Abstract 5588: Epithelial-to-mesenchymal markers of circulating tumor cells for detection of castration-resistant prostate cancer
Authors: Chen Chun-Liang , Osmulski Pawel , Mahalingam Devalingam , Horning Aaron M. , Jadhav Rohit R. , Louie Anna D. , Wang Chiou-Miin , Huang Tim H.-M. 
Published: May 2015

Approximately 30% of prostate cancer (PCa) patients who receive androgen-deprivation therapy experience disease progression, including bone metastasis, in 18-36 months. Clinical diagnosis of this castration resistance is primarily based on a continuous rise in serum PSA levels during therapy. However, the underlying disease progression predates the clinical onset of castration resistance by many months. The clinical challenge is to distinguish indolent vs aggressive cancer for better disease management. During the prostate cancer progression, malignant circulating tumor cells (CTC) shed from the primary site into the bloodstream through epigenetic modifications and an epithelial-to-mesenchymal transition (EMT). Evidence suggests that increased CTCs are present in the blood of castration-resistant PCa patients and can be obtained through routine phlebotomy. EMT changes of CTCs present a great potential for disease prognosis, treatment stratification and subsequent disease monitoring.

We have recently established the combined microfiltration-micromanipulation system (CM2S) to enrich prostate CTCs from blood as published in The Prostate. Our preliminary studies based on a relatively small sample size show that CTCs isolated from advanced PCa patients often lose the typical features of prostate epithelial cells and display incremental EMT-related gene expression signatures, higher elasticity and smoother membrane features. Furthermore, incremental expression of these genes and particular nanomechanical features in CTCs are associated with castration-resistant and metastatic PCa. In this study, we would like expand the sample size to clinically develop this methodology.

CTCs were isolated from patients' blood (∼15 ml) using microfiltration system (ScreenCell). We analyzed castration-sensitive (CS) and castration-resistant (CR) PCa cells and CTCs using high throughput microfluidic single-cell RT-PCR. The data were subject to hierarchical clustering, violin plot and Ingenuity Pathway Analysis (IPA). Increased expression of EMT genes was found in CR PCa cells and CTCs as compared to CS counterparts. IPA indicated that these EMT genes are closely related to AKT, β-catenin, Myc and NFκB pathways. Some of CTCs isolated from patients were subject to nanomechanical and nanochemical analysis using PeakForce QNM Catalyst atomic force microscopy (AFM) (Bruker). CTCs of CR patients are about 3 fold more elastic and 7 fold more adherent than CTCs of CS patients. Additionally, CTCs of CR patients are about 3 fold more deformable than CTCs of CS patients.

In conclusion, we confirmed our previous study with a larger patient sample size. The increased expression of EMT-related genes and nanomechanical and nanochemical phenotypes in CTCs are potential biomarkers for detection of the castration-resistant PCa, treatment stratification and subsequent disease monitoring.

Citation Format: Chun-Liang Chen, Pawel Osmulski, Devalingam Mahalingam, Aaron M. Horning, Rohit R. Jadhav, Anna D. Louie, Chiou-Miin Wang, Tim H.-M. Huang. Epithelial-to-mesenchymal markers of circulating tumor cells for detection of castration-resistant prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5588. doi:10.1158/1538-7445.AM2014-5588