Development of a novel long-lived immunoPET tracer for monitoring lymphoma therapy in a humanized transgenic mouse model.

Abstract

Positron emission tomography (PET) is an attractive imaging tool to localize and quantify tracer biodistribution. ImmunoPET with an intact mAb typically requires two to four days to achieve optimized tumor-to-normal ratios. Thus, a positron emitter with a half-life of two to four days such as zirconium-89 [(89)Zr] (t1/2: 78.4 h) is ideal. We have developed an antibody-based, long-lived immunoPET tracer (89)Zr-Desferrioxamine-p-SCN (Df-Bz-NCS)-rituximab (Zr-iPET) to image tumor for longer durations in a humanized CD20-expressing transgenic mouse model. To optimize the radiolabeling efficiency of (89)Zr with Df-Bz-rituximab, multiple radiolabelings were performed. Radiochemical yield, purity, immunoreactivity, and stability assays were carried out to characterize the Zr-iPET for chemical and biological integrity. This tracer was used to image transgenic mice that express the human CD20 on their B cells (huCD20TM). Each huCD20TM mouse received a 7.4 MBq/dose. One group (n = 3) received a 2 mg/kg predose (blocking) of cold rituximab 2 h prior to (89)Zr-iPET; the other group (n = 3) had no predose (nonblocking). Small animal PET/CT was used to image mice at 1, 4, 24, 48, 72, and 120 h. Quality assurance of the (89)Zr-iPET demonstrated NCS-Bz-Df: antibody ratio (c/a: 1.5 ± 0.31), specific activity (0.44-1.64 TBq/mol), radiochemical yield (>70%), and purity (>98%). The Zr-iPET immunoreactivity was >80%. At 120 h, Zr-iPET uptake (% ID/g) as mean ± STD for blocking and nonblocking groups in spleen was 3.2 ± 0.1% and 83.3 ± 2.0% (p value <0.0013.). Liver uptake was 1.32 ± 0.05% and 0.61 ± 0.001% (p value <0.0128) for blocking and nonblocking, respectively. The small animal PET/CT image shows the spleen specific uptake of Zr-iPET in mice at 120 h after tracer injection. Compared to the liver, the spleen specific uptake of Zr-iPET is very high due to the expression of huCD20. We optimized the radiolabeling efficiency of (89)Zr with Df-Bz-rituximab. These radioimmunoconjugate lots were stable up to 5 days in serum in vitro. The present study showed that (89)Zr is well-suited for mAbs to image cancer over an extended period of time (up to 5 days).

• Authors: Arutselvan Natarajan, Frezghi Habte, Sanjiv S Gambhir
• Journal: Bioconjugate chemistry (Bioconjug Chem) Vol. 23 Issue 6 Pg. 1221-9 (Jun 20 2012) ISSN: 1520-4812 [Electronic] United States
• PMID: 22621257 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• 4-isothiocyanatobenzyl-desferrioxamine
• Antibodies, Monoclonal, Murine-Derived
• Antigens, CD20
• Immunoconjugates
• Isothiocyanates
• Rituximab
• Zirconium
• Deferoxamine

Topics

• Animals
• Antibodies, Monoclonal, Murine-Derived (chemistry, pharmacokinetics)
• Antigens, CD20 (genetics)
• Cell Line, Tumor
• Deferoxamine (analogs & derivatives, chemistry, pharmacokinetics)
• Gene Expression
• Humans
• Immunoconjugates (chemistry, pharmacokinetics)
• Isothiocyanates (chemistry, pharmacokinetics)
• Lymphoma (diagnosis, diagnostic imaging, genetics, therapy)
• Mice
• Mice, Nude
• Mice, Transgenic
• Positron-Emission Tomography (methods)
• Rituximab
• Tissue Distribution
• Zirconium (chemistry, pharmacokinetics)