Small Animal Single Photon Emission Computed Tomography Imaging Research Articles

Synthesis and Evaluation of Radioiodine-Labeled pH (Low) Insertion Peptide Variant 7-Like Peptide as a Noninvasive Tumor Microenvironment Imaging Agent in a Mouse MDA-MB-231 Triple-Negative Breast Cancer Model.

The pH (low) insertion peptide (pHLIP) family can target the tumor microenvironment (TME). If pHLIP can be labeled with radioiodine, the imaging and treatment of tumors can be considered. However, tyrosine and tryptophan can bind with iodine in the insertion region of pHLIP, and radioiodine labeling may affect the formation of α-helix structures in acidic environments; therefore, it is necessary to adjust the structure of pHLIP. This study aims to develop an 125I-labeled pH (low) insertion peptide variant 7-like peptide (pHLIP (Var7) LP) for imaging the TME in MDA-MB-231 triple-negative breast cancer (TNBC) xenograft tumor models. Based on pHLIP (Var7), a new peptide sequence, pHLIP (Var7) LP, was obtained by the sequence modification method and then characterized. The binding of pHLIP (Var7) LP to MDA-MB-231 cells was analyzed. pHLIP (Var7) LP was labeled with 125I by the iodogen iodination method. Serial biodistribution studies and small-animal single photon emission computed tomography (SPECT)/computed tomography (CT) imaging in subcutaneous MDA-MB-231 TNBC-bearing mice were performed using [125I] I-pHLIP (Var7) LP. A novel peptide, pHLIP (Var7) LP, has the characteristics of an α-helix structure, electronegativity, and amphiphilicity. Circular dichroism (CD) spectroscopy showed that the peptide presented a typical pH-dependent transition from an unstructured conformation to an α-helix structure when the pH was reduced from 8.0 to 4.0. The relative fluorescence intensities of 5-carboxytetramethylrhodamine (5-TAMRA)-pHLIP(var7) LP at pH = 6.0, 6.6, and 7.4 were 100.00 ± 5.98%, 72.10 ± 4.65%, and 13.72 ± 1.41%, respectively. The distribution of [125I] I-pHLIP (Var7) LP in tumors reached the highest level (8.7 ± 1.6% ID/g) at 2h after injection, and the tumor-to-muscle ratios and tumor-to-blood ratios increased with time. Of the measured off-target organs, the stomach, kidney, and bladder showed higher uptake levels. SPECT imaging revealed rapid and sustained tumor uptake of [125I] I-pHLIP (Var7) LP in breast cancer-bearing mice. This study showed that [125I]I-pHLIP (Var7)LP had rapid and sustained tumor uptake in MDA-MB-231 TNBC and provided a new method for TNBC imaging and further treatment.

Inhibition of MDA-MB-231 cell proliferation by pHLIP(Var7)-P1AP andSPECT imaging of MDA-MB-231 breast cancer-bearing nude miceusing 125I-pHLIP(Var7)-P1AP.

To observe the effect of pHLIP(Var7)-P1AP on the proliferation of MDA-MB-231 triple-negative breast cancer cells and the small-animal single-photon-emission computed tomography (SPECT) imaging of breast cancer-bearing mice carrying MDA-MB-231 cells. Peptide pHLIP(Var7)-P1AP was synthesized by solid-phase peptide synthesis. The binding of fluorescently labeled pHLIP(Var7)-P1AP to MDA-MB-231 cells under various pH conditions and its effect on MDA-MB-231 cell proliferation were analyzed. pHLIP(Var7)-P1AP was labeled with 125I, and the biological distribution of 125I-pHLIP(Var7)-P1AP in the breast cancer mouse model carrying MDA-MB-231 cells as well as the outcome of small-animal SPECT imaging were evaluated. pHLIP(Var7)-P1AP was successfully synthesized. Under pH 6.0, fluorescently labeled pHLIP(Var7)-P1AP had a higher binding ability to MDA-MB-231 cells and significantly inhibited the proliferation of MDA-MB-231 cells. The labeling efficiency of pHLIP(Var7)-P1AP with 125I was 33.1 ± 2.7 %, and the radiochemical purity was 98.5 ± 1.8 %. 125I-pHLIP(Var7)-P1AP showed a high concentration in tumors. Small-animal SPECT imaging showed clearly visible tumors at 4 h after injection. In the acidic environment, pHLIP(Var7)-P1AP can efficiently target MDA-MB-231 cells and inhibit their growth. Small-animal SPECT of 125I-pHLIP(Var7)-P1AP can clearly image tumors.

Evaluation of Timepix3 based CdTe photon counting detector for fully spectroscopic small animal SPECT imaging

The imaging method of SPECT (Single Photon Emission Computed Tomography) is used in nuclear medicine for diagnostics of various diseases or organs malfunctions. The distribution of medically injected, inhaled, or ingested radionuclides (radiotracers) in the patient body is imaged using gamma-ray sensitive camera with suitable imaging collimator. The 3D image is then calculated by combining many images taken from different observation angles. Most of SPECT systems use scintillator based cameras. These cameras do not provide good energy resolution and do not allow efficient suppression of unwanted signals such as those caused by Compton scattering. The main goal of this work is evaluation of Timepix3 detector properties for SPECT method for functional imaging of small animals during preclinical studies. Advantageous Timepix3 properties such as energy and spatial resolution are exploited for significant image quality improvement. Preliminary measurements were performed on specially prepared plastic phantom with cavities filled by radioisotopes and then repeated with in vivo mouse sample.

Implementation of absolute quantification in small-animal SPECT imaging: Phantom and animal studies.

PurposePresence of photon attenuation severely challenges quantitative accuracy in single‐photon emission computed tomography (SPECT) imaging. Subsequently, various attenuation correction methods have been developed to compensate for this degradation. The present study aims to implement an attenuation correction method and then to evaluate quantification accuracy of attenuation correction in small‐animal SPECT imaging.MethodsImages were reconstructed using an iterative reconstruction method based on the maximum‐likelihood expectation maximization (MLEM) algorithm including resolution recovery. This was implemented in our designed dedicated small‐animal SPECT (HiReSPECT) system. For accurate quantification, the voxel values were converted to activity concentration via a calculated calibration factor. An attenuation correction algorithm was developed based on the first‐order Chang's method. Both phantom study and experimental measurements with four rats were used in order to validate the proposed method.ResultsThe phantom experiments showed that the error of −15.5% in the estimation of activity concentration in a uniform region was reduced to +5.1% when attenuation correction was applied. For in vivo studies, the average quantitative error of −22.8 ± 6.3% (ranging from −31.2% to −14.8%) in the uncorrected images was reduced to +3.5 ± 6.7% (ranging from −6.7 to +9.8%) after applying attenuation correction.ConclusionThe results indicate that the proposed attenuation correction algorithm based on the first‐order Chang's method, as implemented in our dedicated small‐animal SPECT system, significantly improves accuracy of the quantitative analysis as well as the absolute quantification.

A modified TEW approach to scatter correction for In-111 and Tc-99m dual-isotope small-animal SPECT.

In dual-isotope (Tc-99m/In-111) small-animal single-photon emission computed tomography (SPECT), quantitative accuracy of Tc-99m activity measurements is degraded due to the detection of Compton-scattered photons in the Tc-99m photopeak window, which originate from the In-111 emissions (cross talk) and from the Tc-99m emission (self-scatter). The standard triple-energy window (TEW) estimates the total scatter (self-scatter and cross talk) using one scatter window on either side of the Tc-99m photopeak window, but the estimate is biased due to the presence of unscattered photons in the scatter windows. The authors present a modified TEW method to correct for total scatter that compensates for this bias and evaluate the method in phantoms and in vivo. The number of unscattered Tc-99m and In-111 photons present in each scatter-window projection is estimated based on the number of photons detected in the photopeak of each isotope, using the isotope-dependent energy resolution of the detector. The camera-head-specific energy resolutions for the 140 keV Tc-99m and 171 keV In-111 emissions were determined experimentally by separately sampling the energy spectra of each isotope. Each sampled spectrum was fit with a Linear + Gaussian function. The fitted Gaussian functions were integrated across each energy window to determine the proportion of unscattered photons from each emission detected in the scatter windows. The method was first tested and compared to the standard TEW in phantoms containing Tc-99m:In-111 activity ratios between 0.15 and 6.90. True activities were determined using a dose calibrator, and SPECT activities were estimated from CT-attenuation-corrected images with and without scatter-correction. The method was then tested in vivo in six rats using In-111-liposome and Tc-99m-tetrofosmin to generate cross talk in the area of the myocardium. The myocardium was manually segmented using the SPECT and CT images, and partial-volume correction was performed using a template-based approach. The rat heart was counted in a well-counter to determine the true activity. In the phantoms without correction for Compton-scatter, Tc-99m activity quantification errors as high as 85% were observed. The standard TEW method quantified Tc-99m activity with an average accuracy of -9.0% ± 0.7%, while the modified TEW was accurate within 5% of truth in phantoms with Tc-99m:In-111 activity ratios ≥0.52. Without scatter-correction, In-111 activity was quantified with an average accuracy of 4.1%, and there was no dependence of accuracy on the activity ratio. In rat myocardia, uncorrected images were overestimated by an average of 23% ± 5%, and the standard TEW had an accuracy of -13.8% ± 1.6%, while the modified TEW yielded an accuracy of -4.0% ± 1.6%. Cross talk and self-scatter were shown to produce quantification errors in phantoms as well as in vivo. The standard TEW provided inaccurate results due to the inclusion of unscattered photons in the scatter windows. The modified TEW improved the scatter estimate and reduced the quantification errors in phantoms and in vivo.

Optimization of helical acquisition parameters to preserve uniformity of mouse whole body using multipinhole collimator in single-photon emission computed tomography

Focusing on whole-body uniformity in small-animal single-photon emission computed tomography (SPECT), we examined the optimal helical acquisition parameters using five-pinhole collimators for mouse imaging. SPECT images of an 80-mm-long cylindrical phantom with 99mTc solution were acquired using an Inveon multimodality imaging platform. The bed travels used in this study were 0, 30, 60, 90 and 120mm, and the numbers of revolutions traversed during the SPECT scan were 1.0, 2.0, 3.0, 4.0, 5.0 and 7.0, respectively. Artifacts that degrade uniformity in reconstructed images were conspicuous when the bed travel was smaller than the object length. Regarding the distal-to-center ratio (DCR) of SPECT values in the object’s axial direction, the DCR nearest to the ideal ratio of 1.00 was 1.02 in the optimal uniformity with 4.0 revolutions and a bed travel of 120mm. Moreover, the helical acquisition using these parameters suppressed the formation of artifacts. We proposed the optimal parameters in whole-body helical SPECT; the bed travel was sufficiently larger than the object length; the 4.0 or more revolutions were required for a pitch of approximately 30mm/revolution. The optimal acquisition parameters in SPECT to preserve uniformity would contribute to the accurate quantification of whole-body biodistribution.

Iodine-131 imaging using 284 keV photons with a small animal CZT-SPECT system dedicated to low-medium-energy photon detection.

Iodine-131 is widely used for radionuclide therapy because of its β-particle and for diagnostic imaging employing its principal gamma ray. Since that principal gamma ray has the relatively high energy of 364 keV, small animal single-photon emission computed tomography (SPECT) imaging systems may be required to possess the ability to image such higher energy photons. The aim of this study was to investigate the possibility of imaging I-131 using its 284 keV photons instead of its 364 keV photons in a small animal SPECT imaging system dedicated to the detection of low-medium-energy photons (below 300 keV). The imaging system used was a commercially available preclinical SPECT instrument with CZT detectors that was equipped with multi-pinhole collimators and was accompanied by a CT imager. An energy window for I-131 imaging was set to a photopeak of 284 keV with a low abundance compared with 364 keV photons. Small line sources and two mice, one of each of two types, that were injected with NaI-131 were scanned. Although higher counts occurred at the peripheral region of the reconstructed images due to the collimator penetration by the 364 keV photons, the shape of the small line sources could be well visualized. The measured spatial resolution was relatively poor (~1.9 mm for full width at half maximum and ~3.9 mm for full width at tenth maximum). However, a good linear correlation between SPECT values and the level of I-131 radioactivity was observed. Furthermore, the uptake of NaI-131 to the thyroid gland for the two mice was clearly identified in the 3D-SPECT image fused with the X-ray CT image. We conclude that the use of an energy window set on the photopeak of 284 keV and the multi-pinhole collimator may permit I-131 imaging for a preclinical CZT-SPECT system that does not have the ability to acquire images using the 364 keV photons.

Measurement of [123I]FP-CIT binding to the dopamine transporter (DAT) in healthy mouse striatum using dedicated small animal SPECT imaging: feasibility, optimization and validation.

In-vivo imaging of dopamine transporter (DAT), a reliable marker of degeneration of nigrostriatal dopaminergic innervation, has gained increasing interest in preclinical neurodegenerative research for studying disease mechanisms and testing new therapeutic strategies. We assessed the feasibility and the reliability of in vivo and ex vivo quantification of Methyl (3S,4S,5R)-8-(3-fluoropropyl)-3-(4-iodophenyl)-8-azabicyclo[3.2.1]octane-4-carboxylate ([123I]FP-CIT) binding to striatal DAT sites in mouse brain. Dedicated small animal single-photon emission computed tomography (SPECT) images of [123I]FP-CIT binding were obtained in 3 groups of healthy mice: untreated (N.=6), pre-treated with lugol solution (N.=4), and pre-treated with selective dopamine transporter uptake inhibitor GBR12909 (N.=4). Ex-vivo autoradiography studies were performed at the end of SPECT studies with phosphor image system in 4 out of the 6 untreated mice and in all mice pre-treated with lugol. Regions of interest were defined over the striatum. The specific binding (SB) was calculated using the cerebral cortex as reference region. SPECT images in untreated mice showed high [123I]FP-CIT uptake in the striatum and extracerebral regions. Lugol pretreatment improved striatal images quality decreasing salivary and thyroid glands uptake. SB was higher (P<0.0001) in mice pre-treated with lugol (5.97±0.60) than in untreated mice (2.25±0.28). Autoradiography showed similar SB findings in untreated (2.27±0.33) and lugol-treated (4.27±0.57) mice (P<0.0001). In-vivo striatal 123I-FP-CIT SB and ex-vivo striatal 123I-FP-CIT SB were significantly correlated (r=0.87; P<0.0001). SPECT competition studies showed a significant (P<0.0001) reduction of [123I]FP-CIT SB in the striatum after GBR12909. We demonstrated the feasibility of [123I]FP-CIT imaging of the normal mouse brain using small-animal SPECT without pinhole collimators. The reliability of quantitative measurement of striatal [123I]FP-CIT SB is supported by competition studies showing measurable inhibition of uptake induced by GBR12909 and by the strong correlation between in vivo and ex vivo striatal [123I]FP-CIT SB. Our data also demonstrate that pre-treatment with lugol might improve striatal [123I]FP-CIT SB in mice.

MP66-07 TARGETING ANDROGEN RECEPTOR N-TERMINAL DOMAIN FOR PROSTATE CANCER IMAGING AND THERAPY

You have accessJournal of UrologyProstate Cancer: Basic Research V1 Apr 2015MP66-07 TARGETING ANDROGEN RECEPTOR N-TERMINAL DOMAIN FOR PROSTATE CANCER IMAGING AND THERAPY Yusuke Imamura, Amy H. Tien, Nasrin R. Mawji, Jian Kun Zhong, Jinhe Pan, Kuo-Shyan Lin, Raymond J. Andersen, and Marianne D. Sadar Yusuke ImamuraYusuke Imamura More articles by this author , Amy H. TienAmy H. Tien More articles by this author , Nasrin R. MawjiNasrin R. Mawji More articles by this author , Jian Kun ZhongJian Kun Zhong More articles by this author , Jinhe PanJinhe Pan More articles by this author , Kuo-Shyan LinKuo-Shyan Lin More articles by this author , Raymond J. AndersenRaymond J. Andersen More articles by this author , and Marianne D. SadarMarianne D. Sadar More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.2360AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Resistance to therapies that target the androgen receptor (AR) ligand-binding domain (LBD) may be due to expression of constitutively active AR splice variants that lack the LBD. EPI small molecules bind to AR N-terminal domain (NTD) and inhibit transcriptional activity of AR. We developed an analogue of radiolabeled iodine 123- (123I) EPI for imaging of prostate cancer using single-photon emission computed tomography (SPECT). Currently the approach to image AR uses positron emission tomography with 16β-[18F]-fluoro-5α dihydrotestosterone (F18-FDHT) that binds to AR LBD. F18-FDHT cannot detect AR splice variants lacking LBD. Our approach would ultimately employ sequential imaging with F18-FDHT to detect solely full-length (fl)-AR and 123I-EPI to detect NTD of both fl-AR and truncated AR splice variants. A discordant distribution or discordant level of uptake between 18F-FDHT and 123I-EPI may indicate expression of splice variants lacking LBD. METHODS Effects of non-radiolabeled iodine (I)-EPI was evaluated in castrate-resistant prostate cancer (CRPC) model LNCaP95 cells and xenograft-bearing mice. EPI was successfully radiolabeled with the isotope 123I. Binding experiments of 123I-EPI to endogenous AR in cells were evaluated in vitro. 123I-EPI uptake and plasma clearance by quantifying 123I-EPI in harvested organs at various time points as whole-body distribution were evaluated, and small animal SPECT imaging study were conducted with LNCaP95 xenograft-bearing mice. RESULTS I-EPI was specific for blocking AR transcriptional activity. It decreased AR-dependent growth and had no toxicity at therapeutic doses in vivo. Consistent with EPI targeting the AR, I-EPI showed efficacy at 10mg/kg body weight and decreased the tumor size of LNCaP95 xenografts in castrated mice. 123I-EPI showed specific covalent binding to endogenous AR in LNCaP95 cells. Co-treatment with therapeutic doses of EPI significantly blocked the accumulation of 123I-EPI in the LNCaP95 xenograft. Furthermore, 123I-EPI enabled visualization of the LNCaP95 xenograft within 4hrs after injection. The compound appeared to be excreted via the liver rather than the kidney. Low level of uptake was found in the bladder after 1 hour; bone and muscle uptake was also low. CONCLUSIONS An AR NTD-targeted molecular imaging probe such as 123I-EPI would be useful to select patients for subsequent antiandrogen therapies, monitor treatment response, and provide insight into the role of all AR species in resistance mechanisms. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e818 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Yusuke Imamura More articles by this author Amy H. Tien More articles by this author Nasrin R. Mawji More articles by this author Jian Kun Zhong More articles by this author Jinhe Pan More articles by this author Kuo-Shyan Lin More articles by this author Raymond J. Andersen More articles by this author Marianne D. Sadar More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Optimal energy window selection of a CZT-based small-animal SPECT for quantitative accuracy

Cadmium zinc telluride (CZT)-based small-animal single-photon emission computed tomography (SPECT) has desirable characteristics such as superior energy resolution, but data acquisition for SPECT imaging has been widely performed with a conventional energy window. The aim of this study was to determine the optimal energy window settings for technetium-99m (99mTc) and thallium-201 (201Tl), the most commonly used isotopes in SPECT imaging, using CZT-based small-animal SPECT for quantitative accuracy.We experimentally investigated quantitative measurements with respect to primary count rate, contrast-to-noise ratio (CNR), and scatter fraction (SF) within various energy window settings using Triumph X-SPECT. The two ways of energy window settings were considered: an on-peak window and an off-peak window. In the on-peak window setting, energy centers were set on the photopeaks. In the off-peak window setting, the ratios of energy differences between the photopeak from the lower- and higher-threshold varied from 4:6 to 3:7. In addition, the energy-window width for 99mTc varied from 5% to 20%, and that for 201Tl varied from 10% to 30%.The results of this study enabled us to determine the optimal energy windows for each isotope in terms of primary count rate, CNR, and SF. We selected the optimal energy window that increases the primary count rate and CNR while decreasing SF. For 99mTc SPECT imaging, the energy window of 138–145keV with a 5% width and off-peak ratio of 3:7 was determined to be the optimal energy window. For 201Tl SPECT imaging, the energy window of 64–85keV with a 30% width and off-peak ratio of 3:7 was selected as the optimal energy window.Our results demonstrated that the proper energy window should be carefully chosen based on quantitative measurements in order to take advantage of desirable characteristics of CZT-based small-animal SPECT. These results provided valuable reference information for the establishment of new protocol for CZT-based small-animal study, and may offer insights into the potential of multiple-isotope imaging with CZT-based small-animal SPECT.

MP24-02 DEVELOPMENT OF AN IMAGING APPROACH TO DETECT SPLICE VARIANTS OF ANDROGEN RECEPTOR IN PROSTATE CANCER

INTRODUCTION AND OBJECTIVES: Resistance to therapies that target the androgen receptor (AR) ligand binding domain (LBD) may be due to expression of constitutively active AR splice variants that lack the LBD. EPI-001/EPI-002 small molecules bind to AR amino-terminal domain (NTD).Wepropose todevelopananalogueof radiolabeled iodine 123(I) EPI-002 for imaging of prostate cancer using single-photon emission computed tomography (SPECT). Currently the approach to image AR uses positron emission tomography with 16b-[18F]-fluoro-5a dihydrotestosterone (F18-FDHT) that binds to AR LBD. F18-FDHT cannot detect AR splice variants lacking LBD. Our approach would employ sequential imaging with F18-FDHT to detect solely full-length (fl)AR and I-EPI-002 to detect NTD of both fl-AR and variant AR. A discordant distribution or discordant level of uptake between 18F-FDHT and I-EPI-002may indicate expression of splice variants lacking LBD. METHODS: Activity and specificity of non-radiolabeled iodine (I)-EPI-002 for inhibiting AR were confirmed in cells using reporter gene constructs for the AR and related steroid hormone receptors, glucocorticoid receptor (GR) and progesterone receptor (PR). Competitive ligand binding assays to detect the displacement of fluorescently labeled ligand from recombinant LBDs of AR, PR, GR and estrogen receptor (ER) by R1881, MDV3100, EPI-002 or I-EPI-002 were performed. EPI002 was successfully radiolabeled with the isotope I. On-going experiments include: binding experiments of I-EPI-002 to recombinant proteins AR activation function-1 and fl-AR, as well as to endogenous flAR and variant AR in cells; evaluation of I-EPI-002 in whole-body distribution including dosimetry in xenografts by small animal SPECT imaging; and evaluation of I-EPI-002 uptake and plasma clearance by quantifying I in harvested organs at various time points. Biological, physical and effective half-life of I-EPI-002 will be determined. RESULTS: In cell-based assays, I-EPI-002 was specific for AR and blocked the induction of AR-driven reporter activity in response to androgen, but did not inhibit either PR or GR transcriptional activities in response to ligand. I-EPI-002 does not bind to LBDs of AR, PR, GR and ER. Ongoing in vivo experiments will reveal the potential of I-EPI-002 for clinical application. CONCLUSIONS: An AR NTD-targeted molecular imaging probe such as I-EPI-002 may be useful to select patients for subsequent antiandrogen therapies, monitor treatment response, and provide insight into the role of all AR species in resistance mechanisms.

A high resolution SPECT detector based on thin continuous LYSO

Single-photon emission computed tomography (SPECT) detectors with improved spatial resolution can be used to build multi-pinhole SPECT systems that have a higher sensitivity or a higher spatial resolution. In order to improve the spatial resolution we investigate the performance of a 2 mm thick continuous Lutetium Yttrium Orthosilicate (LYSO) scintillator and compare it to the performance of a 5 mm thick continuous NaI(Tl) scintillator. The advantages of LYSO are its high stopping power and its non-hygroscopicity. Drawbacks are the lower light output and the intrinsic radioactivity. The hypothesis of this study is that such a thin LYSO scintillator will have a small light spread and, as a consequence, will also have an improved spatial resolution when coupled to a Hamamatsu H8500 position sensitive photomultiplier tube. To optimize the spatial resolution and the useful detector area we used a mean nearest neighbor event-positioning method. Beam source measurements (99mTc, 140 keV) were done to investigate the energy resolution and the spatial resolution of both detectors. The effect of the intrinsic radioactivity of the LYSO scintillator in the energy window was quantified. The mean energy resolution is 9.3% for the NaI(Tl) scintillator and 21.3% for the LYSO scintillator. The LYSO spectrum shows an X-ray escape peak which decreases the detection efficiency with 9.1%. The spatial resolution of the LYSO detector (0.93 mm full width at half maximum (FWHM)) is superior to the spatial resolution of the NaI(Tl) detector (1.37 mm FWHM). The intrinsic radioactivity in the energy window (42% window centered at 140 keV) is low (125.6 cps, 0.024 cps mm−3). LYSO is a promising scintillator for small-animal SPECT imaging, where spatial resolution is more important than energy resolution.

PEGylation of 99mTc-labeled bombesin analogues improves their pharmacokinetic properties

Radiolabeled bombesin (BN) conjugates are promising radiotracers for imaging and therapy of breast and prostate tumors in which BN(2)/gastrin-releasing peptide (GRP) receptors are overexpressed. However, the low in vivo stability of BN conjugates may limit their clinical application. In an attempt to improve their pharmacokinetics and counteract their rapid enzymatic degradation, we prepared a series of polyethylene glycol (PEG)-ylated BN(7-14) analogues for radiolabeling with (99m)Tc(CO)(3) and evaluated them in vitro and in vivo. Derivatization of a stabilized (N(α)His)Ac-BN(7-14)[Cha(13),Nle(14)] analogue with linear PEG molecules of various sizes [5 kDa (PEG(5)), 10 kDa (PEG(10)) and 20 kDa (PEG(20))] was performed by PEGylation of the ɛ-amino group of a β(3)hLys-βAla-βAla spacer between the stabilized BN sequence and the (N(α)His)Ac chelator. The analogues were then radiolabeled by employing the (99m)Tc-tricarbonyl technique. Binding affinity and internalization/externalization studies were performed in vitro in human prostate carcinoma PC-3 cells. Stability was investigated in vitro in human plasma and in vivo in Balb/c mice. Finally, single photon emission computed tomography (SPECT)/X-ray computed tomography studies were performed in nude mice bearing PC-3 tumor xenografts. PEGylation did not affect the binding affinity of BN analogues, as the binding affinity for BN(2)/GRP receptors remained high (K(d)<0.9 nM). However, in vitro binding kinetics of the PEGylated analogues were slower. Steady-state condition was reached after 4 h, and the total cell binding was 10 times lower than that for the non-PEGylated counterpart. Besides, PEGylation improved the stability of BN conjugates in vitro and in vivo. The BN derivative conjugated with a PEG(5) molecule showed the best pharmacokinetics in vivo, i.e., faster blood clearance and preferential renal excretion. The tumor uptake of the (99m)Tc-PEG(5)-Lys-BN conjugate was slightly higher compared to that of the non-PEGylated analogue (3.91%±0.44% vs. 2.80%±0.28% injected dose per gram 1 h postinjection, p.i.). Tumor retention was also increased, resulting in a threefold higher amount of radioactivity in the tumor at 24 h p.i. Furthermore, decreased hepatobiliary excretion and increased tumor-to-nontarget ratios (tumor-to-blood: 17.1 vs. 2.1; tumor-to-kidney: 1.1 vs. 0.4; tumor-to-liver: 5.8 vs. 1.0, 24 h p.i.) were observed and further confirmed via small-animal SPECT images 1 h p.i. PEGylation proved to be an effective strategy to enhance the tumor-targeting potential of (99m)Tc-labeled BN-based radiopharmaceuticals and probably other radiolabeled peptides.

A clinical gamma camera-based pinhole collimated system for high resolution small animal SPECT imaging

The main objective of the present study was to upgrade a clinical gamma camera to obtain high resolution tomographic images of small animal organs. The system is based on a clinical gamma camera to which we have adapted a special-purpose pinhole collimator and a device for positioning and rotating the target based on a computer-controlled step motor. We developed a software tool to reconstruct the target's three-dimensional distribution of emission from a set of planar projections, based on the maximum likelihood algorithm. We present details on the hardware and software implementation. We imaged phantoms and heart and kidneys of rats. When using pinhole collimators, the spatial resolution and sensitivity of the imaging system depend on parameters such as the detector-to-collimator and detector-to-target distances and pinhole diameter. In this study, we reached an object voxel size of 0.6 mm and spatial resolution better than 2.4 and 1.7 mm full width at half maximum when 1.5- and 1.0-mm diameter pinholes were used, respectively. Appropriate sensitivity to study the target of interest was attained in both cases. Additionally, we show that as few as 12 projections are sufficient to attain good quality reconstructions, a result that implies a significant reduction of acquisition time and opens the possibility for radiotracer dynamic studies. In conclusion, a high resolution single photon emission computed tomography (SPECT) system was developed using a commercial clinical gamma camera, allowing the acquisition of detailed volumetric images of small animal organs. This type of system has important implications for research areas such as Cardiology, Neurology or Oncology.

3-D Rat Brain Phantom for High-Resolution Molecular Imaging

With the steadily improving resolution of novel small-animal single photon emission computed tomography (SPECT) and positron emission tomography devices, highly detailed phantoms are required for testing and optimizing these systems. We present a three-dimensional (3-D) digital and physical phantom pair to represent, e.g., cerebral blood flow, glucose metabolism, or neuroreceptor binding in small regions of the rat brain. The anatomical structures are based on digital photographs of the uncut part of a rat brain cryosection block. The photographs have been segmented into ventricles and gray and white matter and have been stacked afterwards. In the resulting voxelized digital phantom, tracer concentration in gray and white matter can be scaled independently. This is of relevance since, e.g., cerebral blood flow or metabolism are much higher in gray than in white matter. The physical phantom is based on the digital phantom and has been manufactured out of hardened polymer using rapid prototyping, a process in which complicated 3-D objects can be built up layer by layer. X-ray computed tomography and high-resolution SPECT images of the physical phantom are compared with the digital phantom. The detailed physical phantom can be filled bubble-free. Excellent correspondence is shown between details in the digital and physical phantom. Therefore, this newly developed brain phantom will enable the optimization of high-resolution imaging for recovery of complex shaped molecular distributions.

High resolution micro‐SPECT scanning in rats using 125I β‐CIT: Effects of chronic treatment with carbamazepine

Carbamazepine (CBZ) is a first-line antiepileptic agent with mood-stabilizing effects in bipolar disorder. It has been reported to influence extracellular concentrations of serotonin and dopamine, suggesting an interaction with monoamine transporters. We have investigated this effect using in vivo single photon emission computed tomography (SPECT) in rats. Adult male rats received 3 mg/kg/h CBZ via mini-osmotic pump. After 14 days continuous treatment, animals underwent two consecutive SPECT scans, using 125I beta-CIT as a radiotracer to label serotonin transporter (SERT) and dopamine transporter (DAT) sites in the brain. Pharmacologic distinction was enabled by 125I beta-CIT SPECT imaging in rats acutely exposed to the serotonin and dopamine transporter inhibitors, fluoxetine and GBR12909. The interaction between CBZ and 125I beta-CIT binding to SERT and DAT was investigated using in vitro autoradiography. Carbamazepine (10 microm) did not affect binding of 125I beta-CIT to isolated rat brain slices, thereby excluding a direct effect on ligand binding to SERT and DAT. SPECT studies with fluoxetine and GBR12909 highlighted SERT binding in thalamus, hippocampus, centromedial nuclei, and occipital cortex, and DAT binding in the caudate. Prolonged treatment with CBZ failed to influence 125I beta-CIT binding to either SERT or DAT in any of the brain regions examined. This study employed the novel technique of small animal SPECT imaging to investigate the effects of CBZ on monoamine transporters in rat brain. Following prolonged treatment, the drug was without effect on SERT or DAT availability. The mechanism by which CBZ exerts its mood stabilizing effects remains elusive.

Synthesis, Radiolabeling, and Bio-imaging of High-Generation Polyester Dendrimers

A series of aliphatic polyester dendrons, generations 1 through 8, were prepared with a core p-toluenesulfonyl ethyl (TSe) ester as an easily removable protecting group that can be efficiently replaced with a variety of nucleophiles. Using amidation chemistry, a tridentate bis(pyridyl)amine ligand which is known to form stable complexes with both Tc(I) and Re(I) was introduced at the dendrimer core. Metalation of the core ligand with (99m)Tc was accomplished for generations 5 through 7, and resulted in regioselective radiolabeling of the dendrimers. The distribution of the radiolabeled dendrimers was evaluated in healthy adult Copenhagen rats using dynamic small-animal single photon emission computed tomography (SPECT). The labeled dendrimers were cleanly and rapidly eliminated from the bloodstream via the kidneys with negligible nonspecific binding to organs or tissues being observed. These data were corroborated by a quantitative biodistribution study on the generation 7 dendrimer following necropsy. The quantitative biodistribution results were in excellent agreement with the data obtained from the dynamic SPECT images.

A high-sensitivity small animal SPECT system

Medical imaging using single gamma-ray-emitting radionuclides typically makes use of parallel hole collimators or pinholes in order to achieve good spatial resolution. However, a tradeoff in sensitivity is inherent in the use of a collimator, and modern preclinical single photon emission computed tomography (SPECT) systems detect a very small fraction of emitted gamma rays, often less than 0.1%. A system for small animal SPECT imaging which uses no collimators could potentially achieve very high sensitivity—several tens of percent—with reasonably sized detectors. This would allow two significant improvements in preclinical studies: images could be obtained more rapidly, allowing higher throughput for screening applications, or for dynamic processes to be observed with very good time resolution; and images could be obtained with less radioactive tracer, making possible the in vivo imaging of low-capacity receptor systems, aiding research into new tracer compounds, and reducing the cost and easing the regulatory burden of an experiment. Of course, a system with no collimator will not be able to approach the submillimeter spatial resolutions produced by the most advanced pinhole and collimated systems, but a high-sensitivity system with resolution of order 1 cm could nonetheless find significant and new use in the many molecular imaging applications which do not require good spatial resolution—for example, screening applications for drug development or new imaging agents. Rather than as an alternative to high-resolution SPECT systems, the high-sensitivity system is proposed as a radiotracer alternative to optical imaging for small animals. We have developed a prototype system for mouse imaging applications. The scanner consists of two large, thin, closely spaced scintillation detectors. Simulation studies indicate that a FWHM spatial resolution of 7 mm is possible. In an in vivo mouse imaging study using the 99mTc labeled tracer MAG-3, the sensitivity of the system is measured to be 40%. Simple projection images created by analytically combining the two detectors' data show sufficient resolution to observe the dynamic distribution of the radiotracer in the mouse.

Geometric characterization of multi-axis multi-pinhole SPECT

A geometric model and calibration process are developed for single photon emission computed tomography (SPECT) imaging with multiple pinholes and multiple mechanical axes. Unlike the typical situation where pinhole collimators are mounted directly to rotating gamma ray detectors, this geometric model allows for independent rotation of the detectors and pinholes, for the case where the pinhole collimator is physically detached from the detectors. This geometric model is applied to a prototype small animal SPECT device with a total of 22 pinholes and which uses dual clinical SPECT detectors. All free parameters in the model are estimated from a calibration scan of point sources and without the need for a precision point source phantom. For a full calibration of this device, a scan of four point sources with 360 degrees rotation is suitable for estimating all 95 free parameters of the geometric model. After a full calibration, a rapid calibration scan of two point sources with 180 degrees rotation is suitable for estimating the subset of 22 parameters associated with repositioning the collimation device relative to the detectors. The high accuracy of the calibration process is validated experimentally. Residual differences between predicted and measured coordinates are normally distributed with 0.8 mm full width at half maximum and are estimated to contribute 0.12 mm root mean square to the reconstructed spatial resolution. Since this error is small compared to other contributions arising from the pinhole diameter and the detector, the accuracy of the calibration is sufficient for high resolution small animal SPECT imaging.

Coregistration of Magnetic Resonance and Single Photon Emission Computed Tomography Images for Noninvasive Localization of Stem Cells Grafted in the Infarcted Rat Myocardium

This paper demonstrates the application of mutual information based coregistration of radionuclide and magnetic resonance imaging (MRI) in an effort to use multimodality imaging for noninvasive localization of stem cells grafted in the infarcted myocardium in rats. Radionuclide imaging such as single photon emission computed tomography (SPECT) or positron emission tomography (PET) inherently has high sensitivity and is suitable for tracking of labeled stem cells, while high-resolution MRI is able to provide detailed anatomical and functional information of myocardium. Thus, coregistration of PET or SPECT images with MRI will map the location and distribution of stem cells on detailed myocardium structures. To validate this coregistration method, SPECT data were simulated by using a Monte Carlo-based projector that modeled the pinhole-imaging physics assuming nonzero diameter and photon penetration at the edge. Translational and rotational errors of the coregistration were examined with respect to various SPECT activities, and they are on average about 0.50 mm and 0.82 degrees , respectively. Only the rotational error is dependent on activity of SPECT data. Stem cells were labeled with (111)Indium oxyquinoline and grafted in the ischemic myocardium of a rat model. Dual-tracer small-animal SPECT images were acquired, which allowed simultaneous detection of (111)In-labeled stem cells and of [(99m)Tc]sestamibi to assess myocardial perfusion deficit. The same animals were subjected to cardiac MRI. A mutual-information-based coregistration method was then applied to the SPECT and MRIs. By coregistration, the (111)In signal from labeled cells was mapped into the akinetic region identified on cine MRIs; the regional perfusion deficit on the SPECT images also coincided with the akinetic region on the MR image.