EML4-ALK G1202R mutation induces EMT and confers resistance to ceritinib in NSCLC cells via activation of STAT3/Slug signaling

The echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion gene occurs in approximately 5% of non-small-cell lung cancers (NSCLCs). The development of ALK tyrosine kinase inhibitors (ALK-TKIs) is a major advance in treating NSCLC with the ALK fusion gene. Nevertheless, acquired resistance to ALK-TKIs ultimately limits their use. A prevalent mechanism of drug resistance in kinases occurs through the mutation of G1202R in ALK. However, the mechanisms underlying G1202R resistance to ceritinib are not fully understood. Here, we demonstrated that the expression of EML4-ALK G1202R mutation in A549 cells induced an epithelial-mesenchymal transition (EMT) phenotype and significantly increased the migration and invasion abilities. These phenomena may be due to the upregulation of signal transducer and activator of transcription 3 (STAT3), accompanied by the elevated expression of Slug in EML4-ALK G1202R mutant cells. Furthermore, the combination of ALK and STAT3 inhibitors restored the sensitivity of EML4-ALK G1202R mutant cells to ceritinib. In conclusion, these data indicate that the EML4-ALK G1202R mutation mediates the EMT phenotype by activating the STAT3/Slug signaling pathway, resulting in resistance to ceritinib, and that the combination of STAT3 and ALK inhibitors may overcome ALK mutation-driven drug resistance in the clinic. READ ARTICLE

Cellular Signalling DOI:10.1016/j.cellsig.2022.110264

Authors: Jiwei Shen, Yuting Meng, Kunlun Wang, Minghuan Gao, Jianan Du, Junfang Wang, Zengqiang Li, Daiying Zuo, Yingliang Wu

Case StudyKirk SmithApril 1, 2022EML4-ALK G1202R mutation, EMT, STAT3, Slug, Resistance, Ceritinib

Deciphering the Mechanism of Gilteritinib Overcoming Lorlatinib Resistance to the Double Mutant I1171N/F1174I in Anaplastic Lymphoma Kinase

Anaplastic lymphoma kinase (ALK) is validated as a therapeutic molecular target in multiple malignancies, such as non-small cell lung cancer (NSCLC). However, the feasibility of targeted therapies exerted by ALK inhibitors is inevitably hindered owing to drug resistance. The emergence of clinically acquired drug mutations has become a major challenge to targeted therapies and personalized medicines. Thus, elucidating the mechanism of resistance to ALK inhibitors is helpful for providing new therapeutic strategies for the design of next-generation drug. Here, we used molecular docking and multiple molecular dynamics simulations combined with correlated and energetical analyses to explore the mechanism of how gilteritinib overcomes lorlatinib resistance to the double mutant ALK I1171N/F1174I. We found that the conformational dynamics of the ALK kinase domain was reduced by the double mutations I1171N/F1174I. Moreover, energetical and structural analyses implied that the double mutations ..... READ ARTICLE

Frontiers of Cell and Developmental Biology DOI:10.3389/fcell.2021.808864

Authors: Liang Shuai, Wang Qing, Qi Xuesen, Liu Yudi, Li Guozhen, Lu Shaoyong, Mou Linkai, Chen Xiangyu

Pre-Clinical, group1Kirk SmithDecember 23, 2021Gilteritinib, Lorlatinib, Resistance, ALK

Identification of a High-Level MET Amplification in CTCs and cfTNA of an ALK-Positive NSCLC Patient Developing Evasive Resistance to Crizotinib

We report the case of a 47-year-old female patient with stage IV NSCLC harboring an echinoderm microtubule associated protein like 4 (EML4)-ALK E20;A20 fusion who was initially treated with four cycles of platinum/pemetrexed until progressive disease. Thereafter, she was treated with crizotinib for 9 months until development of evasive resistance. Subsequently, the patient was primarily resistant to alectinib and ceritinib.
We obtained low-pass copy number profiles of single isolated CTCs after the patient had experienced development of evasive resistance to crizotinib and shown primary resistance to alectinib. Furthermore, we generated additional copy number profiles of CTCs upon development of primary resistance to ceritinib. These analyses revealed the presence of an approximately eightfold MNNG HOS Transforming gene (MET) amplification in all evaluable CTCs at both time points. MET amplification (∼sevenfold) could also be detected in the cfTNA at comparable time points.
This study ..... READ ARTICLE

Journal of Thoracic Oncology
DOI:10.1016/j.jtho.2018.08.2025

Authors: Lars-Arne Berger, Melanie Janning, Janna-Lisa Velthaus, Isabel Ben-Batalla, Stefanie Schatz, Markus Falk, Peter Iglauer, Ronald Simon, Ru Cao, Claudio Forcato, Nicolò Manaresi, Kelli Bramlett, Genny Buson, Annkathrin Hanssen, Markus Tiemann, Guido Sauter, Carsten Bokemeyer, Sabine Riethdorf, Martin Reck, Klaus Pantel, Harriet Wikman, and Sonja Loges

Case Study, group2Kirk SmithNovember 29, 2021Crizotinib, Resistance, MET amplification

Immunotherapy Treatment Patterns and Outcomes Among ALK-Positive Patients With Non–Small-Cell Lung Cancer

Real-world effectiveness (rwPFS) of ICIs in ALK-positive NSCLC patients, whether provided before or after TKIs, was limited, underscoring the relative lack of efficacy of ICI in this patient population, particularly compared to approved ALK TKIs. READ ARTICLE

Clinical Lung Cancer DOI: 10.1016/j.cllc.2020.08.003

Authors: Mohammad Jahanzeb, Huamao M.Lin, Xiaoyun Pan, Yu Yin, Pia Baumann, Corey J. Langer

MCL-1 inhibitors, fast-lane development of a new class of anti-cancer agents

Cell death escape is one of the most prominent features of tumor cells and closely linked to the dysregulation of members of the Bcl-2 family of proteins. Among those, the anti-apoptotic family member myeloid cell leukemia-1 (MCL-1) acts as a master regulator of apoptosis in various human malignancies. Irrespective of its unfavorable structure profile, independent research efforts recently led to the generation of highly potent MCL-1 inhibitors that are currently evaluated in clinical trials. This offers new perspectives to target a so far undruggable cancer cell dependency. However, a detailed understanding about the tumor and tissue type specific implications of MCL-1 are a prerequisite for the optimal (i.e., precision medicine guided) use of this novel drug class. In this review, we summarize the major functions of MCL-1 with a special focus on cancer, provide insights into its different roles in solid vs. hematological tumors and give an update about the (pre)clinical development p..... READ ARTICLE

Journal of Hematology and Oncology DOI: 10.1186/s13045-020-01007-9

Authors: rnold Bolomsky, Meike Vogler, Murat Cem Köse, Caroline A. Heckman, Grégory Ehx, Heinz Ludwig, Jo Caers

Review PaperKirk SmithDecember 11, 2020MCL-1, Resistance

Development of a Brigatinib degrader (SIAIS117) as a potential treatment for ALK positive cancer resistance

EML4-ALK and NPM-ALK fusion proteins possess constitutively activated ALK (anaplastic lymphoma kinase) activity, which in turn leads to the development of non-small cell lung cancer and anaplastic large-cell lymphomas (ALCLs). FDA-approved ALK inhibitor drugs cause significant cancer regression. However, drug resistance eventually occurs and it becomes a big obstacle in clinic. Novel proteolysis targeting chimera (PROTAC) technology platform provides a potential therapeutic strategy for drug resistance. Herein, we designed and synthesized a series of ALK PROTACs based on Brigatinib and VHL-1 conjunction, and screened SIAIS117 as the best degrader which not only blocked the growth of SR and H2228 cancer cell lines, but also degraded ALK protein. In addition, SIAIS117 also showed much better growth inhibition effect than Brigatinib on 293T cell line that exogenously expressed G1202R-resistant ALK proteins. Furthermore, it also degraded G1202R mutant ALK protein in vitro. At last, it has ..... READ ARTICLE

European Journal of Medicinal Chemistry DOI:10.1016/j.ejmech.2020.112190

Authors: Ning Sun, Chaowei Ren, Ying Kong, Hui Zhong, Jinju Chen, Yan Li, Jianshui Zhang, Yuedong Zhou, Xing Qiu, Haifan Lin, Xiaoling Song, Xiaobao Yang, Biao Jiang

Pre-ClinicalKirk SmithMay 1, 2020ALK, Brigatinib, Degrader, VHL, PROTAC, NSCLC, ALCL, Resistance

Overcoming resistance by ALK compound mutation (I1171S + G1269A) after sequential treatment of multiple ALK inhibitors in non-small cell lung cancer

Background: Anaplastic lymphoma kinase (ALK) fusion genes are found in 3%–5% of non-small cell lung cancers (NSCLCs). ALK inhibitors show a very high response rate to ALK-positive NSCLCs. However, the emergence of acquired resistance is inevitable. In this study, we investigated the drugs for overcoming resistance especially compound mutations after sequential treatment with crizotinib, alectinib, and lorlatinib. Conclusions: With the sequential ALK inhibitors treatment, cancer cells accumulate new mutations in addition to mutations acquired previously. The identified compound mutation (I1171S + G1269A) was found to be sensitive to ceritinib and brigatinib, and indeed the patient's tumor partially responded to ceritinib. READ ARTICLE

Thoracic Cancer DOI:10.1111/1759-7714.13299

Authors: Ken Takahashi, Yosuke Seto, Koutaroh Okada, Shinya Uematsu, Ken Uchibori, Mika Tsukahara, Tomoko Oh-hara, Naoya Fujita, Noriko Yanagitani, Makoto Nishio, Kenichi Okubo, Ryohei Katayama

Stratégies thérapeutiques dans le cancer bronchique non à petites cellules ALK positif de stade IV Therapeutic strategies in advanced ALK positive non-small cell lung cancer

Stratégies thérapeutiques dans le cancer bronchique non à petites cellules ALK positif de stade IVTherapeutic strategies in advanced ALK positive non-small cell lung cancer. READ ARTICLE

Revue des Maladies Respiratoires DOI:Therapeutic strategies in advanced ALK positive non-small cell lung cancer

Authors: A. Tiotiu, Y. Billon, P. Vaillant, O. Menard, P. Hofman, C. Mascaux

Review PaperKirk SmithDecember 1, 2019ALK TKI, Therapeutic strategy, Rebiopsy, Resistance

ALK Mutation Status Before and After Alectinib Treatment in Locally Advanced or Metastatic ALK-Positive NSCLC: Pooled Analysis of Two Prospective Trials

Introduction: The effectiveness of ALK receptor tyrosine kinase (ALK) inhibitors can be limited by the development of ALK resistance mutations. This exploratory analysis assessed the efficacy of alectinib in patients with NSCLC and ALK point mutations using pooled data from two single-arm phase II studies. Conclusions: Alectinib appears clinically active against ALK rearrangements and mutations, as well as several ALK variants that can cause resistance to crizotinib. The use of cell-free DNA in plasma samples may be an alternative noninvasive method for monitoring resistance mutations during therapy. READ ARTICLE

Journal of Thoracic Oncology DOI:10.1016/j.jtho.2019.10.015

Authors: Johannes Noé, Alex Lovejoy, Sai-Hong Ignatius Ou, Stephanie J. Yaung, Walter Bordogna, Daniel M. Klass, Craig A. Cummings, Alice T. Shaw

Review PaperKirk SmithNovember 8, 2019Alectinib, ALK-positive, NSCLC, Mutation analysis, Resistance

P2.14-18 Upregulation of AURKA Leads to Acquired Resistance in EML4-ALK NSCLC Cell Line

Background: Molecular targeted therapies in NSCLC often results in profound initial patient responses, these responses are short-term due to the development of acquired resistance. In an EML4-ALK NSCLC background, acquired resistance can be developed in two ways ALK dependent (ALK secondary mutations) and ALK independent (alternative oncogenic pathways). In our study, we have shown that increased expression of Aurora kinase A (AURKA) leads to acquired resistance upon treatment with ALK TKI crizotinib. Conclusion: Our results indicate a new mechanism to acquire resistance upon treatment with ALK TKI crizotinib. Inhibition of both ALK and AURKA activity might be beneficial for ALK TKI resistant tumors with increased AURKA gene expression/activity. READ ARTICLE

Journal of Thoracic Oncology DOI:10.1016/j.jtho.2019.08.1803

Authors: G. Umapathy, R. Palmer, B. Hallberg

Conference PaperKirk SmithOctober 21, 2019EML4-ALK, AURKA, Resistance

Relevance of Detection of Mechanisms of Resistance to ALK Inhibitors in ALK-Rearranged NSCLC in Routine Practice

Background: Anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitors (TKIs) have shown efficacy in the treatment of ALK-rearranged non–small-cell lung cancer (NSCLC), but the disease eventually progresses in all patients. In many cases, resistance to ALK TKIs arises through ALK mutations. Although clinical and biological data suggest variations in TKI efficacy according to the mechanism of resistance, ALK mutations are still rarely investigated in routine practice. Conclusion: Targeted next-generation sequencing is suitable for detecting ALK resistance mutations in ALK-rearranged NSCLC patients in routine practice. It might help select the best treatment at the time of disease progression during treatment with an ALK TKI. READ ARTICLE

Clinical Lung Cancer DOI:10.1016/j.cllc.2019.02.013

Authors: Philippe Jamme, Clotilde Descarpentries, Radj Gervais, Eric Dansin, Marie Wislez, Valérie Grégoire, Nicolas Richard, Simon Baldacci, Nathalie Rabbe, Maeva Kyheng, Zoulika Kherrouche, Fabienne Escande, Marie Christine Copin, Alexis B. Cortot

MYC Amplification as a Potential Mechanism of Primary Resistance to Crizotinib in ALK-Rearranged Non-Small Cell Lung Cancer: A Brief Report

Translocations of the anaplastic lymphoma kinase (ALK) can be effectively targeted in advanced non-small cell lung cancer by ALK-TKI inhibitors including Crizotinib. However, the development of acquired resistance often limits the duration of these therapies. While several mechanisms of secondary resistance have been already identified, little is known about molecular determinants of primary resistance. In our brief report we investigated the tumor molecular profile of a patient who failed to respond to Crizotinib.
We postulate that the MYC gene may be implicated in the mechanism of primary resistance to ALK inhibitors. We also suggest potential MYC-directed inhibition strategies to overcome primary resistance in advanced ALK-rearranged NSCLC. READ ARTICLE

Translational Oncology DOI:10.1016/j.tranon.2018.09.013

Authors: Karim Rihawi, Roberta Alfieri, Michelangelo Fiorentino, Francesca Fontana, Elisa Capizzi, Andrea Cavazzoni, Mario Terracciano, Silvia La Monica, Alberto Ferrarini, Genny Buson, Pier Giorgio Petronini, Andrea Ardizzoni

Case StudyKirk SmithJanuary 1, 2019ALK, Crizotinib, Resistance, MYC Amplification

Impact of EML4-ALK Variant on Resistance Mechanisms and Clinical Outcomes in ALK-Positive Lung Cancer

Advanced anaplastic lymphoma kinase (ALK) fusion-positive non–small-cell lung cancers (NSCLCs) are effectively treated with ALK tyrosine kinase inhibitors (TKIs). However, clinical outcomes in these patients vary, and the benefit of TKIs is limited as a result of acquired resistance. Emerging data suggest that the ALK fusion variant may affect clinical outcome, but the molecular basis for this association is unknown. We identified 129 patients with ALK-positive NSCLC with known ALK variants. ALK resistance mutations and clinical outcomes on ALK TKIs were retrospectively evaluated according to ALK variant. A Foundation Medicine data set of 577 patients with ALK-positive NSCLC was also examined. Specific ALK variants may be associated with the development of ALK resistance mutations, particularly G1202R, and provide a molecular link between variant and clinical outcome. ALK variant thus represents a potentially important factor in the selection of next-generation ALK inhibitors. READ ARTICLE

Journal of Clinical Oncology DOI:10.1200/JCO.2017.76.2294

Authors: Lin JJ, Zhu VW, Yoda S, Yeap BY, Schrock AB, Dagogo-Jack I, Jessop NA, Jiang GY, Le LP, Gowen K, Stephens PJ, Ross JS, Ali SM, Miller VA, Johnson ML, Lovly CM, Hata AN, Gainor JF, Iafrate AJ, Shaw AT, Ou SI

Real-World OutcomesKirk SmithApril 20, 2018EML4-ALK, Variant, Resistance

Sequential ALK Inhibitors Can Select for Lorlatinib-Resistant Compound ALK Mutations in ALK-Positive Lung Cancer

The cornerstone of treatment for advanced ALK-positive lung cancer is sequential therapy with increasingly potent and selective ALK inhibitors. The third-generation ALK inhibitor lorlatinib has demonstrated clinical activity in patients who failed previous ALK inhibitors. To define the spectrum of ALK mutations that confer lorlatinib resistance, we performed accelerated mutagenesis screening of Ba/F3 cells expressing EML4–ALK. Under comparable conditions, N-ethyl-N-nitrosourea (ENU) mutagenesis generated numerous crizotinib-resistant but no lorlatinib-resistant clones harboring single ALK mutations. In similar screens with EML4–ALK containing single ALK resistance mutations, numerous lorlatinib-resistant clones emerged harboring compound ALK mutations. To determine the clinical relevance of these mutations, we analyzed repeat biopsies from lorlatinib-resistant patients. Seven of 20 samples (35%) harbored compound ALK mutations, including two identified in the ENU screen. Whole-..... READ ARTICLE

Cancer Discovery DOI:10.1158/2159-8290.CD-17-1256

Authors: Yoda S, Lin JJ, Lawrence MS, Burke BJ, Friboulet L, Langenbucher A, Dardaei L, Prutisto-Chang K, Dagogo-Jack I, Timofeevski S, Hubbeling H, Gainor JF, Ferris LA, Riley AK, Kattermann KE, Timonina D, Heist RS, Iafrate AJ, Benes CH, Lennerz JK, Mino-Kenudson M, Engelman JA, Johnson TW, Hata AN, Shaw AT.

Pre-ClinicalKirk SmithApril 12, 2018ALK Inhibitors, Lorlatinib, Resistance, ALK Mutations