Charlotte Fontolliet

Clinical photodynamic therapy for superficial cancer in the oesophagus and the bronchi: 514 nm compared with 630 nm light irradiation after sensitization with Photofrin II.

Photodynamic therapy (PDT) for cancer in the oesophagus and bronchi with red (630 nm) light may occasionally lead to wall perforation and fistula. Therefore, we investigated the clinical use of a less penetrating wavelength (514 nm) for the curative treatment of nine superficial carcinomas in the oesophagus and bronchi after photosensitization with Photofrin II. Tumours without infiltration beyond the submucosa in the oesophagus and beyond the lamina propria in the bronchi were considered as superficial cancers. The outcome and complications were compared with those of 13 superficial cancers treated with PDT and 630 nm light. In addition, we evaluated histologically the extent of the long-term tissue damage and scarring following treatment of six oesophageal cancers with either green or red light. At first endoscopic control, 7-10 days after PDT, tissue necrosis simply matched the illuminated area, without evidence of selective tumour damage. Six of nine tumours treated with 514 nm light had a complete response compared with nine of 13 after 630 nm irradiation. No perforation or fistula occurred in either treatment group. However, severe chest pain and fever with or without pleural effusion, consistent with occult perforation, were observed in three patients after 630 nm illumination in the oesophagus. Histologically, fibrous scarring in the three distinct sites treated with green light was limited to the superficial layers of the oesophagus. After red light treatment, transmural fibrosis with marked thinning of the oesophageal wall was evident in two of the three specimens available for inspection. These results indicate that PDT with 514 nm light has the potential to cure superficial cancer in the oesophagus and bronchi with essentially the same probability of success as red light. In the oesophagus, green light prevents deep tissue damage, thus reducing the risk of perforation.

Photodynamic therapy of early squamous cell carcinoma with tetra(m-hydroxyphenyl)chlorin: optimal drug-light interval.

The optimal drug-light interval for effective photodynamic therapy (PDT) of early squamous cell carcinomas was evaluated with tetra(m-hydroxyphenyl)chlorin (mTHPC) by means of two complementary modalities: irradiation tests and ex vivo fluorescence microscopy. A Syrian hamster cheek pouch tumour model was used in these experiments. Photodynamic therapy on both tumour-bearing and contralateral healthy cheek pouch mucosae was performed at 650 nm and 514 nm. Light doses of 12 J cm(-2) were delivered at a light dose rate of 150 mW cm(-2) and light doses of 80 J cm(-2) were delivered at a light dose rate of 100 mW cm(-2) respectively, at these two wavelengths, between 6 h and 12 days after the injection of 0.5 mg kg(-1) body weight mTHPC. Two histologically different types of tissue damage were observed: first, a non-selective and non-specific ischaemic vascular necrosis for the cases in which PDT took place during the first 48 h after the injection of the dye and, second, tissue-specific PDT damage, as a coagulation necrosis, when PDT took place more than 72 h after injection of the dye. The time-dependent biodistribution of mTHPC investigated by fluorescence microscopy shows a weak and non-significant difference in relative fluorescence intensities between early SCC and healthy mucosae. Up to 2 days after the injection, the drug is mainly localized in the endothelial cells of the blood vessels. After this period, the dye accumulates in the squamous epithelia with a concentration peaking at 4 days. At all time points, a weak fluorescence intensity is observed in the underlying lamina propria and striated muscle. The information obtained from these studies could well be relevant to clinical trials as it suggests that time delays between 4 and 8 days after i.v. injection should be optimal for PDT of early malignancies in hollow organs.

In-vivo fluence rate effect in photodynamic therapy of early cancers with tetra(m-hydroxyphenyl)chlorin

Abstract— Several parameters affect clinical trials in photodynamic therapy and influence the therapeutic outcome. Beside drug dose, light dose, drug‐light interval and other variables, the fluence rate is a parameter that can influence the therapeutic results. In this study we have evaluated the fluence rate effect with a second‐generation photosensitizer, tetra(m‐hydroxyphenyl)chlorin (mTHPC) using a 7,12‐dimethylbenz(a)anthracene induced early squamous cell carcinoma of the Syrian hamster cheek pouch as a tumor model. Following injection of 0.5 mg/kg of mTHPC, irradiation tests were performed at two drug‐light intervals, 4 and 8 days. Wavelength and light dose were adapted from those applied routinely in clinical trials. Irradiations at 652 nm were carried out with fluences ranging from 8 to 20 J/cm2 delivered at fluence rates of 15 and 150 mW/cm2. Similar tests were also performed at 514 nm with a fluence of 80 J/cm2 delivered at fluence rates ranging from 25 to 125 mW/cm2. At both wavelengths and drug‐light intervals for a given fluence, the higher fluence rates resulted in less tissue damage in tumor and healthy mucosae. However, the lower fluence rates yielded slightly less therapeutic selectivity. This study confirms that the fluence rate is of major importance in clinical PDT.

Monoallelic Methylation of the APC Promoter Is Altered in Normal Gastric Mucosa Associated with Neoplastic Lesions

Adenomatous polyposis coli (APC) promoter hypermethylation has been reported frequently in normal gastric mucosa, but it remained to be clarified whether this occurs in every individual. In this study, methylation of the APC promoter was analyzed in histologically normal-appearing gastric mucosa samples by methylation-sensitive single-strand conformation analysis and by a methylation-sensitive dot blot assay. Epithelial cell samples were collected by microdissection from tissue sections. Equal amounts of methylated and unmethylated APC alleles were found in all gastric mucosa samples from patients without any gastric lesions (20 samples). Allele-specific methylation analysis showed that the methylation of the APC promoter was monoallelic; however, which allele was methylated depended on the cell type. Increased or decreased methylation was found in 10 of 36 (28%) normal gastric mucosa samples adjacent to a gastric or esophageal adenocarcinoma. No allelic loss was found at the APC locus. Modification of the methylation status was also found in 3 of 21 (14%) normal-appearing gastric mucosa samples adjacent to intestinal metaplasia. In contrast, all normal mucosa samples in cases with chronic gastritis but without metaplasia or dysplasia showed a monoallelic methylation pattern. Our results indicate the following: (a) In normal gastric mucosa, the APC promoter shows monoallelic methylation, which is not due to imprinting but most likely due to allelic exclusion; (b) the excluded allele differs between foveolar and glandular epithelial cells; (c) the APC methylation pattern is frequently altered in normal-appearing gastric mucosa of gastric or esophageal adenocarcinoma patients; and (d) such alterations also occur in normal gastric mucosa adjacent to intestinal metaplasia.

Photodynamic Therapy for 101 Early Cancers of the Upper Aerodigestive Tract, the Esophagus, and the Bronchi: A Single-Institution Experience

Cancer, when detected at an early stage, has a very good probability of being eradicated by surgery or radiotherapy. However, less aggressive treatments also tend to provide high rates of cure without the side effects of radical therapy. We report on the results of our clinical experience with photodynamic therapy (PDT) for the treatment of early carcinomas in the upper aerodigestive tract, the esophagus, and the tracheobronchial tree. Sixty-four patients with 101 squamous cell carcinomas were treated with three different photosensitizers: hematoporphyrin derivative (HPD), Photofrin II, and tetra (m-hydroxyphenyl)chlorin (mTHPC). Seventy-seven (76%) tumors showed a complete rsponse with no recurrence after a mean follow-up period of 27 months. There was no significant difference in terms of cure rates among the three dyes. However, mTHPC has a stronger phototoxicity and induces a shorter skin photosensitization than either of the other photosensitizers. There were eight major complications: three esophagotracheal fistulae after illumination with red light in the esophagus, two esophageal stenoses following 360° circumferential irradiation, and three bronchial stenoses. Illumination with the less penetrating green light and the use of a 180° or 240° windowed cylindrical light distributor render the risk of complications in the esophagus essentially impossible, without reducing the efficacy of the treatment. Therefore, PDT may be considered as a safe and effective treatment for early carcinomas of the upper aerodigestive tract, the esophagus, and the tracheobronchial tree.

Epidemiology and Natural History of Reflux Esophagitis

Studies describing the natural history of erosive reflux esophagitis in patients presenting with or without symptoms of gastroesophageal reflux are exceedingly rare. This study followed a first group of 759 patients receiving intermittent courses of medical therapy for initial Savary grades 1 to 3 reflux esophagitis (median follow-up period of 4.5 years). In 23% of cases, reflux esophagitis evolved as a recurrent progressive form of the disease, which may potentially cause attendant complications such as ulcer, stenosis, or short esophagus; to avoid unnecessary long-term medical treatment, surgery should be considered in these cases if the patient is fit for surgery. In the remaining 77% of cases, reflux esophagitis evolved as a benign form of the disease, either as an isolated episode of reflux esophagitis or as a recurrent but nonprogressive form of the disease. Another group of 1,022 patients presenting with initial Savary grades 1 to 4 reflux esophagitis (median follow-up period of 3 years) was studied to detect the appearance of columnar epithelium during the healing process. In 18% of all cases, follow-up endoscopies detected islands, tongues, or circumferential forms of columnar epithelium as patterns of healing of the initial erosions; these patients should undergo endoscopic surveillance for the detection of dysplasias, especially if a specialized type of epithelium is present on biopsy specimens.

Selectivity of the photosensitiser Tookad for photodynamic therapy evaluated in the Syrian golden hamster cheek pouch tumour model.

The response to photodynamic therapy (PDT) with the photosensitiser (PS) Tookad® was measured in the Syrian hamster cheek pouch model on normal mucosae and chemically induced squamous cell carcinoma. This PS is a palladium-bacteriopheophorbide presenting absorption peaks at 538 and 762 nm. The light dose, drug dose and drug injection-light irradiation times (DLI), ranging between 100 and 300 J cm−2, 1–5 mg kg−1 and 10–240 min respectively, were varied and the response to PDT was analysed by staging the macroscopic response and by the histological examination of the sections of the irradiated cheek pouch. A fast time decay of the tissular response with drug dose of 1–5 mg kg−1 was observed for DLI ranging from 10 to 240 min and for light doses of 100–300 J cm−2 delivered at a light dose rate of 150 mW cm−2. A significantly higher level of tissular response was observed for squamous cell carcinoma compared to normal tissue. Nevertheless, the threshold level of the drug–light dose for a detectable response was not significantly different in the tumoral vs normal tissue. The highest response at the shortest DLIs and the absence of measurable response at DLI larger than 240 min at light dose of 300 J cm−2 and drug dose of 5 mg kg−1 reveals the predominantly vascular effect of Tookad®. This observation suggests that Tookad® could be effective in PDT of vascularised lesions.

Photodetection of early cancer in the upper aerodigestive tract and the bronchi using photofrin II and colorectal adenocarcinoma with fluoresceinated monoclonal antibodies

The performance of a fluorescence endoscope for the detection of early cancer is clinically evaluated. the apparatus is based on the imaging of the laser-induced fluorescence (LIF) of a dye which localizes in the tumor after IV injection with a higher concentration than in the surrounding normal tissue. The tests are carried out in several of the hollow organs, such as the upper aerodigestive tract, the bronchi, and the colon. In the two former cases the dye used is photofrin II, whereas in the latter case conjugates between monoclonal antibodies (Mab) directed against carcinoembrionic antigen (CEA) and fluorescein molecules are injected. The fluorescence contrast between tumor and surrounding tissue is enhanced by real-time image processing which eliminates most of the tissue autofluorescence as well as the fluorescence due to the relatively small amount of dye localized in the normal tissue. This is done by recording the fluorescence image in two spectral domains, after which these two images are digitized and manipulated with a mathematical operator (lookup-table). The sources of false positives and false negatives are evaluated in terms of the fluorescent dye and tissue optical properties.

Mucosal ablation with photodynamic therapy in the esophagus: optimization of light dosimetry in the sheep model

BACKGROUND Photodynamic therapy is an attractive technique for mucosal ablation in patients with superficial squamous cell carcinoma of the esophagus, or high-grade dysplasia or early stage adenocarcinoma arising in Barretts esophagus. Although illumination with green light is assumed to be safe, choice of the light has been empirical in clinical studies; light dose is often reduced to avoid potential complications. The present study assessed the safety of green and blue lights during photodynamic therapy in the esophagus by progressively administrating increasing doses in an attempt to standardize the dose and determine a safe upper limit. This would considerably simplify photodynamic therapy and improve therapeutic results. METHODS The sheep model was chosen because of similarities with humans regarding the thickness and histologic structure of the esophagus. Irradiation with a 180 degrees windowed cylindrical light distributor was performed in 19 sheep 4 days after injection of 0.15 mg/kg of tetra(m-hydroxyphenyl) chlorin. Light doses ranged from 10 to 500 J/cm(2) at 514 nm (argon ion laser) and from 5 to 250 J/cm(2) at 413 nm (krypton laser). RESULTS Follow-up endoscopies revealed a tissue response with a fibrinous area at almost all light doses, whereas application of extremely high light doses tended to induce circumferential necrosis with subsequent stenosis. Three months after irradiation with green light, histologic examination of the resected specimens revealed transmural scarring at doses higher than 100 J/cm(2). After illumination with blue light, partial or more extensive fibrosis of the muscular layer was observed only at light doses of 175 to 250 J/cm(2). CONCLUSIONS Application of high doses of green light after sensitization with tetra(m-hydroxyphenyl) chlorin led to severe complications in the esophagus of the sheep that are highly likely to occur in humans as well. Blue light causes significantly less damage than green light and may, therefore, be considered as an alternative for photodynamic therapy of early stage superficial esophageal cancer.

Elevated hepatocyte paraffin 1 and neprilysin expression in hepatocellular carcinoma are correlated with longer survival

Hepatocyte paraffin 1 (Hep Par 1) and neprilysin (CD10) are well-known markers of hepatocellular carcinoma (HCC). To assess their potential prognostic role, we conducted a retrospective analysis of 97 formalin-fixed and paraffin-embedded HCC from patients treated by surgery with curative intent, using standard immunohistochemical procedures and semiquantitative analysis. Strong Hep Par 1 expression and canalicular CD10 staining pattern were significantly correlated with smaller tumor size (p=0.007 and 0.04, respectively). On univariate analysis, longer overall survival was observed in patients with strong Hep Par 1 expression (p=0.0005) and in patients with a CD10can staining pattern (p=0.02). On multivariate analysis, the combined immunohistochemical score (CIS) obtained by addition of Hep Par 1 and CD10can scores and subtraction of cytoplasmic CD10 score was retained as the single most important prognostic factor (p=0.001). Patients with a CIS <4 had a 3.5-fold increased risk of death, as compared to those with a CIS ≥4. In conclusion, strong Hep Par 1 expression, presence of CD10can labeling, and absence of CD10cyt staining are favorable prognostic factors in HCC, which can be easily combined into a single immunohistochemical score for routine clinical use.