Characterization of microparticles of iron oxide for magnetic resonance imaging.

Microparticles of iron oxide (MPIOs) are increasingly used for contrast generation in magnetic resonance imaging (MRI). In particular, Dynabeads® MyOne™ Tosylactivated MPIOs have enabled sensitive and targeted molecular imaging, e.g., to detect vascular inflammation. For the first time we measured the relaxivities as well as the molar susceptibility χM of these MPIOs at 7 T in agarose gels. They are r1 = 0.69 ± 0.03 s-1/mM, r2 = 220 ± 6 s-1/mM, r2* = 679 ± 14 s-1/mM, and χM = 0.66 ± 0.05 ppm/mM, when expressed with respect to the iron concentration. These material parameters are essential to optimize MRI protocols and progress toward quantitative imaging. To address the heterogeneous nature of the MPIO distributions over the size of a typical MRI voxel, we coupled the MPIOs to a fluorophore to create a bimodal phantom that can be imaged by both Light Sheet microscopy and MRI. In this phantom, the MPIOs produced contrast similar to that found in vivo . The submicron resolution of Light Sheet microscopy images provided a precise measurement of the MPIO spatial distribution in phantoms also imaged by MRI. MPIO aggregates occupying less than one MRI voxel were responsible for alterations in R2* and magnetic susceptibility χ across several MRI voxels. In these cases, the sum of R2* or χ over the affected MRI volume correlated better with the microscopically determined number of MPIOs. These findings were confirmed with simulations performed in the static dephasing regime. The microscopically determined MPIO distribution was also entered directly into the simulation framework, indicating that the bimodal phantom is a useful tool to test theoretical models against experimental measurements.

Brain irradiation leads to persistent neuroinflammation and long-term neurocognitive dysfunction in a region-specific manner.

Long-term cognitive deficits are observed after treatment of brain tumors or metastases by radiotherapy. Treatment optimization thus requires a better understanding of the effects of radiotherapy on specific brain regions, according to their sensitivity and interconnectivity. In the present study, behavioral tests supported by immunohistology and magnetic resonance imaging provided a consistent picture of the persistent neurocognitive decline and neuroinflammation after the onset of irradiation-induced necrosis in the right primary somatosensory cortex of Fischer rats. Necrosis surrounded by neovascularization was first detected 54 days after irradiation and then spread to 110 days in the primary motor cortex, primary somatosensory region, striatum and right ventricle, resulting in fiber bundle disruption and demyelination in the corpus callosum of the right hemisphere. These structural damages translated into selective behavioral changes including spatial memory loss, disinhibition of anxiety-like behaviors, hyperactivity and pain hypersensitivity, but no significant alteration in motor coordination and grip strength abilities. Concomitantly, activated microglia and reactive astrocytes, accompanied by infiltration of leukocytes (CD45+) and T-cells (CD3+) cooperated to shape the neuroinflammation response. Overall, our study suggests that the slow and gradual onset of cellular damage would allow adaptation in brain regions that are susceptible to neuronal plasticity; while other cerebral structures that do not have this capacity would be more affected. The planning of radiotherapy, adjusted to the sensitivity and adaptability of brain structures, could therefore preserve certain neurocognitive functions; while higher doses of radiation could be delivered to brain areas that can better adapt to this treatment. In addition, strategies to block early post-radiation events need to be explored to prevent the development of long-term cognitive dysfunction.

Cerebrovascular inflammation promotes the formation of brain metastases.

Brain metastases are the most prevalent intracranial malignancy. Patient outcome is poor and treatment options are limited. Hence, new avenues must be explored to identify potential therapeutic targets. Inflammation is a known critical component of cancer progression. Intratumoral inflammation drives progression and leads to the release of circulating tumor cells (CTCs). Inflammation at distant sites promotes adhesion of CTCs to the activated endothelium and then initiates the formation of metastases. These interactions mostly involve cell adhesion molecules expressed by activated endothelial cells. For example, the vascular cell adhesion molecule-1 (VCAM-1) is known to promote transendothelial migration of cancer cells in different organs. However, it is unclear whether a similar mechanism occurs within the specialized environment of the brain. Our objective was therefore to use molecular imaging to assess the potential role of VCAM-1 in promoting the entry of CTCs into the brain. First, magnetic resonance imaging (MRI) and histological analyses revealed that cerebrovascular inflammation induced by intracranial injection of lipopolysaccharide significantly increased the expression of VCAM-1 in the Balb/c mouse brain. Next, intracardiac injection of 4T1 mammary carcinoma cancer cells in animals with cerebrovascular inflammation yielded a higher brain metastasis burden than in the control animals. Finally, blocking VCAM-1 prior to 4T1 cells injection prevented this increased metastatic burden. Here, we demonstrated that by contributing to CTCs adhesion to the activated cerebrovascular endothelium, VCAM-1 improves the capacity of CTCs to form metastatic foci in the brain.

Cerebrovascular MRI in the mouse without an exogenous contrast agent.

PURPOSE: To assess the effect of a variety of anesthetic regimes on T2∗ -weighted MRI of the mouse brain and to determine the optimal regimes to perform T2∗ -weighted MRI of the mouse cerebrovasculature without a contrast agent. METHODS: Twenty mice were imaged with a 3D T2∗ -weighted sequence under isoflurane, dexmedetomidine, or ketamine-xylazine anesthesia with a fraction of inspired oxygen varied between 10% and 95% + 5% CO2 . Some mice were also imaged after an injection of an iron oxide contrast agent as a positive control. For every regime, whole brain vessel conspicuity was visually assessed and the apparent vessel density in the cortex was quantified and compared. RESULTS: The commonly used isoflurane anesthetic leads to poor vessel conspicuity for fraction of inspired oxygen higher or equal to 21%. Dexmedetomidine and ketamine-xylazine enable the visualization of a significantly larger portion of the vasculature for the same breathing gas. Under isoflurane anesthesia, the fraction of inspired oxygen must be lowered to between 10% and 14% to obtain similar vessel conspicuity. Initial results on automatic segmentation of veins and arteries using the iron oxide positive control are also reported. CONCLUSION: T2∗ -weighted MRI in combination with an appropriate anesthetic regime can be used to visualize the mouse cerebrovasculature without a contrast agent. The differences observed between regimes are most likely caused by blood-oxygen level dependent effects, highlighting the important impact of the anesthetic regimes on cerebral blood oxygenation of the mouse brain at rest.

Automated detection and quantification of breast cancer brain metastases in an animal model using democratized machine learning tools.

Advances in digital whole-slide imaging and machine learning (ML) provide new opportunities for automated examination and quantification of histopathological slides to support pathologists and biologists. However, implementation of ML tools often requires advanced skills in computer science that may not be immediately available in the traditional wet-lab environment. Here, we propose a simple and accessible workflow to automate detection and quantification of brain epithelial metastases on digitized histological slides. We leverage 100 Hematoxylin & Eosin (H&E)-stained whole slide images (WSIs) from 25 Balb/c mice with various level of brain metastatic tumor burden. A supervised training of the Trainable Weka Segmentation (TWS) from Fiji was achieved from annotated WSIs. Upon comparison with manually drawn regions, it is apparent that the algorithm learned to identify and segment cancer cell-specific nuclei and normal brain tissue. Our approach resulted in a robust and highly concordant correlation between automated metastases quantification of brain metastases and manual human assessment (R2 = 0.8783; P < 0.0001). This simple approach is amenable to other similar analyses, including that of human tissues. Widespread adoption of these tools aims to democratize ML and improve precision in traditionally qualitative tasks in histopathology-based research.

Determination of an Optimal Pharmacokinetic Model of 18F-FET for Quantitative Applications in Rat Brain Tumors.

O-(2-18F-fluoroethyl)-l-tyrosine (18F-FET) is a radiolabeled artificial amino acid used in PET for tumor delineation and grading. The present study compares different kinetic models to determine which are more appropriate for 18F-FET in rats. Methods: Rats were implanted with F98 glioblastoma cells in the right hemisphere and scanned 9-15 d later. PET data were acquired during 50 min after a 1-min bolus of 18F-FET. Arterial blood samples were drawn for arterial input function determination. Two compartmental pharmacokinetic models were tested: the 2-tissue model and the 1-tissue model. Their performance at fitting concentration curves from regions of interest was evaluated using the Akaike information criterion, F test, and residual plots. Graphical models were assessed qualitatively. Results: Metrics indicated that the 2-tissue model was superior to the 1-tissue model for the current dataset. The 2-tissue model allowed adequate decoupling of 18F-FET perfusion and internalization by cells in the different regions of interest. Of the 2 graphical models tested, the Patlak plot provided adequate results for the tumor and brain, whereas the Logan plot was appropriate for muscles. Conclusion: The 2-tissue-compartment model is appropriate to quantify the perfusion and internalization of 18F-FET by cells in various tissues of the rat, whereas graphical models provide a global measure of uptake.

Understanding the continuum of radionecrosis and vascular disorders in the brain following gamma knife irradiation: An MRI study.

PURPOSE: The radiation dose delivered to brain tumors is limited by the possibility to induce vascular damage and necrosis in surrounding healthy tissue. In the present study, we assessed the ability of MRI to monitor the cascade of events occurring in the healthy rat brain after stereotactic radiosurgery, which could be used to optimize the radiation treatment planning. METHODS: The primary somatosensory forelimb area (S1FL) and the primary motor cortex in the right hemisphere of Fischer rats (n = 6) were irradiated with a single dose of Gamma Knife radiation (Leksell Perfexion, Elekta AG, Stockholm, Sweden). Rats were scanned with a small-animal 7 Tesla MRI scanner before treatment and 16, 21, 54, 82, and 110 days following irradiation. At every imaging session, T2 -weighted (T2 w), Gd-DTPA dynamic contrast-enhanced MRI (DCE-MRI), and T2*-weighted ( T2* w) images were acquired to measure changes in fluid content, blood vessel permeability, and structure, respectively. At days 10, 110, and 140, histopathology was performed on brain sections. Locomotion and spatial memory ability were assessed longitudinally by behavioral tests. RESULTS: No vascular changes were initially observed. After 54 days, a small necrotic volume in the white matter below the S1FL, surrounded by an area presenting significant vascular permeability, was revealed. Between 54 and 110 days, the necrotic volume increased and was accompanied by the formation of a ring-like region, where a mixture of necrosis and permeable blood vessels were observed, as confirmed by histology. Behavioral changes were only observed after day 82. CONCLUSION: Together, DCE-MRI and T2* w images supported by histology provided a coherent picture of the phenomena involved in the formation of new, leaky blood vessels, which was followed by the detection of radionecrosis in a preclinical model of brain irradiation. Magn Reson Med 78:1420-1431, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

MRI-Guided Derivation of the Input Function for PET Kinetic Modeling.

Blood samples obtained by arterial cannulation are the gold standard to measure the input function for PET pharmacokinetic modeling. There is interest in less invasive methods, such as image-derived input functions (IDAIF). MRI can be used to segment and correct partial volume effects of the PET images, improving IDAIF extraction. Preclinical studies have shown that the input function of PET tracers, namely fluorodeoxyglucose and [(18)F]fluoroethyl-l-tyrosine, can be derived from the Gd-DTPA input function. Noninvasive, MRI-guided, PET input function derivation is a promising avenue to reduce or eliminate the need for arterial plasma samples in preclinical and clinical settings.