Praveen Gulaka

Proton imaging of siloxanes to map tissue oxygenation levels (PISTOL): A tool for quantitative tissue oximetry

Hexamethyldisiloxane (HMDSO) has been identified as a sensitive proton NMR indicator of tissue oxygenation (pO2) based on spectroscopic spin‐lattice relaxometry. A rapid MRI approach has now been designed, implemented, and tested. The technique, proton imaging of siloxanes to map tissue oxygenation levels (PISTOL), utilizes frequency‐selective excitation of the HMDSO resonance and chemical‐shift selective suppression of residual water signal to effectively eliminate water and fat signals and pulse‐burst saturation recovery 1H echo planar imaging to map T1 of HMDSO and hence pO2. PISTOL was used here to obtain maps of pO2 in rat thigh muscle and Dunning prostate R3327 MAT‐Lu tumor‐implanted rats. Measurements were repeated to assess baseline stability and response to breathing of hyperoxic gas. Each pO2 map was obtained in 3½ min, facilitating dynamic measurements of response to oxygen intervention. Altering the inhaled gas to oxygen produced a significant increase in mean pO2 from 55 Torr to 238 Torr in thigh muscle and a smaller, but significant, increase in mean pO2 from 17 Torr to 78 Torr in MAT‐Lu tumors. Thus, PISTOL enabled mapping of tissue pO2 at multiple locations and dynamic changes in pO2 in response to intervention. This new method offers a potentially valuable new tool to image pO2 in vivo for any healthy or diseased state by 1H MRI. Copyright

Synthesis and Characterization of a Hypoxia‐Sensitive MRI Probe

Tissue hypoxia occurs in pathologic conditions, such as cancer, ischemic heart disease and stroke when oxygen demand is greater than oxygen supply. An imaging method that can differentiate hypoxic versus normoxic tissue could have an immediate impact on therapy choices. In this work, the gadolinium(III) complex of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) with a 2-nitroimidazole attached to one carboxyl group via an amide linkage was prepared, characterized and tested as a hypoxia-sensitive MRI agent. A control complex, Gd(DO3A-monobutylamide), was also prepared in order to test whether the nitroimidazole side-chain alters either the water proton T(1) relaxivity or the thermodynamic stability of the complex. The stabilities of these complexes were lower than that of Gd(DOTA)(-) as expected for mono-amide derivatives. The water proton T(1) relaxivity (r(1)), bound water residence lifetime (τ(M)) and rotational correlation time (τ(R)) of both complexes was determined by relaxivity measurements, variable temperature (17) O NMR spectroscopy and proton nuclear magnetic relaxation dispersion (NMRD) studies. The resulting parameters (r(1) =6.38 mM(-1)  s(-1) at 20 MHz, τ(M) =0.71 μs, τ(R) =141 ps) determined for the nitroimidazole derivative closely parallel to those of other Gd(DO3A-monoamide) complexes of similar molecular size. In vitro MR imaging experiments with 9L rat glioma cells maintained under nitrogen (hypoxic) versus oxygen (normoxic) gas showed that both agents enter cells but only the nitroimidazole derivative was trapped in cells maintained under N(2) as evidenced by an approximately twofold decrease in T(1) measured for hypoxic cells versus normoxic cells exposed to this agent. These results suggest that the nitroimidazole derivative might serve as a molecular reporter for discriminating hypoxic versus normoxic tissues by MRI.

Hexamethyldisiloxane-based nanoprobes for 1H MRI oximetry

Quantitative in vivo oximetry has been reported using 19F MRI in conjunction with reporter molecules, such as perfluorocarbons, for tissue oxygenation (pO2). Recently, hexamethyldisiloxane (HMDSO) has been proposed as a promising alternative reporter molecule for 1H MRI‐based measurement of pO2. To aid biocompatibility for potential systemic administration, we prepared various nanoemulsion formulations using a wide range of HMDSO volume fractions and HMDSO to surfactant ratios. Calibration curves (R1 versus pO2) for all emulsion formulations were found to be linear and similar to neat HMDSO for low surfactant concentrations (< 10% v/v). A small temperature dependence in the calibration curves was observed, similar to previous reports on neat HMDSO, and was characterized to be approximately 1 Torr/ °C under hypoxic conditions. To demonstrate application in vivo, 100 µL of this nanoemulsion was administered to healthy rat thigh muscle (Fisher 344, n = 6). Dynamic changes in mean thigh tissue pO2 were measured using the PISTOL (proton imaging of siloxanes to map tissue oxygenation levels) technique in response to oxygen challenge. Changing the inhaled gas to oxygen for 30 min increased the mean pO2 significantly (p < 0.001) from 39 ± 7 to 275 ± 27 Torr. When the breathing gas was switched back to air, the tissue pO2 decreased to a mean value of 45 ± 6 Torr, not significantly different from baseline (p > 0.05), in 25 min. A first‐order exponential fit to this part of the pO2 data (i.e. after oxygen challenge) yielded an oxygen consumption‐related kinetic parameter k = 0.21 ± 0.04 min−1. These results demonstrate the feasibility of using HMDSO nanoemulsions as nanoprobes of pO2 and their utility to assess oxygen dynamics in vivo, further developing quantitative 1H MRI oximetry. Copyright

Free-Breathing Phase Contrast MRI with Near 100% Respiratory Navigator Efficiency using k-space Dependent Respiratory Gating

To investigate the efficacy of a novel respiratory motion scheme, where only the center of k‐space is gated using respiratory navigators, versus a fully respiratory‐gated acquisition for three‐dimensional flow imaging.

Novel S-Gal® analogs as 1H MRI reporters for in vivo detection of β-galactosidase

The quantitative assessment of gene expression and related enzyme activity in vivo could be important for the characterization of gene altering diseases and therapy. The development of imaging techniques, based on specific reporter molecules may enable routine non-invasive assessment of enzyme activity and gene expression in vivo. We recently reported the use of commercially available S-Gal(®) as a β-galactosidase reporter for (1)H MRI, and the synthesis of several S-Gal(®) analogs with enhanced response to β-galactosidase activity. We have now compared these analogs in vitro and have identified the optimal analog, C3-GD, based on strong T1 and T2 response to enzyme presence (ΔR1 and ΔR2~1.8 times S-Gal(®)). Moreover, application is demonstrated in vivo in human breast tumor xenografts. MRI studies in MCF7-lacZ tumors implanted subcutaneously in athymic nude mice (n=6), showed significant reduction in T1 and T2 values (each~13%) 2h after intra-tumoral injection of C3-GD, whereas the MCF7 (wild type) tumors showed slight increase. Thus, C3-GD successfully detects β-galactosidase activity in vivo and shows promise as a lacZ gene (1)H MR reporter molecule.

Dual-modality, dual-functional nanoprobes for cellular and molecular imaging.

An emerging need for evaluation of promising cellular therapies is a non-invasive method to image the movement and health of cells following transplantation. However, the use of a single modality to serve this purpose may not be advantageous as it may convey inaccurate or insufficient information. Multi-modal imaging strategies are becoming more popular for in vivo cellular and molecular imaging because of their improved sensitivity, higher resolution and structural/functional visualization. This study aims at formulating Nile Red doped hexamethyldisiloxane (HMDSO) nanoemulsions as dual modality (Magnetic Resonance Imaging/Fluorescence), dual-functional (oximetry/detection) nanoprobes for cellular and molecular imaging. HMDSO nanoprobes were prepared using a HS15-lecithin combination as surfactant and showed an average radius of 71±39 nm by dynamic light scattering and in vitro particle stability in human plasma over 24 hrs. They were found to readily localize in the cytosol of MCF7-GFP cells within 18 minutes of incubation. As proof of principle, these nanoprobes were successfully used for fluorescence imaging and for measuring pO2 changes in cells by magnetic resonance imaging, in vitro, thus showing potential for in vivo applications.

Improved efficiency for respiratory motion compensation in three-dimensional flow measurements

Background Phase contrast (PC) CMR is clinically used for in-vivo assessment of blood flow in cardiovascular disease [1]. Typically, a through-plane 2D acquisition is performed for evaluating the blood flow. Recently, 3D time-resolved PC CMR has been used for quantification and visualization of the blood flow in all three directions of a volume [2]. However, such acquisitions require long scan times, which are further prolonged by the need for respiratory motion compensation, typically using respiratory navigators. In this study, we hypothesized that respiratory gating the center of k-space only will yield similar measurements to a fully respiratory-gated acquisition, since the phase information is mainly coming from the central k-space, and evaluated these two gating approaches.