Anaplastic Lymphoma Kinase Gene Research Articles

Abstract B40: A novel activating I1171T ALK mutation in neuroblastoma responds better to ceritinib compare to the first generation inhibitor crizotinib

Abstract Mutations in the kinase domain of Anaplastic Lymphoma Kinase (ALK) have been suggested to be important players in the genetics and progression of the childhood tumor neuroblastoma. Here we report the appearance of a novel ALK mutation in neuroblastoma together with other chromosomal aberrations that mediate neuroblastoma initiation and progression. Analyses of genomic tumor DNA from biopsy samples initially showed an 11q minus, 17q gain with a mutation of the ALK gene at protein position 1171, which mediate an amino acid change from isoleucine to threonine. We show that mutation of I1171 to threonine generates a potent gain-of-function mutant, as observed in two independent systems. Firstly, in PC12 cell lines expressing ALKI1171T display ligand independent activation of ALK, neurite outgrowth and further downstream signaling activation. Secondly, ALKI1171T meditate foci formation in a NIH3T3 transformation analysis. Finally, pharmacological inhibition of ALKI1171T employing ceritinib, an FDA approved ALK inhibitor show 14-fold better ability to abrogate ALKI1171T compared with crizotinib. Citation Format: Joachim T. Siaw, Jikui Guan, Tommy Martinsson, Magnus Hansson, Per Kogner, Ruth Palmer. A novel activating I1171T ALK mutation in neuroblastoma responds better to ceritinib compare to the first generation inhibitor crizotinib [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr B40.

Evolving concepts in lung cancer pathology and its impact on thoracic oncology practice

Lung cancer is the commonest cause of death worldwide but unfortunately, most patients present in an advanced stage of the disease. The classification of lung cancer has evolved since the first WHO publication in 1981, and the present edition was published in March 2015. A major change in 2015 was the introduction of the new terms adenocarcinoma in situ and minimally invasive adenocarcinoma to describe early adenocarcinoma of lung and abolition of the previous term bronchioloalveolar carcinoma. Tumors with micropapillary pattern and those showing spread through alveolar spaces have significant clinical implications with an increased propensity for recurrence. Tissue specimens require proper fixation in 10% buffered formalin which is an essential but an often neglected basic step; proper fixation is a prerequisite for good immunohistochemistry (IHC) and molecular test outcomes. Needle core biopsies should be handled judiciously to ensure that sufficient material is prioritized for molecular studies which are needed to guide personalized treatment. The WHO 2015 classification recommends the use of a single squamous cell marker (p40, or CK5/6) and a single adenocarcinoma marker (TTF1, or napsin) to distinguish squamous cell carcinoma and adenocarcinoma in difficult biopsies. The non-committal designation of non-small cell lung carcinoma (NSCLC) is discouraged. The exciting ongoing research findings on molecular biology of lung cancer necessitate constant expansion of laboratory test menu. Testing and treating lung cancer patients with epidermal growth factor (EGFR) mutation and anaplastic lymphoma kinase (ALK) gene fusion is now a standard of care, and ROS1 has been approved recently. The updated guidelines from the College of American Pathologists (CAP) and approval notifications from the Federal Drug Authority (FDA) are invaluable to justify initiation of new tests by pathology laboratories. Recently, FDA has approved testing for programmed death ligand (PDL1) with 22C3 antibody by IHC and EGFR testing on plasma by a non-invasive liquid biopsy test approach. To conclude, a comprehensive pathology report is vital for staging, comparing outcomes, and developing better strategies for patient care in the future.

Novel method for rapid fluorescence in-situ hybridization of ALK rearrangement using non-contact alternating current electric field mixing

Echinoderm microtubule-associated protein-like 4 gene and anaplastic lymphoma kinase gene (EML4-ALK) rearrangement is a key driver mutation in non-small cell lung cancer (NSCLC). Although Break-Apart ALK fluorescence in situ hybridization (FISH) is a reliable diagnostic method for detecting ALK gene rearrangement, it is too costly and time-consuming for use as a routine screening test. Our aim was to evaluate the clinical utility of a novel rapid FISH (RaFISH) method developed to facilitate hybridization. RaFISH takes advantage of the non-contact mixing effect of an alternating current (AC) electric field. Eighty-five specimens were used from patients diagnosed with NSCLC identified immunohistochemically as ALK 0, (1/2+) or (3+). With RaFISH, the ALK test was completed within 4.5 h, as compared to 20 h needed for the standard FISH. Although RaFISH produced results more promptly, the staining and accuracy of the ALK evaluation with RaFISH was equal to the standard. We found 97.6% agreement between FISH and RaFISH based on the status of the ALK signals. These results suggest RaFISH could be used as a clinical tool to promptly determine ALK status.

Clinical characteristics and sequence complexity of anaplastic lymphoma kinase gene fusions in Chinese lung cancer patients

ObjectivesTo investigate the clinical characteristics of anaplastic lymphoma kinase (ALK) rearrangements and sequence complexity of the ALK fusion gene in Chinese lung cancer patients. MethodsWe prospectively screened ALK rearrangements in 1474 lung cancer specimens, including 1387 cases of non-small cell lung cancer (NSCLC), 54 cases of small cell lung cancer (SCLC), and 33 cases of cancer with lung metastasis from other organs by both standard polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE)-coupled PCR. Fifteen cases of ALK-positive RACE-coupled PCR products were transformed into Escherichia coli for molecular cloning and sequenced for complexity analysis. ResultsThe overall frequency of ALK rearrangements was 5.1% (71/1387) in NSCLC. In 71 positive cases, the coexistence of epidermal growth factor receptor (EGFR) and ALK variations was found in 6 cases (8.5%), and the coexistence of different ALK variants was found in 2 cases (2.8%) (1 case with variants 1 and 9; the other case with variants 3 and 2) by PCR analysis. Furthermore, through sequence cloning analysis of 15 cases of non-selective ALK-positive samples, two cases with variants 1 and 3 harbored the coexistence of three subtypes (variant 1 subtypes: E13; A20, E13del63; A20 and E7E12E13; A20 and variant 3 subtypes: E6; A20, E6ins33; A20 and E3E6; A20). Variant 3a and 3b subtypes were always coexistent and had the same proportion of ALK variant 3 rearrangements. ALK rearrangement was associated with young age, female gender, never-smokers, those with adenocarcinoma, advanced stage, and EGFR mutations. No ALK fusion was detected in 54 cases of SCLC or 33 cases of cancer with lung metastasis from other organs. ConclusionsThe identification of novel ALK variants, the coexistence of EGFR mutations and ALK fusions, the coexistence of ALK variants, and the coexistence of subtypes reveal the diversity and sequence complexity of ALK fusions.

MA 09.05 Evidence in Favor of or against the Use of TKIs with Concurrent Cranial Radiotherapy in Patients with NSCLC and CNS Involvement

Central nervous system (CNS) metastases, including brain metastases (BM) and leptomeningeal metastases (LM) represent a frequent complication of non-small cell lung cancer (NSCLC). Patients with CNS metastases comprise a heterogeneous group, with a median survival that ranges from 3 to 14 months. Local therapies, such as whole brain radiotherapy (WBRT), stereotactic radiosurgery (SRS) or surgical resection, are available treatment strategies for BM. Introduction of tyrosine kinase inhibitors (TKIs) in clinical practice has led to individualization of therapy based upon the presence of the exact abnormality, resulting in a major therapeutic improvement in patients with NSCLC who harbor epidermal growth factor receptor (EGFR) activating mutations or anaplastic lymphoma kinase (ALK) gene rearrangements, respectively, but their role in combination with radiotherapy is controversial. We performed a systematic literature review of published data in PUBMED, SCOPUS and COCHRANE databases, as well as a manual search of reported data in major congresses, using the terms “EGFR-TKIs”, “ALK-TKIs”, “CNS”, “BM” or ““LM” and “concurrent radiotherapy” or “stereotactic radiotherapy” from January 2000 through May 2017. Only prospective or retrospective clinical trials and meta-analyses reported in English language were included. See Table. At this time, concurrent use of TKIs with radiotherapy, although appearing to be safe, is not recommended outside of a clinical trial. Interestingly, data in EGFR mutant patients treated with an EGFR-TKI alone prompt the question whether this could be a front line approach in patients with asymptomatic BM, reserving WBRT for symptomatic cases. In clinical practice, burden of extracranial disease might also guide treatment decisions; physicians might select not to discontinue a TKI during WBRT in case of extensive extracranial organ involvement Ongoing clinical trials are currently evaluating the effectiveness of concomitant use of radiotherapy and TKIs.

P1.03-010 Efficacy and Safety of Anaplastic Lymphoma Kinase (ALK) Tyrosine Kinase Inhibitors in ALK-Positive Non-Small Cell Lung Cancer

Anaplastic lymphoma kinase (ALK) gene rearrangements occur in 3-5% of non-small cell lung cancer (NSCLC) cases. ALK tyrosine kinase inhibitors (ALK-TKIs), such as crizotinib and alectinib, are recommended for the treatment of ALK-positive NSCLC. However, acquired resistance to the ALK-TKIs develops after treatment with these agents. Therefore, it is necessary to consider the alteration of the therapeutic approach against ALK-positive NSCLC. This investigation, reviewed patients with ALK-positive NSCLC treated at Hiratsuka Kyosai Hospital in 2016, to determine the efficacy and safety of ALK-TKIs in this setting. The medical data of 11 patients who had been diagnosed with ALK-positive advanced NSCLC and treated with ALK-TKIs at Hiratsuka Kyosai Hospital in 2016 were reviewed retrospectively. A total of 11 patients (3 males and 8 females; mean age: 62 years) with ALK-positive NSCLC were investigated. All the pathological types were adenocarcinomas. Eight patients were treated with crizotinib as first-line therapy, and 3 out of those patients were treated with alectinib as second-line therapy. The remaining 3 patients were treated with alectinib as first-line therapy. The overall response rate was 87.5%, and the median progression-free survival was not reached. Four patients had developed PD while receiving crizotinib. Two out of those patients have developed brain metastasis, and were administered local radiotherapy to the brain. Patients who progressed following treatment with ALK-TKIs, were treated with pemetrexed-based chemotherapy. Although adverse events (AEs) of crizotinib were more than those of alectinib, most of them were of Grade 1 to 2 severity. Most common AEs of crizotinib included vision disorder (62.5%), diarrhea (50%), elevated aminotransferases (50%), and nausea (37.5%). Grade 3 to 4 adverse events were reported in 4 cases. However, all of them were controlled by withdrawal of treatment or reduction of dosage. ALK-TKIs demonstrate good efficacy and safety in patients with ALK-positive NSCLC.

The Presence of ALK Alterations and Clinical Relevance of Crizotinib Treatment in Pediatric Solid Tumors.

Soft tissue sarcomas (STS) and neuroblastomas (NBL), arechildhood malignancies still associated with poor prognoses despite the overall improvement in childhood tumor survival of the past decades. Anaplastic lymphoma kinase (ALK) inhibition is promising new strategy to improve the outcome of these pediatric tumors.Eighteen histologic samples of pediatric STS and 19 NBL patients were analyzed for ALK abnormalities using fluorescent in situ hybridization (FISH) with break-apart probes and immunohistochemistry (IHC). ALK alterations were presented in 20 of the 37 sections. The presence of ALK alteration in NBL samples were detected using IHC in 84,2% of all cases compared to 21,1% FISH positivity. In STS cases the results were less different (IHC 16,7% vs FISH 22,2%). The difference can be explained by the different type of molecular alterations. FISH method detected translocation and amplification, but not the point mutation of ALK gene. IHC confirmed the diagnosis by detecting the expression of ALK protein.After ALK positivity was proven, the effectiveness and safety of the crizotinib therapy was examined in 4 patients (1 alveolar rhabdomyosarcoma (RMA), 1 embryonal rhabdomyosarcoma (RME), 1 inflammatory myofibroblastic tumor (IMT), 1 NBL). We observed continuous remission of the IMT patient, all other cases the inhibitor treatment was not curative.Our findings underline the importance of screening the ALK status parallel with both IHC and FISH. Crizotinib treatment had a long-term effect in ALK positive IMT patients, however itwas only temporary efficient in relapsed, progressive STS and NBL.

ROS1 fusion gene in targeted therapy of non-small cell lung cancer

The molecular target therapy of non-small cell lung cancer (NSCLC) has become a hot research direction in the field of medicine. Following the discovery of well-known tumor-driven genes such as epidermal growth factor receptor (EGFR), Kirsten rat sarcoma viral oncogene (KRAS) and anaplastic lymphoma kinase (ALK) genes, more and more scholars have shifted focus to ROS1 fusion gene. The protein encoded by ROS1 is a member of the receptor tyrosine kinase super family, and it plays an important role in cell growth and cell survival. ROS1 fusion gene plays a vital role in the occurrence, development and clinical treatment of NSCLC. Key words: Carcinoma, non-small-cell lung; ROS1 fusion gene; Crizotinib; Molecular targeted the-rapy

Detection of known and novel ALK fusion transcripts in lung cancer patients using next-generation sequencing approaches

Rearrangements of the anaplastic lymphoma kinase (ALK) gene in non-small cell lung cancer (NSCLC) represent a novel molecular target in a small subset of tumors. Although ALK rearrangements are usually assessed by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH), molecular approaches have recently emerged as relevant alternatives in routine laboratories. Here, we evaluated the use of two different amplicon-based next-generation sequencing (NGS) methods (AmpliSeq and Archer®FusionPlex®) to detect ALK rearrangements, and compared these with IHC and FISH. A total of 1128 NSCLC specimens were screened using conventional analyses, and a subset of 37 (15 ALK-positive, and 22 ALK-negative) samples were selected for NGS assays. Although AmpliSeq correctly detected 25/37 (67.6%) samples, 1/37 (2.7%) and 11/37 (29.7%) specimens were discordant and uncertain, respectively, requiring further validation. In contrast, Archer®FusionPlex® accurately classified all samples and allowed the correct identification of one rare DCTN1-ALK fusion, one novel CLIP1-ALK fusion, and one novel GCC2-ALK transcript. Of particular interest, two out of three patients harboring these singular rearrangements were treated with and sensitive to crizotinib. These data show that Archer®FusionPlex® may provide an effective and accurate alternative to FISH testing for the detection of known and novel ALK rearrangements in clinical diagnostic settings.

Homogeneity and High Concordance of ALK Translocation in Primary Lung Adenocarcinoma and Paired Lymph Node Metastasis

Translocation of anaplastic lymphoma kinase (ALK) gene is an important determinator for the response to ALK tyrosine kinase inhibitor (TKI) in non-small-cell lung cancer (NSCLC) patients. The existence of genetic heterogeneity will affect the results of molecular testing, especially in biopsy samples from primary or metastatic sites of patients with advanced stage NSCLC. We intended to explore the heterogeneity of ALK gene translocation in excision specimens and to examine the existence of discordance of ALK status between primary tumours and corresponding lymph node metastases. A total of 106 ALK positive lung adenocarcinoma cases were collected for assessment of intratumour heterogeneity of ALK gene translocation, which were stained by the fully automated Ventana ALK D5F3 immunohistochemistry (IHC) analysis. In addition, the ALK gene translocations were evaluated in a series of 53 primary tumours and their paired lymph node metastases using ALK D5F3 IHC staining. The concordance rate between primary tumours and paired metastatic lymph nodes was 100%. ALK status was homogeneous in lung adenocarcinoma samples and was generally stable during metastasis. Therefore, ALK gene translocation can be measured reliably in material from either primary or metastatic tumours in lung adenocarcinoma patients.

Baseline and On-Treatment Characteristics of Serum Tumor Markers in Stage IV Oncogene-Addicted Adenocarcinoma of the Lung.

The role of serum tumor markers in the modern management of advanced NSCLC remains poorly described. A single-center retrospective analysis of available carcinoembryonic antigen, CA125, CA19.9, and CA27.29 levels at baseline and during treatment of stage IV lung adenocarcinoma by oncogenic driver was conducted. A total of 142 patients were analyzed (60 with anaplastic lymphoma kinase gene [ALK] rearrangement, 50 with EGFR mutation, four with ROS1 rearrangement, and 29 with KRAS mutation). Of these, 82% had at least one marker (95% if all four markers were measured), with CA27.29 being the most commonly increased and CA19.9 the rarest. Only CA27.29 differed significantly by oncogene (it was less common in KRAS) (p= 0.016). The median times to nadir during tyrosine kinase inhibitor (TKI) therapy in EGFR and ALK cases were 16.4 and 20 weeks, respectively. Of the 41 patients with EGFR mutation or ALK or ROS1 rearrangement, 24 (59%) demonstrated an initial increase within the first 4 weeks of TKI therapy, 58% of whom then had their levels fall below baseline. An increase in marker level of 10% or more from nadir occurred in 53% of systemic and 22% of central nervous system-only progression. Serum tumor markers are frequently increased in lung adenocarcinoma regardless of driver oncogene. Changes within the first 4 weeks of therapy may be misleading. Progression is associated with marker increases, especially in sites other than the central nervous system.

The role of anaplastic lymphoma kinase in pediatric cancers.

The anaplastic lymphoma kinase (ALK) gene was initially identified as a fusion partner of the nucleophosmin gene in anaplastic large‐cell lymphoma with t(2;5)(p23;q35) translocation, and then described with different genetic abnormalities in a number of tumors. Although ALK is known to be involved in the pathogenesis of neuroblastoma through activating mutations or gene amplification, its role in the pathogenesis of other pediatric cancers is still elusive. In addition to neuroblastoma, the high‐grade amplification of ALK has been described in a subset of rhabdomyosarcoma cases. Normal ALK protein expression is restricted to the nervous systems of adult mammals, but the aberrant expression of ALK has been observed in a variety of pediatric cancers, including glioma and Ewing sarcoma. The discovery of oncogenic activation of ALK in neuroblastoma suggests that this cancer could be potentially treated with an ALK inhibitor, as could other cancers, such as non‐small‐cell lung cancer and anaplastic large‐cell lymphoma. However, cellular responses to mutant ALK are complex when compared to rearranged ALK, and treatment remains a challenge. This review focuses on the biology of ALK in pediatric cancers and possible therapeutic strategies for ALK‐associated tumors.

ALK signaling cascade confers multiple advantages to glioblastoma cells through neovascularization and cell proliferation.

Anaplastic lymphoma kinase (ALK), which is a receptor tyrosine kinase, is essentially and transiently expressed in the developing nervous system. Here we examined the functional role of the ALK gene in glioblastomas (GBMs). In clinical samples of GBMs, high ALK expression without gene rearrangements or mutations was frequently observed in perivascular lesions, in contrast to the relatively low expression in the perinecrotic areas, which was positively correlated with N-myc and phosphorylated (p) Stat3 scores and Ki-67 labeling indices. ALK immunoreactivity was also found to be associated with neovascular features including vascular co-option and vascular mimicry. In astrocytoma cell lines, cells stably overexpressing full-length ALK showed an increase in expression of pStat3 and pAkt proteins, as well as hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor-A (VEGF-A) mRNAs, in contrast to cells with knockdown of endogenous ALK which showed decreased expression of these molecules. Transfection of the constitutively active form of Stat3 induced an increase in HIF-1α promoter activity, and the overexpression of HIF-1α in turn resulted in enhancement of VEGF-A promoter activity. In addition, cells with overexpression or knockdown of ALK also showed a tendency toward increased and decreased proliferation, respectively, through changes in expression of pAkt and pStat3. Finally, ALK promoter was significantly activated by transfection of Sox4 and N-myc, which are known to contribute to neuronal properties. These findings therefore suggest that N-myc/Sox4-mediated ALK signaling cascades containing Stat3, Akt, HIF-1α, and VEGF-A confer multiple advantages to tumor growth through alterations in neovascularization and cell proliferation in GBMs.

Association between Charlson comorbidity index score and outcome in patients with stage IIIB-IV non-small cell lung cancer

BackgroundThis retrospective study investigated the association between the Charlson comorbidity index (CCI) score and the survival of patients with stage IIIB-IV (advanced, non-resectable) non-small cell lung cancer (NSCLC) who also did not have gene mutations in epidermal growth factor receptor (EGFR) or anaplastic lymphoma kinase (ALK).MethodsThe records of 165 patients (28–80 y, median 61 y) who met the above criteria and were admitted to Beijing Friendship Hospital Capital Medical University from 1 May 2010 to 1 October 2014were reviewed. Associations between baseline variables and the CCI score were assessed via univariate and multivariate logistic regression analysis. Overall survival was defined as the time from the first clinic visit to death from any cause, or to the end of follow-up. Survival curves were estimated via the Kaplan-Meier method and compared with the log-rank test.ResultsLogistic regression analyses indicated that smoking and performance status were independently associated with the CCI score. Smoking was associated with an increased risk of mortality (odds ratio (OR) 4.12 (95% confidence interval [CI] 1.92–8.84) compared to non-smokers), as was performance status 2 (ambulatory, capable of self-care, unable to perform any work activities; active for >50% of waking hours) (OR 2.22 (95% CI, 1.14–4.33) compared to performance status 1). Univariate Cox’s regression analyses showed that the hazard ratios were significantly associated with the CCI score (P = 0.009), smoking (P = 0.042), and male gender (P = 0.021).ConclusionThe CCI score is an important prognostic factor for the prediction of overall survival in patients with stage IIIB-IV NSCLC who are negative for EGFR and ALK gene mutations.

MET Exon 14 Alterations and New Resistance Mutations to Tyrosine Kinase Inhibitors: Risk of Inadequate Detection with Current Amplicon-Based NGS Panels

IntroductionTargeted therapies such as tyrosine kinase inhibitors (TKIs) have dramatically improved the treatment of lung adenocarcinoma, and detection of activating mutations of genes such as EGFR or anaplastic lymphoma kinase gene (ALK) is now mandatory in the clinical setting. However, additional targetable alterations are continuously being described and forcing us to adapt our detection methods. Here we have evaluated the ability of eight amplicon-based next-generation sequencing (NGS) panels to detect the recently described mesenchymal epithelial transition factor (MET) exon 14 (METex14) alterations or new mutations conferring resistance to TKIs. MethodsA total of 191 tumor samples from patients with NSCLC were screened for METex14 mutations by Sanger sequencing, and 62 additional cases were screened by Sanger sequencing and two amplicon-based NGS panels. In silico comparison of eight commercially available targeted NGS panels was also performed for the detection of METex14 alterations or ALK, ROS1, or EGFR resistance mutations. ResultsNGS analysis of the positive METex14 cases revealed a false-negative case because of amplicon design. Moreover, in silico analysis revealed that none of the eight panels considered would be able to detect more than 63% of literature-reported cases of METex14 mutations and similar limitations would be expected with new ALK, ROS1, or EGFR resistance mutations. ConclusionsWe have illustrated major limitations of commercially available amplicon-based DNA NGS panels for detection of METex14 and recently described resistance mutations to TKIs. Documented choice of available panels and their frequent reevaluation are mandatory to deliver the most accurate data to the clinician for therapeutic decisions.

Treatment of ALK-rearranged non-small cell lung cancer: A review of the landscape and approach to emerging patterns of treatment resistance in the Australian context.

Since the identification of anaplastic lymphoma kinase (ALK) gene rearrangements in non-small cell lung cancer (NSCLC) in 2005, the treatment of ALK-rearranged NSCLC (ALK+ NSCLC) has evolved at a rapid pace. This molecularly distinct subset of NSCLC has uniquely important biology, clinicopathologic features and mechanisms of drug resistance which impact on the choice of treatment for a patient with this disease. There are multiple ALK tyrosine kinase inhibitors now available in clinical practice with efficacy data continuing to emerge and guide the optimal treatment algorithm. A detailed search of medical databases and clinical trial registries was conducted to capture all relevant articles on this topic enabling an updated detailed overview of the landscape of management of ALK-rearranged NSCLC.

Non-Small-Cell Lung Cancer (NSCLC) Harboring ALK Translocations: Clinical Characteristics and Management in a Real-Life Setting: a French Retrospective Analysis (GFPC 02-14 Study).

Chromosomal translocations involving the anaplastic lymphoma kinase gene (ALK) are rare oncogenic events found in 3-5% of non-small-cell lung cancers (NSCLC). Limited data have been published on the management of these patients outside clinical trials. To investigate the clinical characteristics and management of patients with NSCLC harboring ALK translocations (ALK+) in a real-life setting in France. This multicenter, observational, retrospective study included all NSCLC patients harboring ALK translocations diagnosed in participating centers between January 2012 and December 2014. Patient data include clinical characteristics, disease management, and outcomes [progression-free survival (PFS) and overall survival (OS)]. The 31 participating centers reported data on 132 patients, of whom 51% (n=67) were male. The median age was 60.1 ±14.5 (standard deviation)years; 89% (n=106/119) had performance status 0/1 at diagnosis; 79% (n=103/130) were non- or former smokers; 93% (n=120/129) had adenocarcinomas and 74%(n=97)/19%(n=25)/7%(n=10) had disease stages IV/III/I-II at diagnosis, respectively; co-mutations included EGFR (n=2), BRAF (n =2), KRAS (n=1), and HER2 (n=1). Of the patients with stage IV NSCLC (n=97), 96% received first-line treatment [75% chemotherapy-based, 21% ALK tyrosine kinase inhibitor (TKI)], with an associated response rate (RR), disease-control rate (DCR), and PFS of 42%, 64%, and 7.5 [95% confidence interval (CI) 5.9-9.5] months, respectively; 62% received second-line treatment (28% chemotherapy, 72% ALK TKI) with an associated RR, DCR, and PFS of 43.4%, 70%, and 4.7 (95% CI 4.0-8.1) months, respectively. The 2-year OS was 56.7% (95% CI 45.5-70.4%); median OS was not reached. The results of this real-life analysis suggest that the prognosis of NSCLC patients with theALK translocation may be better than that of the overall NSCLC population, but the outcomes were poorer than those of ALK+ NSCLC patients included in clinical studies.

Anaplastic lymphoma kinase gene rearrangement predicts better prognosis in NSCLC patients: A meta-analysis

ObjectiveFor non-small cell lung cancer (NSCLC), anaplastic lymphoma kinase (ALK) rearrangement and epidermal growth factor receptor (EGFR) mutations are predictive markers of the treatment benefit from selective tyrosine kinase inhibitors (TKI). However, their prognostic roles remained uncertain. Thus, we conducted this meta-analysis to evaluate the prognosis of ALK+ NSCLC patients in the treatment of surgery, chemotherapy, and/or EGFR-TKI. Materials and methodsPubMed, Embase and Cochrane databases were thoroughly searched to identify relevant studies. Primary endpoints of this study included overall survival (OS), recurrence/progression free survival (RFS/PFS) and objective response rate (ORR). All statistical analyses were performed via REVIEW MANAGER (version 5.3). ResultsIn total, 15 studies involving 4981 NSCLC patients were included. This study demonstrated that smoking status profoundly influenced prognosis of ALK related NSCLC. In the general population with NSCLC, compared with ALK- arm, ALK+ arm obtained a significantly better prognosis (HR=0.81 for OS, 95% CI=0.72-0.91; 0.80 for RFS/PFS, 95% CI=0.70-0.90), even after further stratification analysis according to disease stage. However, in the non-smoking population with NSCLC, compared with ALK- arm, those in the ALK+ arm had a worse prognosis (HR=1.65 for OS, 95% CI=1.28-2.12; 1.23 for RFS/PFS, 95% CI=1.05-1.44). Furthermore, ALK+ patients experienced a significantly higher ORR in pemetrexed-based chemotherapy but not in EGFR-TKI. ConclusionsSmoking status had a profound influence on the ALK-related prognosis of NSCLC. ALK rearrangement predicted a better prognosis in the general population with NSCLC, but a poor survival in the non-smoking population. Therefore, stratification according to smoking status is strongly recommended for future studies exploring ALK-related prognosis.

Ventana immunohistochemistry assay for anaplastic lymphoma kinase gene rearrangement detection in patients with non-small cell lung cancer: A meta-analysis.

BackgroundThe aim of this study was to evaluate the diagnostic value of Ventana immunohistochemistry (IHC) assay for anaplastic lymphoma kinase (ALK) gene rearrangement screening in patients with non‐small cell lung cancer (NSCLC).MethodsOpen published studies that reported the diagnostic performance of Ventana IHC assay for ALK gene rearrangement detection in NSCLC patients were extracted from PubMed, Embase, Google scholar, Wanfang, and China National Knowledge Infrastructure. The general information and number of true positive (tp), false positive (fp), false negative (fn), and true negative (tn) cases identified by Ventana IHC assay were extracted. The diagnostic sensitivity, specificity, positive likelihood ratio (+lr), negative likelihood ratio (−lr), diagnostic odds ratio (dor) and the summary receiver operating characteristic (ROC) curve were calculated using Stata 11.0 software.ResultsTen studies, including 240 ALK positive and 1973 ALK negative NSCLC patients were included in this meta‐analysis. The pooled diagnostic sensitivity, specificity, +lr, −lr, and dor were 0.94 (95% confidence interval [CI] 0.85–0.98), 1.00 (95% CI 0.99–1.00), 859.61 (95% CI 60.81–1200.00), 0.06 (95% CI 0.03–0.16), and 1400.00 (95% CI 813.29–23 000.00), respectively. The area under the ROC curve was 0.996 for Ventana IHC assay in detecting ALK gene rearrangement in NSCLC patients.ConclusionThe sensitivity and specificity of Ventana IHC assay for the detection of ALK gene rearrangement were high, thus Ventana IHC could substitute fluorescence in situ hybridization for the screening of ALK+ NSCLC patients.

ALK and ROS1 testing on lung cancer cytologic samples: Perspectives.

Cytologic sampling is the mainstay of diagnosing advanced lung cancer. Moreover, to select patients for personalized first-line or second-line treatment, epidermal growth factor receptor (EGFR) mutations and anaplastic lymphoma kinase (ALK) and c-ros oncogene 1 (ROS1) rearrangements are tested on cytologic preparations. Commercially available fluorescence in situ hybridization (FISH) and immunocytochemistry (ICC) assays have primarily been used for the identification of cells harboring ALK or ROS1 gene fusions on histologic rather than cytologic preparations. However, it is now recognized that FISH and ICC also can be applied on cytologic samples provided the cytopathologist is aware that FISH and ICC results are not always concordant and that the performance of ICC largely depends on antibody clones, signal detection systems, and scoring systems. Notably, the routine clinical use of FISH and ICC may be replaced by emerging next-generation sequencing and digital, color-coded barcode technologies, which have the advantage of simultaneously evaluating ALK, ROS1, and EGFR alterations in a single analysis. Although their use in clinical cytologic practice remains to be fully established, it is conceivable that this technology will replace both FISH and ICC analyses in future diagnostic algorithms. Here, the authors review studies devoted to testing ALK and ROS1 on cytology specimens in an attempt to provide an update for the cytopathologist regarding current and evolving practice. Cancer Cytopathol 2017;125:817-30. © 2017 American Cancer Society.