Assessment of the Tumor Redox Status in Head and Neck Cancer by 62Cu-ATSM PET.

PURPOSE: Tumor redox is an important factor for cancer progression, resistance to treatments, and a poor prognosis. The aim of the present study was to define tumor redox (over-reduction) using 62Cu-diacetyl-bis(N4-methylthiosemicarbazone) (62Cu-ATSM) PET and compare its prognostic potential in head and neck cancer (HNC) with that of 2-deoxy-2-[18F]fluoro-D-glucose (18F-FDG). METHODS: Thirty HNC patients (stage II-IV) underwent pretreatment 62Cu-ATSM and 18F-FDG PET scans. Maximum standardized uptake values (SUVATSM and SUVFDG) and tumor-to-muscle activity concentration ratios (TMRATSM and TMRFDG) were measured. Reductive-tumor-volume (RTV) was then determined at four thresholds (40%, 50%, 60%, and 70% SUVATSM), and total-lesion-reduction (TLR) was calculated as the product of the mean SUV and RTV for 62Cu-ATSM. In 18F-FDG, metabolic-tumor-volume (MTV) and total-lesion-glycolysis (TLG) were obtained at a threshold of 40%. A ROC analysis was performed to determine % thresholds for RTV and TLR showing the best predictive performance, and these were then used to determine the optimal cut-off values to stratify patients for each parameter. Progression-free-survival (PFS) and cause-specific-survival (CSS) were evaluated by the Kaplan-Meier method. RESULTS: The means ± standard deviations of PFS and CSS periods were 16.4±13.4 and 19.2±12.4 months, respectively. A ROC analysis determined that the 70% SUVATSM threshold for RTV and TLR was the best for predicting disease progression and cancer death. Optimal cut-offs for each index were SUVATSM = 3.6, SUVFDG = 7.9, TMRATSM = 3.2, TMRFDG = 5.6, RTV = 2.9, MTV = 8.1, TLR = 14.0, and TLG = 36.5. When the cut-offs for TMRATSM and TLR were set as described above in 62Cu-ATSM PET, patients with higher TMRATSM (p = 0.03) and greater TLR (p = 0.02) showed significantly worse PFS, while patients with greater TLR had significantly worse CSS (p = 0.02). Only MTV in 18F-FDG PET predicted differences in PSF and CSS (p = 0.03 and p = 0.03, respectively). CONCLUSION: Tumor redox parameters measured by 62Cu-ATSM PET may be determinants of HNC patient outcomes and help define optimal patient-specific treatments.

Assessing tumor hypoxia in head and neck cancer by PET with 6²Cu-diacetyl-bis(N4-methylthiosemicarbazone).

PURPOSE: The aim of this study was to investigate the potential of PET imaging with a hypoxia-selective tracer 6²Cu-diacetyl-bis(N4-methylthiosemicarbazone) (6²Cu-ATSM) for evaluating the prognosis of patients with head and neck cancer (HNC). METHODS: Twenty-five patients with HNC including stage II to IV underwent both 6²Cu-ATSM and ¹8F-FDG PET before the initiation of treatment. Volumes of interest were placed on the tumor and sternocleidomastoid muscles to obtain SUVmax and to calculate the tumor-to-muscle activity ratio (TMR). The PET results were correlated with clinical follow-up, and the receiver operating characteristic analysis was used to determine the optimal cutoff values. Progression-free survival (PFS) and cause-specific survival (CSS) were statistically analyzed. RESULTS: Patients were followed up for periods ranging from 4 to 32 months. Twelve patients died from local recurrence or metastasis of a primary cancer, and 2 had recurrence of the 13 remaining patients. Mean (SD) periods of PFS and CSS were 15.5 (12.5) and 18.6 (11.0) months, respectively. Optimal cutoff values for each PET index were as follows: SUVs of 6²Cu-ATSM (SUVATSM) and FDG were 3.6 and 7.9; TMRs of ATSM (TMRATSM) and FDG were 3.2 and 5.6. When the cutoff for TMRATSM was set at 3.2, patients with a greater TMRATSM had significantly worse PFS (P = 0.014) and CSS (P = 0.031). Two-year PFS and CSS rates were 73% and 80% for patients with a lower TMRATSM (≤3.2); however, they were 20% and 33% for those with hypoxic tumors (TMRATSM, >3.2), respectively. F-FDG-related indices did not show any significant difference in either PFS or CSS. CONCLUSIONS: Pretreatment 6²Cu-ATSM PET is useful for predicting the prognosis of patients with HNC.

Different distribution of (62) Cu ATSM and (18)F-FDG in head and neck cancers.

PURPOSE: [6²Cu]-diacetyl-bis(N4-methlythiosemicarbazone) (Cu-ATSM) was used to delineate hypoxic tissue in head-and-neck cancer, and its distribution was compared with that of ¹8F-FDG. MATERIALS AND METHODS: Thirty patients with head-and-neck cancer underwent Cu-ATSM and FDG PET within a 1 week interval. Accumulation of tracer for each PET image was converted to SUV. After coregisteration of PET images with individual anatomic images, multiple small ROIs were drawn on the tumor mass and applied to both PET images. SUV values were obtained for all ROIs (SUV(roi)), and the SUV(roi) regression lines between Cu-ATSM and FDG of each tumor were determined. RESULTS: The SUV mean of Cu-ATSM was lower than that of FDG for both squamous cell carcinoma (SCC) and adenocarcinoma (P < 0.05). In 27 patients with SCC, Cu-ATSM accumulated higher in the peripheral region than in the center of the tumor, and FDG showed the other tendency. Thus, the relationship of the SUV(roi) for Cu-ATSM and FDG showed a negative correlation in SCC. However, 3 adenocarcinoma cases showed similar and homogenous accumulation in the tumor mass with a positive SUV(roi) correlation for the 2 tracers. The regression slope means were -0.12 ± 0.08 for SCC (n = 27) and 0.28 ± 0.12 (n = 3) for adenocarcinoma. CONCLUSION: In patients with head-and-neck cancer, intratumoral distribution of Cu-ATSM and FDG showed a negative correlation in SCC and a positive correlation in adenocarcinoma. The 2 tracers represented different pathophysiological microenvironments in different tumors, suggesting that noninvasive hypoxic tissue imaging with Cu-ATSM would be beneficial in the pretreatment evaluation of head-and-neck cancer.

Platelet derived endothelial cell growth factor/thymidine phosphorylase enhanced human IgE production.

BACKGROUND: Angiogenesis is one pathogenesis of allergic airway disease. METHODS: A potent angiogenic factor is platelet-derived endothelial cell growth factor (PD-ECGF), also known as thymidine phosphorylase (TP) in the field of cancer-associated research. Vascular endothelial growth factor (VEGF) is another representative angiogenic factor. Both factors were added to the culture system of human peripheral blood mononuclear cells (PBMC) with IL-4 and anti-CD40 monoclonal antibody (mAb). Total IgE levels in the supernatants and signal transduction of stimulated PBMC were evaluated. RESULTS: Addition of PD-ECGF enhances in vitro IgE production by PBMC in the presence of IL-4 and anti-CD40 mAb, but VEGF does not enhance IgE production. Although PD-ECGF catalyzes the reversible phosphorolysis of thymidine to 2-deoxy-D-ribose-1-phosphate (2DDR), treatment of 2DDR has no effect on IgE production by human PBMC. Both IL-4 and anti-CD40 mAb induce PD-ECGF by human PBMC. Thymidine phosphorylase inhibitor (TPI), 5-chloro-6-[1- (2-iminopyrrolidinyl) methyl] uracil hydrochloride reduce IgE production via blocking of STAT6- phosphorylation. CONCLUSIONS: Taken together, these results suggest TP involvement in the enhancement of IgE production and suggest that TPI is a novel strategy against IgE-related allergic disease.

Copper-64-diacetyl-bis (N4-methylthiosemicarbazone) accumulates in rich regions of CD133+ highly tumorigenic cells in mouse colon carcinoma.

INTRODUCTION: (64)Cu-diacetyl-bis (N(4)-methylthiosemicarbazone) ((64)Cu-ATSM) is a potential imaging agent of hypoxic tumor for use with PET. Recent literature demonstrated that cancer cells expressing CD133, which is a frequently used marker for so-called cancer stem cells or cancer stem cell-like cells (collectively referred to here as CSCs), contribute to tumor's therapeutic resistance and metastasis ability. Culturing under hypoxia is also reported to enlarge the proportion of CD133(+) cells, which would indicate survival advantage of CD133(+) cells under hypoxia. Here, we investigated the relationships between (64)Cu-ATSM accumulation and existence of CD133(+) cells using mouse colon carcinoma (colon-26) tumor. METHODS: Intratumor distribution of (64)Cu-ATSM and (18)F-fluorodeoxyglucose ((18)FDG) was compared with immunohistochemical staining for CD133 with a colon-26 model. In vitro characterization of CD133(+) colon-26 cells was also performed. RESULTS: In colon-26 tumors, (64)Cu-ATSM localized preferentially in regions with a high density of CD133(+) cells. The percentage of CD133(+) cells was 11-fold higher in (64)Cu-ATSM high-uptake regions compared with (18)FDG high- (but (64)Cu-ATSM low-) uptake regions. CD133(+) colon-26 cells showed characteristics previously linked with CSCs in other cancer cell lines, such as high colony-forming ability, high tumor-initiating ability and enrichment under hypoxic cultivation. The proportion of CD133(+) cells was enlarged by culturing under glucose starvation as well as hypoxia, and (64)Cu-ATSM uptake was increased under such conditions. CONCLUSIONS: Our findings showed that, in colon-26 tumors, (64)Cu-ATSM accumulates in rich regions of CD133(+) cells with characteristics of CSCs. Therefore (64)Cu-ATSM could be a potential imaging agent for rich regions of CD133(+) cells, associated with CSCs, within tumors.

Comparison of the transcellular transport of FDG and D-glucose by the kidney epithelial cell line, LLC-PK1.

OBJECTIVES: Almost all D-glucose (GLU) filtered through the glomerulus is reabsorbed by the renal proximal tubules, whereas a high portion of 2-[18F]fluoro-2-deoxy-D-glucose [(18F)FDG] is excreted through the urine. However, [18F]FDG is not entirely excreted in the urine suggesting that it may be partially reabsorbed by the proximal tubules. The purpose of this study was to compare the time course of transcellular transport of administered [14C] labeled FDG ([14C]FDG) with that of [14C] labeled GLU ([14C]GLU) using the kidney epithelial cell line, LLC-PK1. METHODS: Transcellular transport of [14C]FDG and [14C]GLU by LLC-PK1 cells was measured in Na+-containing or Na+-free Dulbecco's phosphate-buffered saline [PBS(+) and PBS(-), respectively] in the presence or absence of phlorizin, phloretin, probenecid, or tetraethylammonium bromide inhibitors that predominantly inhibit sodium-dependent glucose transporters (SGLTs), sodium-independent glucose transporters, organic anion transporters, and organic cation transporters, respectively. RESULTS: When assayed in PBS(+), less [14C]FDG than [14C]GLU was reabsorbed by the proximal tubular cells over the entire incubation time. Reabsorption of [14C]FDG was mediated mainly by SGLT at early time points in the incubation, whereas high reabsorption of [14C]GLU was mediated by both SGLT and glucose transporter over 90 min of incubation. Secretion of [14C]FDG also tended to be slightly higher than that of [14C]GLU over 90 min of incubation. CONCLUSION: Transcellular transport of [14C]FDG over time by LLC-PK1 cells was clarified. The polarized distribution of transcellular transporters of [14C]FDG and [14C]GLU in LLC-PK1 cells differs.

Radio-copper-labeled Cu-ATSM: an indicator of quiescent but clonogenic cells under mild hypoxia in a Lewis lung carcinoma model.

UNLABELLED: The purpose of this study is to reveal characteristics of (64)Cu-labeled diacetyl-bis(N(4)-methylthiosemicarbazone) ([(64)Cu]Cu-ATSM) during cell proliferation and hypoxia by autoradiography imaging and immunohistochemical staining. METHODS: The intratumoral distributions of [(64)Cu]Cu-ATSM and [(18)F]-2-fluoro-2-deoxy-D-glucose ([(18)F]FDG) in mice implanted with Lewis lung carcinoma (LLC1) tumor cells according to dual autoradiography were compared with the immunohistochemical staining patterns of proliferating markers [Ki-67 and 5-bromo-2'-deoxyuridine (BrdU)] and a hypoxic marker (pimonidazole). A clonogenic assay was performed using the cells of LLC1 tumor-implanted mice, and it was compared with the distribution of [(64)Cu]Cu-ATSM. RESULTS: [(64)Cu]Cu-ATSM mainly accumulated at the edge of tumors, whereas [(18)F]FDG was distributed inside the tumor and inside the [(64)Cu]Cu-ATSM accumulation. The number of Ki-67-positive cells/area tended to increase with [(18)F]FDG accumulation and decrease with [(64)Cu]Cu-ATSM accumulation. On the other hand, the number of BrdU-positive cells/area was negatively correlated with [(18)F]FDG accumulation and positively correlated with [(64)Cu]Cu-ATSM accumulation. High [(64)Cu]Cu-ATSM accumulation was found outside the high-[(18)F]FDG-accumulation and pimonidazole-positive regions. Colony formation ability was significantly higher in the tumor cells obtained from high-[(64)Cu]Cu-ATSM-accumulation regions than the cells from the intermediate- and the low-accumulation regions. CONCLUSION: [(64)Cu]Cu-ATSM accumulation regions in tumor cells indicate quiescent but clonogenic tumor cells under mild hypoxia. [(64)Cu]Cu-ATSM could play an important role in planning appropriate tumor radiotherapy.

Infection of Haemophilus parainfluenzae in tonsils is associated with IgA nephropathy.

We investigated the functions of tonsillar mononuclear cells (TMC) regarding whether a Haemophilus parainfluenzae (HP) outer membranes antigen (HPOM) enhances IgA-related cytokine (IFN-gamma, IL-10. and TGF-beta) production in vitro by TMC in IgA nephropathy (IgAN) patients. In addition, we examined the effect of synthetic oligodeoxynucleotide and HPOM stimulation by TMC on IgA production, whether the constant region antisense to IgA inhibits the production of IgA in vitro by TMC. Eighteen patients with IgAN and 25 patients with chronic tonsillitis (CT) from 6 to 45 years (mean age of 20.9 years) participated in this study. TMC were obtained from resected tonsils, and total and HP-specific IgA levels, along with the concentration of TGF-beta, IL-10 and IFN-gamma in the supernatant of stimulated TMC were measured by ELISA. Isolated TMC were cultured with HPOM in the presence of 23 oligodeoxynucleotides (ODNs), and the induction of total IgA and HP-specific IgA in the supernatant was also measured using ELISA. To investigate the inhibition of IgA production, TMC were cultured with HPOM and antisence to IgA. We found that IgA-related cytokine (IFN-gamma, IL-10, and TGF-beta) production by unstimulated or stimulated TMC was higher in IgAN patients than CT patients. Two types of synthetic oligodeoxynucleotides produced higher HP-specific IgA than with HPOM stimulation alone. HPOM and antisence IgA inhibited the production of total IgA and HP-specific IgA in dose-depend manner. In conclusion, IFN-gamma, TGF-beta, and IL-10 influence each other in the pathogenesis of IgAN, and infection by not only HP but other bacteria or viruses which possess specific DNA sequences such as CpG motifs induce the production of HP-specific IgA by TMC.

[Treatment strategy for T4N1 thyroid papillary carcinoma based upon our clinical experience].

We retrospectively evaluated clinical profiles and prognoses in 152 patients with thyroid papillary carcinoma treated at Fukui Medical University between 1986 and 2000. As standard treatment, 106 (70%) underwent hemithyroidectomy to preserve the normal thyroid lobe. Subtotal thyroidectomy or total thyroidectomy was conducted on 40 cases (23%). Regional lymphnodes were extirpated in 104 (68%) with pathological N0, and radical or conservative neck dissection for 46 cases (30%) with pathological N1. Overall survival for 10 years, estimated using the Kaplan-Meier method, was 100% in both stage I and II, and 95% in stage III. Of 152 thyroid papillary cases, 22 (14%) had tumor recurrence. Of 51 in stage III, 14 (27%) had tumor recurrence. The 14 recurrent in stage III showed local extrathyroidal invasion. Note that 5 of 10 (50%) T4N1 treated with hemithyroidectomy had tumor recurrence in the residual thyroid lobe. Of 11 T4N0 cases who underwent hemithyroidectomy, none had tumor recurrence in the residual thyroid lobe. Results suggest that patients with T4N1 should be treated by total thyroidectomy and neck dissection at initial treatment. Tumor size, cervical lymphnodal metastasis, and distant metastasis may be prognostic factors for thyroid papillary carcinoma.