常规免疫组织化学初筛 ALK 阳性非小细胞肺癌专家共识

肺癌是全球范围内发病率和病死率最高的恶性肿瘤，其中非小细胞肺癌（ NSCLC ）约占80％。间变性淋巴瘤激酶（ ALK）阳性NSCLC是肺癌的一个特定分子亚型，约占全部NSCLC的3％～7％［1-2］。临床研究显示，ALK抑制剂克唑替尼（ crizotinib ）能显著延长晚期ALK阳性NSCLC患者的无进展生存期（ PFS），一线、二线单药治疗患者的中位PFS分别为10.9个月和7.7个月［3-4］。2013年1月中国食品和药品管理局（ CFDA）批准克唑替尼用于治疗局部晚期或转移性ALK阳性NSCLC患者。     

克唑替尼对ALK阳性非小细胞肺癌脑转移的疗效分析

目的：非小细胞肺癌(non-small cell lung cancer,NSCLC)脑转移是影响NSCLC患者生存期及生存质量的重要因素,本研究将探讨克唑替尼在间变性淋巴瘤激酶(anaplastic lymphoma kinase, ALK)阳性NSCLC脑转移患者中的疗效。方法：筛选20例ALK阳性NSCLC脑转移患者,给予克唑替尼治疗,观察临床疗效。结果：克唑替尼治疗前基线有脑转移患者,颅内疗效部分缓解(partial response,PR)12例,疾病稳定(stable disease,SD)7例,疾病进展(progressive disease,PD)1例。客观缓解率(objective response rate,ORR)为60%,疾病控制率(disease control rate,DCR)为95%。中位无进展生存期(progression free survival,PFS)为6个月(95%CI：3.92~8.08)。结论：克唑替尼治疗ALK阳性NSCLC脑转移患者有效。     

克唑替尼在ALK阳性中晚期非小细胞肺癌中的疗效观察

目的：探讨克唑替尼治疗晚期间变淋巴瘤激酶(anaplastic lymphoma kinase,ALK)融合基因阳性中晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)的近期疗效及毒副反应。方法：回顾性分析43例ALK阳性的中晚期NSCLC患者,服用克唑替尼治疗,服用至病情进展或出现不可耐受的毒副反应,随访12个月,观察疗效。结果：克唑替尼治疗ALK阳性NSCLC的疾病控制率(disease control rate,DCR)为93%(3/43),客观缓解率(objective response rate,ORR)为62%(26/43),中位无进展生存时间(progression free survival,PFS)为7.0个月(95% CI,6.0~8.0月),不良反应主要为消化道症状,其次是谷丙转氨酶升高,视觉障碍,大部分为1~2级。结论：克唑替尼作为NSCLC患者的多靶点靶向治疗,具有良好的疗效及安全性,不良反应轻微。     

ALK和ROS1：肺癌治疗的联合靶点

间变性淋巴瘤激酶（anaplasticlymphomakinase，ALK）基因重排可见于包括非小细胞肺癌non-smallcelllungcancer，NSCLC）在内的多种恶性肿瘤中。ALK融合基因使激酶具有异常活性，而野生型ALK激酶域突变也可使它被激活。ALK基因重排使得NSCLC中出现了新的分子亚型，该亚型对ALK抑制剂高度耐药。克唑替尼（crizotinib）是一个口服小分子ATP模拟化合物，它最初作为MET抑制剂被开发，随后被发现具有抗ALK活性的脱靶效应（off-target），并被美国FDA批准用于治疗ALK阳性的NSCLC患者。近来在NSCLC患者中还发现了ROSl受体酪氨酸激酶染色体重排，而克唑替尼正处于治疗该分子亚型NSCLC患者的临床试验中。任何计算机辅助药物设计都是依赖其分子结构和配体的药物设计方法，每种方法的详细信息中均应重点强调利用这二者，以开发多靶点小分子激酶抑制剂。此类多靶点小分子激酶抑制剂均可对ROS1和ALK重排的NSCLC有抑制增殖作用。因此，本综述重点强调了关于靶向这些激酶的重要性，以及在优化出效能更佳、选择性更强的ROS1和ALK激酶抑制剂中所取得的进步。     

ALK免疫组化切片重复利用进行FISH检测的可行性

正目前肺癌是全球发病率和病死率增长较快的恶性肿瘤之一,其中非小细胞肺癌(non-small cell lung cancer,NSCLC)占80%以上,约5%的NSCLC出现间变性淋巴瘤激酶(anaplastic lymphoma kinase,ALK)重排[1]。近年来克唑替尼等ALK基因靶向药物临床发展迅速,对晚期NSCLC患者疗效显著。晚期肺癌患者通常无法进行手术,临床活检组织标本、细胞学标本的珍贵性不言而喻。免疫组化是病理科常用的检测方法,经济实用,方便快捷,适合ALK基因重排检测的初筛;荧光原位杂交(fluorescence in situ hybridization,     

杂交时间对非小细胞肺癌中c-MET与ROS1 FISH实验的影响

正肺癌是全球病死率最高的恶性肿瘤之一。随着分子医学进展和靶向药物的不断涌现,晚期非小细胞肺癌(nonsmallcell lung cancer,NSCLC)的治疗已经进入到个体化治疗的时代。针对ALK基因靶点的小分子抑制剂克唑替尼(Crizotinib)是一种ATP竞争性酪氨酸酶抑制剂(tyrosine kinase inhibitor,TKI),可特异性靶向抑制ALK,也可抑制c-MET和ROS1等信号通路。FISH技术日益成为检测患者是否适合这些靶向治疗的重要手段。然而目前相对比较成熟的ALK     

活化的RET和ROS：肺腺癌新的驱动基因

ROS1和RET的基因重排是新发现的肺腺癌驱动基因，各自突变频率约为1%。肺腺癌中RET和ROS1基因重排与其他已知的驱动基因几乎没有重叠，成为独立的新的肺腺癌分子亚型。肺腺癌中ROS1和RET突变基因的临床病理学特征已经阐明，体外实验也证实数种多靶点受体激酶抑制剂能抑制含ROS1和RET突变基因的非小细胞肺癌（non-small cell lung cancer，NSCLC）细胞系。而临床研究显示MET/ALK/ROS抑制剂克唑替尼在ROS1突变阳性的肺癌患者中疗效显著。目前，正开展一些针对肺腺癌ROS1和RET基因的各种激酶抑制剂的早期临床试验。针对这些基因突变的新的肺腺癌亚群的个体化治疗极有可能在不久的将来作为常规的临床治疗措施。     

非小细胞肺癌个体化治疗的靶向分子检测

非小细胞肺癌(non-small cell lung cancer, NSCLC)的个体化治疗是目前肿瘤研究领域的热点之一.以EGFR体系突变活化和ALK重排为分子学特征的NSCLC对酪氨酸激酶抑制剂(tyrosine kinase inhibitors, TKIs)如吉非替尼(gefitinib)、埃洛替尼(erlotinib)以及可卓替尼(crizotinib)有良好的反应性,而EGFR突变和ALK重排也成为筛选TKIs治疗对象和预测治疗反应性的最有价值指标.近年来,欧洲、北美和亚洲地区先后公布EGFR突变检测的共识,我国也于2011年达成了EGFR突变检测的专家共识[1-4].     

Mir-30a抑制自噬及上皮细胞间质化增强肺腺癌细胞对克唑替尼的敏感性

目的探索上调肺腺癌细胞中Mir-30a的表达水平能否增强细胞对克唑替尼的敏感性,观察细胞增殖率的变化及克唑替尼IC50等的变化,并探索相关的机制。方法使用Lipofectamine2000携带Mir-30a对H3122细胞进行瞬时转染,并与克唑替尼共同作用。将肺腺癌H3122细胞分为对照组、克唑替尼与联合组。MTT法检测各组细胞增殖率变化,Transwell检测细胞侵袭性。Western blot检测ALK、c-MET、Beclin-1、E-cadherin及Vimentin蛋白的表达。结果在H3122细胞中Mir-30a转染与克唑替尼共同作用较克唑替尼单独作用有更显著的细胞杀伤作用。转染后的细胞较单独应用克唑替尼组侵袭力减弱。转染与克唑替尼联合组Beclin1、ALK、c-MET及Vimentin的蛋白的表达有明显下降,E-cadherin稍有增强。结论 Mir-30a增强肺腺癌细胞对克唑替尼的敏感性,可能与过量表达的Mir-30a抑制癌细胞的自噬及上皮细胞间质化相关。     

吉非替尼联合培美曲塞治疗老年NSCLC患者的效果及其对肿瘤生物学指标、血流动力学的影响

目的:探究吉非替尼与培美曲塞联合应用于老年非小细胞肺癌(Non-small cell lung cancer,NSCLC)患者的效果观察.方法:选取本院我科85例老年中晚期NSCLC患者作为研究对象,采用抽签法分成两组,对照组42例采用培美曲塞治疗,观察组43例加用吉非替尼治疗,观察两组患者的临床疗效,肿瘤生物学指标、...     

ALK-rearranged squamous cell lung cancer: a case report

ALK rearrangement is a very rare subset of squamous cell cancer of lung and the efficacy of crizotinib treatment for these patients is lack of data. Here we report a case with squamous cell cancer of lung that harbored the ALK rearrangement was given crizotinib in the second-line therapy. A 55-year-old female patient was diagnosed with squamous cell carcinoma of lung by bronchoscopy biopsy with stage IV. The patient was given two cycles of chemotherapy and the response was progressive disease. After failure to chemotherapy, genotype testing showed wild-type EGFR/KRAS and ALK rearrangement positive. The patient was administered with crizotinib and had a partial response, and the progression-free survival was 6 months. The side effects were tolerable.     

Screening for ALK abnormalities in central nervous system metastases of non‐small‐cell lung cancer

Anaplastic lymphoma kinase (ALK) gene rearrangement was reported in 3%–7% of primary non‐small‐cell lung cancer (NSCLC) and its presence is commonly associated with adenocarcinoma (AD) type and non‐smoking history. ALK tyrosine kinase inhibitors (TKIs) such as crizotinib, alectinib and ceritinib showed efficiency in patients with primary NSCLC harboring ALK gene rearrangement. Moreover, response to ALK TKIs was observed in central nervous system (CNS) metastatic lesions of NSCLC. However, there are no reports concerning the frequency of ALK rearrangement in CNS metastases. We assessed the frequency of ALK abnormalities in 145 formalin fixed paraffin embedded (FFPE) tissue samples from CNS metastases of NSCLC using immunohistochemical (IHC) automated staining (BenchMark GX, Ventana, USA) and fluorescence in situ hybridization (FISH) technique (Abbot Molecular, USA). The studied group was heterogeneous in terms of histopathology and smoking status. ALK abnormalities were detected in 4.8% (7/145) of CNS metastases. ALK abnormalities were observed in six AD (7.5%; 6/80) and in single patients with adenosuqamous lung carcinoma. Analysis of clinical and demographic factors indicated that expression of abnormal ALK was significantly more frequently observed (P = 0.0002; χ² = 16.783) in former‐smokers. Comparison of IHC and FISH results showed some discrepancies, which were caused by unspecific staining of macrophages and glial/nerve cells, which constitute the background of CNS tissues. Their results indicate high frequency of ALK gene rearrangement in CNS metastatic sites of NSCLC that are in line with prior studies concerning evaluation of the presence of ALK abnormalities in such patients. However, they showed that assessment of ALK by IHC and FISH methods in CNS tissues require additional standardizations.     

Differential inhibitor sensitivity of anaplastic lymphoma kinase variants found in neuroblastoma

Activating mutations in the anaplastic lymphoma kinase (ALK) gene were recently discovered in neuroblastoma, a cancer of the developing autonomic nervous system that is the most commonly diagnosed malignancy in the first year of life. The most frequent ALK mutations in neuroblastoma cause amino acid substitutions (F1174L and R1275Q) in the intracellular tyrosine kinase domain of the intact ALK receptor. Identification of ALK as an oncogenic driver in neuroblastoma suggests that crizotinib (PF-02341066), a dual-specific inhibitor of the ALK and Met tyrosine kinases, will be useful in treating this malignancy. Here, we assessed the ability of crizotinib to inhibit proliferation of neuroblastoma cell lines and xenografts expressing mutated or wild-type ALK. Crizotinib inhibited proliferation of cell lines expressing either R1275Q-mutated ALK or amplified wild-type ALK. In contrast, cell lines harboring F1174L-mutated ALK were relatively resistant to crizotinib. Biochemical analyses revealed that this reduced susceptibility of F1174L-mutated ALK to crizotinib inhibition resulted from an increased adenosine triphosphate-binding affinity (as also seen in acquired resistance to epidermal growth factor receptor inhibitors). Thus, this effect should be surmountable with higher doses of crizotinib and/or with higher-affinity inhibitors.     

Clinical observation of crizotinib in the treatment of ALK-positive advanced non-small cell lung cancer

BackgroundALK is a prognostic and predictive tumor marker in non-small cell lung carcinoma (NSCLC), and is more often found in lung adenocarcinomas.MethodsThe clinical and pathological data of 87 patients confirmed to have NSCLC by pathology or cytology were selected from April 2014 to January 2017 at the Tumor Hospital of Hebei Province.ResultsOf the 87 ALK-positive-patients, 47 patients were treated with oral administration of crizotinib. The objective response rate (ORR) was 61.7%, the disease control rate (DCR) was 93.6%, and the mPFS was 19 months. In an analysis of the number of metastatic sites, the patients who had more than three metastatic sites, the ORR, DCR, and mPFS were 63.9%, 94.5%, and 19 months, compared with the 45.5%, 91%, and 11 months in the patients with less sites (P = 0.040). For patients of 60 years or older, ORR and DCR were 40% and 100%, the other group was 71.9% and 90.6%, respectively(P = 0.036). The timing of treatment was analyzed. At the first application of crizotinib, ORR and DCR were 78.2% and 100% corresponding 45.8% and 87.5% at the second and final application of crizotinib group (P = 0.022). Baseline brain metastases were present in 25.5% (12/47) of patients in this study. 9 of the patients who developed disease progression during crizotinib treatment had new brain metastases or increased preexisting cranial foci. Most of them took the treatment strategy of continuing crizotinib or replacing the second/third generation ALK-TKI treatment combined with local radiotherapy for brain metastases.ConclusionsThe efficacy of crizotinib in patients with advanced NSCLC is related to the number of metastatic organs, age and timing of treatment. The use of crizotinib is prone to intracranial progression, and progression of simple brain metastases is not an indication that crizotinib is discontinued. Patients will continue to benefit from combination of local radiotherapy.     

Different histopathology but the same clonality: ALK rearrangement in a patient with metastatic non-small-cell lung cancer

EML4-ALK rearrangement is detected in 2% to 7% of lung adenocarcinomas, these tumors are sensitive to crizotinib. The histologic feature of ALK translocated non-small-cell lung cancer (NSCLC) has been studied, presence of signet-ring cells was a powerful histologic indicator of ALK rearrangement, and this characteristic histology was present both in primary sites and metastases. However, the case we discribed here has different histomorphology in primary sites and metastases, but has the same genotype which both present ALK rearrangement, while absent of EGFR mutation, KRAS mutation and ROS1 rearrangement. This histologic heterogeneity may be a supplement of the histologic feature of ALK rearranged tumor. Moreover, genomic analysis can help distinguish clonal tumors from independent primaries.     

What's new in non-small cell lung cancer for pathologists: the importance of accurate subtyping, EGFR mutations and ALK rearrangements

In the past, the only critical point of distinction in the pathological diagnosis of lung cancer was between small cell and non-small cell lung cancer (NSCLC). The emergence of new targeted therapies and clinical trials demonstrating differing efficacy and toxicity of treatments according to specific histological subtypes of NSCLC, has resulted in an increasing need for improvements in pathological diagnosis. Accurate distinction between adenocarcinoma and squamous cell carcinoma is now critical as histological subtyping has the potential to influence clinical decision making and impact on patient outcome. While morphological criteria remain the most important feature to distinguish NSCLC subtypes, use of mucin and immunohistochemical stains (TTF-1, p63 and CK5/6) can be of assistance in difficult small biopsy cases. With the emergence of selective kinase inhibitors targeting epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK), there is a corresponding need to identify the subset of NSCLCs harbouring specific genetic mutations associated with sensitivity to these agents, almost all of which are found in adenocarcinomas. In this review, the importance of accurately subtyping NSCLC is discussed, along with a suggested approach for distinguishing histological subtypes in small biopsy specimens. The significance of EGFR and ALK mutations in NSCLC and the impact of these genotypes on pathology and clinical practice are also reviewed.     

非小细胞肺癌胸水细胞块ALK融合基因检测分析

目的：探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)胸水细胞块在间变性淋巴瘤激酶(anaplastic lymphoma kinase,ALK)融合基因检测中的临床价值。方法：采用突变扩增系统PCR(ARMS-PCR)法检测215例NSCLC细胞块和404例NSCLC组织块中ALK基因的三类融合类型,并检测细胞块同时送检组织块的患者74例的一致性。结果：细胞块ALK基因融合阳性26例,阳性率12.09%(26/215);组织块ALK基因融合阳性25例,阳性率6.19%(25/404);74例有组织块对照的细胞块ALK融合基因结果一致性有67例,一致率达90.54%(67/74),其中细胞块ALK融合基因的阳性率14.86%(11/74),组织块阳性率18.92%(14/74)。结论：NSCLC胸水细胞块ALK融合基因的阳性率略高于组织块;有恶性胸水的NSCLC患者原发灶组织发生ALK融合基因阳性的概率较高。     

Targeting anaplastic lymphoma kinase in neuroblastoma

Over the last decade, anaplastic lymphoma kinase (ALK), a receptor tyrosine kinase (RTK), has been identified as a fusion partner in a diverse variety of translocation events resulting in oncogenic signaling in many different cancer types. In tumors where the full‐length ALK RTK itself is mutated, such as neuroblastoma, the picture regarding the role of ALK as an oncogenic driver is less clear. Neuroblastoma is a complex and heterogeneous tumor that arises from the neural crest derived peripheral nervous system. Although high‐risk neuroblastoma is rare, it often relapses and becomes refractory to treatment. Thus, neuroblastoma accounts for 10–15% of all childhood cancer deaths. Since most cases are in children under the age of 2, understanding the role and regulation of ALK during neural crest development is an important goal in addressing neuroblastoma tumorigenesis. An impressive array of tyrosine kinase inhibitors (TKIs) that act to inhibit ALK have been FDA approved for use in ALK‐driven cancers. ALK TKIs bind differently within the ATP‐binding pocket of the ALK kinase domain and have been associated with different resistance mutations within ALK itself that arise in response to therapeutic use, particularly in ALK‐fusion positive non‐small cell lung cancer (NSCLC). This patient population has highlighted the importance of considering the relevant ALK TKI to be used for a given ALK mutant variant. In this review, we discuss ALK in neuroblastoma, as well as the use of ALK TKIs and other strategies to inhibit tumor growth. Current efforts combining novel approaches and increasing our understanding of the oncogenic role of ALK in neuroblastoma are aimed at improving the efficacy of ALK TKIs as precision medicine options in the clinic.