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Open Access 01-12-2014 | Research

Quantitative characterization of androgen receptor protein expression and cellular localization in circulating tumor cells from patients with metastatic castration-resistant prostate cancer

Authors: Edwin E Reyes, David J VanderWeele, Masis Isikbay, Ryan Duggan, Alexa Campanile, Walter M Stadler, Donald J Vander Griend, Russell Z Szmulewitz

Published in: Journal of Translational Medicine | Issue 1/2014

Abstract

Background

Many current therapies for metastatic castration-resistant prostate cancer (mCRPC) are aimed at AR signaling; however, resistance to these therapies is inevitable. To personalize CRPC therapy in an individual with clinical progression despite maximal AR signaling blockade, it is important to characterize the status of AR activity within their cancer. Biopsies of bone metastases are invasive and frequently fail to yield sufficient tissue for further study. Evaluation of circulating tumor cells (CTCs) offers an alternative, minimally invasive mechanism to characterize and study late-stage disease. The goal of this study was to evaluate the utility of CTC interrogation with respect to the AR as a potential novel therapeutic biomarker in patients with mCRPC.

Methods

Fifteen mL of whole blood was collected from patients with progressive, metastatic mCRPC, the mononuclear cell portion was isolated, and fluorescence-activated cell sorting (FACS) was used to isolate and evaluate CTCs. A novel protocol was optimized to use ImageStreamX to quantitatively analyze AR expression and subcellular localization within CTCs. Co-expression of AR and the proliferation marker Ki67 was also determined using ImageStreamX.

Results

We found inter-patient and intra-patient heterogeneity in expression and localization of AR. Increased AR expression and nuclear localization are associated with elevated co-expression of Ki-67, consistent with the continued role for AR in castration-resistant disease. Despite intra-patient heterogeneity, CTCs from patients with prior exposure to abiraterone had increased AR expression compared to CTCs from patients who were abiraterone-naïve.

Conclusions

As our toolbox for targeting AR function expands, our ability to evaluate AR expression and function within tumor samples from patients with late-stage disease will likely be a critical component of the personalized management of advanced prostate cancer. AR expression and nuclear localization varies within patients and between patients; however it remains associated with markers of proliferation. This supports a molecularly diverse AR-centric pathobiology imparting castration-resistance.

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Siegel R, Ma J, Zou Z, Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 2014, 64: 9-29. 10.3322/caac.21208.CrossRefPubMed

Huggins C, Hodges CV: Studies on prostatic cancer. I. The effect of castration, of estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate. Cancer Res. 1941, 1: 293-297.

Zhu M-L, Horbinski CM, Garzotto M, Qian DZ, Beer TM, Kyprianou N: Tubulin-targeting chemotherapy impairs androgen receptor activity in prostate cancer. Cancer Res. 2010, 70: 7992-8002. 10.1158/0008-5472.CAN-10-0585.PubMedCentralCrossRefPubMed

Visakorpi T, Hyytinen E, Koivisto P, Tanner M, Keinänen R, Palmberg C, Palotie A, Tammela T, Isola J, Kallioniemi O-P: In vivo amplification of the androgen receptor gene and progression of human prostate cancer. Nat Genet. 1995, 9: 401-406. 10.1038/ng0495-401.CrossRefPubMed

Koivisto P, Kononen J, Palmberg C, Tammela T, Hyytinen E, Isola J, Trapman J, Cleutjens K, Noordzij A, Visakorpi T, Kallioniemi O-P: Androgen receptor gene amplification: a possible molecular mechanism for androgen deprivation therapy failure in prostate cancer. Cancer Res. 1997, 57: 314-319.PubMed

Taplin M-E, Bubley GJ, Shuster TD, Frantz ME, Spooner AE, Ogata GK, Keer HN, Balk SP: Mutation of the androgen-receptor gene in metastatic androgen-independent prostate cancer. N Engl J Med. 1995, 332: 1393-1398. 10.1056/NEJM199505253322101.CrossRefPubMed

Dehm SM, Schmidt LJ, Heemers HV, Vessella RL, Tindall DJ: Splicing of a novel androgen receptor exon generates a constitutively active androgen receptor that mediates prostate cancer therapy resistance. Cancer Res. 2008, 68: 5469-5477. 10.1158/0008-5472.CAN-08-0594.PubMedCentralCrossRefPubMed

Hu R, Dunn TA, Wei S, Isharwal S, Veltri RW, Humphreys E, Han M, Partin AW, Vessella RL, Isaacs WB, Bova GS, Luo J: Ligand-independent androgen receptor variants derived from splicing of cryptic exons signify hormone-refractory prostate cancer. Cancer Res. 2009, 69: 16-22. 10.1158/0008-5472.CAN-08-2764.PubMedCentralCrossRefPubMed

Guo Z, Yang X, Sun F, Jiang R, Linn DE, Chen H, Chen H, Kong X, Melamed J, Tepper CG, Kung H-J, Brodie AMH, Edwards J, Qiu Y: A novel androgen receptor splice variant is up-regulated during prostate cancer progression and promotes androgen depletion-resistant growth. Cancer Res. 2009, 69: 2305-2313. 10.1158/0008-5472.CAN-08-3795.PubMedCentralCrossRefPubMed

10.

Ross RW, Halabi S, Ou S-S, Rajeshkumar BR, Woda BA, Vogelzang NJ, Small EJ, Taplin M-E, Kantoff PW: Cancer and leukemia group B: predictors of prostate cancer tissue acquisition by an undirected core bone marrow biopsy in metastatic castration-resistant prostate cancer-a cancer and leukemia group B study. Clin Cancer Res. 2005, 11: 8109-8113. 10.1158/1078-0432.CCR-05-1250.CrossRefPubMed

11.

Rubin MA, Putzi M, Mucci N, Smith DC, Wojno K, Korenchuk S, Pienta KJ: Rapid (“warm”) autopsy study for procurement of metastatic prostate cancer. Clin Cancer Res. 2000, 6: 1038-1045.PubMed

12.

Miyamoto DT, Lee RJ, Stott SL, Ting DT, Wittner BS, Ulman M, Smas ME, Lord JB, Brannigan BW, Trautwein J, Bander NH, Wu C-L, Sequist LV, Smith MR, Ramaswamy S, Toner M, Maheswaran S, Haber DA: Androgen receptor signaling in circulating tumor cells as a marker of hormonally responsive prostate cancer. Cancer Discov. 2012, 2: 995-1003. 10.1158/2159-8290.CD-12-0222.PubMedCentralCrossRefPubMed

13.

Parkinson DR, Dracopoli N, Petty BG, Compton C, Cristofanilli M, Deisseroth A, Hayes DF, Kapke G, Kumar P, Lee JS, Liu MC, McCormack R, Mikulski S, Nagahara L, Pantel K, Pearson-White S, Punnoose EA, Roadcap LT, Schade AE, Scher HI, Sigman CC, Kelloff GJ: Considerations in the development of circulating tumor cell technology for clinical use. J Transl Med. 2012, 10: 138-10.1186/1479-5876-10-138.PubMedCentralCrossRefPubMed

14.

Allard WJ, Matera J, Miller MC, Repollet M, Connelly MC, Rao C, Tibbe AGJ, Uhr JW, Terstappen LWMM: Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin Cancer Res. 2004, 10: 6897-6904. 10.1158/1078-0432.CCR-04-0378.CrossRefPubMed

15.

Harb W, Fan A, Tran T, Danila DC, Keys D, Schwartz M, Ionescu-Zanetti C: Mutational analysis of circulating tumor cells using a novel microfluidic collection device and qPCR assay. Transl Oncol. 2013, 6: 528-538. 10.1593/tlo.13367.PubMedCentralCrossRefPubMed

16.

Danila DC, Anand A, Schultz N, Heller G, Wan M, Sung CC, Dai C, Khanin R, Fleisher M, Lilja H, Scher HI: Analytic and clinical validation of a prostate cancer-enhanced messenger RNA detection assay in whole blood as a prognostic biomarker for survival. Eur Urol. 2014, 65: 1191-1197. 10.1016/j.eururo.2013.07.006.PubMedCentralCrossRefPubMed

17.

Leversha MA, Han J, Asgari Z, Danila DC, Lin O, Gonzalez-Espinoza R, Anand A, Lilja H, Heller G, Fleisher M, Scher HI: Fluorescence in situ hybridization analysis of circulating tumor cells in metastatic prostate cancer. Clin Cancer Res. 2009, 15: 2091-2097. 10.1158/1078-0432.CCR-08-2036.PubMedCentralCrossRefPubMed

18.

Ozkumur E, Shah AM, Ciciliano JC, Emmink BL, Miyamoto DT, Brachtel E, Yu M, Chen P, Morgan B, Trautwein J, Kimura A, Sengupta S, Stott SL, Karabacak NM, Barber TA, Walsh JR, Smith K, Spuhler PS, Sullivan JP, Lee RJ, Ting DT, Luo X, Shaw AT, Bardia A, Sequist LV, Louis DN, Maheswaran S, Kapur R, Haber DA, Toner M: Inertial focusing for tumor antigen-dependent and -independent sorting of rare circulating tumor cells. Sci Transl Med. 2013, 5: 179ra47-10.1126/scitranslmed.3005616.PubMedCentralCrossRefPubMed

19.

Saliba A-E, Saias L, Psychari E, Minc N, Simon D, Bidard F-C, Mathiot C, Pierga J-Y, Fraisier V, Salamero J, Saada V, Farace F, Vielh P, Malaquin L, Viovy J-L: Microfluidic sorting and multimodal typing of cancer cells in self-assembled magnetic arrays. Proc Natl Acad Sci U S A. 2010, 107: 14524-14529. 10.1073/pnas.1001515107.PubMedCentralCrossRefPubMed

20.

Saucedo-Zeni N, Mewes S, Niestroj R, Gasiorowski L, Murawa D, Nowaczyk P, Tomasi T, Weber E, Dworacki G, Morgenthaler NG, Jansen H, Propping C, Sterzynska K, Dyszkiewicz W, Zabel M, Kiechle M, Reuning U, Schmitt M, Lücke K: A novel method for the in vivo isolation of circulating tumor cells from peripheral blood of cancer patients using a functionalized and structured medical wire. Int J Oncol. 2012, 41: 1241-1250.PubMedCentralPubMed

21.

Talasaz AH, Powell AA, Huber DE, Berbee JG, Roh K-H, Yu W, Xiao W, Davis MM, Pease RF, Mindrinos MN, Jeffrey SS, Davis RW: Isolating highly enriched populations of circulating epithelial cells and other rare cells from blood using a magnetic sweeper device. Proc Natl Acad Sci U S A. 2009, 106: 3970-3975. 10.1073/pnas.0813188106.PubMedCentralCrossRefPubMed

22.

Mitra R, Chao O, Urasaki Y, Goodman OB, Le TT: Detection of lipid-rich prostate circulating tumour cells with coherent anti-Stokes Raman scattering microscopy. BMC Cancer. 2012, 12: 540-10.1186/1471-2407-12-540.PubMedCentralCrossRefPubMed

23.

Hou HW, Warkiani ME, Khoo BL, Li ZR, Soo RA, Tan DS-W, Lim W-T, Han J, Bhagat AAS, Lim CT: Isolation and retrieval of circulating tumor cells using centrifugal forces. Sci Rep. 2013, 3: 1259-PubMedCentralPubMed

24.

Vona G, Estepa L, Béroud C, Damotte D, Capron F, Nalpas B, Mineur A, Franco D, Lacour B, Pol S, Bréchot C, Paterlini-Bréchot P: Impact of cytomorphological detection of circulating tumor cells in patients with liver cancer. Hepatol Baltim Md. 2004, 39: 792-797. 10.1002/hep.20091.CrossRef

25.

López-Riquelme N, Minguela A, Villar-Permuy F, Ciprian D, Castillejo A, Álvarez-López M-R, Soto J-L: Imaging cytometry for counting circulating tumor cells: comparative analysis of the CellSearch vs ImageStream systems. APMIS. 2013, 121: 1139-1143. 10.1111/apm.12061.CrossRefPubMed

26.

Reyes EE, Kunovac SK, Duggan R, Kregel S, Griend DJV: Growth kinetics of CD133-positive prostate cancer cells. Prostate. 2013, 73: 724-733. 10.1002/pros.22616.PubMedCentralCrossRefPubMed

27.

Brickley DR, Agyeman AS, Kopp RF, Hall BA, Harbeck MC, Belova L, Volden PA, Wu W, Roe MW, Conzen SD: Serum- and glucocorticoid-induced protein kinase 1 (SGK1) is regulated by store-operated Ca2+ entry and mediates cytoprotection against necrotic cell death. J Biol Chem. 2013, 288: 32708-32719. 10.1074/jbc.M113.507210.PubMedCentralCrossRefPubMed

28.

Korenchuk S, Lehr JE, MClean L, Lee YG, Whitney S, Vessella R, Lin DL, Pienta KJ: VCaP, a cell-based model system of human prostate cancer. Vivo Athens Greece. 2001, 15: 163-168.

29.

Lin HK, Zheng S, Williams AJ, Balic M, Groshen S, Scher HI, Fleisher M, Stadler W, Datar RH, Tai Y-C, Cote RJ: Portable filter-based microdevice for detection and characterization of circulating tumor cells. Clin Cancer Res. 2010, 16: 5011-5018. 10.1158/1078-0432.CCR-10-1105.PubMedCentralCrossRefPubMed

30.

Hosokawa M, Hayata T, Fukuda Y, Arakaki A, Yoshino T, Tanaka T, Matsunaga T: Size-selective microcavity array for rapid and efficient detection of circulating tumor cells. Anal Chem. 2010, 82: 6629-6635. 10.1021/ac101222x.CrossRefPubMed

31.

Yap TA, Lorente D, Omlin A, Olmos D, de Bono JS: Circulating tumor cells: a multifunctional biomarker. Clin Cancer Res. 2014, 20: 2553-2568. 10.1158/1078-0432.CCR-13-2664.CrossRefPubMed

32.

Van Bokhoven A, Varella-Garcia M, Korch C, Johannes WU, Smith EE, Miller HL, Nordeen SK, Miller GJ, Lucia MS: Molecular characterization of human prostate carcinoma cell lines. Prostate. 2003, 57: 205-225. 10.1002/pros.10290.CrossRefPubMed

33.

Isikbay M, Otto K, Kregel S, Kach J, Cai Y, Vander Griend DJ, Conzen SD, Szmulewitz RZ: Glucocorticoid receptor activity contributes to resistance to androgen-targeted therapy in prostate cancer. Horm Cancer. 2014, 5: 72-89. 10.1007/s12672-014-0173-2.PubMedCentralCrossRefPubMed

34.

Heemers HV, Tindall DJ: Androgen receptor (AR) coregulators: a diversity of functions converging on and regulating the AR transcriptional complex. Endocr Rev. 2007, 28: 778-808. 10.1210/er.2007-0019.CrossRefPubMed

35.

Sasaki K, Murakami T, Kawasaki M, Takahashi M: The cell cycle associated change of the Ki-67 reactive nuclear antigen expression. J Cell Physiol. 1987, 133: 579-584. 10.1002/jcp.1041330321.CrossRefPubMed

36.

Coumans FAW, Doggen CJM, Attard G, de Bono JS, Terstappen LWMM: All circulating EpCAM + CK + CD45- objects predict overall survival in castration-resistant prostate cancer. Ann Oncol. 2010, 21: 1851-1857. 10.1093/annonc/mdq030.CrossRefPubMed

37.

Cai C, Chen S, Ng P, Bubley GJ, Nelson PS, Mostaghel EA, Marck B, Matsumoto AM, Simon NI, Wang H, Chen S, Balk SP: Intratumoral de novo steroid synthesis activates androgen receptor in castration-resistant prostate cancer and is upregulated by treatment with CYP17A1 inhibitors. Cancer Res. 2011, 71: 6503-6513. 10.1158/0008-5472.CAN-11-0532.PubMedCentralCrossRefPubMed

38.

Mostaghel EA, Marck BT, Plymate SR, Vessella RL, Balk S, Matsumoto AM, Nelson PS, Montgomery RB: Resistance to CYP17A1 inhibition with abiraterone in castration-resistant prostate cancer: induction of steroidogenesis and androgen receptor splice variants. Clin Cancer Res. 2011, 17: 5913-5925. 10.1158/1078-0432.CCR-11-0728.PubMedCentralCrossRefPubMed

39.

Armstrong AJ, Marengo MS, Oltean S, Kemeny G, Bitting RL, Turnbull JD, Herold CI, Marcom PK, George DJ, Garcia-Blanco MA: Circulating tumor cells from patients with advanced prostate and breast cancer display both epithelial and mesenchymal markers. Mol Cancer Res. 2011, 9: 997-1007. 10.1158/1541-7786.MCR-10-0490.PubMedCentralCrossRefPubMed

40.

Bitting RL, Boominathan R, Rao C, Kemeny G, Foulk B, Garcia-Blanco MA, Connelly M, Armstrong AJ: Development of a method to isolate circulating tumor cells using mesenchymal-based capture. Meth San Diego Calif. 2013, 64: 129-136. 10.1016/j.ymeth.2013.06.034.CrossRef

41.

Attard G, Swennenhuis JF, Olmos D, Reid AHM, Vickers E, A'Hern R, Levink R, Coumans F, Moreira J, Riisnaes R, Oommen NB, Hawche G, Jameson C, Thompson E, Sipkema R, Carden CP, Parker C, Dearnaley D, Kaye SB, Cooper CS, Molina A, Cox ME, Terstappen LWMM, de Bono JS: Characterization of ERG, AR and PTEN gene status in circulating tumor cells from patients with castration-resistant prostate cancer. Cancer Res. 2009, 69: 2912-2918. 10.1158/0008-5472.CAN-08-3667.CrossRefPubMed

42.

Lohr JG, Adalsteinsson VA, Cibulskis K, Choudhury AD, Rosenberg M, Cruz-Gordillo P, Francis JM, Zhang C-Z, Shalek AK, Satija R, Trombetta JJ, Lu D, Tallapragada N, Tahirova N, Kim S, Blumenstiel B, Sougnez C, Lowe A, Wong B, Auclair D, Van Allen EM, Nakabayashi M, Lis RT, Lee G-SM, Li T, Chabot MS, Ly A, Taplin M-E, Clancy TE, Loda M: Whole-exome sequencing of circulating tumor cells provides a window into metastatic prostate cancer. Nat Biotechnol. 2014, 32: 479-484. 10.1038/nbt.2892.PubMedCentralCrossRefPubMed

43.

Yu M, Bardia A, Wittner BS, Stott SL, Smas ME, Ting DT, Isakoff SJ, Ciciliano JC, Wells MN, Shah AM, Concannon KF, Donaldson MC, Sequist LV, Brachtel E, Sgroi D, Baselga J, Ramaswamy S, Toner M, Haber DA, Maheswaran S: Circulating breast tumor cells exhibit dynamic changes in epithelial and mesenchymal composition. Science. 2013, 339: 580-584. 10.1126/science.1228522.PubMedCentralCrossRefPubMed

44.

Cummings J, Sloane R, Morris K, Zhou C, Lancashire M, Moore D, Elliot T, Clarke N, Dive C: Optimisation of an immunohistochemistry method for the determination of androgen receptor expression levels in circulating tumour cells. BMC Cancer. 2014, 14: 226-10.1186/1471-2407-14-226.PubMedCentralCrossRefPubMed

45.

Antonarakis ES, Lu C, Wang H, Luber B, Nakazawa M, Roeser JC, Chen Y, Mohammad TA, Chen Y, Fedor HL, Lotan TL, Zheng Q, De Marzo AM, Isaacs JT, Isaacs WB, Nadal R, Paller CJ, Denmeade SR, Carducci MA, Eisenberger MA, Luo J: AR-V7 and resistance to Enzalutamide and Abiraterone in prostate cancer. N Engl J Med. 2014, 0: null-

46.

Taplin M-E, Rajeshkumar B, Halabi S, Werner CP, Woda BA, Picus J, Stadler W, Hayes DF, Kantoff PW, Vogelzang NJ, Small EJ: Androgen receptor mutations in androgen-independent prostate cancer: cancer and leukemia group B study 9663. J Clin Oncol. 2003, 21: 2673-2678. 10.1200/JCO.2003.11.102.CrossRefPubMed

47.

Carreira S, Romanel A, Goodall J, Grist E, Ferraldeschi R, Miranda S, Prandi D, Lorente D, Frenel J-S, Pezaro C, Omlin A, Rodrigues DN, Flohr P, Tunariu N, de Bono JS, Demichelis F, Attard G: Tumor clone dynamics in lethal prostate cancer. Sci Transl Med. 2014, 6: 254ra125-10.1126/scitranslmed.3009448.PubMedCentralCrossRefPubMed

48.

Darshan MS, Loftus MS, Thadani-Mulero M, Levy BP, Escuin D, Zhou XK, Gjyrezi A, Chanel-Vos C, Shen R, Tagawa ST, Bander NH, Nanus DM, Giannakakou P: Taxane-induced blockade to nuclear accumulation of the androgen receptor predicts clinical responses in metastatic prostate cancer. Cancer Res. 2011, 71: 6019-6029. 10.1158/0008-5472.CAN-11-1417.PubMedCentralCrossRefPubMed

Title: Quantitative characterization of androgen receptor protein expression and cellular localization in circulating tumor cells from patients with metastatic castration-resistant prostate cancer
Authors: Edwin E Reyes
David J VanderWeele
Masis Isikbay
Ryan Duggan
Alexa Campanile
Walter M Stadler
Donald J Vander Griend
Russell Z Szmulewitz
Publication date: 01-12-2014
Publisher: BioMed Central
Published in: Journal of Translational Medicine / Issue 1/2014
Electronic ISSN: 1479-5876
DOI: https://doi.org/10.1186/s12967-014-0313-z

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