ALK alteration is a frequent event in aggressive breast cancers

IntroductionBreast cancer is the most common female malignancy worldwide and, despite improvements in treatment modalities, there are increased chances of recurrence and metastasis in a substantial number of cases and it remains one of the major causes of mortality among female cancer patients. Anaplastic lymphoma kinase (ALK) gene has been found to be altered in several solid and hematologic tumors. We aimed to comprehensively study the prevalence of ALK expression, and changes in copy number and translocation in a large cohort of breast cancer cases in a Middle Eastern population.MethodsALK protein expression was investigated by immunohistochemistry and numerical and structural variations of the ALK gene were analyzed by fluorescence in situ hybridization (FISH) in a tissue microarray format in a cohort of more than 1000 Middle Eastern breast cancers. The data were correlated with clinicopathologic parameters and other important molecular biomarkers.ResultsImmunohistochemical analysis showed ALK overexpression in 36.0 % of the breast cancer patients and gene amplification was present in 13.3 % of cases, seen by FISH analyses. ALK overexpression was significantly associated with ALK gene amplification (p = 0.0031). ALK-overexpressing tumors showed significant association with high-grade tumors (p = 0.0039), ductal histologic subtype (p = 0.0076), triple-negative phenotype (p = 0.0034), and high Ki-67 (p = 0.0001) and p-AKT (p <0.0001).ConclusionsImmunohistochemical analysis showed ALK is overexpressed in a substantial proportion of breast cancers and possibly plays a significant role in the aggressive behavior of this cancer. Gene amplification is hypothesized to be a possible cause for a significant proportion of this overexpression. Based on these findings, a potential role for an ALK inhibitor, as a therapeutic agent targeting aggressive subtypes of breast cancer, merits further investigation.

[1]  W. Miller,et al.  Molecular characterization of WDCP, a novel fusion partner for the anaplastic lymphoma tyrosine kinase ALK. , 2015, Biomedical reports.

[2]  A. Iafrate,et al.  A Novel, Highly Sensitive Antibody Allows for the Routine Detection of ALK-Rearranged Lung Adenocarcinomas by Standard Immunohistochemistry , 2010, Clinical Cancer Research.

[3]  J. D. de Wilt,et al.  Inflammatory breast cancer: an overview. , 2015, Critical reviews in oncology/hematology.

[4]  D. Ekwueme,et al.  Health and economic impact of breast cancer mortality in young women, 1970-2008. , 2014, American journal of preventive medicine.

[5]  J. Stagg,et al.  Immunotherapeutic approaches in triple-negative breast cancer: latest research and clinical prospects , 2013, Therapeutic advances in medical oncology.

[6]  W. Han,et al.  Ki-67 Expression Gives Additional Prognostic Information on St. Gallen 2007 and Adjuvant! Online Risk Categories in Early Breast Cancer , 2009, Annals of Surgical Oncology.

[7]  Gudrun Schleiermacher,et al.  Somatic and germline activating mutations of the ALK kinase receptor in neuroblastoma , 2008, Nature.

[8]  D. Gary Gilliland,et al.  Activating mutations in ALK provide a therapeutic target in neuroblastoma , 2008, Nature.

[9]  F. Speleman,et al.  Activated Alk triggers prolonged neurogenesis and Ret upregulation providing a therapeutic target in ALK-mutated neuroblastoma , 2014, Oncotarget.

[10]  C. Perou,et al.  The Triple Negative Paradox: Primary Tumor Chemosensitivity of Breast Cancer Subtypes , 2007, Clinical Cancer Research.

[11]  Steven J. M. Jones,et al.  Comprehensive molecular portraits of human breast tumours , 2013 .

[12]  K. Al-Kuraya,et al.  Prognostic significance of alterations in KRAS isoforms KRAS‐4A/4B and KRAS mutations in colorectal carcinoma , 2009, The Journal of pathology.

[13]  A. Iafrate,et al.  Unique Clinicopathologic Features Characterize ALK-Rearranged Lung Adenocarcinoma in the Western Population , 2009, Clinical Cancer Research.

[14]  Philippe Autier,et al.  Disparities in breast cancer mortality trends between 30 European countries: retrospective trend analysis of WHO mortality database , 2010, BMJ : British Medical Journal.

[15]  Mitch Dowsett,et al.  Proliferation marker Ki-67 in early breast cancer. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[16]  E. Grande,et al.  Targeting Oncogenic ALK: A Promising Strategy for Cancer Treatment , 2011, Molecular Cancer Therapeutics.

[17]  A. Rosenwald,et al.  TRK-fused gene (TFG) is a new partner of ALK in anaplastic large cell lymphoma producing two structurally different TFG-ALK translocations. , 1999, Blood.

[18]  Wei Zhang,et al.  Anaplastic lymphoma kinase is activated through the pleiotrophin/receptor protein-tyrosine phosphatase beta/zeta signaling pathway: an alternative mechanism of receptor tyrosine kinase activation. , 2007, The Journal of biological chemistry.

[19]  G. Sauter,et al.  Predominance of high-grade pathway in breast cancer development of Middle East women , 2005, Modern Pathology.

[20]  Q. Zhan,et al.  Proteomic profiling of proteins dysregulted in Chinese esophageal squamous cell carcinoma , 2007, Journal of Molecular Medicine.

[21]  Ewan Birney,et al.  Ensembl Genome Browser , 2010 .

[22]  S. Serrano,et al.  Increased ALK Gene Copy Number and Amplification are Frequent in Non-small Cell Lung Cancer , 2011, Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer.

[23]  F. Hirsch,et al.  Anaplastic lymphoma kinase (ALK): A potential oncogenic driver in triple-negative breast cancer? , 2013 .

[24]  F. Hirsch,et al.  A dramatic response to crizotinib in a non-small-cell lung cancer patient with IHC-positive and FISH-negative ALK. , 2012, Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer.

[25]  Ravi Salgia,et al.  Targeting ALK: a promising strategy for the treatment of non-small cell lung cancer, non-Hodgkin’s lymphoma, and neuroblastoma , 2012, Targeted Oncology.

[26]  Jeffrey W. Clark,et al.  Anaplastic lymphoma kinase inhibition in non-small-cell lung cancer. , 2010, The New England journal of medicine.

[27]  Aleix Prat Aparicio.  Comprehensive molecular portraits of human breast tumours , 2012 .

[28]  R. Tuma.  ALK gene amplified in most inflammatory breast cancers. , 2012, Journal of the National Cancer Institute.

[29]  S. Sleijfer,et al.  Anaplastic lymphoma kinase aberrations in rhabdomyosarcoma: clinical and prognostic implications. , 2012, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[30]  R. Walker,et al.  World Health Organization Classification of Tumours. Pathology and Genetics of Tumours of the Breast and Female Genital Organs , 2005 .

[31]  Y. Ishikawa,et al.  KIF5B-ALK, a Novel Fusion Oncokinase Identified by an Immunohistochemistry-based Diagnostic System for ALK-positive Lung Cancer , 2009, Clinical Cancer Research.

[32]  Z. Jehan,et al.  Frequent PIK3CA gene amplification and its clinical significance in colorectal cancer , 2009, The Journal of pathology.

[33]  G. Horgan,et al.  Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR , 2002 .

[34]  R. George,et al.  Emerging importance of ALK in neuroblastoma. , 2011, Seminars in cancer biology.

[35]  D. Rimm,et al.  X-Tile , 2004, Clinical Cancer Research.

[36]  Steven J. M. Jones,et al.  Comprehensive molecular portraits of human breast tumors , 2012, Nature.

[37]  Li Li,et al.  Exon Array Profiling Detects EML4-ALK Fusion in Breast, Colorectal, and Non–Small Cell Lung Cancers , 2009, Molecular Cancer Research.

[38]  M. Dietel,et al.  EML4-ALK testing in non-small cell carcinomas of the lung: a review with recommendations , 2012, Virchows Archiv.

[39]  H. Aburatani,et al.  Identification of the transforming EML4–ALK fusion gene in non-small-cell lung cancer , 2007, Nature.

[40]  P. Pérez-Piñera,et al.  Anaplastic lymphoma kinase is expressed in different subtypes of human breast cancer. , 2007, Biochemical and biophysical research communications.

[41]  H. Bush,et al.  Breast Cancer Research , 1978, British Journal of Cancer.

[42]  M. Abd-Elazeem,et al.  Claudin 4 expression in triple-negative breast cancer: correlation with androgen receptors and Ki-67 expression. , 2015, Annals of diagnostic pathology.

[43]  A. Mehrjardi,et al.  Expression of Anaplastic Lymphoma Kinase Protein in Human Breast Cancer , 2013 .

[44]  S. Ogawa,et al.  Oncogenic mutations of ALK kinase in neuroblastoma , 2008, Nature.

[45]  C. Powers,et al.  Pleiotrophin Signaling through Anaplastic Lymphoma Kinase Is Rate-limiting for Glioblastoma Growth* , 2002, The Journal of Biological Chemistry.

[46]  M. Link,et al.  ALK+ anaplastic large cell lymphoma exhibits phosphatidylinositol‐3 kinase/akt activity with retained but inactivated PTEN—A report from the Children's Oncology Group , 2012, Pediatric blood & cancer.

[47]  S. Raimondi,et al.  Renal cell carcinoma with novel VCL–ALK fusion: new representative of ALK-associated tumor spectrum , 2011, Modern Pathology.

[48]  G. Sauter,et al.  Rare oncogenic mutations of predictive markers for targeted therapy in triple-negative breast cancer , 2012, Breast Cancer Research and Treatment.

[49]  Craig D. Shriver,et al.  Effect of ASCO/CAP Guidelines for Determining ER Status on Molecular Subtype , 2012, Annals of Surgical Oncology.

[50]  F. Kelleher,et al.  The emerging pathogenic and therapeutic importance of the anaplastic lymphoma kinase gene. , 2010, European journal of cancer.

[51]  T. Utsunomiya,et al.  EML4–ALK fusion transcript is not found in gastrointestinal and breast cancers , 2008, British Journal of Cancer.

[52]  Robin L. Jones,et al.  Relationship between oestrogen receptor status and proliferation in predicting response and long-term outcome to neoadjuvant chemotherapy for breast cancer , 2009, Breast Cancer Research and Treatment.

[53]  W. Han,et al.  Prognostic impact of clinicopathologic parameters in stage II/III breast cancer treated with neoadjuvant docetaxel and doxorubicin chemotherapy: paradoxical features of the triple negative breast cancer , 2007, BMC Cancer.

[54]  Yuki Togashi,et al.  RET, ROS1 and ALK fusions in lung cancer , 2012, Nature Medicine.

[55]  Z. Jehan,et al.  Clinicopathological analysis of papillary thyroid cancer with PIK3CA alterations in a Middle Eastern population. , 2008, The Journal of clinical endocrinology and metabolism.

[56]  Jiri Polivka,et al.  Molecular targets for cancer therapy in the PI3K/AKT/mTOR pathway. , 2014, Pharmacology & therapeutics.

[57]  F. Bertucci,et al.  Presence of anaplastic lymphoma kinase in inflammatory breast cancer , 2013, SpringerPlus.

[58]  D N Shapiro,et al.  Fusion of a kinase gene, ALK, to a nucleolar protein gene, NPM, in non-Hodgkin's lymphoma. , 1994, Science.

[59]  Ayesha Ahmed,et al.  Protein expression profile and prevalence pattern of the molecular classes of breast cancer - a Saudi population based study , 2010, BMC Cancer.

[60]  B. Brown,et al.  Prognostic significance of Akt, phospho-Akt and BAD expression in primary breast cancer. , 2009, European journal of cancer.

[61]  Marc Ladanyi,et al.  Crizotinib in ALK-rearranged inflammatory myofibroblastic tumor. , 2010, The New England journal of medicine.

[62]  K. Ross,et al.  Molecular rationale for the use of PI3K/AKT/mTOR pathway inhibitors in combination with crizotinib in ALK-mutated neuroblastoma , 2014, Oncotarget.