Abstract

Purpose

Current screening and monitoring of prostate cancer (PCa) is insufficient, producing inaccurate diagnoses. Presence of the receptor for advanced glycation end-products (RAGE) is associated with signature characteristics of PCa development such as cell proliferation, anchorage-independent growth, angiogenesis, migration, invasion, and poor patient survival. Therefore, we developed a preclinical multimodal imaging strategy targeted at RAGE to diagnose and monitor PCa.

Methods

In this work, RAGE-targeted multimodal nanoparticles (64Cu-Cy5-G4-CML) were synthesized and rendered functional for nuclear and optical imaging using previously established methods. The probe’s binding affinity and targeting specificity was assessed in androgen-dependent (LNCaP) and androgen-independent (DU145) prostate cancer cells using flow cytometry and confocal microscopy. In vivo PET-CT imaging was used to evaluate RAGE levels in DU145 and LNCaP xenograft models in mice. Then, tumors were excised post-imaging for histological staining and autoradiography to further assess RAGE levels and targeting efficiency of the tracer. Finally, RAGE levels from human PCa samples of varying Gleason Scores were evaluated using Western blot and immunohistochemical staining.

Results

PCa cell culture studies confirmed adequate RAGE-targeting with 64Cu-Cy5-G4-CML with K_D between 360 and 540 nM as measured by flow cytometry. In vivo PET-CT images of PCa xenografts revealed favorable kinetics, rapid blood clearance, and a non-homogenous, enhanced uptake in tumors, which varied based on cell type and tumor size with mean uptake between 0.5 and 1.4%ID/g. RAGE quantification of human samples confirmed increased RAGE uptake corresponding to increased Gleason scoring.

Conclusions

Our study has shown that RAGE-targeted cancer imaging is feasible and could significantly impact PCa management.

中文翻译：

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前列腺癌晚期糖基化终产物受体的定量成像。

摘要

目的

当前对前列腺癌（PCa）的筛查和监测不足，导致诊断不准确。晚期糖基化终产物（RAGE）受体的存在与PCa发育的特征相关，例如细胞增殖，不依赖锚定的生长，血管生成，迁移，侵袭和患者生存差。因此，我们开发了针对RAGE的临床前多峰成像策略，以诊断和监测PCa。

方法

在这项工作中，使用先前建立的方法合成了靶向RAGE的多峰纳米颗粒（64Cu-Cy5-G4-CML），使其具有核和光学成像功能。使用流式细胞术和共聚焦显微镜在雄激素依赖性（LNCaP）和雄激素非依赖性（DU145）前列腺癌细胞中评估了探针的结合亲和力和靶向特异性。体内PET-CT成像用于评估小鼠DU145和LNCaP异种移植模型中的RAGE水平。然后，在成像后切除肿瘤以进行组织学染色和放射自显影，以进一步评估RAGE水平和示踪剂的靶向效率。最后，使用蛋白质印迹和免疫组化染色评估了来自不同Gleason评分的人PCa样品的RAGE水平。

结果

的PCa细胞培养研究证实足够RAGE靶向与64Cu-Cy5的-G4-CML具有K _d如通过流式细胞术测量的360和NM 540之间。PCa异种移植物的体内PET-CT图像显示出良好的动力学，快速的血液清除和非均一的，增强的肿瘤吸收，其随细胞类型和肿瘤大小的不同而变化，平均吸收在0.5％至1.4％ID / g之间。人类样品的RAGE定量证实了相应于Gleason评分增加的RAGE摄取。

结论

我们的研究表明，针对RAGE的癌症影像学是可行的，并且可能显着影响PCa管理。