PET imaging of a 68 Ga labeled modified HER2 affibody in breast cancers: from xenografts to patients.

Objective: Overexpression of human epidermal growth factor receptor-2 (HER2) in breast cancers provides promising opportunities for imaging and targeted therapy. Developing HER2 targeted positron emission tomography (PET) probes might be benefit for management of the disease. Small high-affinity scaffold proteins, affibodies, are ideal vectors for imaging HER2 overexpressed tumors. Despite of the initial success on development of F-18 labeled ZHER(2:342) affibody, the tedious synthesis producers, low yields and unfavorable pharmacokinetics may hinder the clinical use. Ga-68 is an attractive positron emitter for PET imaging. A simple preparation of Ga-68 labeled ZHER(2:342) analog, Ga-68-NOTA-MAL-Cys-MZHER(2:342), was reported in the study. The in vivo performances of the tracer for assessing HER2 status in breast cancers were also evaluated. Methods: NOTA-MAL conjugated Cys-MZHER(2:342) was radiolabeled with Ga-68. The probe was evaluated by in vitro tests including stability and cell binding studies in breast cancer cells with different HER2 levels. In vivo evaluation was performed in mice bearing tumors using microPET imaging and biodistribution experiments. A PET/CT imaging study was initially performed in patients with breast cancers. Results: The tracer was synthesized in a straightforward chelation method with satisfactory non-decay corrected yield (81 +/- 5%) and radiochemical purity (>95%). In vivo micro-PET imaging showed that HER2 high levels expressed BT474 xenografts were more clear visualized than HER2 low levels expressed MCF-7 tumors (16.12 +/- 2.69 ID%/g vs 1.32 +/- 0.19 ID%/g at 1 h post-injection). The outcome was consistent with the immunohistochemical analysis. No significant radioactivity was accumulated in healthy tissues (less than 2% I D/g) except kidneys. In a preliminary clinical study, Ga-68-NOTA-MAL-CysMZHER(2:342) PET imaging allowed more high-contrast detection of HER2 positive primary tumors (maximum standardized uptake value = 2.16 +/- 0.27) than those in HER2 negative primary focus (maximum standardized uptake value = 0.32 +/- 0.05). No detectable side-effects were found. Conclusion: In summary, this study indicates the significant efficiency of the Ga-68 labeled HER2 affibody. Preclinical and clinical studies support the possibility of monitoring HER2 levels in breast cancers using 68 Ga-NOTA-MAL-Cys-MZHER(2:342). Advances In knowledge: The research investigated the feasibility of a Ga-68 labeled HER2 affibody modified with a hydrophilic linker for breast cancer PET imaging. Favorable outcomes showed that the probe might be valuable for determining HER2 status of the disease.