Analytic validation of a clinical-grade PTEN immunohistochemistry assay in prostate cancer by comparison with PTEN FISH

Tamara L. Lotan, Wei Wei, Olga Ludkovski, Carlos L. Morais, Liana B. Guedes, Tamara Jamaspishvili, Karen Lopez, Sarah T. Hawley, Ziding Feng, Ladan Fazli, Antonio Hurtado-Coll, Jesse K. McKenney, Jeffrey Simko, Peter R. Carroll, Martin Gleave, Daniel W. Lin, Peter S. Nelson, Ian M. Thompson, Lawrence D. True, James D. BrooksRaymond Lance, Dean Troyer, Jeremy A. Squire

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

PTEN loss is a promising prognostic and predictive biomarker in prostate cancer. Because it occurs most commonly via PTEN gene deletion, we developed a clinical-grade, automated, and inexpensive immunohistochemical assay to detect PTEN loss. We studied the sensitivity and specificity of PTEN immunohistochemistry relative to four-color fluorescence in situ hybridization (FISH) for detection of PTEN gene deletion in a multi-institutional cohort of 731 primary prostate tumors. Intact PTEN immunostaining was 91% specific for the absence of PTEN gene deletion (549/602 tumors with two copies of the PTEN gene by FISH showed intact expression of PTEN by immunohistochemistry) and 97% sensitive for the presence of homozygous PTEN gene deletion (absent PTEN protein expression by immunohistochemistry in 65/67 tumors with homozygous deletion). PTEN immunohistochemistry was 65% sensitive for the presence of hemizygous PTEN gene deletion, with protein loss in 40/62 hemizygous tumors. We reviewed the 53 cases where immunohistochemistry showed PTEN protein loss and FISH showed two intact copies of the PTEN gene. On re-review, there was ambiguous immunohistochemistry loss in 6% (3/53) and failure to analyze the same tumor area by both methods in 34% (18/53). Of the remaining discordant cases, 41% (13/32) revealed hemizygous (n=8) or homozygous (n=5) PTEN gene deletion that was focal in most cases (11/13). The remaining 19 cases had two copies of the PTEN gene detected by FISH, representing truly discordant cases. Our automated PTEN immunohistochemistry assay is a sensitive method for detection of homozygous PTEN gene deletions. Immunohistochemistry screening is particularly useful to identify cases with heterogeneous PTEN gene deletion in a subset of tumor glands. Mutations, small insertions, or deletions and/or epigenetic or microRNA-mediated mechanisms may lead to PTEN protein loss in tumors with normal or hemizygous PTEN gene copy number.Modern Pathology advance online publication, 13 May 2016; doi:10.1038/modpathol.2016.88.

Original languageEnglish (US)
JournalModern Pathology
DOIs
StateAccepted/In press - May 13 2016

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Gene Deletion
Fluorescence In Situ Hybridization
Prostatic Neoplasms
Immunohistochemistry
PTEN Phosphohydrolase
Neoplasms
Genes
Gene Dosage
Sequence Deletion
Insertional Mutagenesis
MicroRNAs
Epigenomics
Publications
Prostate
Color
Biomarkers
Pathology
Sensitivity and Specificity

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Lotan, T. L., Wei, W., Ludkovski, O., Morais, C. L., Guedes, L. B., Jamaspishvili, T., ... Squire, J. A. (Accepted/In press). Analytic validation of a clinical-grade PTEN immunohistochemistry assay in prostate cancer by comparison with PTEN FISH. Modern Pathology. https://doi.org/10.1038/modpathol.2016.88

Analytic validation of a clinical-grade PTEN immunohistochemistry assay in prostate cancer by comparison with PTEN FISH. / Lotan, Tamara L.; Wei, Wei; Ludkovski, Olga; Morais, Carlos L.; Guedes, Liana B.; Jamaspishvili, Tamara; Lopez, Karen; Hawley, Sarah T.; Feng, Ziding; Fazli, Ladan; Hurtado-Coll, Antonio; McKenney, Jesse K.; Simko, Jeffrey; Carroll, Peter R.; Gleave, Martin; Lin, Daniel W.; Nelson, Peter S.; Thompson, Ian M.; True, Lawrence D.; Brooks, James D.; Lance, Raymond; Troyer, Dean; Squire, Jeremy A.

In: Modern Pathology, 13.05.2016.

Research output: Contribution to journalArticle

Lotan, TL, Wei, W, Ludkovski, O, Morais, CL, Guedes, LB, Jamaspishvili, T, Lopez, K, Hawley, ST, Feng, Z, Fazli, L, Hurtado-Coll, A, McKenney, JK, Simko, J, Carroll, PR, Gleave, M, Lin, DW, Nelson, PS, Thompson, IM, True, LD, Brooks, JD, Lance, R, Troyer, D & Squire, JA 2016, 'Analytic validation of a clinical-grade PTEN immunohistochemistry assay in prostate cancer by comparison with PTEN FISH', Modern Pathology. https://doi.org/10.1038/modpathol.2016.88
Lotan, Tamara L. ; Wei, Wei ; Ludkovski, Olga ; Morais, Carlos L. ; Guedes, Liana B. ; Jamaspishvili, Tamara ; Lopez, Karen ; Hawley, Sarah T. ; Feng, Ziding ; Fazli, Ladan ; Hurtado-Coll, Antonio ; McKenney, Jesse K. ; Simko, Jeffrey ; Carroll, Peter R. ; Gleave, Martin ; Lin, Daniel W. ; Nelson, Peter S. ; Thompson, Ian M. ; True, Lawrence D. ; Brooks, James D. ; Lance, Raymond ; Troyer, Dean ; Squire, Jeremy A. / Analytic validation of a clinical-grade PTEN immunohistochemistry assay in prostate cancer by comparison with PTEN FISH. In: Modern Pathology. 2016.
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abstract = "PTEN loss is a promising prognostic and predictive biomarker in prostate cancer. Because it occurs most commonly via PTEN gene deletion, we developed a clinical-grade, automated, and inexpensive immunohistochemical assay to detect PTEN loss. We studied the sensitivity and specificity of PTEN immunohistochemistry relative to four-color fluorescence in situ hybridization (FISH) for detection of PTEN gene deletion in a multi-institutional cohort of 731 primary prostate tumors. Intact PTEN immunostaining was 91{\%} specific for the absence of PTEN gene deletion (549/602 tumors with two copies of the PTEN gene by FISH showed intact expression of PTEN by immunohistochemistry) and 97{\%} sensitive for the presence of homozygous PTEN gene deletion (absent PTEN protein expression by immunohistochemistry in 65/67 tumors with homozygous deletion). PTEN immunohistochemistry was 65{\%} sensitive for the presence of hemizygous PTEN gene deletion, with protein loss in 40/62 hemizygous tumors. We reviewed the 53 cases where immunohistochemistry showed PTEN protein loss and FISH showed two intact copies of the PTEN gene. On re-review, there was ambiguous immunohistochemistry loss in 6{\%} (3/53) and failure to analyze the same tumor area by both methods in 34{\%} (18/53). Of the remaining discordant cases, 41{\%} (13/32) revealed hemizygous (n=8) or homozygous (n=5) PTEN gene deletion that was focal in most cases (11/13). The remaining 19 cases had two copies of the PTEN gene detected by FISH, representing truly discordant cases. Our automated PTEN immunohistochemistry assay is a sensitive method for detection of homozygous PTEN gene deletions. Immunohistochemistry screening is particularly useful to identify cases with heterogeneous PTEN gene deletion in a subset of tumor glands. Mutations, small insertions, or deletions and/or epigenetic or microRNA-mediated mechanisms may lead to PTEN protein loss in tumors with normal or hemizygous PTEN gene copy number.Modern Pathology advance online publication, 13 May 2016; doi:10.1038/modpathol.2016.88.",
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AU - Hawley, Sarah T.

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AU - Hurtado-Coll, Antonio

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AU - Simko, Jeffrey

AU - Carroll, Peter R.

AU - Gleave, Martin

AU - Lin, Daniel W.

AU - Nelson, Peter S.

AU - Thompson, Ian M.

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