First-in-human uPAR PET: Imaging of Cancer Aggressiveness

Morten Persson, Dorthe Skovgaard, Malene Brandt-Larsen, Camilla Christensen, Jacob Madsen, Carsten H Nielsen, Tine Thurison, Thomas Levin Klausen, Søren Holm, Annika Loft, Anne Kiil Berthelsen, Michael Ploug, Helle Pappot, Klaus Brasso, Niels Kroman, Liselotte Højgaard, Andreas Kjaer, Morten Persson, Dorthe Skovgaard, Malene Brandt-Larsen, Camilla Christensen, Jacob Madsen, Carsten H Nielsen, Tine Thurison, Thomas Levin Klausen, Søren Holm, Annika Loft, Anne Kiil Berthelsen, Michael Ploug, Helle Pappot, Klaus Brasso, Niels Kroman, Liselotte Højgaard, Andreas Kjaer

Abstract

A first-in-human clinical trial with Positron Emission Tomography (PET) imaging of the urokinase-type plasminogen activator receptor (uPAR) in patients with breast, prostate and bladder cancer, is described. uPAR is expressed in many types of human cancers and the expression is predictive of invasion, metastasis and indicates poor prognosis. uPAR PET imaging therefore holds promise to be a new and innovative method for improved cancer diagnosis, staging and individual risk stratification. The uPAR specific peptide AE105 was conjugated to the macrocyclic chelator DOTA and labeled with (64)Cu for targeted molecular imaging with PET. The safety, pharmacokinetic, biodistribution profile and radiation dosimetry after a single intravenous dose of (64)Cu-DOTA-AE105 were assessed by serial PET and computed tomography (CT) in 4 prostate, 3 breast and 3 bladder cancer patients. Safety assessment with laboratory blood screening tests was performed before and after PET ligand injection. In a subgroup of the patients, the in vivo stability of our targeted PET ligand was determined in collected blood and urine. No adverse or clinically detectable side effects in any of the 10 patients were found. The ligand exhibited good in vivo stability and fast clearance from plasma and tissue compartments by renal excretion. In addition, high uptake in both primary tumor lesions and lymph node metastases was seen and paralleled high uPAR expression in excised tumor tissue. Overall, this first-in-human study therefore provides promising evidence for safe use of (64)Cu-DOTA-AE105 for uPAR PET imaging in cancer patients.

Keywords: PET; clinical trial; uPAR.

Conflict of interest statement

Competing interests: MPE, JM & AK are inventor of a patent on the composition of matter of uPAR PET. MPE, JM, CHN & AK are co-founders of a start-up company (Curasight) that has licensed the uPAR PET patent and is currently raising funds to commercialize uPAR PET technology. Pending patent for MPE, JM & AK: Positron Emitting Radionuclides Labeled Peptides for Human uPAR PET Imaging (WO 2014/086364 A1). The authors declare that their spouses, partners, or children have no financial relationships relevant to the submitted work. Inquiries should be directed to the corresponding author.

Figures

Fig 1
Fig 1
uPAR PET imaging and overview of first-in-human uPAR PET study design. (A) Schematic of the uPAR PET ligand 64Cu-DOTA-AE105 showing the chemical structure, a chromatogram of the final product, a transverse PET/CT image from a prostate cancer patient with tumor uptake of 64Cu-DOTA-AE105 and a Pymol visualization of uPAR (surface representation) in complex with the targeting peptide shown as a cartoon representation. (B) Clinical trial events after single dose injection of 64Cu-DOTA-AE105. Timeline denotes injection, acquisition of serial PET/CT imaging, and collection of blood and tissue specimens. (C) Patient characteristics.
Fig 2
Fig 2
Whole-body distribution and PK of 64Cu-DOTA-AE105. (A) Maximum intensity projection PET images at 1, 3 and 24 hours following injection of 64Cu-DOTA-AE105 (patient 10). The highest accumulation of activity was in the liver, bowel and bladder. (B) Decay corrected SUV values in blood and major organs plotted individually for n = 10 patients. For each patient ROIs were drawn on selected organ/tissue of interest at all three consecutive PET scans.
Fig 3
Fig 3
Metabolic analysis of 64Cu-DOTA-AE105 in plasma and urine. Data are shown for patient 10. (A) Relative time-dependent activity concentrations in plasma. Plasma half life was estimated to 8.5 min. (B) Chromatograms of 64Cu-DOTA-AE105 in plasma. Data are shown for standard (Rt=11.8 min) and for the time points: 1, 10, 30 and 120 minutes. A single metabolite was formed (Rt=11.1 min) Vertical lines discriminate peaks corresponding to the uPAR PET ligand 64Cu-DOTA-AE105. (C) Urine collection data. Time-dependent excretion of activity (top) and accumulated activity (bottom) are displayed. (D) Chromatograms of 64Cu-DOTA-AE105 in urine. Data are shown for standard (Rt=11.8 min) and for the time points 20, 30 and 50 minutes. A single metabolite was found (Rt=11.1 min), corresponding to the single metabolite formed in plasma.
Fig 4
Fig 4
Dosimetry and whole-body PET/CT imaging of 64Cu-DOTA-AE105. (A) Mean absorbed dose per unit administrated (mGy/MBq) of major organs and tissues (table S4) were derived from 6 patients (patient 3,4,5,6,8 and 10) from serial whole-body PET scans acquired over 24 hours time interval following injection of 64Cu-DOTA-AE105 using VOI-based time activity data (table S5). (B). Coronal whole-body PET/CT images (patient 5) show time dependent biodistribution at 1,3 and 24 hours and demonstrates accumulation of activity mainly in the liver and bowel.
Fig 5
Fig 5
uPAR PET imaging in bladder cancer. (A) Representative transverse CT, PET and co-registered PET/CT images of a primary tumor lesion (blue circle), top images, with intense uptake of 64Cu-DOTA-AE105 (patient 7). (B) Bottom images show a uPAR positive inguinal lymph node metastasis (blue arrow) with high uptake (patient 1). (C) Tumor-to-liver, tumor-to-blood, tumor-to-kidney and tumor-to-muscle ratios as a function of time post injection. Data are averages ± SD (n = 1 ROI per time point, n = 3 patients).
Fig 6
Fig 6
uPAR PET imaging in breast cancer. (A) Representative transverse CT, PET and co-registered PET/CT images of a primary tumor lesion (blue arrow), top images, with intense uptake of 64Cu-DOTA-AE105 (patient 8). (B) Bottom images show an uPAR positive axillary lymph node metastasis (blue arrow) with high uptake in the same patient. (C) Tumor-to-liver, tumor-to-blood, tumor-to-kidney and tumor-to-muscle ratios as a function of time post injection. Data are averages ± SD (n = 1 ROI per time point, n = 3 patients).
Fig 7
Fig 7
uPAR PET imaging in prostate cancer. (A) Representative transverse CT, PET and co-registered PET/CT images of a primary tumor lesion (blue arrow), top images, with high uptake of 64Cu-DOTA-AE105 (patient 4). (B) Bottom images show an uPAR positive regional lymph node metastasis (blue arrow) with high uptake (patient 9). (C) Tumor-to-liver, tumor-to-blood, tumor-to-kidney and tumor-to-muscle ratios as a function of time post injection. Data are averages ± SD (n = 1 ROI per time point, n = 3 patients).

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