Development of a real-time RT-PCR assay for detecting EGFRvIII in glioblastoma samples

被引:83
作者
Yoshimoto, Koji [1 ,2 ]
Dang, Julie [1 ,2 ]
Zhu, Shaojun [1 ,2 ]
Nathanson, David [1 ,2 ]
Huang, Tiffany [1 ,2 ]
Dumont, Rebecca [1 ,2 ]
Seligson, David B. [1 ,2 ,3 ]
Yong, William H. [1 ,2 ]
Xiong, Zhenggang [1 ,2 ]
Rao, Nagesh
Winther, Henrik [9 ]
Chakravarti, Arnab [10 ]
Bigner, Darell D. [11 ]
Mellinghoff, Ingo K. [3 ,7 ,8 ]
Horvath, Steve [4 ,5 ]
Cavenee, Webster K. [12 ]
Cloughesy, Timothy F. [6 ,7 ,8 ]
Mischel, Paul S. [1 ,2 ,3 ,6 ,7 ,8 ]
机构
[1] Univ Calif Los Angeles, David Greffen Sch Med, Dept Pathol, Los Angeles, CA 90095 USA
[2] Univ Calif Los Angeles, David Greffen Sch Med, Lab Med, Los Angeles, CA USA
[3] Univ Calif Los Angeles, David Greffen Sch Med, Dept Mol & Med Pharmacol, Los Angeles, CA USA
[4] Univ Calif Los Angeles, David Greffen Sch Med, Dept Human Genet, Los Angeles, CA USA
[5] Univ Calif Los Angeles, David Greffen Sch Med, Dept Biostat, Los Angeles, CA USA
[6] Univ Calif Los Angeles, David Greffen Sch Med, Dept Neurol, Los Angeles, CA USA
[7] Univ Calif Los Angeles, Jonsson Comprehens Canc Ctr, David Greffen Sch Med, Los Angeles, CA 90024 USA
[8] Univ Calif Los Angeles, David Greffen Sch Med, Henry E Singleton Brain Tumor Program, Los Angeles, CA 90024 USA
[9] DAKO A S, Glostrup, Denmark
[10] Harvard Univ, Massachusetts Gen Hosp, Sch Med, Dept Radiat Oncol, Boston, MA USA
[11] Preston Robert Tisch Brain Tumor Ctr Duke, Dept Pathol, Durham, NC USA
[12] Univ Calif San Diego, Ludwig Inst Canc Res, San Diego, CA 92103 USA
关键词
D O I
10.1158/1078-0432.CCR-07-1966
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Purpose: Epidermal growth factor receptor variant III (EGFRvIII) is an oncogenic, constitutively active mutant form of the EGFR that is commonly expressed in glioblastoma and is also detected in a number of epithelial cancers. EGFRvIII presents a unique antigenic target for anti-EGFRvIII vaccines and it has been shown to modulate response to EGFR kinase inhibitor therapy. Thus, detection in clinical samples may be warranted. Existing patents preclude the use of anti-EGFRvIII antibodies for clinical detection. Further, frozen tissue is not routinely available, particularly for patients treated in the community. Thus, detection of EGFRvIII in formalin-fixed paraffin-embedded (FFPE) clinical samples is a major challenge. Experimental Design: We developed a real-time reverse transcription-PCR (RT-PCR) assay for detecting EGFRvIII in FFPE samples and analyzed 59 FFPE glioblastoma clinical samples with paired frozen tissue from the same surgical resection. We assessed EGFRvIII protein expression by immunohistochemistry using two distinct specific anti-EGFRvIII antibodies and examined EGFR gene amplification by fluorescence in situ hybridization. Results: The FFPE RT-PCR assay detected EGFRvIII in 16 of 59 (27%) samples, exclusively in cases with EGFR amplification, consistent with the expected frequency of this alteration. The FFPE RT-PCR assay was more sensitive and specific for detecting EGFRvIII than either of the two antibodies alone, or in combination, with a sensitivity of 93% (95% confidence interval, 0.78-1.00) and a specificity of 98% (95% confidence interval, 0.93-1.00). Conclusion: This assay will facilitate accurate assessment of EGFRvIII in clinical samples and may aid in the development of strategies for stratifying patients for EGFRvIII-directed therapies.
引用
收藏
页码:488 / 493
页数:6
相关论文
共 23 条
  • [1] Immunohistochemical detection of EGFRvIII in high malignancy grade astrocytomas and evaluation of prognostic significance
    Aldape, KD
    Ballman, K
    Furth, A
    Buckner, JC
    Giannini, C
    Burger, PC
    Scheithauer, BW
    Jenkins, RB
    James, CD
    [J]. JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY, 2004, 63 (07) : 700 - 707
  • [2] Choe G, 2003, CANCER RES, V63, P2742
  • [3] Frederick L, 2000, CANCER RES, V60, P1383
  • [4] GARCIA DP, 1993, CANCER RES, V53, P3217
  • [5] Taming a mutinous mutant: An errant receptor becomes a prime cancer target
    Goldman, Bruce
    [J]. JOURNAL OF THE NATIONAL CANCER INSTITUTE, 2007, 99 (07) : 504 - 505
  • [6] Prognostic effect of epidermal growth factor receptor and EGFRvIII in glioblastoma multiforme patients
    Heimberger, AB
    Hlatky, R
    Suki, D
    Yang, D
    Weinberg, J
    Gilbert, M
    Sawaya, R
    Aldape, K
    [J]. CLINICAL CANCER RESEARCH, 2005, 11 (04) : 1462 - 1466
  • [7] HEIMBERGER AB, 2006, EPIDERMAL GROWTH FAC
  • [8] Epidermal growth factor receptor variant III mutations in lung tumorigenesis and sensitivity to tyrosine kinase inhibitors
    Ji, Hongbin
    Zhao, Xiaojun
    Yuza, Yuki
    Shimamura, Takeshi
    Li, Danan
    Protopopov, Alexei
    Jung, Boonim L.
    McNamara, Kate
    Xia, Huili
    Glatt, Karen A.
    Thomas, Roman K.
    Sasaki, Hidefumi
    Horner, James W.
    Eck, Michael
    Mitchell, Albert
    Sun, Yangping
    Al-Hashem, Ruqayyah
    Bronson, Roderick T.
    Rabindran, Sriclhar K.
    Discafani, Carolyn M.
    Maher, Elizabeth
    Shapiro, Geoffrey I.
    Meyerson, Matthew
    Wong, Kwok-Kin
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (20) : 7817 - 7822
  • [9] Therapeutic anti-EGFR antibody 806 generates responses in murine de novo EGFR mutant-dependent lung carcinomas
    Li, Danan
    Ji, Hongbin
    Zaghlul, Sara
    McNamara, Kate
    Liang, Mei-Chih
    Shimamura, Takeshi
    Kubo, Shigeto
    Takahashi, Masaya
    Chirieac, Lucian R.
    Padera, Robert F.
    Scott, Andrew M.
    Jungbluth, Achim A.
    Cavenee, Webster K.
    Old, Lloyd J.
    Demetri, George D.
    Wong, Kwok-Kin
    [J]. JOURNAL OF CLINICAL INVESTIGATION, 2007, 117 (02) : 346 - 352
  • [10] Glioma-associated antigen expression in oligodendroglial neoplasms: Tenascin and epidermal growth factor receptor
    McLendon, RE
    Wikstrand, CJ
    Matthews, MR
    Al-Baradei, R
    Bigner, SH
    Bigner, DD
    [J]. JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 2000, 48 (08) : 1103 - 1110