OJPathology  Vol.4 No.3 , July 2014
Automated Detection and Quantification of Prostate Cancer in Needle Biopsies by Digital Image Analysis
ABSTRACT
Introduction: Triple immunohistochemical (IHC) stains including antibodies specific for alpha-methylacyl-CoA-racemase and basal cell markers have been a valuable aid in accurate identification of prostate carcinoma. However, accurate quantification of minuscule areas of prostate carcinoma in biopsy specimens can often be a challenge. Here we assessed the diagnostic value and quantitative use of automated digital image analysis on triple IHC stained prostate needle biopsies. Methods: Twelve cases of prostate needle biopsy material including 75 needle cores were stained with triple-antibody cocktail (P504S + 34βE12 + p63). Slides were digitally scanned with the APERIO digital image analyzer and evaluated with the GENIE pattern and color recognition digital image analysis that we developed. A slide with known areas of adenocarcinoma, high grade prostatic intraepithelial neoplasia (PIN), benign glands and stroma was used as a training set for the automated digital image analysis platform. Results: Among 75 needle biopsy cores, 19 (25.33%) contained adenocarcinoma by histology. Digital image analysis recognized adenocarcinoma in 95% of these needle biopsies. The average area of the needle biopsy was 7.63 mm2 and overall the average area of tumor was 0.196 mm2. The smallest area of tumor recognized by the program was 0.0022 mm2 (0.0363% of the core) and the largest was 0.62 mm2 (8.17% of the core) among needle core biopsies. False positives resulted from areas of high grade PIN with patchy basal cells. The false negative was caused by uneven AMACR staining in one area of adenocarcinoma. Digital recognition of areas of interest was improved by three successive image analysis training which increased the sensitivity and specificity from 83% and 89% to 90% and 93%, respectively. Conclusions: Digital image analysis in concert with IHC triple staining may be useful for accurate detection and quantitative analysis of small foci of prostatic adenocarcinoma. Defining methods to increase the sensitivity and specificity of quantitative automated digital image analysis will likely evolve as an area of investigation. Future automated digital scanning and innovative pattern and color recognition technologies may open avenues for classifying a variety of prostate lesions.

Cite this paper
Parimi, V. , Eisengart, L. and Yang, X. (2014) Automated Detection and Quantification of Prostate Cancer in Needle Biopsies by Digital Image Analysis. Open Journal of Pathology, 4, 138-150. doi: 10.4236/ojpathology.2014.43020.
References
[1]   DiGiuseppe, J.A., Sauvageot, J. and Epstein, J.I. (1997) Increasing Incidence of Minimal Residual Cancer in Radical Prostatectomy Specimens. American Journal of Surgical Pathology, 21, 174-178. http://dx.doi.org/10.1097/00000478-199702000-00006

[2]   Jemal, A., et al. (2011) Global Cancer Statistics. CA: A Cancer Journal for Clinicians, 61, 69-90. http://dx.doi.org/10.3322/caac.20107

[3]   Epstein, J.I. (1995) Diagnostic Criteria of Limited Adenocarcinoma of the Prostate on Needle Biopsy. Human Pathology, 26, 223-229. http://dx.doi.org/10.1016/0046-8177(95)90041-1

[4]   Epstein, J.I. and Potter, S.R. (2001) The Pathological Interpretation and Significance of Prostate Needle Biopsy Findings: Implications and Current Controversies. Journal of Urology, 166, 402-410. http://dx.doi.org/10.1016/S0022-5347(05)65953-8

[5]   Epstein, J.I., Walsh, P.C., Carmichael, M. and Brendler, C.B. (1994) Pathologic and Clinical Findings to Predict Tumor Extent of Nonpalpable (Stage T1c) Prostate Cancer. JAMA, 271, 368-374.
http://dx.doi.org/10.1001/jama.1994.03510290050036

[6]   Magi-Galluzzi, C., Luo, J., Isaacs, W.B., Hicks, J.L., de Marzo, A.M. and Epstein, J.I. (2003) Alpha-Methylacyl-CoA Racemase: A Variably Sensitive Immunohistochemical Marker for the Diagnosis of Small Prostate Cancer Foci on Needle Biopsy. The American Journal of Surgical Pathology, 27, 1128-1133. http://dx.doi.org/10.1097/00000478-200308000-00010

[7]   Heidenreich, A., et al. (2008) EAU Guidelines on Prostate Cancer. European Urology, 53, 68-80. http://dx.doi.org/10.1016/j.eururo.2007.09.002

[8]   Epstein, J.I. and Y. XJ. (2002) Prostate Biopsy Interpretation. 3rd Edition, 320p.

[9]   Algaba, F., et al. (1996) Assessment of Prostate Carcinoma in Core Needle Biopsy—Definition of Minimal Criteria for the Diagnosis of Cancer in Biopsy Material. Cancer, 78, 376-381.

[10]   Thorson, P. and Humphrey, P.A. (2000) Minimal Adenocarcinoma in Prostate Needle Biopsy Tissue. American Journal of Clinical Pathology, 114, 896-909. http://dx.doi.org/10.1309/KVPX-C1EM-142L-1M6W

[11]   Iczkowski, K.A., Chen, H.M., Yang, X.J. and Beach, R.A. (2002) Prostate Cancer Diagnosed after Initial Biopsy with Atypical Small Acinar Proliferation Suspicious for Malignancy Is Similar to Cancer Found on Initial Biopsy. Urology, 60, 851-854. http://dx.doi.org/10.1016/S0090-4295(02)01981-7

[12]   Molinie, V., Hervé, J.M., Lugagne, P.M., Lebret, T. and Botto, H. (2006) Diagnostic Utility of a p63/Alpha-Methyl Coenzyme A Racemase (p504s) Cocktail in Ambiguous Lesions of the Prostate upon Needle Biopsy. BJU International, 97, 1109-1115.
http://dx.doi.org/10.1111/j.1464-410X.2006.06069.x

[13]   Shah, R.B., Zhou, M., LeBlanc, M., Snyder, M. and Rubin, M.A. (2002) Comparison of the Basal Cell-Specific Markers, 34betaE12 and p63, in the Diagnosis of Prostate Cancer. The American Journal of Surgical Pathology, 26, 1161-1168. http://dx.doi.org/10.1111/j.1464-410X.2006.06069.x

[14]   Srigley, J.R. (2004) Benign Mimickers of Prostatic Adenocarcinoma. Modern Pathology, 17, 328-348. http://dx.doi.org/10.1038/modpathol.3800055

[15]   Troxel, D.B. and Sabella, J.D. (1994) Problem Areas in Pathology Practice. Uncovered by a Review of Malpractice Claims. The American Journal of Surgical Pathology, 18, 821-831.
http://dx.doi.org/10.1097/00000478-199408000-00009

[16]   Jiang, Z., Woda, B.A., Wu, C.L. and Yang, X.J. (2004) Discovery and Clinical Application of a Novel Prostate Cancer Marker: Alpha-Methylacyl CoA Racemase (P504S). American Journal of Clinical Pathology, 122, 275-289. http://dx.doi.org/10.1309/EJUYUQPEX1MG68MK

[17]   Brawer, M.K., Peehl, D.M., Stamey, T.A. and Bostwick, D.G. (1985) Keratin Immunoreactivity in the Benign and Neoplastic Human Prostate. Cancer Research, 45, 3663-3667.

[18]   Ouyang, B., et al. (2011) Alpha-Methylacyl-CoA Racemase Spliced Variants and Their Expression in Normal and Malignant Prostate Tissues. Urology, 77, 249 e1-7.

[19]   Gown, A.M. and Vogel, A.M. (1984) Monoclonal Antibodies to Human Intermediate Filament Proteins. II. Distribution of Filament Proteins in Normal Human Tissues. American Journal of Pathology, 114, 309-321.

[20]   Henneberry, J.M., Kahane, H., Humphrey, P.A., Keetch, D.W. and Epstein, J.I. (1997) The Significance of Intraluminal Crystalloids in Benign Prostatic Glands on Needle Biopsy. The American Journal of Surgical Pathology, 21, 725-728. http://dx.doi.org/10.1097/00000478-199706000-00014

[21]   Wojno, K.J. and Epstein, J.I. (1995) The Utility of Basal Cell-Specific Anti-Cytokeratin Antibody (34 Beta E12) in the Diagnosis of Prostate Cancer. A review of 228 cases. The American Journal of Surgical Pathology, 19, 251-260. http://dx.doi.org/10.1097/00000478-199503000-00002

[22]   Bostwick, D.G. and Brawer, M.K. (1987) Prostatic Intra-Epithelial Neoplasia and Early Invasion in Prostate Cancer. Cancer, 59, 788-794.
http://dx.doi.org/10.1002/1097-0142(19870215)59:4<788::AID-CNCR2820590421>3.0.CO;2-I

[23]   Signoretti, S., et al. (2000) p63 Is a Prostate Basal Cell Marker and Is Required for Prostate Development. American Journal of Pathology, 157, 1769-1775. http://dx.doi.org/10.1016/S0002-9440(10)64814-6

[24]   Hedrick, L. and Epstein, J.I. (1989) Use of Keratin 903 as an Adjunct in the Diagnosis of Prostate Carcinoma. The American Journal of Surgical Pathology, 13, 389-396.
http://dx.doi.org/10.1097/00000478-198905000-00006

[25]   Martens, M.B. and Keller, J.H. (2006) Routine Immunohistochemical Staining for High-Molecular Weight Cytokeratin 34-Beta and Alpha-Methylacyl CoA Racemase (P504S) in Postirradiation Prostate Biopsies. Modern Pathology, 19, 287-290. http://dx.doi.org/10.1038/modpathol.3800531

[26]   Rubin, M.A., Zhou, M., Dhanasekaran, S.M., Varambally, S., Barrette, T.R., Sanda, M.G., Pienta, K.J., Ghosh, D. and Chinnaiyan, A.M. (2002) α-Methylacyl Coenzyme A Racemase as a Tissue Biomarker for Prostate Cancer. JAMA, 287, 1662-1670. http://dx.doi.org/10.1001/jama.287.13.1662

[27]   Xu, J., Stolk, J.A., Zhang, X., Silva, S.J., Houghton, R.L., Matsumura, M., Vedvick, T.S., Leslie, K.B., Badaro, R. and Reed, S.G. (2000) Identification of Differentially Expressed Genes in Human Prostate Cancer Using Subtraction and Microarray. Cancer Research, 60, 1677-1682.

[28]   Yang, X.J., Wu, C.L., Woda, B.A., Dresser, K., Tretiakova, M., Fanger, G.R. and Jiang, Z. (2002) Expression of α-Methylacyl-CoA Racemase (P504S) in Atypical Adenomatous Hyperplasia of the Prostate. American Journal of Surgical Pathology, 26, 921-925. http://dx.doi.org/10.1097/00000478-200207000-00011

[29]   Jiang, Z., Iczkowski, K.A., Woda, B.A., Tretiakova, M. and Yang, X.J. (2004) P504S Immunostaining Boosts Diagnostic Resolution of “Suspicious” Foci in Prostatic Needle Biopsy Specimens. American Journal of Clinical Pathology, 121, 99-107. http://dx.doi.org/10.1309/7T7RJCCL84JGXH3L

[30]   Beach, R., Gown, A.M., De Peralta-Venturina, M.N., Folpe, A.L., Yaziji, H., Salles, P.G., Grignon, D.J., Fanger, G.R. and Amin, M.B. (2002) P504S Immunohistochemical Detection in 405 Prostatic Specimens Including 376 18-Gauge Needle Biopsies. American Journal of Surgical Pathology, 26, 1588-1596. http://dx.doi.org/10.1097/00000478-200212000-00006

[31]   Kuefer, R., Varambally, S., Zhou, M., Lucas, P.C., Loeffler, M., Wolter, H., Mattfeldt, T., Hautmann, R.E., Gschwend, J.E., Barrette, T.R., Dunn, R.L., Chinnaiyan, A.M. and Rubin, M.A. (2002) α-Methylacyl-CoA Racemase: Expression Levels of this Novel Cancer Biomarker Depend on Tumor Differentiation. American Journal of Pathology, 161, 841-848. http://dx.doi.org/10.1016/S0002-9440(10)64244-7

[32]   Zhou, M., Chinnaiyan, A.M., Kleer, C.G., Lucas, P.C. and Rubin, M.A. (2002) Alpha-Methylacyl-CoA Racemase: A Novel Tumor Marker Over-Expressed in Several Human Cancers and Their Precursor Lesions. American Journal of Surgical Pathology, 26, 926-931. http://dx.doi.org/10.1097/00000478-200207000-00012

[33]   Luo, J., Zha, S., Gage, W.R., Dunn, T.A., Hicks, J.L., Bennett, C.J., Ewing, C.M., Platz, E.A., Ferdinandusse, S., Wanders, R.J., Trent, J.M., Isaacs, W.B. and De Marzo, A.M. (2002) Alpha-methylacyl-CoA Racemase: A New Molecular Marker for Prostate Cancer. Cancer Research, 62, 2220-2226.

[34]   Zheng, S.L., Chang, B.L., Faith, D.A., Johnson, J.R., Isaacs, S.D., Hawkins, G.A., Turner, A., Wiley, K.E., Bleecker, E.R., Walsh, P.C., Meyers, D.A., Isaacs, W.B. and Xu, J. (2002) Sequence Variants of Alpha-methylacyl-CoA Racemase Are Associated with Prostate Cancer Risk. Cancer Research, 62, 6485-6488.

[35]   Leav, I., McNeal, J.E., Ho, S.M. and Jiang, Z. (2003) α-methylacyl-CoA Racemase (P504S) Expression in Evolving Carcinomas within Benign Prostatic Hyperplasia and in Cancers of the Transition Zone. Human Pathology, 34, 228-233. http://dx.doi.org/10.1053/hupa.2003.42

[36]   Ferdinandusse, S., Denis, S., IJlst, L., Dacremont, G., Waterham, H.R. and Wanders, R.J. (2000) Subcellular Localization and Physiological Role of Alpha-methylacyl-CoA Racemase. Journal of Lipid Research, 41, 1890-1896.

[37]   Aboseif, S., Shinohara, K., Weidner, N., Narayan, P. and Carroll, P.R. (1995) The Significance of Prostatic Intra-Epithelial Neoplasia. British Journal of Urology, 76, 355-359.
http://dx.doi.org/10.1111/j.1464-410X.1995.tb07714.x

[38]   Davidson, D., Bostwick, D.G., Qian, J., Wollan, P.C., Oesterling, J.E., Rudders, R.A., Siroky, M. and Stilmant, M. (1995) Prostatic Intraepithelial Neoplasia Is a Risk Factor for Adenocarcinoma: Predictive Accuracy in Needle Biopsies. Journal of Urology, 154, 1295-1299. http://dx.doi.org/10.1016/S0022-5347(01)66840-X

[39]   Keetch, D.W., Humphrey, P., Stahl, D., Smith, D.S. and Catalona, W.J. (1995) Morphometric Analysis and Clinical Followup of Isolated Prostatic Intraepithelial Neoplasia in Needle Biopsy of the Prostate. Journal of Urology, 154, 347-351. http://dx.doi.org/10.1016/S0022-5347(01)67044-7

[40]   Kronz, J.D., Allan, C.H., Shaikh, A.A. and Epstein, J.I. (2001) Predicting Cancer following a Diagnosis of High-Grade Prostatic Intraepithelial Neoplasia on Needle Biopsy: Data on Men with More than One Follow-Up Biopsy. American Journal of Surgical Pathology, 25, 1079-1085.
http://dx.doi.org/10.1097/00000478-200108000-00014

[41]   O’Dowd, G.J., Miller, M.C., Orozco, R. and Veltri, R.W. (2000) Analysis of Repeated Biopsy Results within 1 Year after a Noncancer Diagnosis. Urology, 55, 553-559. http://dx.doi.org/10.1016/S0090-4295(00)00447-7

[42]   Raviv, G., Janssen, T., Zlotta, A.R., Louis, L., Descamps, F., Verhest, A. and Schulman, C.C. (1996) High-Grade Intraepithelial Prostatic Neoplasms: Diagnosis and Association with Prostate Cancer. Acta Urologica Belgica, 64, 11-15.

[43]   Shepherd, D., Keetch, D.W., Humphrey, P.A., Smith, D.S. and Stahl, D. (1996) Repeat Biopsy Strategy in Men with Isolated Prostatic Intraepithelial Neoplasia on Prostate Needle Biopsy. Journal of Urology, 156, 460-463. http://dx.doi.org/10.1016/S0022-5347(01)65881-6

[44]   Weinstein, M.H. and Epstein, J.I. (1993) Significance of High-Grade Prostatic Intraepithelial Neoplasia on Needle Biopsy. Human Pathology, 24, 624-629. http://dx.doi.org/10.1016/0046-8177(93)90242-9

[45]   Zhou, M., Jiang, Z. and Epstein, J.I. (2003) Expression and Diagnostic Utility of Alpha-Methylacyl-CoA-Racemase (P504S) in Foamy Gland and Pseudohyperplastic Prostate Cancer. American Journal of Surgical Pathology, 27, 772-778. http://dx.doi.org/10.1097/00000478-200306000-00007

[46]   Jiang, Z., Woda, B.A., Rock, K.L., Xu, Y., Savas, L., Khan, A., Pihan, G., Cai, F., Babcook, J.S., Rathanaswami, P., Reed, S.G., Xu, J. and Fanger, G.R. (2001) P504S: A New Molecular Marker for the Detection of Prostate Carcinoma. American Journal of Surgical Pathology, 25, 1397-1404. http://dx.doi.org/10.1097/00000478-200111000-00007

[47]   Yang, X.J., Laven, B., Tretiakova, M., Blute Jr., R.D., Woda, B.A., Steinberg, G.D. and Jiang, Z. (2003) Detection of Alpha-Methylacyl-Coenzyme A Racemase in Postradiation Prostatic Adenocarcinoma. Urology, 62, 282-286. http://dx.doi.org/10.1016/S0090-4295(03)00259-0

[48]   Zha, S., Ferdinandusse, S., Denis, S., Wanders, R.J., Ewing, C.M., Luo, J., De Marzo, A.M. and Isaacs, W.B. (2003) Alpha-Methylacyl-CoA Racemase as an Androgen-Independent Growth Modifier in Prostate Cancer. Cancer Research, 63, 7365-7376.

[49]   He, L., Long, L.R., Antani, S. and Thoma, G.R. (2012) Histology Image Analysis for Carcinoma Detection and Grading. Computer Methods and Programs in Biomedicine, 107, 538-556. http://dx.doi.org/10.1016/j.cmpb.2011.12.007

[50]   Diamond, J., Anderson, N.H., Bartels, P.H., Montironi, R. and Hamilton, P.W. (2004) The Use of Morphological Characteristics and Texture Analysis in the Identification of Tissue Composition in Prostatic Neoplasia. Human Pathology, 35, 1121-1131.
http://dx.doi.org/10.1016/j.humpath.2004.05.010

[51]   Stotzka, R., Manner, R., Bartels, P.H. and Thompson, D. (1995) A Hybrid Neural and Statistical Classifier System for Histopathologic Grading of Prostatic Lesions. Analytical and Quantitative Cytology and Histology, 17, 204-218.

[52]   Epstein, J.I., Allsbrook Jr., W.C., Amin, M.B., Egevad, L.L. and ISUP Grading Committee (2005) The 2005 International Society of Urological Pathology (ISUP) Consensus Conference on Gleason Grading of Prostatic Carcinoma. American Journal of Surgical Pathology, 29, 1228-1242.
http://dx.doi.org/10.1097/01.pas.0000173646.99337.b1

[53]   Rubin, M.A., Zerkowski, M.P., Camp, R.L., Kuefer, R., Hofer, M.D., Chinnaiyan, A.M. and Rimm, D.L. (2004) Quantitative Determination of Expression of the Prostate Cancer Protein Alpha-methylacyl-CoA Racemase Using Automated Quantitative Analysis (AQUA): A Novel Paradigm for Automated and Continuous Biomarker Measurements. American Journal of Pathology, 164, 831-840.
http://dx.doi.org/10.1016/S0002-9440(10)63171-9

[54]   Bailar III, J.C., Mellinger, G.T. and Gleason, D.F. (1966) Survival Rates of Patients with Prostatic Cancer, Tumor Stage, and Differentiation—Preliminary Report. Cancer Chemotherapy Reports, 50, 129-136.

[55]   Gleason, D.F. (1966) Classification of Prostatic Carcinomas. Cancer Chemotherapy Reports, 50, 125-128.

[56]   Gleason, D.F. and Mellinger, G.T. (1974) Prediction of Prognosis for Prostatic Adenocarcinoma by Combined Histological Grading and Clinical Staging. Journal of Urology, 111, 58-64.

[57]   Jiang, Z., Wu, C.L., Woda, B.A., Dresser, K., Xu, J., Fanger, G.R. and Yang, X.J. (2002) P504S/α-Methylacyl-CoA Racemase: A Useful Marker for Diagnosis of Small Foci of Prostatic Carcinoma on Needle Biopsy. American Journal of Surgical Pathology, 26, 1169-1174.
http://dx.doi.org/10.1097/00000478-200209000-00007

[58]   Lin, F., Brown, R.E., Shen, T., Yang, X.J. and Schuerch, C. (2004) Immunohistochemical Detection of P504S in Primary and Metastatic Renal Cell Carcinomas. Applied Immunohistochemistry & Molecular Morphology, 12, 153-159. http://dx.doi.org/10.1097/00129039-200406000-00010

 
 
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