PO.CL01.22 · 临床研究
监测mCRPC患者CTCs中雄激素受体及神经内分泌标志物表达动态变化
Monitoring androgen receptor and neuroendocrine marker expression dynamics in CTCs in patients with mCRPC
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
雄激素受体(AR)仍是前列腺癌的重要治疗靶点，雄激素受体信号抑制剂(ARSI)，如醋酸阿比特龙(AA)或enzalutamide已在日常实践中广泛应用。然而，约30%的患者表现为原发难治，且几乎所有患者最终在数年内对ARSI产生耐药。耐药机制之一是AR扩增和/或突变，可在ARSI治疗后通过ctDNA检测到。另一种机制是在ARSI治疗压力下由腺癌向神经内分泌前列腺癌(NEPC)的表型转化。本研究通过纵向采集循环肿瘤细胞(CTCs)，探究阿比特龙治疗期间AR及神经内分泌标志物表达的动态变化。我们开发了一种利用分子成像流式细胞术检测CTCs及其AR和突触素(SYP)表达的方法。为从全血中富集CTC，采用了基于细胞尺寸的微流控技术方法。我们的重点是转移性去势抵抗性前列腺癌(mCRPC)患者CTCs的AR核定位状态。为验证我们方法的分析性能，将检测到CTCs的AR及SYP表达与血液采样前后28天内获得的组织活检病理结果进行比较。89.4%(17/19)的患者检测到一个或多个CTCs，17例患者中有13例有可用的病理结果。CTC检测相对于病理结果的敏感性及特异性，AR分别为67%及100%，SYP分别为75%及89%。此外，SYP显示出相对较高的阳性预测值75%及阴性预测值89%。此外，我们研究了AA治疗开始前及开始一个月后AR阳性CTCs的AR核定位变化。通过计算核染色与AR免疫荧光图像之间的相似性，量化每个CTC的AR核定位程度。我们的结果显示，对AA有反应的患者AR核定位程度较低的趋势。在AA治疗队列中，PSA反应者平均AR核定位下降，而无反应者则无下降(治疗一个月后AR核定位阳性率为0% vs 33.0%，p=0.206，Fisher精确检验)。相比之下，在对照队列(非AA治疗组)中，PSA反应者及无反应者的AR核定位评分均保持不变或升高(33.0% vs 33.3%，p=1.0)。我们的观察结果提示，CTC SYP可指导组织再活检以检测NEPC，而CTCs中的AR核定位可作为AA治疗mCRPC的药效学标志物。这对于预测治疗疗效、监测治疗及抗前列腺癌药物选择等临床应用而言是一项重要发现。
查看英文原文 English abstract
Androgen receptor (AR) remains important treatment target for prostate cancer and androgen receptor signaling inhibitor (ARSI), such as abiraterone acetate (AA) or enzalutamide have already been widely used in daily practice. However, approximately 30% of patients show primary refractory and almost all patients eventually resist ARSI within several years. One of the resistant mechanisms is AR amplification and/or mutation which is detected by ctDNA after ARSI treatment. Another one is phenotypic transformation from adenocarcinoma to neuroendocrine prostate cancer (NEPC) by treatment pressure with ARSI. This study investigated dynamic changes of AR and neuroendocrine markers expression during abiraterone treatment by longitudinal Circulating Tumor Cells (CTCs) collection. We have developed a method using molecular imaging flow cytometry to detect CTCs and their AR and Synaptophysin (SYP) expression. For CTC enrichment from whole blood, cell size-based approach using microfluidics technology was adapted. Our focus was on AR nuclear localization status of CTCs in patients with metastatic castration resistant prostate cancer (mCRPC). To validate the analytical performance of our method, the AR and SYP expression of detected CTCs was compared with the pathological results of tissue biopsy obtained within 28 days before or after blood sampling. One or more CTCs were detected in 89.4% (17/19) of patients, and 13 of 17 patients had available pathological results. The sensitivity and specificity of CTC detection for the pathological results were 67% and 100% in AR, and 75% and 89% in SYP, respectively. Additionally, SYP showed relatively high positive predictive value of 75% and negative predictive value of 89%. Furthermore, we have studied the change in AR nuclear localization of AR-positive CTCs before and one month after the start of AA treatment. The degree of AR nuclear localization of each CTC was quantified by calculating the similarity between the nucleus staining and AR immunofluorescent image. Our results demonstrated the tendency with lower degree of AR nuclear localization in patients who responded to AA. In the AA-treated cohort, PSA responders showed a decrease in AR nuclear localization in average and non-responders did not (AR nuclear localization positive rate one month after treatment (0% vs 33.0%, p=0.206, Fisher's exact tests). In contrast, in the control cohort (non-AA-treated group), AR nuclear localization scores remained unchanged or increased in both PSA responders and non-responders (33.0% vs 33.3%, p=1.0). Our observations suggest that CTC SYP may guide indication for tissue re-biopsy to detect NEPC and the AR nuclear localization in CTCs may serve as a pharmacodynamic marker of AA for mCRPC. It represents an important finding for clinical applications such as predicting treatment efficacy, monitoring treatment and selection for anti-prostate cancer drug.
利益披露 Disclosure
N. Matsubara,
Bayer Yakuhin, Ltd ).
MSD ).
Chugai Pharmaceutical Co., Ltd ).
Eisai Co., Ltd. ).
Astellas Pharma Inc. ).
Janssen Pharmaceutical K.K. ).
Pfizer Japan Inc. ), Travel.
Amgen K.K. ).
Takeda Pharmaceutical Co., Ltd. ).
Novartis Pharma K.K. ).
Eli Lilly Japan K.K. ).
Seagen Inc. ).
BicycleTx Limited. ).
K. Shirai,
Sysmex Employment.
H. Obinata,
Sysmex Employment.
H. Nakajima,
Merk Biopharma ).
Grants-in-Aid for Scientific Research ).
C. Funasaka,
Daiichi Sankyo Other, Honoraria.
Eisai Other, Honoraria.
Pfizer Other, Honoraria.
Chugai Other, Honoraria.
MSD Other, Honoraria.
Eli Lilly Other, Honoraria.
C. Kondoh,
Astellas Pharma ).
Daiichi-Sankyo ).
Takeda ).
K. Kawasaki,
Sysmex Employment.
E. Katsumata,
Sysmex Employment.
T. Ajiri,
Sysmex Employment.
M. Bishnu,
Sysmex Employment.
F. Kato,
Sysmex Employment.
M. Yanagida,
Sysmex Employment.
R. Watanabe, None.
T. Mukohara,
Sysmex ).
Sanofi ).
MSD ).
Pfizer ).
Novartis ).
Chugai ).
AstraZeneca ).
Ono ).
Daiichi-Sankyo ).
Gilead Sciences ).