Targeting of radio-enhancing drugs.

OBJECTIVE: Toxicity to normal tissue is frequently the dose-limiting factor in the chemotherapy and mixed modality treatments of cancer. If the radio-enhancing drug could be localized at the disease site and released slowly over time, then systemic drug toxicities could be decreased while simultaneously maintaining high drug concentrations in the tumor. These considerations support a role for a sustained release intra-tumoral delivery systems for the delivery of radio-enhancing drugs. METHODS: Two approaches aimed at achieving the end of localizing the radio-enhancing drug to the tumor are described. First, nanoparticles, which have a prolonged circulation time and facility for enhanced tumor targeting. Structural defects in the walls of the tumor vasculature allow the passage of particles too large to pass through the walls of normal blood vessels. This characteristic of tumor blood vessels, referred to as the enhanced permeability and retention (EPR) effect, allows relatively large entities (typically liposomes, nanoparticles, and macromolecular drugs) to pass from the blood vessels to tumor tissue and as a result nanoparticles accumulate in the tumor while being excluded from normal tissue. Second, biodegradable implanted polymers. In these devices, the radio-enhancing drug is physically trapped within the polymer matrix which is implanted in the tumor. The drug is released as the polymer degrades in response to its local environment. The degradation rate of the polymer device can be adjusted to control the rate of drug release. By this means, the level of radio-enhancing drug can be maintained at the tumor site for the duration of radiation treatment. RESULTS AND CONCLUSIONS: Results of experiments indicate that for both methods tumor control could be optimized by maintaining the radio-enhancing drug at a useful concentration in the tumor over a period of time compatible with the duration of fractionated radiation treatment. These studies have provided proof of principle support for the further development of this approach. To date, while some of the methods and devices for drug delivery described in this paper have been involved in clinical trials, none have so far been developed for routine clinical application.

FDG-PET-based differential uptake volume histograms: a possible approach towards definition of biological target volumes.

OBJECTIVE: Integration of fluorine-18 fludeoxyglucose ((18)F-FDG)-positron emission tomography (PET) functional data into conventional anatomically based gross tumour volume delineation may lead to optimization of dose to biological target volumes (BTV) in radiotherapy. We describe a method for defining tumour subvolumes using (18)F-FDG-PET data, based on the decomposition of differential uptake volume histograms (dUVHs). METHODS: For 27 patients with histopathologically proven non-small-cell lung carcinoma (NSCLC), background uptake values were sampled within the healthy lung contralateral to a tumour in those image slices containing tumour and then scaled by the ratio of mass densities between the healthy lung and tumour. Signal-to-background (S/B) uptake values within volumes of interest encompassing the tumour were used to reconstruct the dUVHs. These were subsequently decomposed into the minimum number of analytical functions (in the form of differential uptake values as a function of S/B) that yielded acceptable net fits, as assessed by χ(2) values. RESULTS: Six subvolumes consistently emerged from the fitted dUVHs over the sampled volume of interest on PET images. Based on the assumption that each function used to decompose the dUVH may correspond to a single subvolume, the intersection between the two adjacent functions could be interpreted as a threshold value that differentiates them. Assuming that the first two subvolumes spread over the tumour boundary, we concentrated on four subvolumes with the highest uptake values, and their S/B thresholds [mean ± standard deviation (SD)] were 2.88 ± 0.98, 4.05 ± 1.55, 5.48 ± 2.06 and 7.34 ± 2.89 for adenocarcinoma, 3.01 ± 0.71, 4.40 ± 0.91, 5.99 ± 1.31 and 8.17 ± 2.42 for large-cell carcinoma and 4.54 ± 2.11, 6.46 ± 2.43, 8.87 ± 5.37 and 12.11 ± 7.28 for squamous cell carcinoma, respectively. CONCLUSION: (18)F-FDG-based PET data may potentially be used to identify BTV within the tumour in patients with NSCLC. Using the one-way analysis of variance statistical tests, we found a significant difference among all threshold levels among adenocarcinomas, large-cell carcinoma and squamous cell carcinomas. On the other hand, the observed significant variability in threshold values throughout the patient cohort (expressed as large SDs) can be explained as a consequence of differences in the physiological status of the tumour volume for each patient at the time of the PET/CT scan. This further suggests that patient-specific threshold values for the definition of BTVs could be determined by creation and curve fitting of dUVHs on a patient-by-patient basis. ADVANCES IN KNOWLEDGE: The method of (18)F-FDG-PET-based dUVH decomposition described in this work may lead to BTV segmentation in tumours.

Mesenchymal Stem Cells Adopt Lung Cell Phenotype in Normal and Radiation-induced Lung Injury Conditions.

Lung tissue exposure to ionizing irradiation can invariably occur during the treatment of a variety of cancers leading to increased risk of radiation-induced lung disease (RILD). Mesenchymal stem cells (MSCs) possess the potential to differentiate into epithelial cells. However, cell culture methods of primary type II pneumocytes are slow and cannot provide a sufficient number of cells to regenerate damaged lungs. Moreover, effects of ablative radiation doses on the ability of MSCs to differentiate in vitro into lung cells have not been investigated yet. Therefore, an in vitro coculture system was used, where MSCs were physically separated from dissociated lung tissue obtained from either healthy or high ablative doses of 16 or 20 Gy whole thorax irradiated rats. Around 10±5% and 20±3% of cocultured MSCs demonstrated a change into lung-specific Clara and type II pneumocyte cells when MSCs were cocultured with healthy lung tissue. Interestingly, in cocultures with irradiated lung biopsies, the percentage of MSCs changed into Clara and type II pneumocytes cells increased to 40±7% and 50±6% at 16 Gy irradiation dose and 30±5% and 40±8% at 20 Gy irradiation dose, respectively. These data suggest that MSCs to lung cell differentiation is possible without cell fusion. In addition, 16 and 20 Gy whole thorax irradiation doses that can cause varying levels of RILD, induced different percentages of MSCs to adopt lung cell phenotype compared with healthy lung tissue, providing encouraging outlook for RILD therapeutic intervention for ablative radiotherapy prescriptions.

Mechanisms of radiation-induced sensorineural hearing loss and radioprotection.

Patients that receive radiotherapy are at risk of late sensorineural hearing loss when the inner ear is included within the radiation field. Preclinical and human temporal bone studies have shown that there is differential damage to cochlear structures depending on the amount of dose delivered to the inner ear. In vitro studies have suggested that reactive oxygen species (ROS) are the main initial actors in radiation-induced damage. The interaction of ROS with different cellular components can result in different apoptotic pathways. Therefore, approaches to radioprotection are mainly aimed to reduce ROS production through antioxidants. This review summarizes recent research in the field that can improve the understanding and boost preventive efforts of this adverse effect.

Defining radiotherapy target volumes using 18F-fluoro-deoxy-glucose positron emission tomography/computed tomography: still a Pandora's box?

PURPOSE: We discuss the effect of (18)F-fluoro-deoxy-glucose (FDG) positron emission tomography (PET)/computed tomography (CT) data on target volume definition for radiotherapy planning. We compared the effect of various thresholding methods on the PET-based target volume vs. the standard CT-based tumor volume. METHODS AND MATERIALS: Different thresholding methods were reviewed and compared to our PET-based gross tumor volume data obtained from a cohort of 31 non-small-cell lung carcinoma patients who had undergone preoperative PET/CT scans for staging. The feasibility and limitations of FDG-based PET/CT data on target volume delineation in radiotherapy planning have been demonstrated with frequently used approaches for target outlining such as the qualitative visual method and the fixed 15% or 40% of the maximal iso-uptake value threshold methods. RESULTS: The relationship between PET-based and CT-based volumes generally suffers from poor correlation between the two image data sets, expressed in terms of a large statistical variation in gross tumor volume ratios, irrespective of the threshold method used. However, we found that the maximal signal/background ratios in non-small-cell lung carcinoma patients correlated well with the pathologic results, with an average ratio for adenocarcinoma, large cell carcinoma, and squamous cell carcinoma of 10.5 ± 3.5, 12.6 ± 2.8, and 14.1 ± 5.9, respectively. CONCLUSION: The fluctuations in tumor volume using different quantitative PET thresholding approaches did not depend on the thresholding method used. They originated from the nature of functional imaging in general and PET imaging in particular. Functional imaging will eventually be used for biologically tailored target radiotherapy volume definition not as a replacement of CT- or magnetic resonance imaging-based anatomic gross tumor volumes but with the methods complementing each other in a complex mosaic of distinct biologic target volumes.

Relative biologic effectiveness in terms of tumor response of 125I implants compared with 60Co gamma rays.

PURPOSE: To measure the relative biologic effectiveness (RBE) for 125I seeds compared with external beam radiotherapy using a clinically relevant in vivo system. METHODS AND MATERIALS: Photon emission from a detailed source model was simulated using the Monte Carlo code MCNP4C, sampling from a 125I spectrum. The mouse RIF-1 tumor was treated with either temporary implant of an 125I seed or with 60Co gamma rays. The tumors were always the same size at the initiation of treatment, and the endpoint was growth inhibition. RESULTS: The dose-response curve for both modalities was close to linear and was independent of the initial 125I activity (dose rate) for the range investigated. Calculation of the RBE for tumor response requires assigning a unique value for the tumor dose that is not homogenous but depends on the distance from the 125I source. Because tumor regrowth will depend on the subpopulation of cells that have the greatest probability of survival (i.e., those at the greatest distance from the 125I source), one approach is to use the dose to this population. On this basis, the RBE for 125I compared with 60Co gamma rays is 1.5. If the 125I dose is computed as the average dose to the tumor, corrected for the dose that is wasted as overkill in the cell population closest to the center of the 125I seed, the RBE is 1.4. CONCLUSION: The result, an RBE of 1.4-1.5 is similar to findings obtained by other methods, supporting the validity of this approach to derive an RBE with validity in a clinical context.

Silver leaf nylon dressing to prevent radiation dermatitis in patients undergoing chemotherapy and external beam radiotherapy to the perineum.

PURPOSE: Silver-leaf nylon dressing (SLND) has been shown to have effective antimicrobial activity and to enhance healing in burns and skin grafts. The purpose of this study was to evaluate the value of SLND in preventing radiation dermatitis in patients undergoing radiotherapy to target volumes that included the perineum and concurrent chemotherapy. METHODS AND MATERIALS: Fifteen consecutive patients with either anal canal or gynecologic cancer were offered the SLND as a preventive intervention. The evaluation was based on standardized photographs taken at the end of treatment. A historical control group of 15 patients with the same neoplasms who received standard skin care was assessed in the same fashion. Ten observers unaware of the treatment intervention were enrolled in the evaluation of the skin changes. The Mann-Whitney U test was used to assess the statistical significance of the differences in the dermatitis scores between the two patient groups. RESULTS: The mean dermatitis score for controls was 2.62 (standard deviation, 0.48). The mean score for the SLND group was significantly lower at 1.16 (standard deviation, 0.40; p <0.0001). CONCLUSION: The results of this study suggest that SLND is effective in reducing radiation dermatitis, apparently because of its antibacterial properties.

Sensitization to radiation from an implanted 125I source by sustained intratumoral release of chemotherapeutic drugs.

We have investigated tumor response to low-dose-rate irradiation from an implanted 125I source alone or in conjunction with intratumoral drug administration. The drug (cis-DDP or 5-FU) was incorporated homogeneously into the co-polymer CPP-SA, 20:80, and the polymer/drug rods were implanted in the RIF-1 fibrosarcomas growing subcutaneously in C3H mice. Twenty-four hours later, the tumor was implanted with an 125I seed. Tumor growth time was the end point in these experiments. For implanted 125I sources of different dose rates and implant times giving a range of total doses, a consistent dose-response relationship was shown between tumor growth time and total dose. In other experiments, 125I sources of different specific activities were implanted for periods of time adjusted so that the total dose to the tumor was always the same. When the 125I implant was combined with 5-FU, greater than additive responses were seen for both short (30 h) and long (96 h) 125I treatment times. In contrast, a short-duration (30 h) 125I implant combined with cis-DDP was the least effective treatment, giving a combined response that was no better than additive, whereas 96 h exposure to 125I combined with cis-DDP was the most effective combined treatment. It is conjectured that this inverse dose-rate effect seen when cis-DDP is combined with low-dose rate radiation is related to a cell cycle effect and/or to inhibition of repair of radiation damage by cis-DDP.

Treatment of intracranial rat glioma model with implant of radiosensitizer and biomodulator drug combined with external beam radiotherapy.

PURPOSE: To evaluate an intracranial polymer implant containing bromodeoxyuridine (BrdUrd) and N-(phosphonacetyl)-L-aspartic acid (PALA) in combination with external beam radiotherapy (EBRT) in the treatment of a rat glioma. METHODS AND MATERIALS: Combinations of the biomodulators 5-fluorouracil, methotrexate, or PALA with BrdUrd were evaluated as radiosensitizers in vitro by clonogenic assay. In in vivo experiments, BrdUrd and PALA were incorporated into a polyanhydride-based polymer, bis(p-carboxyphenoxy)propane sebacic acid, and implanted in the C6 rat glioma growing intracranially. The effectiveness of treatment was evaluated on the basis of survival. EBRT was given as 10-MV X-rays. RESULTS: In tissue culture experiments, C6 cells were refractory to radiosensitization by BrdUrd even when the thymidine analog was combined with a biomodulator intended to reduce de novo thymidine synthesis. The most effective compound in vitro was PALA. When PALA and BrdUrd in a polymer formulation were implanted intracranially and combined with 10-Gy EBRT, the treatment was highly effective, with 83% of treated rats surviving 180 days. CONCLUSION: Although the in vitro results were not encouraging, the combination of intratumoral BrdUrd and PAL with 10-Gy EBRT was highly effective in treating a rat glioma. These results indicate the clinical potential of combined and mixed modality treatments involving intratumoral sustained-release drug delivery.

Inducible nitric oxide synthase and nitrotyrosine in mice with radiation-induced lung damage.

The purpose of this study was to determine if radiation-induced lung damage is associated with induction of nitric oxide synthase (NOS) II and nitrotyrosine in an irradiated lung mouse model. The thorax of BALBc mice were exposed to 14 Gy radiation (experimental) or no radiation (control) and killed after at 1, 3, 6, 12, and 24 hours; 3, 15, and 30 days; and 3 and 6 months after treatment. Lung sections were processed for immunohistochemistry using NOS II and nitrotyrosine polyclonal antisera and in situ hybridization using 35S labeled probes for mouse NOS II. Quantitative analysis of experimental and control sections showed significant induction of NOS II and nitrotyrosine in alveolar macrophages from 6 hours to 30 days postirradiation, which was diminished by 3 months. The airway and alveolar epithelium and vascular endothelium showed strong NOS II expression at 15 to 30 days postirradiation. Nitrotyrosine immunostaining was also strongly evident in the alveolar epithelium and vascular endothelium during this period. There was little or no NOS II or nitrotyrosine in the sham control lungs throughout the study. These findings demonstrate increased formation of both NO and nitrotyrosine after radiation treatment and suggest a role for these molecules in the pathogenesis of radiation-induced lung damage.

Tumor treatment by sustained intratumoral release of 5-fluorouracil: effects of drug alone and in combined treatments.

PURPOSE: To evaluate an intratumoral polymer implant for sustained delivery of 5-fluorouracil (5-FU) in a mouse tumor model. METHODS AND MATERIALS: 5-FU was incorporated into a polyanhydride-based polymer, bis(p-carboxyphenoxy)propane sebacic acid (CPP:SA) and implanted in RIF-1 mouse fibrosarcoma growing s.c. The effectiveness of treatment was evaluated by tumor growth delay. External beam radiation was 60Co gamma rays, and the source of interstitial radiation was implanted 125I seeds. A second drug, cis-diamminedichloroplatinum (cis-DDP), was administered by intraperitoneal injection or by osmotic pump. RESULTS: For drug/polymer implant alone, the tumor growth delay was proportional to the amount of drug in the implant. The 5-FU polymer implant was most effective when combined with cis-DDP or with acute or fractionated radiation, and in some cases, the effects of combined treatments were greater than additive. The most effective combination was intratumoral 5-FU and low-dose-rate radiation delivered from an interstitial radiation source. CONCLUSION: Results indicate that 5-FU can be effectively delivered by polymer implant and that this mode of delivery is particularly appropriate for combined treatments.