A Genetically-Encoded Reporter for In Vivo Imaging in Deep Tissues

Nivin N. Nasri, The University of Western Ontario

Abstract

Introduction. The ability to track cells in living organisms with sensitivity, accuracy and high spatial resolution would revolutionize the way we study disease. Reporter genes are valuable tools as they encode detectible products, allowing for quantitative “reporting” of cells that express them. Previously, a gene encoding Organic anion-transporting polypeptide 1a1 (Oatp1a1) was established as a magnetic resonance imaging (MRI) reporter based on its ability to take up the paramagnetic contrast agent gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA). Our objective was to assess, characterize and further develop this system for whole-body tracking of cells in vivo. Methods. Cancer cells were engineered to synthetically express Oatp1a1, or Oatp1b3, a closely related human transporter protein. In our first study, T₁-weighted images of Oatp1a1-expressing primary tumours in preclinical animals were acquired before and after administration of 0.1-mmol/kg Gd-EOB-DTPA at 3-Tesla. At endpoint, heterogenous contrast enhancement patterns within the primary tumour architecture were compared to whole-tumour fluorescent histology. In the next study, T₁-weighted images of Oatp1b3-expressing primary tumours, and their spontaneous metastases to the lymph nodes and lungs, were acquired before and after administration of 1-mmol/kg Gd-EOB-DTPA at 3-Tesla. In the final study, the feasibility of Oatp1b3 as a photoacoustic reporter gene was assessed by acquiring full-spectrum near infrared photoacoustic images of primary tumours in preclinical animals before and after administration of 8-mg/kg indocyanine green. Results. We were able to demonstrate the feasibility of imaging cancer cells with Oatp1a1 at 3-Tesla and 0.1 mmol/kg Gd-EOB-DTPA. Importantly, as primary tumours grew over time, heterogeneous contrast enhancement patterns that emerged near-endpoint strongly correlated to viable cell distributions on whole-tumour histology. Oatp1b3 was also shown to operate as a MRI reporter gene at 3-Tesla, based on the same principle as Oatp1a1. Impressively, single lymph node metastases and the formation of micrometastases in the lungs of preclinical animals were detected with Oatp1b3-MRI. Finally, we also demonstrated the ability of Oatp1b3 to operate as a photoacoustic reporter gene based on its ability to take up indocyanine green. Conclusion. The Oatp1 reporter gene system is a versatile imaging tool for longitudinal tracking of engineered cells in vivo with sensitivity, high resolution, and 3-dimensional spatial information.