Erica Markiewicz

High-resolution magnetic resonance colonography and dynamic contrast-enhanced magnetic resonance imaging in a murine model of colitis

Inflammatory bowel disease, including ulcerative colitis, is characterized by persistent or recurrent inflammation and can progress to colon cancer. Colitis is difficult to detect and monitor noninvasively. The goal of this work was to develop a preclinical imaging method for evaluating colitis. Herein, we report improved MRI methods for detecting and characterizing colitis noninvasively in mice, using high‐resolution in vivo MR images and dynamic contrast‐enhanced MRI studies, which were confirmed by histologic studies in a murine model of colitis. C57Bl6/J male mice were treated with 2.5% dextran sulfate sodium in their drinking water for 5 days to induce colitis. MR images were acquired using a 9.4‐T Bruker scanner from 5–25 days following dextran sulfate sodium treatment. In dynamic contrast‐enhanced MRI studies, Gd uptake (Ktrans) and its distribution (ve) were measured in muscle and normal and inflamed colons after administering Gd‐diethyltriaminepentaacetic acid (Gd‐DTPA). T2‐weighted MR images distinguished normal colon from diffusely thickened colonic wall occurring in colitis (P <0.0005) and correlated with histologic features. Values of Ktrans and ve obtained from dynamic contrast‐enhanced MRI were also significantly different in inflamed colons compared to normal colon (P < 0.0005). The results demonstrate that both T2‐weighted anatomic imaging and quantitative analysis of dynamic contrast‐enhanced MRI data can successfully distinguish colitis from normal colon in mice. Magn Reson Med 63:922–929, 2010.

Sensitivity to tumor microvasculature without contrast agents in high spectral and spatial resolution MR images

Contrast‐enhanced (CE)‐MRI is sensitive to cancers but can produce adverse reactions and suffers from insufficient specificity and morphological detail. This research investigated whether high spectral and spatial resolution (HiSS) MRI detects tumor vasculature without contrast agents, based on the sensitivity of the water resonance line shape to tumor blood vessels. HiSS data from AT6.1 tumors inoculated in the hind legs of rats (N = 8) were collected pre‐ and post–blood pool contrast agent (iron‐oxide particles) injection. The waterline in small voxels was significantly more asymmetric at the tumor rim compared to the tumor center and normal muscle (P < 0.003). Composite images were synthesized, with the intensity in each voxel determined by the Fourier component (FC) of the water resonance having the greatest relative image contrast at that position. We tested whether regions with high contrast in FC images (FCIs) contain vasculature by comparing FCIs with CE‐MRI as the “gold standard” of vascular density. The FCIs had 75% ± 13% sensitivity, 74% ± 10% specificity, and 91% ± 4% positive predictive value (PPV) for vasculature detection at the tumor rim. These results suggest that tumor microvasculature can be detected using HiSS imaging without the use of contrast agents. Magn Reson Med 61:291–298, 2009.

Use of a reference tissue and blood vessel to measure the arterial input function in DCEMRI

Accurate measurement of the arterial input function is critical for quantitative evaluation of dynamic contrast enhanced magnetic resonance imaging data. Use of the reference tissue method to derive a local arterial input function avoided large errors associated with direct arterial measurements, but relied on literature values for Ktrans and ve. We demonstrate that accurate values of Ktrans and ve in a reference tissue can be measured by comparing contrast media concentration in a reference tissue to plasma concentrations measured directly in a local artery after the 1–2 passes of the contrast media bolus—when plasma concentration is low and can be measured accurately. The values of Ktrans and ve calculated for the reference tissue can then be used to derive a more complete arterial input function including the first pass of the contrast bolus. This new approach was demonstrated using dynamic contrast enhanced magnetic resonance imaging data from rodent hind limb. Values obtained for Ktrans and ve in muscle, and the shape and amplitude of the derived arterial input function are consistent with published results. Magn Reson Med, 2010.

Magnetic resonance imaging of the natural history of in situ mammary neoplasia in transgenic mice: a pilot study

IntroductionBecause of the small size of in situ mammary cancers in mouse models, high-resolution imaging techniques are required to effectively observe how lesions develop, grow and progress over time. The purpose of this study was to use magnetic resonance (MR) imaging to track in vivo the transition from in situ neoplasia to invasive cancer in a transgenic mouse model of human cancer.MethodsMR images of 12 female C3(1) SV40 Tag mice that develop mammary intraepithelial neoplasia (MIN) were obtained. MIN is believed to be similar to human ductal carcinoma in situ (DCIS) and is considered a precursor of invasive tumors. Images were serially obtained from 10-21 weeks of age at 2-3 week intervals. MIN lesions were identified based on their morphology on MR images. Lesions were followed over time and several lesion features were measured including volume, growth rate and morphology. For those MIN lesions that progressed to invasive cancer the progression time was measured.ResultsOverall, 21 MIN lesions were initially detected at an average initial volume of 0.3 ± 0.2 mm3 with an average growth rate of -0.15 ± 0.66 week-1. Even though all mice were inbred to express the SV40 Tag transgene in the mammary epithelium and expected to develop invasive carcinoma, the individual MIN lesions took vastly different progression paths: (i) 9 lesions progressed to invasive tumors with an average progression time of 4.6 ± 1.9 weeks; (ii) 2 lesions regressed, i.e., were not detected on future images; and (iii) 5 were stable for over 8 weeks, and were demonstrated by a statistical model to represent indolent disease.ConclusionsTo our knowledge, the results reported here are the first measurements of the timescale and characteristics of progression from in situ neoplasia to invasive carcinoma and provide image-based evidence that DCIS may be a non-obligate precursor lesion with highly variable outcomes. In addition, this study represents a first step towards developing methods of image acquisition for identifying radiological characteristics that might predict which in situ neoplasias will become invasive cancers and which are unlikely to progress.

Comparison and evaluation of mouse cardiac MRI acquired with open birdcage, single loop surface and volume birdcage coils

Although the quality and speed of MR images have vastly improved with the development of novel RF coil technologies, the engineering expertise required to implement them is often not available in many animal in vivo MR laboratories. We present here an open birdcage coil design which is easily constructed with basic RF coil expertise and produces high quality images. The quality and advantages of mouse cardiac MR images acquired with open birdcage coils were evaluated and compared to images acquired with a bent single loop surface, and standard birdcage coils acquired at 4.7 Tesla. Two low pass open birdcage coils, two single loop surface coils, and a low pass volume birdcage coil were constructed and their B(1) distributions were evaluated and compared. The calculated average signal-to-noise ratio for the left ventricular wall was 10, 23 and 32 for the volume birdcage coil, single loop surface coil and open birdcage coil, respectively. The results demonstrate that the open birdcage coil provides greater sensitivity than the volume coil and a higher signal/contrast-to-noise ratio and B(1) homogeneity than the single loop surface coil. The open birdcage coil offers easy access and better quality mouse cardiac imaging than both the single loop surface coil and volume birdcage coil and does not require extensive RF engineering expertise to construct.

High spectral and spatial resolution MRI of age related changes in murine prostate

The purpose of this work was to evaluate high‐resolution echo‐planar spectroscopic MRI of normal and precancerous prostatic changes in a transgenic mouse line. Simian virus large T‐antigen transgenic male mice (N = 7, age = 34 ± 3.7 weeks) with prostatic hyperplasia and intraepithelial neoplasia (PIN) were studied. High spectral and spatial resolution (HiSS) MRI of the water proton signal was compared to the free induction decay (FID) integral image and conventional gradient‐echo and spin‐echo imaging. Water peak‐height images of the prostate produced from HiSS datasets showed improved contrast‐to‐noise ratio (CNR) (P < 0.03), and greater morphological detail (P < 0.004) based on texture analysis. Despite the high spectral resolution of the HiSS datasets, signal‐to‐noise ratio (SNR) compared favorably with that of the FID integral and conventional images. Lobular features in HiSS images of older mice were consistent with hyperplasia seen on histology. A partially deuterated water‐filled catheter was inserted in the mouse rectum for susceptibility matching between the colon interior and exterior to minimize image artifacts. These preliminary results suggest that HiSS MRI provides detailed morphology of the murine prostate and can detect early changes associated with the development of cancer. HiSS MRI of patients may have similar advantages. Magn Reson Med 60:575–581, 2008.

In vivo MRI of early stage mammary cancers and the normal mouse mammary gland

Since the advent of screening mammography, approximately one‐quarter of newly diagnosed breast cancers are at the earliest preinvasive stage of ductal carcinoma in situ (DCIS). Concomitant with this improvement in early detection has been a growing clinical concern that distinguishing aggressive from indolent DCIS is necessary to optimize patient management. Genetically engineered mouse models offer an appealing experimental framework in which to investigate factors that influence and predict progression of preinvasive neoplasias. Because of the small size of early stage carcinomas in mice, high‐resolution imaging techniques are required to effectively observe longitudinal progression. The purpose of the present study was to evaluate the feasibility of MRI for assessment of in situ mammary neoplasias and early invasive mammary cancers that stochastically arise in mammary glands of C3(1) SV40 Tag transgenic mice. Additionally, images of normal mammary glands from wild‐type FVB/N mice were acquired and compared with those from transgenic mice. Sixteen mice underwent MR examinations employing axial two‐dimensional multi‐slice gradient recalled echo scans (TR/TE =∼1000/5.5 ms) with fat suppression in a two‐step process targeting both the upper and lower mammary glands. MRI successfully detected in situ and early invasive neoplasias in transgenic mice with high sensitivity and specificity. The average signal‐to‐noise ratio (SNR) of in situ lesions on fat‐suppressed high‐resolution T1‐weighted images was 22.9, which was lower than that of invasive tumors, lymph nodes and muscle (average SNR of 29.5–34.9, p < 0.0001) but significantly higher than that of normal mammary tissue (average SNR = 5.5, p < 0.0001). Evaluation of wild‐type mammary glands revealed no cancerous or benign lesions, and comparable image contrast characteristics (average SNR = 5.2) as compared with normal tissue areas of transgenic mice. This present study demonstrates that MRI is an excellent candidate for performing longitudinal assessment of early stage mammary cancer disease progression and response to therapy in the transgenic model system. Copyright

MRI accurately identifies early murine mammary cancers and reliably differentiates between in situ and invasive cancer: correlation of MRI with histology

MRI methods that accurately identify various stages of mouse mammary cancer could provide new knowledge that may have a direct impact on the management of breast cancer in patients. This research investigates whether we can accurately follow the progression from in situ to invasive cancer by the evaluation of in vivo and ex vivo MRI, and in comparison with histology as the gold standard for the diagnosis and staging of cancer. Six C3(1)SV40Tag virgin female mice, aged 12–16 weeks, were studied. At this age, these mice develop in situ cancer that resembles human ductal carcinoma in situ (DCIS). Fast spin‐echo images of inguinal mammary glands were acquired at 9.4 T. After in vivo MRI, mice were sacrificed; inguinal mammary glands were excised and fixed in formalin for ex vivo MRI. Three‐dimensional, volume‐rendered, in vivo and ex vivo MR images were then correlated with histology. High‐resolution ex vivo scans facilitated the comparison of in vivo scans with histology. The sizes of mammary cancers classified as in situ on the basis of histology ranged from 150 to 400 µm in largest diameter, and the average signal intensity relative to muscle was 1.40 ± 0.18 on T2‐weighted images. Cancers classified as invasive on the basis of histology were >400 µm in largest diameter, and the average intensity relative to muscle on T2‐weighted images was 2.34 ± 0.26. Using a cut‐off of 400 µm in largest diameter to distinguish between in situ and invasive cancers, a T2‐weighted signal intensity of at least 1.4 times that of muscle for in situ cancer, and at least 2.3 times that of muscle for invasive cancer, 96% of in situ and 100% of invasive cancers were correctly identified on in vivo MRI, using histology as the gold standard. Precise MRI–histology correlation demonstrates that MRI reliably detects early in situ cancer and differentiates in situ from invasive cancers in the SV40Tag mouse model of human breast cancer. Copyright

Mammary cancer initiation and progression studied with magnetic resonance imaging.

IntroductionPrevious work from this laboratory demonstrated that magnetic resonance imaging (MRI) detects early murine mammary cancers and reliably differentiates between in situ and invasive cancer. Based on this previous work, we used MRI to study initiation and progression of murine mammary cancer, and monitor the transition from the in situ to the invasive phase.MethodsIn total, seven female C3(1) SV40 Tag mice were imaged every two weeks between the ages of 8 to 23 weeks. Lesions were identified on T2-weighted images acquired at 9.4 Tesla based on their morphology and growth rates. Lesions were traced manually on MR images of each slice. Volume of each lesion was calculated by adding measurements from individual slices. Plots of lesion volume versus time were analyzed to obtain the specific growth rate (SGR). The time at which in situ cancers (referred to as ‘mammary intraepithelial neoplasia (MIN)’) and invasive cancers were first detected; and the time at which in situ cancers became invasive were recorded.ResultsA total of 121 cancers (14 to 25 per mouse) were identified in seven mice. On average the MIN lesions and invasive cancers were first detected when mice were 13 and 18 weeks old, respectively. The average SGR was 0.47 ± 0.18 week-1 and there were no differences (P >0.05) between mice. 74 lesions had significantly different tumor growth rates before and after ~17 weeks of age; with average doubling times (DT) of 1.88 and 1.27 weeks, respectively. The average DT was significantly shorter (P <0.0001) after 17 weeks of age. However, the DT for some cancers was longer after 17 weeks of age, and about 10% of the cancers detected did not progress to the invasive stage.ConclusionsA wide range of growth rates were observed in SV40 mammary cancers. Most cancers transitioned to a more aggressive phenotype at approximately 17 weeks of age, but some cancers became less aggressive. The results suggest that the biology of mammary cancers is extremely heterogeneous. This work is a first step towards use of MRI to improve understanding of factors that control and/or signal the development of aggressive breast cancer.

MRI of neonatal necrotizing enterocolitis in a rodent model

Neonatal necrotizing enterocolitis (NEC) is a poorly understood life‐threatening illness afflicting premature infants. Research is hampered by the absence of a suitable method to monitor disease progression noninvasively. The primary goal of this research was to test in vivo MRI methods for the noninvasive early detection and staging of inflammation in the ileum of an infant rat model of NEC. Neonatal rats were delivered by cesarean section at embryonic stage of day 20 after the beginning of pregnancy and stressed with formula feeding, hypoxia and bacterial colonization to induce NEC. Naturally born and dam‐fed neonatal rats were used as healthy controls. In vivo MRI studies were performed using a Bruker 9.4‐T scanner to obtain high‐resolution anatomical MR images using both gradient echo and spin echo sequences, pixel‐by‐pixel T2 maps using a multi‐slice–multi‐echo sequence, and maps of the apparent diffusion coefficient (ADC) of water using a spin echo sequence, to assess the degree of ileal damage. Pups were sacrificed at the end of the MRI experiment on day 2 or 4 for histology. T2 measured by MRI was increased significantly in the ileal regions of pups with NEC by histology (106.3 ± 6.1 ms) compared with experimentally stressed pups without NEC (85.2 ± 6.8 ms) and nonstressed, control rat pups (64.9 ± 2.3 ms). ADC values measured by diffusion‐weighted MRI were also increased in the ileal regions of pups with NEC by histology [(1.98 ± 0.15) × 10–3 mm2/s] compared with experimentally stressed pups without NEC [(1.43 ± 0.16) × 10–3 mm2/s] and nonstressed control pups [(1.10 ± 0.06) × 10–3 mm2/s]. Both T2 and ADC values between these groups were found to be significantly different (p < 0.03). The correlation of MRI results with histologic images of the excised ileal tissue samples strongly suggests that MRI can noninvasively identify NEC and assess intestinal injury prior to clinical symptoms in a physiologic rat pup model of NEC.