Measuring the Pharmacokinetics of 123I-Albumin and 18F-FDG in Rats Administered via Intrathecal Injection at Different Volumes.

Jack Hoppin1, Jacob Y Hesterman1, Kelly Orcutt1, William Cupelo1, Mary Germino1, Eli White1, Mark Lane2, Dustin Kentala2, Tracie Reindfield2, Ajay Verma3
1inviCRO, LLC, Boston, MA, 2MPI Research, Mattawan, MI, 3Biogen-Idec, Cambridge, MA

Objective
The purpose of this work was to develop a novel method for assessing the kinetics of intrathecally administered radiopharmaceuticals in the cerebrospinal fluid using highresolution microSPECT/CT and microPET/CT. To this effort, two well-characterized radiopharmaceuticals were imaged longitudinally following intrathecal injection and novel region-of-interest (ROI) analysis were implemented to measure the spatial and temporal distribution of said radiopharmaceuticals within and outside of the central nervous system.

Atlas Analysis Methodology

Quantitative Results

Data Collection and Analysis Strategy

Collection: 123I-Albumin and 18F-FDG (fludeoxyglucose) were imaged following intrathecal injection between the L5-L6 vertebrae with one of three volumes: 10, 50 or 100L. Imaging was also performed following an intravascular injection (100L) for the sake of comparison. Whole-body 123I-Albumin (figure 1) SPECT images (NanoSPECT/CT, Bioscan Inc.) were collected for the first hour post-injection and again at 2 and 6 hours (36x2.5mm diameter pinholes, 24 angles/rotation, 2.4cm helical pitch). Whole-body 18F-FDG PET images (figure 6) were collected (MicroPET Focus 120, Concorde Microsystems) for the first hour post-injection and again at 3 hours using a continuous bed motion (350-650 keV energy window, 2D OSEM reconstruction). Both SPECT and PET image acquisitions were followed by a whole-body x-ray CT scan for use as an anatomical reference.

CSF-Space Atlas Region Creation: In addition to the analysis of the spinal column a novel atlas region was created using 111In-DTPA rat brain images and incorporated into our anatomical rat atlas. The process for creation was as follows: Four rat brain images from multiple time points (figure 2a) were co-registered and averaged (figure 2b). The CSF-Space was classified in this average image (figure 2c). This CSF-Space region was merged with our standard atlas represented in figure 2d as the Intrathecal region. Note that the volume of this space is made artificially larger intentionally to support blur and partial-volume effects present in nuclear medicine imaging. We store and employ both a dilated (figure 2e) and constricted version of the region.

123I-Albumin IV and IT: Results from SPECT/CT imaging at 15min, 2 and 6 hours following IV (figure 5a IV only) or a 10 uL IT injection (figures 4b). Images and plots (figures 5b and 5d) indicate a delayed delivery to the brain for the small-volume injection. The total brain uptake in these experiments across the various injection volumes and routes ranged between 0.4-1.1% of the injected dose. Plots were generated using analysis methods presented in Figures 1 and 2, extended to include changes over time (figure 5c and 5d). Images across time points were scaled to a fixed max (99.9th percentile of the first time point). Injected doses were 580 +/- 50 Ci.

Figure 3a is a focused image of 123I-Albumin in the rat head. Clearance through the nasal lymphatics was noted along with free iodine metabolism in the nasal epithelium. A nasal atlas was developed (figure 3b) in order to quantify the nasal clearance from the CSF.

Quantitative Results

Analysis of the Spinal Column: Figure 1b above shows a maximum-intensity projection image of 123I-Albumin at 15 minutes following the 100L intrathecal injection. A 14-region rat brain atlas (figure 2c) and a dedicated CSF atlas (figure 2d) were fit to the skull in the x-ray CT image at each time point enabling an estimate of uptake and concentration in the primary regions of the brain as a function of time. For analysis of the kinetics of the relevant radiopharmaceutical in the spinal column of the rat, the vertebrae were segmented using a connected thresholding algorithm. Following the classification of the vertebrae, this ROI (shown in red) was made immutable and the spinal column of the animal was classified manually (shown in green, blue and light blue for lumbar, thoracic and cervical sections, respectively). An additional cylindrical ROI or sheath was created around the spinal column (not shown) to measure penetration away from the spinal column and for assessment of the partial volume effect in the SPECT and PET reconstructions. Time activity curves (figure 1a) are presented for these three ROIs (vertebrae, spinal column and sheath).

18F-FDG ITH: Whole-body (Figure 3a) and focused brain (Figure 3b) PET/CT images 5, 35 and 120 min following IT injection of 50L. Time-activity curve for the entire brain (Figure 6d) across all volumes are shown. Uptake amounts were around 0.1% of the injected dose. A plot was generated using analysis methods presented in Figures 1 and 2 extended to include changes over time (figure 6c). The low values were at or below the sensitivity of the microPET. Injected doses were ~100Ci.

Summary
The top two figures from left-to-right display estimates of the %ID/g of 123IAlbumin in the ventricle space (figure 2d and 2e). The bottom left plot displays a comparison of in vivo and ex vivo estimates of radiotracer concentration in the brain tissue for different volumes and routes. A novel method of assessing the kinetics of radiopharmaceuticals within the cerebrospinal fluid of the rat has been developed. Classification of the spinal column and spinal cord was achieved (Figure 1) and a CSF Space region was created and merged with an anatomical rat brain atlas (Figure 2). The results of this novel approach were presented for three well-characterized commercial radiopharmaceuticals. Early efforts to classify the effect of volume on CSF pharmacokinetics showed promise and will guide future experiments. This effort lays the foundation for a new approach to studying the kinetics of radiolabeled molecules within the CSF space. Please contact Jack Hoppin at hoppin@invicro.com for references and/or questions.