Usefulness of asialoglycoprotein receptor imaging for the evaluation of liver metastasis of neuroblastoma

Neuroblastoma, derived from the neural crest ectoderm, is the most common type of solid abdominal mass seen in infancy. The diagnosis, staging, and follow-up of neuroblastoma are often performed using metaiodobenzylguanidine (MIBG) imaging. However, the evaluation of liver metastasis by this method is complicated by the normal physiological uptake of MIBG by the liver. The asialoglycoprotein receptor is a hepatic cell-surface receptor specific for galactose-terminated glycoprotein, and 99mTc-DTPA-galactosyl human serum albumin (GSA) accumulates selectively in hepatic cells. Here, we report a case of congenital neuroblastoma with liver metastasis in which GSA scans were useful for differentiation between normal and metastatic sites in the liver.     

Functional mapping of regional liver asialoglycoprotein receptor amount from single blood sample and SPECT.

The objective of this study was to validate a method for estimating regional liver asialoglycoprotein (ASGP) receptor amount from single blood samples using static SPECT with (99m)Tc-diethylenetriaminepentaacetic acid galactosyl human serum albumin ((99m)Tc-GSA). METHODS: Based on a 2-compartment nonlinear model, regional ASGP receptor amount could be calculated from total liver ASGP receptor amount (Ro) and regional GSA uptake at a specific time. Because Ro could be estimated from single blood samples using an empiric formula, regional GSA uptake obtained as a SPECT voxel count could be converted to regional ASGP receptor amount by solving a nonlinear model equation. To validate this method, data from 62 patients with chronic liver disease underwent dynamic SPECT (30 rotations per 30 min) and simultaneous multiblood sampling and were analyzed by this method. Ro was calculated as the sum of voxel values of parametric receptor images generated from plasma concentration of GSA at 20 min and of static SPECT images generated by merging dynamic SPECT data (12-20 min). Ro was also estimated by fitting time-activity curves (4-30 min) of plasma and whole liver to the nonlinear model using the nonlinear regression method. Ro obtained from the receptor image was compared with that from curve fitting in relation to the results of hepatic function tests (indocyanine green test, hepaplastin test, and branched-chain amino acids/tyrosine plasma concentration ratio) and Child's classification. RESULTS: Ros from the 2 methods showed a significant linear correlation (r(2) = 0.938; P < 0.0001; slope = 0.90; y-intercept = 1.5). Both Ros had significant correlations with the results of hepatic function tests (P < 0.001) and differed significantly among the 3 groups of Child's classification (P < 0.0001). CONCLUSION: The present method could provide a quantitative ASGP receptor image without dynamic data acquisition. This approach could be useful for quantitative evaluation of regional liver function and estimation of residual liver function in hepatectomy.     

[99mTc]O2-AMD3100 as a SPECT tracer for CXCR4 receptor imaging

PurposeCXCR4 plays an important role in HIV infection, tumor progression, neurogenesis, and inflammation. In-vivo imaging of CXCR4 could provide more insight in the role of this receptor in health and disease. The aim of this study was to investigate [^99mTc]O₂-AMD3100 as a potential SPECT tracer for imaging of CXCR4.MethodAMD3100 was labelled with [^99mTc]pertechnetate. A cysteine challenge assay was performed to test the tracer stability. Heterologous and homologous receptor binding assay and internalization assay were performed in CXCR4 expressing Jurkat-T cells. Ex vivo biodistribution was studied in healthy mice at 30, 60, and 120 min after tracer injection. Tumor uptake of the tracer was determined by microSPECT imaging in nude mice xenografted with human PC-3 prostate tumor. Specificity of tracer uptake was determined by blocking studies using an excess of unlabelled AMD3100.ResultsAMD3100 was labelled with technetium-99 m with a radiochemical yield of > 98%. The tracer was stable in PBS and mouse plasma for at least 6 h at 37 °C. Heterologous and homologous binding assays with AMD3100 showed IC₅₀ values of 240 ± 10 μM, and 92 ± 5 μM for [¹²⁵I]SDF-1α and [^99mTc]O₂-AMD3100 respectively, with negligible receptor internalisation. The tracer showed high uptake in liver, lungs, spleen, thymus, intestine and bone. Blocking dose of AMD3100.8HCl (20 mg/kg) decreased the uptake in these organs (p < 0.05). [^99mTc]O₂-AMD3100 showed specific tumor accumulation in mice bearing PC-3 xenografts model. Time activity curves (TAC) in AMD3100 pre-treated animals tracer showed 1.7 times less tumor uptake as compared to control animals (p < 0.05).Conclusion[^99mTc]O₂-AMD3100 is readily labelled, is stable in plasma and displays a favourable binding affinity for the CXCR4 receptors. [^99mTc O₂-AMD3100 shows specific binding in organs with high CXCR4 expression and in CXCR4 positive tumors. These results justify further evaluation of this radiopharmaceutical as a potential biomarker for the non-invasive imaging of CXCR4 receptors.     

MR receptor imaging: ultrasmall iron oxide particles targeted to asialoglycoprotein receptors

Previously we have reported that ultrasmall superparamagnetic iron oxide (USPIO) particles migrate across capillary endothelium, a prerequisite for the design of particulate pharmaceuticals for MR receptor imaging. In the current study, USPIO particles are directed specifically to asialoglycoprotein (ASG) receptors by coupling galactose terminals in the form of arabinogalactan (AG) to these particles. Biodistribution data showed that ASG-directed, AG-coated USPIO (AG-USPIO) particles selectively accumulate in the liver but not in other organs. Electron microscopy of liver showed electron-dense iron oxide particles bound to hepatocyte cell-surface membranes and in large numbers within intracellular lysosomes. The specificity of AG-USPIO for asialoglycoprotein receptors was confirmed by incubation experiments with and without ASG-blocking agents such as D(+)galactose and asialofetuin. In vivo MR imaging in rats showed a significant decrease in liver signal intensity at low doses (2 mumol Fe/kg); no significant changes were observed in the spleen. This decrease in signal intensity is larger than that observed with conventional iron oxides at equal doses. These initial data suggest that, for the first time, superparamagnetic agents can be directed to specific sites for MR imaging by strategies such as receptor targeting.     

Glycosylated RGD-containing peptides: tracer for tumor targeting and angiogenesis imaging with improved biokinetics.

The alpha(v)beta3 integrin plays an important role in metastasis and tumor-induced angiogenesis. Targeting with radiolabeled ligands of the alpha(v)beta3 integrin may provide information about the receptor status and enable specific therapeutic planning. Previous studies from our group resulted in tracers that showed alpha(v)beta3-selective tumor uptake. However, these first-generation compounds predominantly revealed hepatobiliary excretion with high radioactivity found in the liver. In this report, the synthesis and biological evaluation of the first glycosylated RGD-containing peptide (RGD-peptide) for the noninvasive imaging of alpha(v)beta3 expression are described. METHODS: Peptides were assembled on a solid support using fluorenylmethoxycarbonyl-coupling protocols. The precursor cyclo(-Arg-Gly-Asp-D-Tyr-Lys(SAA)-) GP1 was synthesized by coupling 3-acetamido-2,6-anhydro-4,5,7-tri-O-benzyl-3-deoxy-beta-D-glycero-D-gulo-heptonic acid (SAA(Bn3)) with cyclo(-Arg(Mtr)-Gly-Asp(OtBu)-D-Tyr(tBu)-Lys-) and subsequent removal of the protection groups. Iodine labeling was performed by the Iodo-Gen method (radiochemical yield > 50%). The in vitro binding assays were performed using purified immobilized alpha(IIb)beta3, alpha(v)beta5, and alpha(v)beta3 integrins. For in vivo experiments, nude mice bearing xenotransplanted melanomas and mice with osteosarcomas were used. RESULTS: The glycosylated peptide 3-iodo-Tyr4-cyclo(-Arg-Gly-Asp-D-Tyr-Lys(SAA)-) GP2 showed high affinity and selectivity for alpha(v)beta3 in vitro (50% inhibitory concentration = 40 nmol/L). Pretreatment studies indicate specific binding of [125I]GP2 on alpha(v)beta3-expressing tumors in vivo. Comparison of the pharmacokinetics of [125I]GP2 and [125I]-3-iodo-Tyr4-cyclo(-Arg-Gly-Asp-D-Tyr-Val-) [125I]P2 revealed for [125I]GP2 an increased activity concentration in the blood (e.g., 3.59 +/- 0.35 percentage injected dose [%ID]/g vs. 1.72 +/- 0.44 %ID/g at 10 min postinjection) and a significantly reduced uptake in the liver (e.g., 2.59 +/- 0.24 %ID/g vs. 21.96 +/- 2.78 %ID/g at 10 min postinjection). Furthermore, a clearly increased activity accumulation in the tumor was found (e.g., 3.05 +/- 0.31 %ID/g vs. 0.92 +/- 0.16 %ID/g at 240 min postinjection), which remained almost constant between 60 and 240 min postinjection. This resulted in good tumor-to-organ ratios for the glycosylated tracer (e.g., 240-min postinjection osteosarcoma model: tumor-to-blood = 16; tumor-to-muscle = 7; tumor-to-liver = 2.5), which were confirmed by the first gamma-camera images of osteosarcoma-bearing mice at 240 min postinjection. CONCLUSION: This study demonstrates that the introduction of a sugar moiety improves the pharmakokinetic behavior of a hydrophobic peptide-based tracer. Additionally, this alpha(v)beta3-selective glycosylated radioiodinated second-generation tracer GP2 shows high tumor uptake and good tumor-to-organ ratios that allow noninvasive visualization of alpha(v)beta3-expressing tumors and monitoring therapy with alpha(v)beta3 antagonists. Finally, the favorable biokinetics make the glycosylated RGD-peptide a promising lead structure for tracers to quantify the alpha(v)beta3 expression using PET.     

Compartmental analysis of asialoglycoprotein receptor scintigraphy for quantitative measurement of liver function: A multicentre study

A multicentre study on multicompartmental analysis of hepatic scintigraphy using technetium-99m labelled galactosyl serum albumin (GSA), which binds to the asialoglycoprotein receptor, was carried out at seven institutions in Japan. Seventy-four patients with liver disease received 3 mg (185 MBq) of^99mTc-GSA by intravenous injection. Sequential scanning was performed 30 min after injection to obtain anterior images of the heart and liver, followed by single-photon emission tomography (SPET). The indices included in this analysis were hepatic blood flow (Q) and maximal receptor binding rate (R _max), which showed a good correlation with semiquantitative ratio indices for^99mTc-GSA, namely the retention rate in blood (HH15) and the hepatic uptake rate (LHL15).Q andR _max also showed a significant correlation with other measures of hepatic function. When patients were grouped according to the severity of chronic liver damage (hepatocellular functional damage),Q was reduced in the moderate and severe groups, whileR _max was reduced in proportion to the functional stage. Both parameters showed no inter-institution difference using analysis of co-variance with the functional stage as a co-variant. With regard to the hepatic uptake rate, anterior planar images and SPET images gave similar results forQ andR _max. Acquisition times of 15 or 30 min provided the same results. The multicopartmental model analysis permitted comparable results to be obtained at institutions using different gamma cameras, and is therefore considered a universally applicable method. These results indicate thatQ andR _max are useful general indices for evaluating the function reserve capacity of the liver.     

Evaluation of asialoglycoprotein receptor imaging agent as a marker of hepatic ischemia-reperfusion injury and recovery

Protection of hepatocytes from ischemia-reperfusion injury is a clinically important issue. The purpose of this study was to evaluate changes in acute liver damage and recovery after ischemia-reperfusion in rats with asialoglycoprotein receptor (ASGP-R) ligand. Ischemia was induced by clamping the hepatoduodenal ligament for 90 min. At 1, 3, 24, 48 hr, 1 and 2 wk after reperfusion, I-125-GSA was injected. Five min after injection, blood samples were obtained and the liver was removed. Several regions from each lobe were dissected, weighed and counted. Mean uptakes (% dose/g) in the liver and blood samples were calculated. Histologic sections stained with hematoxylin-eosin (H-E) stain showed ischemic damage at 1 and 3 hr, and focal hepatocyte necrosis at 24 hr. Predominant massive necrosis was not seen. The mitotic index with H-E stain and proliferating cell nuclear antigen (PCNA) labeling index were highest at 1 wk, indicating liver regeneration. At 1 and 3 hr, liver uptake was significantly decreased, and blood uptake was significantly increased, indicating decreased tissue blood flow and ischemic damage. Liver uptake showed significant increases at 48 hr and 1 wk, and was the highest at 1 wk, indicating liver regeneration during the convalescence stage. ASGP-R binding may provide valuable information on ischemia-reperfusion injury and recovery.     

Synthesis and biological evaluation of 99mTc-DMP-NGA as a novel hepatic asialoglycoprotein receptor imaging agent

A novel bifunctional coupling agents-biomolecular compound DMP-NGA was prepared by coupling the SATP with galactosyl-neoglycoalbumin (NGA). The DMP-NGA was labeled with technetium-99 m, and the radiochemical purity in excess of 98% after purified with HPLC. In vivo biodistribution showed that ^99mTc-DMP-NGA had very high initial liver uptake with good retention. The liver accumulated 99.35±9.77%, 74.25±3.03%, 52.47±7.58% of the injected dose per gram at 5, 30 and 120 min after injection, respectively. It had relative higher initial liver uptake and much lower blood uptake than that of ^99mTc-GSA. The liver/blood ratio reached 83.4 at 30 min post-injection, while the ratio of liver/kidney was 14.4. The uptakes in other organs in the abdomen were also slightly low. In addition, the hepatic uptake of ^99mTc-DMP-NGA was blocked by preinjecting free GSA as blocking agent. The result indicates that ^99mTc-DMP-NGA has specific binding to ASGP receptor. Images acquired with Kodak In-Vivo Imaging System FX Pro showed significant difference before and after inhibition. The promising biological properties of ^99mTc-DMP-NGA afford potential applications in liver receptor imaging for assessment of hepatocyte function.     

A novel dual-modality imaging agent targeting folate receptor of tumor for molecular imaging and fluorescence-guided surgery

Annals of Nuclear Medicine - Folate receptor (FR) is an ideal target for cancer imaging because it is frequently overexpressed in major types of human tumor, whereas its expression in normal organs...     

Benzodiazepine receptor imaging with iomazenil SPECT in aphasic patients with cerebral infarction

To investigate the relationship between prognosis of aphasia and neuronal damage in the cerebral cortex, we evaluated the distribution of central-type benzodiazepine receptor (BZR) binding in post-stroke aphasics with [123I]iomazenil and SPECT. We performed iomazenil SPECT in six aphasic patients (aged from 45 to 75 years; all right-handed) with unilateral left cerebral infarction. Three patients showed signs of Broca's aphasia and the other three Wernicke's aphasia. Cerebral blood flow (CBF) imaging was performed with [123I]iodoamphetamine (IMP). The regions of interest (ROIs) on both images were set in the cerebral cortex, cerebellar cortex and language-relevant area in both hemispheres. Three patients were classified in the mild prognosis group and the other three in the moderate prognosis group. The left language-relevant area was more closely concerned with the difference in aphasic symptoms than the right one in both BZR and CBF distribution, but the ipsilateral to the contralateral ratio (I/C ratio) in the language-relevant areas in the BZR distribution was significantly lower in the moderate prognosis group than in the mild prognosis group, although no difference was seen for these values between the two groups in the CBF distribution. These results suggest that BZR imaging, which makes possible an increase in neuronal cell viability in the cerebral cortex, is useful not only for clarifying the aphasic symptoms but also for evaluating the prognosis of aphasia in patients with cerebral infarction.     

D2 receptor imaging in neonates using I-123 iodobenzamide brain SPECT

PURPOSE: Hypoxic-ischemic injury induces early changes in cerebral energy that later lead to the presence and extension of brain damage and subsequently to severe neurodevelopmental impairments such as the dyskinetic form of cerebral palsy, which is associated with damage to the striatum. The purpose of the current study was to evaluate the viability of D2 receptors in the perinatal period using I-123 iodobenzamide brain SPECT and to correlate this with early neurologic status. METHODS: After obtaining informed parental consent, 12 full-term neonates with hypoxic-ischemic events were included. I-123 iodobenzamide brain SPECT was performed 1 week after birth, corresponding to a gestational age of 39.2+/-1.7 weeks. Images were acquired using a brain-dedicated gamma camera 1 hour after intravenous injection of 30 MBq (0.8 mCi) I-123 iodobenzamide. Magnetic resonance images (T2 weighted sequence: repetition time/echo time: 2,000/30 to 150) of the brains of the same neonates were acquired on the same day. RESULTS: The right and left striatum:cerebellum activity ratios were between 1.28 and 2.25, with the greatest concentration of I-123 iodobenzamide occurring in the striatum area. A tendency of the striatum:cerebellum ratio to decrease was observed as the severity of the perinatal hypoxic-ischemic event increased despite striatal hypersignal on magnetic resonance imaging in only two neonates. CONCLUSIONS: This study, which confirms that I-123 iodobenzamide could be used in the neonatal period, shows the biochemical maturation of D2 receptors as early as 1 week after birth and also suggests the deleterious effect of perinatal hypoxic-ischemic events on D2 receptors.     

Technetium-99m spiperone dithiocarbamate: A potential radiopharmaceutical for dopamine receptor imaging with SPECT

Spiperone dithiocarbamate (SPDC) was prepared by reacting spiperone with carbon disulfide followed by sodium hydroxide. SPDC was labelled with ^99mTc by reduction of pertechnetate with formamidine sulfinic acid or sodium dithionite at alkaline pH, resulting in ∼40% incorporation of ^99mTc. The lipophilic complex was conveniently isolated at high specific activity and high radiochemical purity by extraction into dichloromethane, which was then evaporated and the residue was redissolved in a 1:3 mixture of ethanol and saline containing 0.1 mg/mL gentisic acid. Biodistribution studies following i.p. injection in rats showed low uptake of radioactivity in the brain, but striatum/cortex and striatum/cerebellum ratios were reduced by pretreatment with haloperidol. This agent may allow imaging of dopamine D-2 receptors using single-photon emission computed tomography (SPECT).     

Improvements in SPECT technology for cerebral imaging

Advancement in three major areas of SPECT (single photon emission computed tomography) technology have resulted in improved image quality for cerebral studies. In the first area, single-crystal camera electronics, extensive use of microprocessors, custom digital circuitry, an data bus architecture have allowed precise external control of all gantry motions and improved signal processing. The new digital circuitry permits energy, uniformity, and linearity corrections to be an integral part of the processing electronics. Calibration of these correlations is controlled by algorithms stored in the camera's memory. In addition, digital signals can be routed directly to interface circuitry of auxiliary computer systems without analog-to-digital conversion. Look-up tables, downloaded to the interface from the central processing unit (CPU), permit computer-controlled real-time processing of coordinate signals, including truncation, magnification, and spatial calibration. The second area of improved SPECT technology is camera collimation and related imaging techniques. In this area, system resolution has been improved without loss of sensitivity by decreasing the air gap between patient and collimator surface. Rotating the detector in close apposition to the head has required various stratagems to avoid detector-shoulder contact: the selective reduction of camera shielding, the use of long bore collimators, and the 30 degrees angulation of the camera head for slant hole collimation. Since cerebral studies characteristically image high-contrast regions less than 1 cm in size, image quality has been improved by increasing collimator resolution even at the expense of sensitivity. Increased resolution also improved image contrast for studies using 123I-labeled pharmaceuticals with 3% to 4% 124I contamination. Such studied acquired with low energy or medium energy collimation and a window centered on the 159 keV 123I photopeak contain appreciable septal breakthrough signals originating from Compton scatter of high energy photons primarily from 124I. The third area of advancements in technology, multidetector instrumentation, offers the promise of increased sensitivity and resolution. For the dynamic computer-assisted tomograph (DCAT) system, which was especially designed for regional blood flow studies with 133Xe or 127Xe, a count rate of 170,000 counts per microCi/cc for three slices has been achieved. This system consists of four detector banks each with 16 rectangular NaI crystals. An alternative system at Harvard uses an array of 12 moving detectors with focused collimators to acquire a single slice.(ABSTRACT TRUNCATED AT 400 WORDS)     

Dual tracer imaging for localization of parathyroid lesions

During the period from July 1983 through October 1984, a group of 38 patients with elevated serum calcium, parathormone (PTH) and/or clinical suspicion of hyperparathyroidism were studied by TI-201 Tc-99m dual tracer parathyroid imaging (DTPI). Seventeen of 18 parathyroid lesions were identified correctly. There was one false-negative, and the size of the adenoma missed by DTPI was less than 1.0 cm in diameter (1.0 x 0.5 x 0.2). There was one true-negative case. The other fifteen with negative scans are being followed clinically. Because of the small population studied, statistical analysis was not ascertained. However, this simple, noninvasive procedure has become a very useful diagnostic tool for the detection and localization of parathyroid lesions causing hyperparathyroidism, and the DTPI should be used in conjunction with ultrasonography and CT scanning in the preoperative evaluation in primary and secondary hyperparathyroidism.     

Quantitative evaluation of the tracer distribution in dopamine transporter SPECT for objective interpretation

Purpose

Quantification of the tracer distribution would add objectivity to the visual assessments of dopamine transporter (DAT) single photon emission computed tomography (SPECT) data. Our study aimed to evaluate the diagnostic utility of fractal dimension (FD) as a quantitative indicator of tracer distribution and compared with the conventional quantitative value: specific binding ratio (SBR). We also evaluated the utility of the combined index SBR/FD (SBR divided by FD).

Materials and methods

We conducted both clinical and phantom studies. In the clinical study, 150 patients including 110 patients with Parkinsonian syndrome (PS) and 40 without PS were enrolled. In the phantom study, we used a striatal phantom with the striatum chamber divided into two spaces, representing the caudate nucleus and putamen. The SBR, FD, and SBR/FD were calculated and compared between datasets for evaluating the diagnostic utility. Mann–Whitney test and receiver-operating characteristics (ROC) analysis were used for analysis.

Results

ROC analysis revealed that the FD value had high diagnostic performance [the areas under the curve (AUC)?=?0.943] and the combined use of SBR and FD (SBR/FD) delivered better results than the SBR alone (AUC, 0.964 vs 0.899; p?<?0.001). The sensitivity, specificity, and accuracy, respectively, were 79.1, 85.0, and 80.7% with SBR, 84.5, 97.5, and 88.0% with FD, and 92.7, 87.5, and 91.3% with SBR/FD.

Conclusion

Our results confirmed that the FD value is a useful diagnostic index, which reflects the tracer distribution in DAT SPECT images. The combined use of SBR and FD was more useful than either used alone.