Comparison of BOLD contrast and Gd-DTPA dynamic contrast-enhanced imaging in rat prostate tumor.

The microcirculation and oxygenation of a tumor play important roles in its responsiveness to cytotoxic treatment, and noninvasive assessments of its vascular properties may have prognostic value. Dynamic contrast-enhanced (DCE) (1)H MRI based on infusion of Gd-DTPA, and blood oxygen level-dependent (BOLD) contrast based on altering inhaled gas are both sensitive to vascular characteristics. This study compares the effects observed in eight Dunning prostate R3327-AT1 rat tumors imaged sequentially at 4.7 Tesla by echo-planar imaging (EPI). Both interventions generated a significant response, and each revealed significant differences between the center and periphery of the tumors. On a voxel-by-voxel basis across the whole tumor population, there was a close correlation between the maximum rate of signal response and the magnitude of response to each intervention (R(2) >or= 0.6, P < 0.0001). However, when the data were analyzed separately for each individual tumor, some showed a weak correlation (R(2) < 0.4), particularly for DCE, and the nature (slope) varied between separate tumors. Generally, there was a weak correlation (N = 7, R(2) < 0.5) between responses to the two interventions on a tumor-by-tumor basis, which emphasizes that the techniques are not equivalent. Both techniques revealed intra- and intertumor heterogeneity, but the BOLD response was more rapidly reversible than the DCE response. This suggests that the BOLD technique may be a useful tool for investigating interventions (such as drugs) that cause vascular disruption.

Biodistribution of phosphodiester and phosphorothioate siRNA.

Short interfering RNAs (siRNAs) are valuable tools for analyzing protein function in mammalian cell culture. This success has led to high expectations for in vivo and therapeutic applications. However, the pharmacokinetic properties of siRNA are not known. Here we report the biodistribution of a phosphodiester (PO) siRNA duplex and examine the effect of phosphorothioate (PS) linkages. Our findings indicate that biodistribution of siRNA is similar to that for single-stranded antisense oligonucleotides and offer insights for use of siRNA in vivo.

The birth of a quasiparticle in silicon observed in time-frequency space.

The concept of quasiparticles in solid-state physics is an extremely powerful tool for describing complex many-body phenomena in terms of single-particle excitations. Introducing a simple particle, such as an electron, hole or phonon, deforms a many-body system through its interactions with other particles. In this way, the added particle is 'dressed' or 'renormalized' by a self-energy cloud that describes the response of the many-body system, so forming a new entity--the quasiparticle. Using ultrafast laser techniques, it is possible to impulsively generate bare particles and observe their subsequent dressing by the many-body interactions (that is, quasiparticle formation) on the time and energy scales governed by the Heisenberg uncertainty principle. Here we describe the coherent response of silicon to excitation with a 10-femtosecond (10(-14) s) laser pulse. The optical pulse interacts with the sample by way of the complex second-order nonlinear susceptibility to generate a force on the lattice driving coherent phonon excitation. Transforming the transient reflectivity signal into frequency-time space reveals interference effects leading to the coherent phonon generation and subsequent dressing of the phonon by electron-hole pair excitations.

Tumor oximetry: comparison of 19F MR EPI and electrodes.

We recently described a novel approach to measuring regional tumor oxygen tension. This approach is based on 19F pulse burst saturation recovery NMR echo planar imaging relaxometry of hexafluorobenzene or "FREDOM" (Fluorocarbon Relaxometry using Echo planar imaging for Dynamic Oxygen Mapping). We have now compared oxygen tension measurements using FREDOM with a traditional polarographic method (the Eppendorf Histograph) in a group of size matched Dunning prostate rat tumors R3327-AT1. We also compare MR and electrode approaches to monitoring dynamic changes with respect to interventions and demonstrate extension of the MR technique to rat breast tumors.

Oxygenation in a human tumor xenograft: manipulation through respiratory challenge and antibody-directed infarction.

We recently demonstrated the use of 19F NMR relaxometry of hexafluorobenzene to monitor regional tumor oxygen tension dynamics in rats. We have now extended the application to human tumors implanted in immunocompromised (SCID) mice. This has allowed us both to investigate dynamic response to respiratory challenge (carbogen) and to probe the mechanisms of a new anti-vascular therapy designed to produce tumor-specific infarction.

Tumor oxygen dynamics: comparison of 19F MR EPI and frequency domain NIR spectroscopy.

Oxygen plays a key role in tumor therapy and may be related to tumor development: e.g., angiogenesis and metastasis. Using noninvasive techniques to accurately measure tumor oxygenation could assist in developing novel therapies. Here, we have used the FREDOM (Fluorocarbon Relaxometry using Echo planar imaging for Dynamic Oxygen Mapping) approach based on hexafluorobenzene (HFB) to monitor tissue oxygen tension (pO2) of rat breast and prostate tumors and compared the results with changes in tumor vascular hemoglobin saturation (sO2) and concentration observed using a new dual wavelength homodyne near-infrared (NIR) system. The dynamic changes in pO2 and sO2 were assessed while rats were breathing various gases. NIR showed significant changes in vascular oxygenation accompanying respiratory interventions. 19F MR-EPI also showed significant changes in tissue pO2 and revealed considerable regional heterogeneity in both absolute values and rate of change accompanying interventions. Generally, changes in vascular sO2 preceded tissue pO2, particularly for smaller tumors.

Tumor oxygen dynamics: correlation of in vivo MRI with histological findings.

Tumor oxygenation has long been recognized as a significant factor influencing cancer therapy. We recently established a novel magnetic resonance in vivo approach to measuring regional tumor oxygen tension, FREDOM (Fluorocarbon Relaxometry Using Echo Planar Imaging for Dynamic Oxygen Mapping), using hexafluorobenzene (HFB) as the reporter molecule. We have now investigated oxygen dynamics in the two Dunning prostate R3327 rat tumor sublines, AT1 and H. FREDOM revealed considerable intratumoral heterogeneity in the distribution of pO(2) values in both sublines. The anaplastic faster-growing AT1 tumors were more hypoxic compared with the size-matched, well-differentiated, and slower-growing H tumors. Respiratory challenge with oxygen produced significant increases in mean and median pO(2) in all the H tumors (P<.001), but no response in half of the larger AT1 tumors (>3 cm(3)). Immunohistochemical studies using the hypoxia marker, pimonidazole, and the vascular endothelial cell marker, CD31, confirmed that the H tumors had more extensive vasculature and less hypoxia than the AT1 tumors. These results further validate the utilization of FREDOM to monitor tumor oxygenation and concur with the hypothesis that the level of hypoxia is related to tumor growth rate and poor vascularity.

Dynamic response of breast tumor oxygenation to hyperoxic respiratory challenge monitored with three oxygen-sensitive parameters.

The simultaneous measurement of three oxygen-sensitive parameters [arterial hemoglobin oxygen saturation (SaO2), tumor vascular-oxygenated hemoglobin concentration ([HbO2]), and tumor oxygen tension (pO2)] in response to hyperoxic respiratory challenge is demonstrated in rat breast tumors. The effects of two hyperoxic gases [oxygen and carbogen (5% CO2 and 95% O2)] were compared, by use of two groups of Fisher rats with subcutaneous 13762NF breast tumors implanted in pedicles on the foreback. Two different gas-inhalation sequences were compared, i.e., air-carbogen-air-oxygen-air and air-oxygen-air-carbogen-air. The results demonstrate that both of the inhaled, hyperoxic gases significantly improved the tumor oxygen status. All three parameters displayed similar dynamic response to hyperoxic gas interventions, but with different response times: the fastest for arterial SaO2, followed by biphasic changes in tumor vascular [HbO2], and then delayed responses for pO2. Both of the gases induced similar changes in vascular oxygenation and regional tissue pO2 in the rat tumors, and changes in [HbO2] and mean pO2 showed a linear correlation with large standard deviations, which presumably results from global versus local measurements. Indeed, the pO2 data revealed hetergeneous regional response to hyperoxic interventions. Although preliminary near-infrared measurements had been demonstrated previously in this model, the addition of the pO2 optical fiber probes provides a link between the noninvasive relative measurements of vascular phenomena based on endogenous reporter molecules, with the quantitative, albeit, invasive pO2 determinations.

Correlation of tumor oxygen dynamics with radiation response of the dunning prostate R3327-HI tumor.

Our previous studies have shown that oxygen inhalation significantly reduces tumor hypoxia in the moderately well-differentiated HI subline of the Dunning prostate R3327 rat carcinoma. To test our hypothesis that modifying hypoxia could improve the radiosensitivity of these tumors, we performed experimental radiotherapy to compare the tumor response to ionizing radiation alone or in combination with oxygen inhalation. Tumor pO(2) measurements were performed on size-selected tumors several hours before radiotherapy using (19)F nuclear magnetic resonance echo planar imaging relaxometry (FREDOM) of the reporter molecule hexafluorobenzene. In common with our previous findings, the larger tumors (>3.5 cm(3)) exhibited greater hypoxia than the smaller tumors (<2 cm(3); P < 0.001), and oxygen inhalation reduced the hypoxic fraction (<10 Torr): In the larger tumors, hypoxic fraction dropped significantly from a mean baseline value of 80% to 17% (P < 0.001). The effect of oxygen administered 30 min before and during irradiation on tumor response to a single 30-Gy dose of photons was evaluated by growth delay. For the smaller tumors, no difference in growth delay was found when treatment was given with or without oxygen breathing. By contrast, breathing oxygen before and during irradiation significantly enhanced the growth delay in the larger tumors (additional 51 days). The differential behavior may be attributed to the low baseline hypoxic fraction (<10 Torr) in small tumors (20%) as a target for oxygen inhalation. There was a strong correlation between the estimated initial pO(2) value and the radiation-induced tumor growth delay (R > 0.8). Our histological studies showed a good match between the perfused vessels marked by Hoechst 33342 dye and the total vessels immunostained by anti-CD31 and indicated extensive perfusion in this tumor line. In summary, the present results suggest that the ability to detect modulation of tumor pO(2), in particular, the residual hypoxic fraction, with respect to an intervention, could have prognostic value for predicting the efficacy of radiotherapy.

Interplay of tumor vascular oxygenation and tumor pO2 observed using near-infrared spectroscopy, an oxygen needle electrode, and 19F MR pO2 mapping.

This study investigates the correlation of tumor blood oxygenation and tumor pO(2) with respect to carbogen inhalation. After having refined and validated the algorithms for calculating hemoglobin concentrations, we used near-infrared spectroscopy (NIRS) to measure changes of oxygenated hemoglobin concentration (delta[HbO(2)]) and used an oxygen needle electrode and (19)F MRI for pO(2) measurements in tumors. The measurements were taken from Dunning prostate R3327 tumors implanted in rats, while the anesthetized rats breathed air or carbogen. The NIRS results from tumor measurements showed significant changes in tumor vascular oxygenation in response to carbogen inhalation, while the pO(2) electrode results showed an apparent heterogeneity for tumor pO(2) response to carbogen inhalation, which was also confirmed by (19)F MR pO(2) mapping. Furthermore, we developed algorithms to estimate hemoglobin oxygen saturation, sO(2), during gas intervention based on the measured values of delta[HbO(2)] and pO(2). The algorithms have been validated through a tissue-simulating phantom and used to estimate the values of sO(2) in the animal tumor measurement based on the NIRS and global mean pO(2) values. This study demonstrates that the NIRS technology can provide an efficient, real-time, noninvasive approach to monitoring tumor physiology and is complementary to other techniques, while it also demonstrates the need for an NIR imaging technique to study spatial heterogeneity of tumor vasculature under therapeutic interventions.

Differential oxygen dynamics in two diverse Dunning prostate R3327 rat tumor sublines (MAT-Lu and HI) with respect to growth and respiratory challenge.

PURPOSE: Since hypoxia may influence tumor response to therapy and prognosis, we have compared oxygenation of tumors known to exhibit differential growth rate and tissue differentiation. METHODS AND MATERIALS: Regional tumor oxygen tension was measured using 19F nuclear magnetic resonance echo planar imaging relaxometry of hexafluorobenzene, which provided dynamic maps with respect to respiratory intervention. Investigations used two Dunning prostate R3327 rat tumor sublines: the fast growing, highly metastatic MAT-Lu and the moderately well-differentiated, slower growing HI. RESULTS: Both sublines showed significantly higher oxygen tension in smaller tumors (<2 cm(3)) than in larger tumors (>3.5 cm(3)). Pooled data showed that MAT-Lu tumors exhibited greater hypoxia compared with the size-matched HI tumors (p < 0.0001). Respiratory challenge (oxygen or carbogen) produced significant increases in mean pO(2) for tumors of both sublines (p < 0.0001). However, initially hypoxic regions displayed very different behavior in each subline: those in the HI tumors responded rapidly with significant elevation in pO(2), while those in the MAT-Lu tumors showed little response to respiratory intervention. CONCLUSIONS: These results concur with hypotheses that hypoxia is related to tumor growth rate and degree of differentiation. Under baseline conditions, the differences were subtle. However, response to respiratory intervention revealed highly significant differences, which, if held valid in the clinic, could have prognostic value.

Dynamic breast tumor oximetry: the development of prognostic radiology.

A novel pre clinical approach to evaluating tumor oxygen dynamics was recently introduced (Am. J. Clin. Oncol. 24, 462-466 (2001)). FREDOM (Fluorocarbon Relaxometry using Echo planar imaging for Dynamic Oxygen Mapping) allows maps of tumor pO(2) including 50 - 150 individual locations simultaneously to be produced with typical in plane resolution of 1.25 mm in 6.5 mins. The technique has been applied extensively in rat prostate tumors and is now demonstrated in the rat breast 13762NF adenocarcinoma. When anesthetized rats breathed 33% oxygen, mean baseline pO(2) was in the range 17 +/- 2 (se) torr to 74 +/- 4 torr with mean value for nine tumors 46 +/- 8 torr. However, small tumors (< 2.2 cm(3)) were significantly better oxygenated with mean pO(2) = 63 +/- 7 torr than large tumors (> 2.4 cm(3)) with mean pO(2) 24 +/- 5 torr (p < 0.002). Switching the inhaled gas to oxygen or carbogen produced a significant and rapid increase in mean pO(2) for both small and larger tumors (p < 0.05). Given the increasing evidence that tumor oxygenation is related to therapeutic outcome, we believe this approach to measuring tumor oxygen dynamics can be of value in predicting response to therapy, evaluating adjuvant interventions designed to modulate response to therapy, and in providing "Prognostic Radiology".