Imaging and treating tumor vasculature with targeted radiolabeled carbon nanotubes

Alessandro Ruggiero^1*, Carlos H Villa^1*, Jason P Holland¹, Shanna R Sprinkle¹, Chad May², Jason S Lewis¹, David A Scheinberg¹, Michael R McDevitt^1
1Departments of Medicine and Radiology, Pharmacology and Molecular Therapeutics, Memorial Sloan-Kettering Cancer Center, New York, USA; ²ImClone Systems, New York, USA; *Ruggiero and Villa contributed equally to this work

Abstract: Single wall carbon nanotube (SWCNT) constructs were covalently appended with radiometal-ion chelates (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid [DOTA] or desferrioxamine B [DFO]) and the tumor neovascular-targeting antibody E4G10. The E4G10 antibody specifically targeted the monomeric vascular endothelial-cadherin (VE-cad) epitope expressed in the tumor angiogenic vessels. The construct specific activity and blood compartment clearance kinetics were significantly improved relative to corresponding antibody-alone constructs. We performed targeted radioimmunotherapy with a SWCNT-([²²⁵Ac]DOTA)(E4G10) construct directed at the tumor vasculature in a murine xenograft model of human colon adenocarcinoma (LS174T). The specific construct reduced tumor volume and improved median survival relative to controls. We also performed positron emission tomographic (PET) radioimmunoimaging of the tumor vessels with a SWCNT-([⁸⁹Zr]DFO)(E4G10) construct in the same murine LS174T xenograft model and compared the results to appropriate controls. Dynamic and longitudinal PET imaging of LS174T tumor-bearing mice demonstrated rapid blood clearance (<1 hour) and specific tumor accumulation of the specific construct. Incorporation of the SWCNT scaffold into the construct design permitted us to amplify the specific activity to improve the signal-to-noise ratio without detrimentally impacting the immunoreactivity of the targeting antibody moiety. Furthermore, we were able to exploit the SWCNT pharmacokinetic (PK) profile to favorably alter the blood clearance and provide an advantage for rapid imaging. Near-infrared three-dimensional fluorescent-mediated tomography was used to image the LS174T tumor model, collect antibody-alone PK data, and calculate the number of copies of VE-cad epitope per cell. All of these studies were performed as a single administration of construct and were found to be safe and well tolerated by the murine model. These data have implications that support further imaging and radiotherapy studies using a SWCNT-based platform and focusing on the tumor vessels as the target.

Keywords: actinium-225 (²²⁵Ac), zirconium-89 (⁸⁹Zr), angiogenesis, vascular endothelial-cadherin, radioimmunotherapy (RIT), radioimmunoPET