Molecular profiling of cholangiocarcinoma shows potential for targeted therapy treatment decisions

doi:10.1016/j.humpath.2012.11.006

Human Pathology

Volume 44, Issue 7, July 2013, Pages 1216-1222

https://doi.org/10.1016/j.humpath.2012.11.006 Get rights and content

Summary

Cholangiocarcinoma is a highly lethal cancer of the biliary tract. The intrahepatic subtype of cholangiocarcinoma is increasing in incidence globally. Despite technologic advancements over the past decade, little is known about the somatic changes that occur in these tumors. The goal of this study was to determine the frequency of common oncogenes in resected cholangiocarcinoma specimens that could provide potential therapeutic targets for patients diagnosed with cholangiocarcinoma. Formalin-fixed, paraffin-embedded tissue blocks from 94 resected cholangiocarcinomas were used to extract DNA from areas comprising more than 20% tumor. Specimens were evaluated using the Sequenom MassARRAY OncoCarta Mutation Profiler Panel (San Diego, CA). This matrix-assisted laser desorption/ionization–time of flight mass spectrometry single genotyping panel evaluates 19 oncogenes for 238 somatic mutations. Twenty-five mutations were identified in 23 of the 94 cholangiocarcinomas within the following oncogenes: KRAS (n = 12), PIK3CA (n = 5), MET (n = 4), EGFR (n = 1), BRAF (n = 2), and NRAS (n = 1). Mutations were identified in 7 (26%) of 27 extrahepatic cholangiocarcinomas and 16 (24%) of 67 intrahepatic cholangiocarcinomas. When combined with IDH1/2 testing, 40 (43%) of the 94 cholangiocarcinomas had a detectable mutation. MassARRAY technology can be used to detect mutations in a wide variety of oncogenes using paraffin-embedded tissue. Clinical testing for somatic mutations may drive personalized therapy selection for cholangiocarcinomas in the future. The variety of mutations detected suggests that a multiplexed mutation detection approach may be necessary for managing patients with biliary tract malignancy.

Introduction

Cholangiocarcinoma represents a heterogeneous group of malignancies that are often classified as intrahepatic or extrahepatic based on their anatomical location. Recent advancements in imaging technologies and new molecular markers have improved algorithms for diagnosing cholangiocarcinoma. However, most patients with cholangiocarcinoma still present with advanced-stage disease and have a median survival of less than 2 years [1]. Although progress has been made on better understanding the pathophysiology of these tumors and their subclassification, this knowledge has not led to curative therapies [2]. As a result, there continues to be a need for therapies that improve survival in patients with cholangiocarcinoma, especially advanced-stage patients not amenable to surgical intervention.

Somatic mutation analysis has become a useful tool in selecting personalized therapy for many solid tumors. As a result, new broad-based molecular techniques are being used to molecularly profile tumors to identify potential therapeutic targets. One mass spectrometry technique, matrix-assisted laser desorption/ionization–time of flight, has been used to assess tumors for point mutations and small deletions in commonly altered oncogenes [3]. One of the more commonly used matrix-assisted laser desorption/ionization–time of flight instruments, the Sequenom (San Diego, CA) MassARRAY, has been used in conjunction with a commercial kit (OncoCarta v 1.0 kit; Sequenom) to assess tumors for 238 somatic mutations across 19 oncogenes [4]. The relevance of using this gene panel is that it interrogates oncogenes with known companion targeted therapies (eg, EGFR, BRAF) or assesses for mutations in genes that are known to decrease the efficacy of specific personalized therapies (eg, KRAS, NRAS, HRAS). Lastly, this gene panel includes oncogenes (eg, PIK3CA, MET) that are members of pathways that are currently targeted in clinical trials. The goal of this study was to use the OncoCarta kit to determine the frequency of somatic mutations in common oncogenes in resected extrahepatic and intrahepatic cholangiocarcinomas. The findings of this study will help elucidate the molecular pathogenesis of these tumors and identify potential therapeutic targets for patients diagnosed with cholangiocarcinoma.

Section snippets

Materials and methods

Following approval by the Mayo Clinic Institutional Review Board, 94 (67 intrahepatic and 27 extrahepatic cholangiocarcinomas) surgically resected, formalin-fixed, paraffin-embedded primary cholangiocarcinoma specimens were retrieved from laboratory archives. Retrieved hematoxylin and eosin slides were reviewed by a pathologist (S. E. K., J. Z., or K. C. H.) to select blocks for subsequent DNA testing. Eight serial 10-μm sections were then cut from selected tissue blocks, and areas containing

Results

Twenty-nine mutations were identified by MassARRAY within the 94 cholangiocarcinomas. Twenty-five of these mutations were confirmed by follow-up testing, resulting in 24% of the cases having a pathogenic mutation in 1 or more of the targeted oncogenes. The specific mutations identified are listed in the Table. KRAS mutations were the most commonly identified mutation (n = 12, 13%) and were identified in both extrahepatic (n = 6, 22%) and intrahepatic (n = 6, 9%) cholangiocarcinomas. Only one

Discussion

The results of this study demonstrate that formalin-fixed, paraffin-embedded cholangiocarcinoma specimens can be successfully analyzed for numerous oncogene mutations using the Sequenom OncoCarta multiplex assay. Many of the genes assessed for in this study encode for proteins that are important components of tyrosine kinase pathways, which are attractive drug targets for specific monoclonal antibodies and small molecule tyrosine kinase inhibitors [6], [7], [8], [9]. Therapies targeting

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^☆: Disclosure of potential conflicts of interest: L. M. Holtegaard and W. E. Highsmith own shares of stock in Sequenom, Inc. No other potential conflict of interest exists among the remaining coauthors.

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Original contributionMolecular profiling of cholangiocarcinoma shows potential for targeted therapy treatment decisions☆

Summary

Introduction

Section snippets

Materials and methods

Results

Discussion

Hum Pathol

Lancet Oncol

Hum Pathol

Biomed Pharmacother

J Mol Diagn

Lancet Oncol

HPB (Oxford)

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J Surg Oncol

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BMC Cancer

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BMC Cancer

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Nat Rev Cancer

Prognosis of women with metastatic breast cancer by HER2 status and trastuzumab treatment: an institutional-based review

J Clin Oncol

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N Engl J Med

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Original contribution
Molecular profiling of cholangiocarcinoma shows potential for targeted therapy treatment decisions☆