial prominences, midline cranial neural crest cells require Kif3a function in order to integrate and respond to Wnt signals from the surrounding epithelia.