Dimethylamino-fluorenes: ligands for detecting beta-amyloid plaques in the brain

Formation of beta-amyloid (Abeta) plaques in the brain is a major contributing factor in the pathogenesis of Alzheimer's disease (AD). Detection of Abeta plaques in the brain will be potentially useful in early diagnosis and monitoring the progression of the disease. A series of novel Abeta aggregate-specific ligands based on fluorenes, which are simple and rigid tricyclic molecules, are synthesized and characterized. Starting with 2- or 3-aminofluorenes, 1a-1f, the amino group was converted to the N,N-dimethylamino group (2a-2f) in excellent yield. It was found that 7-iodo-2-N,N-dimethylaminofluorene (2f) showed an extremely high binding affinity to preformed Abeta40 aggregates (K(i) = 0.9 nM). In vitro autoradiography study using brain sections obtained from transgenic mice (Tg2576) with [(125)I]2f showed exquisitely high specific binding to Abeta plaques. The same section also displayed an equivalent labeling when stained by Thioflavin-S, a commonly used fluorescent dye for Abeta plaques. When [(125)I]2f was injected intravenously into normal mice, it exhibited an excellent brain uptake. Taken together the data suggest that [(125)I]2f may be useful as an in vivo imaging agent to detect Abeta plaques in the brain.     

Labeling of cerebral amyloid beta deposits in vivo using intranasal basic fibroblast growth factor and serum amyloid P component in mice.

There is currently no method for noninvasive imaging of amyloid beta (Abeta) deposition in Alzheimer's disease (AD). Because Abeta plaques are characteristic of AD and Abeta deposits contain abundant heparan sulfate proteoglycans that can bind basic fibroblast growth factor (bFGF) and serum amyloid P component (SAP), we investigated a novel route of ligand delivery to the brain to assess Abeta deposition in a transgenic (Tg) mouse model overexpressing Abeta-protein precursor. METHODS: The biodistribution of bFGF injected intranasally was studied using (125)I-bFGF in Tg and wild-type control mice and by unlabeled bFGF and SAP immunocytochemistry with light and electron microscopy. RESULTS: Three- to 5-fold higher amounts of (125)I-bFGF were found in the brain of Tg mice than that of wild-type mice (P < 0.05). bFGF or SAP given intranasally labeled cerebral Abeta plaques in the cortex and microvessels of Tg mice but not in wild-type mice. Weak bFGF staining and no SAP staining were detected in Tg mice without intranasal injection of the ligands. bFGF and SAP stained neurons around the rim of Abeta deposits and throughout the cortex in Tg mice. There was only weak staining of neurons in Tg mice without intranasal injection of bFGF and no staining of SAP in Tg mice without intranasal injection of SAP. No bFGF or SAP staining was evident in wild-type control mice. CONCLUSION: We report a novel noninvasive method for labeling Abeta plaques. This method may be modified for human studies using intranasal injection of radiolabeled ligands and imaging with SPECT or PET.     

T2-weighted MR imaging of focal hepatic lesions: Comparison of various RARE and fat-suppressed spin-echo sequences

The authors prospectively compared four T2-weighted magnetic resonance (MR) sequences, including high-resolution 512 × 512 (matrix size) RARE (rapid acquisition with relaxation enhancement), 256 × 256 RARE, 128 × 256 breath-hold RARE, and 192 × 256 fat-suppressed spin-echo (T2FS) sequences, in the evaluation of 16 patients with focal hepatic masses. MR images were evaluated by quantitative lesion-liver signal difference-to-noise ratios (SDNRs) and subjective evaluation of image artifact and image quality. No significant differences were observed between RARE sequences in SDNR values. The T2FS sequence had a significantly higher SDNR than the 512 × 512 RARE sequence (24.6 ± 15.0 vs 14.5 ± 9.7) (P =.008). Image quality was rated highest for the 512 × 512 RARE and T2FS sequences (P =.006). The inherent advantage of high spatial resolution suggests that the 512 × 512 RARE sequence may be of value in detecting hepatic lesions.     

In vivo visualization of Alzheimer's amyloid plaques by magnetic resonance imaging in transgenic mice without a contrast agent.

One of the cardinal pathologic features of Alzheimer's disease (AD) is the formation of senile, or amyloid, plaques. Transgenic mice have been developed that express one or more of the genes responsible for familial AD in humans. Doubly transgenic mice develop "human-like" plaques, providing a mechanism to study amyloid plaque biology in a controlled manner. Imaging of labeled plaques has been accomplished with other modalities, but only MRI has sufficient spatial and contrast resolution to visualize individual plaques noninvasively. Methods to optimize visualization of plaques in vivo in transgenic mice at 9.4 T using a spin echo sequence based on adiabatic pulses are described. Preliminary results indicate that a spin echo acquisition more accurately reflects plaque size, while a T2* weighted gradient echo sequence reflects plaque iron content, not plaque size. In vivo MRI-ex vivo MRI-in vitro histologic correlations are provided. Histologically verified plaques as small as 50 microm in diameter were visualized in living animals. To our knowledge this work represents the first demonstration of noninvasive in vivo visualization of individual AD plaques without the use of a contrast agent.     

Detection of Alzheimer's amyloid in transgenic mice using magnetic resonance microimaging.

The presence of amyloid-beta (Abeta) plaques in the brain is a hallmark pathological feature of Alzheimer's disease (AD). Transgenic mice overexpressing mutant amyloid precursor protein (APP), or both mutant APP and presenilin-1 (APP/PS1), develop Abeta plaques similar to those in AD patients, and have been proposed as animal models in which to test experimental therapeutic approaches for the clearance of Abeta. However, at present there is no in vivo whole-brain imaging method to detect Abeta plaques in mice or men. A novel method is presented to detect Abeta plaques in the brains of transgenic mice by magnetic resonance microimaging (muMRI). This method uses Abeta1-40 peptide, known for its high binding affinity to Abeta, magnetically labeled with either gadolinium (Gd) or monocrystalline iron oxide nanoparticles (MION). Intraarterial injection of magnetically labeled Abeta1-40, with mannitol to transiently open the blood-brain barrier (BBB), enabled the detection of many Abeta plaques. Furthermore, the numerical density of Abeta plaques detected by muMRI and by immunohistochemistry showed excellent correlation. This approach provides an in vivo method to detect Abeta in AD transgenic mice, and suggests that diagnostic MRI methods to detect Abeta in AD patients may ultimately be feasible.     

In vivo measurement of plaque burden in a mouse model of Alzheimer's disease

PURPOSE: To demonstrate an MRI method for directly visualizing amyloid-beta (Abeta) plaques in the APP/PS1 transgenic (tg) mouse brain in vivo, and show that T1rho relaxation rate increases progressively with Alzheimer's disease (AD)-related pathology in the tg mouse brain. MATERIALS AND METHODS: We obtained in vivo MR images of a mouse model of AD (APP/PS1) that overexpresses human amyloid precursor protein, and measured T1rho via quantitative relaxometric maps. RESULTS: A significant decrease in T1rho was observed in the cortex and hippocampus of 12- and 18-month-old animals compared to their age-matched controls. There was also a correlation between changes in T1rho and the age of the animals. CONCLUSION: T1rho relaxometry may be a sensitive method for noninvasively determining AD-related pathology in APP/PS1 mice.     

T2-weighted thin-section imaging with the multislab three-dimensional RARE technique.

A novel three-dimensional (3D) RARE (rapid acquisition with relaxation enhancement) sequence was implemented on a clinical imager. In this technique, multiple slabs are excited in the same way as in the multisection spin-echo sequence, and each slab is further phase encoded into eight sections along the section-slab direction. With a 16-echo RARE sequence, 128 excitations cover the 256 X 256 X 8 3D k space. With a TR of 2,500 msec, 10 slabs can be excited sequentially at each TR, yielding 80 sections in 5 minutes. Slabs were overlapped to give contiguous sections after discarding of the aliased sections at slab edges. This relatively fast sequence makes contiguous thin-section T2-weighted imaging possible, an impractical achievement with the much longer spin-echo method. Compared with 3D Fourier transform gradient-echo imaging, the sensitivity of 3D RARE sequences to magnetic susceptibility is reduced. The clinical potential of T2-weighted 3D imaging is illustrated with high-resolution brain, spine, and temporomandibular joint images.     

Determination of multiple sclerosis plaque size with diffusion-tensor MR Imaging: comparison study with healthy volunteers

PURPOSE: To use diffusion-tensor magnetic resonance (MR) imaging to measure involvement of normal-appearing white matter (WM) immediately adjacent to multiple sclerosis (MS) plaques and thus redefine actual plaque size on diffusion-tensor images through comparison with T2-weighted images of equivalent areas in healthy volunteers. MATERIALS AND METHODS: Informed consent was not required given the retrospective nature of the study on an anonymized database. The study complied with requirements of the Health Insurance Portability and Accountability Act. Twelve patients with MS (four men, eight women; mean age, 35 years) and 14 healthy volunteers (six men, eight women; mean age, 25 years) were studied. The authors obtained fractional anisotropy (FA) values in MS plaques and in the adjacent normal-appearing WM in patients with MS and in equivalent areas in healthy volunteers. They placed regions of interest (ROIs) around the periphery of plaques and defined the total ROIs (ie, plaques plus peripheral ROIs) as abnormal if their mean FA values were at least 2 standard deviations below those of equivalent ROIs within equivalent regions in healthy volunteers. The combined area of the plaque and the peripheral ROI was compared with the area of the plaque seen on T2-weighted MR images by means of a Student paired t test (P = .05). RESULTS: The mean plaque size on T2-weighted images was 72 mm2 +/- 21 (standard deviation). The mean plaque FA value was 0.285 +/- 0.088 (0.447 +/- 0.069 in healthy volunteers [P < .001]; mean percentage reduction in FA in MS plaques, 37%). The mean plaque size on FA maps was 91 mm2 +/- 35, a mean increase of 127% compared with the size of the original plaque on T2-weighted images (P = .03). CONCLUSION: A significant increase in plaque size was seen when normal-appearing WM was interrogated with diffusion-tensor MR imaging. This imaging technique may represent a more sensitive method of assessing disease burden and may have a future role in determining disease burden and activity.     

Thoracic aorta: rapid black-blood MR imaging with half-Fourier rapid acquisition with relaxation enhancement with or without electrocardiographic triggering.

PURPOSE: To evaluate and compare findings for thoracic aortic disease with three black-blood magnetic resonance (MR) pulse sequences: half-Fourier rapid acquisition with relaxation enhancement (RARE), with and without electrocardiographic (ECG) triggering, and ECG-triggered turbo spin echo (SE). MATERIALS AND METHODS: Axial black-blood MR images of the chest acquired at 1.5 T with a phased-array coil were obtained in 38 consecutive patients referred for evaluation of thoracic aortic disease. ECG-triggered and nontriggered half-Fourier RARE images were compared with T1-weighted ECG-triggered turbo SE images. Two readers independently scored images for each of the following parameters: ghosting artifacts; clarity of the mediastinum, cardiac chambers, and aortic wall; conspicuity of abnormality; intraluminal signal void uniformity; and overall image quality. RESULTS: Both half-Fourier RARE sequences outperformed the turbo SE sequence for all measured parameters. Scores for the ECG-triggered half-Fourier RARE sequence were significantly (P < .05) higher than those for the nontriggered version for clarity of the mediastinum and aortic wall, conspicuity of any abnormality other than aortic dissection, and overall image quality. Mean acquisition times for the ECG-triggered (48 seconds) and nontriggered (30 seconds) sequences were significantly shorter than that for the turbo SE sequence (2 minutes 20 seconds). CONCLUSION: Rapid black-blood half-Fourier RARE sequences, with or without ECG triggering, can replace ECG-triggered turbo SE sequences for evaluation of thoracic aortic disease.     

Transverse relaxation time reflects brain amyloidosis in young APP/PS1 transgenic mice.

Amyloid deposits are one of the hallmarks of Alzheimer's disease (AD), one of the most devastating neurodegenerative disorders. In transgenic mice modeling Alzheimer's pathology, the MR transverse relaxation time (T(2)) has been described to be modulated by amyloidosis. This modification has been attributed to the age-related iron deposition that occurs within the amyloid plaques of old animals. In the present study, young APP/PS1 transgenic mice without histochemically detectable iron in the brain were specifically studied. In vivo measurements of T(2) in the hippocampus, at the level of the subiculum, were shown to reflect the density of amyloid plaques. This suggests that T(2) variations can be induced solely by aggregated amyloid deposits in the absence of associated histologically-detectable iron. Thus T(2) from regions with high amyloid load, such as the subiculum, is particularly well suited for following plaque deposition in young animals, i.e., at the earliest stages of the pathological process.     

High-resolution magnetic resonance imaging at 2 Tesla: potential for atherosclerotic lesions exploration in the apolipoprotein E knockout mouse

INTRODUCTION: The aim of the present study was to evaluate the potential of high-resolution MRI at 2 Tesla (T) for direct noninvasive imaging of the aortic wall in a mouse model of atherosclerosis. MATERIAL AND METHODS: A specific mouse antenna was developed and sequence parameters were adjusted. T(1)- and T2-weighted images of abdominal aorta were obtained at 2 T with a spatial resolution of 86 x 86 x 800 microm3 in vivo. With a dedicated small coil, ex vivo MRI of the aorta was performed with a spatial resolution of 54 x 54 x 520 microm3. RESULTS: In vivo, the aortic wall was clearly defined on T(2)-weighted images in 15 of 16 mice: along the aorta the lumen circumference ranged from 1.07 to 3.61 mm and mean wall thickness from 0.11 to 0.67 mm. In vivo measurements of plaque distribution were confirmed by ex vivo MR imaging and by histology, with a good correlation with histology regarding lumen circumference (r = 0.94) and wall thickness (r = 0.97). CONCLUSION: Magnetic resonance imaging at 2 T to analyze in vivo atherosclerotic lesions in mice is possible with a spatial resolution of 86 x 86 x 800 microm3 and thus can be used for noninvasive follow-up in evaluation of new drugs.     

T1- and T2-weighted imaging at 8 Tesla

In this work, both T1- and T2-weighted fast imaging methods at 8 T are presented. These include the modified driven equilibrium Fourier transform (MDEFT) and rapid acquisition with relaxation enhancement (RARE) methods, respectively. Axial MDEFT images were acquired with large nutation angles, both partially suppressing gray and white matter and permitting the visualization of vascular structures rich in unsaturated spins. Sagittal RARE images, acquired from the same volunteer, were highly T2-weighted, thus highlighting the CSF. At the same time, they provided good visualization of the corpus callosum, cerebellum, and gray and white matter structures. Importantly, both MDEFT and RARE images could be acquired without violating specific absorption rate guidelines.     

Redefinition of multiple sclerosis plaque size using diffusion tensor MRI

OBJECTIVE: We used diffusion tensor MRI to redefine the size of multiple sclerosis (MS) plaques on fractional anisotropy (FA) maps. MATERIALS AND METHODS: Thirty-six white matter (WM) plaques were identified in 20 patients with MS. Plaque FA was measured by placing regions of interest (ROIs) on plaques on diffusion tensor images. We compared FA values in identical mirror-image ROIs placed on normal-appearing WM in the contralateral hemisphere. This comparison showed a mean decrease in FA of 41% in plaques, serving as the threshold for outlining abnormal regions in normal-appearing WM surrounding plaques. ROIs were placed around each plaque and FA values were compared with those in the mirror-image ROIs. Combined areas of perilesional normal-appearing WM with 40% or more FA reduction plus plaque were compared with the areas of abnormality on T2-weighted images using a paired Student's t test. A p value of 0.05 or less was considered significant. RESULTS: Mean plaque area was 60 mm(2) (range, 15-103 mm(2)), mean plaque FA was 0.251 (range, 0.133-0.436), and mean FA of contralateral normal-appearing WM was 0.429 (range, 0.204-0.712). Applying a threshold of 40% FA reduction, mean combined area of abnormal WM (including plaque seen on T2-weighted sequences) was 87 mm(2) (range, 30-251 mm(2)) or 145% of the mean plaque area that was seen on T2-weighted images (p < 0.001). CONCLUSION: Using an operator-defined threshold of abnormal FA values based on plaque anisotropy characteristics, we saw a statistically significant increase in plaque size.     

Characterization of aortic root atherosclerosis in ApoE knockout mice: high-resolution in vivo and ex vivo MRM with histological correlation.

In vivo, cardiac-gated, black-blood, and ex vivo magnetic resonance microscopy (MRM) images of the aortic root, and histopathology data were obtained from 12 transgenic and wild-type (WT) mice. MRM was performed using a black-blood imaging spin-echo sequence with upstream and downstream in-flow saturation pulses to obtain aortic root images in three contrast techniques: proton density-weighted (PDW), T(1)- (T(1)W), and T(2)-weighted (T(2)W). Aortic wall thickness and area were measured and correlated with histopathology data (R > 0.90). Ex vivo lesion components (lipid core, fibrous tissue, and cell tissue) were identified and characterized by differing image contrast in PDW, T(1)W, and T(2)W MRM, and by histopathology. The differences between WT and transgenic mice for maximal wall thickness and area were statistically significant (P < 0.05). This study demonstrates the feasibility of in vivo murine aortic root lesion assessment and ex vivo plaque characterization by MRM.     

Noninvasive in vivo MRI detection of neuritic plaques associated with iron in APP[V717I] transgenic mice, a model for Alzheimer's disease.

Transgenic mice overexpressing the London mutant of human amyloid precursor protein (APP[V717I]) in neurons develop amyloid plaques in the brain, thus demonstrating the most prominent neuropathological hallmark of Alzheimer's disease. In vivo 3D T2*-weighted MRI on these mice (24 months of age) revealed hypointense brain inclusions that affected the thalamus almost exclusively. Upon correlating these MRI observations with a panel of different histologic staining techniques, it appeared that only plaques that were positive for both thioflavin-S and iron were visible on the MR images. Numerous thioflavin-S-positive plaques in the cortex that did not display iron staining remained invisible to MRI. The in vivo detection of amyloid plaques in this mouse model, using the intrinsic MRI contrast arising from the iron associated with the plaques, creates an unexpected opportunity for the noninvasive investigation of the longitudinal development of the plaques in the same animal. Thus, this work provides further research opportunities for analyzing younger APP[V717I] mouse models with the knowledge of the final outcome at 24 months of age.     

Multicontrast-weighted magnetic resonance imaging of atherosclerotic plaques at 3.0 and 1.5 Tesla: ex-vivo comparison with histopathologic correlation

The purpose was to analyze magnetic resonance (MR) plaque imaging at 3.0 Tesla and 1.5 Tesla in correlation with histopathology. MR imaging (MRI) of the abdominal aorta and femoral artery was performed on seven corpses using T1-weighted, T2-weighted, and PD-weighted sequences at 3.0 and 1.5 Tesla. Cross-sectional images at the branching of the inferior mesenteric artery and the profunda femoris were rated with respect to image quality. Corresponding cross sections of the imaged vessels were obtained at autopsy. The atherosclerotic plaques in the histological slides and MR images were classified according to the American Heart Association (AHA) and analyzed for differences. MRI at 3.0 Tesla offered superior depiction of arterial wall composition in all contrast weightings, rated best for T2-weighted images. Comparing for field strength, the highest differences were observed in T1-weighted and T2-weighted techniques (both P< or =0.001), with still significant differences in PD-weighted sequence (P< or =0.005). The majority of plaques were histologically classified as calcified plaques. In up to 21% of the cases, MRI at both field strengths detected signal loss characteristic of calcification although calcified plaque was absent in histology. MRI at 3.0 Tesla offers superior plaque imaging quality compared with 1.5 Tesla, but further work is necessary to determine whether this translates in superior diagnostic accuracy.     

Passive staining: a novel ex vivo MRI protocol to detect amyloid deposits in mouse models of Alzheimer's disease.

Amyloid plaques are one of the hallmarks of Alzheimer's disease (AD). This study evaluated a novel microMRI strategy based on "passive staining" of brain samples by gadoteric acid. The protocol was tested at 4.7 T on control animals and APP/PS1 mice modeling AD lesions. T(1) was strongly decreased in passively stained brains. On high-resolution 3D gradient echo images, the contrast between the cortex and subcortical structures was highly improved due to a T2* effect. The brains of APP/PS1 mice revealed plaques as hypo-intense spots. They appeared larger in long compared to short TE images. This suggests that, after passive staining, plaques caused a susceptibility effect. This easily performed protocol is a complementary method to classic histology to detect the 3D location of plaques. It may also be used for the validation of in vivo MRI protocols for plaque detection by facilitating registration with histology via post mortem MRI.     

High-resolution, multicontrast three-dimensional-MRI characterizes atherosclerotic plaque composition in ApoE-/- mice ex vivo

PURPOSE: To systematically investigate intrinsic MR contrast mechanisms that would facilitate plaque characterization and quantification in the aortic root and brachiocephalic artery of ApoE-/- mice ex vivo. MATERIALS AND METHODS: To establish unambiguous MR parameters for routinely analyzing atherosclerotic plaque ex vivo at 11.7 T, relaxation times of plaque components were quantitatively assessed. Magnetization transfer and lipid-proton three-dimensional MR imaging was investigated for visualization of collagen- and lipid-rich plaque regions, respectively. A three-dimensional multiecho sequence with a spatial resolution of 47 x 47 x 63 microm was implemented providing a variable degree of T2-weighting. RESULTS: Relaxation time measurements showed clear tissue heterogeneity between atherosclerotic plaque components in the T2-values, but similar T1-values at 11.7 T (T1/T2 mean +/- SD; cellular plaque component: 1.2 +/- 0.3 seconds/26.3 +/- 0.4 msec; fibrofatty plaque component: 1.1 +/- 0.2 seconds/13.7 +/- 2.0 msec). The three-dimensional multiecho sequence allowed the calculation of the intrinsic proton density and T2-maps. The sum of the multiecho data provided strong T2-weighting that facilitated quantification of various components of atherosclerotic plaque in the mouse aortic root and correlated well with histology (P < 0.0001). CONCLUSION: High-resolution MRI allows for accurate classification and quantification of atherosclerotic plaque components in the aortic root of mice.     

Volume T1-weighted gradient echo MRI in multiple sclerosis patients.

Three-dimensional (3D) volume turbo fast low angle shot (FLASH) techniques have become available which produce heavily T1-weighted images, similar to inversion recovery scans, utilizing the appropriate flip angle and inversion time. The purpose of this study was to compare the sensitivity of a rapid volume gradient echo technique [3D magnetization prepared rapid acquisition gradient echo (MP RAGE)] in identifying multiple sclerosis (MS) plaques with a conventional T2-weighted spin echo (SE) sequence. Ten patients with clinical MS were evaluated. Patients underwent a routine examination consisting of an axial T2-weighted SE sequence (2,500,22/90) and a coronal 3D MP RAGE, 10/4/10, acquired as 128 two mm partitions. In six patients, area measurements of 22 plaques were determined on both the axial T2-weighted SE examinations and the axial reformatted MP RAGE examinations. The overall number of plaques utilizing each technique was approximately the same. One hundred twenty-two plaques were visualized for the 3D MP RAGE sequence, and 128 plaques for the T2-weighted SE sequence. There were differences in detection of plaques in different regions, with plaques in gray matter better demonstrated utilizing the conventional T2-weighted SE sequence. Plaques in the corpus callosum, pons, and brachia pontis were better demonstrated utilizing 3D MP RAGE. No significant difference was found between the areas measured on the MP RAGE sequence and on the T2-weighted SE sequence. Three-dimensional MP RAGE provides a sensitive and complementary method to conventional T2-weighted SE sequences in the evaluation of patients with MS.     

Hybrid RARE: Implementations for abdominal MR imaging

Hybrid RARE (rapid acquisition with relaxation enhancement) is a family of magnetic resonance (MR) imaging techniques whereby a set of images is phase encoded with more than one spin echo per excitation pulse. This increases the efficiency of obtaining T2-weighted images, allowing greater flexibility regarding acquisition time, resolution, signal-to-noise ratio, and tissue contrast. Hybrid RARE techniques involve several important new user-selectable parameters such as effective TE, echo train length, and echo spacing. Choices of other parameters, such as TR, sampling bandwidth, and acquisition matrix, may be different from those of comparable conventional T2-weighted spin-echo images. Different hybrid RARE implementations can be used for abdominal screening, with T2-weighted or T2-weighted and inversion-recovery contrast, or for characterizing liver lesions or imaging the biliary system with an extremely long TE. High-resolution images may be obtained by averaging multiple signals during quiet breathing, or images may be acquired more rapidly during suspended respiration. In this review, the authors discuss the basic principles of hybrid RARE techniques and how various imaging parameters can be manipulated to increase the quality and flexibility of abdominal T2-weighted MR imaging.