A theoretical model for prescription of the patient-specific therapeutic activity for radioiodine therapy of Graves' disease

A fundamental function of the thyroid is to extract iodine from the blood, synthesize it into thyroid hormones, and release it into the circulation under feedback control by pituitary-secreted hormones. This capability of the thyroid, termed as functionality, can in principle be related to the severity of hyperthyroidism in individual patients. In this paper the uptake and release of 131I by the thyroid following the administration of 131I therapy for Graves' disease has been theoretically studied. The kinetics of iodine in the thyroid and blood have been evaluated using a two-compartment model. This simplified model appears to be adequate for dosimetry purposes and allows one to correlate levels of increased thyroid functionality (hyperthyroidism) with clinically measurable kinetic parameters. An expression has been derived for the rate of change of thyroid mass following therapy; this has the same form as an empirical relationship described in an earlier work. A method is presented for calculation of the amount of radioiodine activity to be administered to individual patients in order to achieve the desired final functionality of the gland. The activity to be administered is based on measurements of 131I kinetics after the administration of a 'low-activity' (1850 kBq) tracer for treatment planning.     

Influence of thyroid volume reduction on calculated dose in radioiodine therapy of Graves' hyperthyroidism

Administration of radioactive iodine (131I) is an effective treatment for hyperthyroidism due to Graves' disease. Recently several investigators have shown that the success of this therapy may depend on the absorbed dose to the thyroid. Thyroid dose varies inversely with the mass of the gland. Much experimental evidence demonstrates that a reduction of the thyroid volume (mass) may occur after radioiodine therapy. In this work we evaluate the influence of the volume reduction on the calculation of the absorbed dose to the thyroid. A mathematical model of thyroid mass reduction after 131I therapy is presented, based on masses evaluated with ultrasonography of ten patients treated in the endocrinology department of our hospital. This model was applied to the general formula for calculation of the thyroid doses in these patients. The dose values obtained considering a reduction of thyroid mass after the treatment are often quite different from those obtained without considering change in mass (from 9% to 30% greater). We conclude that the consideration of thyroid mass reduction is important for an accurate estimation of the calculated dose.     

Clinicopathologic features and treatment outcomes in differentiated thyroid carcinoma patients with concurrent Graves' disease

The clinical behaviors and treatment outcomes of thyroid carcinomas in patients with Graves' disease is a matter of controversy. This study aimed to identify the clinicopathologic features, treatment outcome, and the indicators for predicting recurrence, and to suggest the optimal extent of surgery in these patients. We retrospectively analyzed data of 58 patients who underwent surgical treatment for differentiated thyroid cancer and concurrent Graves' disease. The follow-up period ranged from 23 to 260 months (mean+/-standard deviation, 116.8+/-54.0). In our series, the mean age was 40.8+/-12.7 yr (range, 15-70), with a male-to-female ratio of 1: 6.25. The mean tumor size was 13+/-9 mm (range, 3-62). The surgical methods included 19 cases of total thyroidectomy, 38 cases of subtotal thyroidectomy, and 1 case of completion total thyroidectomy. Locoregional recurrence occurred in four patients (6.9%). The 10-yr overall survival and disease-free survival of patients were 95.8% and 91.1%, respectively. Age over 45 yr (p=0.031), tumor size over 10 mm (p=0.049), multiplicity (p=0.007), extracapsular invasion (p=0.021), and clinical cancer (p=0.035) were significantly more prevalent in patients with locoregional recurrence than in those without recurrence. We recommend that Graves' disease patients should undergo regular ultrasonography screening for early detection of thyroid carcinoma. We also suggest that the choice of extent of surgery should depend on the diagnostic timing (clinical or incidental) and factors for predicting recurrence.     

A dosimetric algorithm for patient-specific 131I therapy of thyroid cancer based on a prescribed target-mass reduction

(131)I therapy is used in the treatment of differentiated thyroid cancer both to ablate the post-surgical thyroid remnant and to treat recurrent or metastatic cancer. The optimum administered activity for ablation remains controversial: the most commonly used method is the administration of a fixed radioiodine activity (1110-3700 MBq or more); an alternative is the administration of an activity individually calculated to deliver a prescribed absorbed dose (usually 300 Gy for remnant ablation and 80 Gy for treatment of metastasis). Neither of these two approaches is based on a theoretical model and for this reason the debate on the optimization of (131)I therapy of thyroid cancer could have a weak grounding. In this paper, the meaning of the fixed value of target absorbed dose (Gy) is discussed and a mathematical model for remnant/metastasis optimum absorbed dose calculation is presented. This model is based on the desired reduction of the volume of the target (remnant or metastasis) and allows one to calculate individually the value of the optimum target absorbed dose (Gy) and consequently the optimum therapeutic activity to administer to the patient.     

A systematic approach to the assessment of known TNF-alpha polymorphisms in Graves' disease

Single-nucleotide polymorphisms (SNPs) within the tumour necrosis factor alpha (TNF-alpha) gene on chromosome 6p21.3 have been associated with many autoimmune diseases; however, results have been conflicting and accurate allele frequencies have never been established in a UK Caucasian population. The aim of this study was to assess the frequency of 22 known TNF-alpha SNPs in a UK Caucasian control population and investigate association of all polymorphisms with >5% minor allele frequency in a large case-control data set of patients with Graves' disease (GD). Eight of the 22 SNPs had minor allele frequencies >5% and were investigated further. The other 14 SNPs were present in the UK population at frequencies ranging from 0 to 4.7%. A significant increase of the A allele of the -238 SNP was seen in GD patients when compared with control subjects (9.6 vs 6.8%, respectively; P=0.003) and mirrored in the genotype distribution (P=0.009). Furthermore, association of the -238 SNP appears not to be due to linkage disequilibrium of the known HLA-DRB1(*)03 associations with GD. This study has established accurate allele frequencies of TNF-alpha SNPs in a UK population and provides preliminary evidence for association of the TNF-alpha gene with GD.     

Levels of cytokines and thyroid autoantibodies in Omani patients with Graves' disease

An altered balance of pro- and anti-inflammatory cytokines is thought to play an important role in the pathogenesis of autoimmune thyroiditis. The aim of the present study is to assess the cytokine and autoantibody profiles in Omani patients with Graves' disease (GD). Cytokines and autoantibodies including interleukin (IL)-2, IL-4, tumour necrosis factor (TNF)alpha, interferon (IFN)gamma, thyroid stimulating hormone receptor antibody (TRA) and thyroid peroxidase antibody (TPO) are measured in GD patients (n=59) before treatment (n=23) and after treatment (n=36) with 131I-labelled iodine, and compared with normal controls (n=20). Patients with GD showed comparable serum levels of IL-2 but significantly higher levels of IL-4, TNFalpha, IFNgamma, TRA and TPO, compared with the normal controls. There was also a significant increase in serum levels of IL-4 and TNFalpha, and a decrease in TRA in the treated group, compared to the untreated group. IL-4, TNFalpha, IFNgamma, TRA and TPO showed a high prevalence in Omani patients with GD. Thus, cytokines and autoantibodies may prove useful in the diagnosis of GD and in assessing prognosis.     

Accuracy and optimal timing of activity measurements in estimating the absorbed dose of radioiodine in the treatment of Graves' disease

Calculation of the therapeutic activity of radioiodine (131)I for individualized dosimetry in the treatment of Graves' disease requires an accurate estimate of the thyroid absorbed radiation dose based on a tracer activity administration of (131)I. Common approaches (Marinelli-Quimby formula, MIRD algorithm) use, respectively, the effective half-life of radioiodine in the thyroid and the time-integrated activity. Many physicians perform one, two, or at most three tracer dose activity measurements at various times and calculate the required therapeutic activity by ad hoc methods. In this paper, we study the accuracy of estimates of four 'target variables': time-integrated activity coefficient, time of maximum activity, maximum activity, and effective half-life in the gland. Clinical data from 41 patients who underwent (131)I therapy for Graves' disease at the University Hospital in Pisa, Italy, are used for analysis. The radioiodine kinetics are described using a nonlinear mixed-effects model. The distributions of the target variables in the patient population are characterized. Using minimum root mean squared error as the criterion, optimal 1-, 2-, and 3-point sampling schedules are determined for estimation of the target variables, and probabilistic bounds are given for the errors under the optimal times. An algorithm is developed for computing the optimal 1-, 2-, and 3-point sampling schedules for the target variables. This algorithm is implemented in a freely available software tool. Taking into consideration (131)I effective half-life in the thyroid and measurement noise, the optimal 1-point time for time-integrated activity coefficient is a measurement 1 week following the tracer dose. Additional measurements give only a slight improvement in accuracy.     

A NTCP approach for estimating the outcome in radioiodine treatment of hyperthyroidism

Radioiodine has been in use for over 60 years as a treatment for hyperthyroidism. Major changes in clinical practice have led to accurate dosimetry capable of avoiding the risks of adverse effects and the optimization of the treatment. The aim of this study was to test the capability of a radiobiological model, based on normal tissue complication probability (NTCP), to predict the outcome after oral therapeutic 131I administration. Following dosimetric study, 79 patients underwent treatment for hyperthyroidism using radioiodine and then 67 had at least a one-year follow up. The delivered dose was calculated using the MIRD formula, taking into account the measured maximum uptake of administered iodine transferred to the thyroid, U0, and the effective clearance rate, Teff and target mass. The dose was converted to normalized total dose delivered at 2 Gy per fraction (NTD2). Furthermore, the method to take into account the reduction of the mass of the gland during radioiodine therapy was also applied. The clinical outcome and dosimetric parameters were analyzed in order to study the dose-response relationship for hypothyroidism. The TD50 and m parameters of the NTCP model approach were then estimated using the likelihood method. The TD50, expressed as NTD2, resulted in 60 Gy (95% C.I.: 45-75 Gy) and 96 Gy (95% C.I.: 86-109 Gy) for patients affected by Graves or autonomous/multinodular disease, respectively. This supports the clinical evidence that Graves' disease should be characterized by more radiosensitive cells compared to autonomous nodules. The m parameter for all patients was 0.27 (95% C.I.: 0.22-0.36). These parameters were compared with those reported in the literature for hypothyroidism induced after external beam radiotherapy. The NTCP model correctly predicted the clinical outcome after the therapeutic administration of radioiodine in our series.     

Distribution of lymphocytic subpopulations infiltrated in thyroid glands of Graves' disease

We studied ninety cases of thyroid glands both histopathologically and by immunohistochemical methods in patients with Graves' disease using B and T cell markers to evaluate the role of lymphocytic subpopulation. Females were affected more frequently than males with a ratio of 6.5:1, and usually the females were younger than the males at the time of surgery. The heavier the lymphocytic infiltration, the higher was the percentage of germinal center formation or fibrosis. The degree of lymphocytic infiltration was also related to the titers of antithyroglobulin or antimicrosomal antibodies. T cells were mostly scattered individually or in small groups between the follicles; however, in the severely infiltrated group, the major pattern was in clusters. T8 positive cells were more abundant than T4 positive cells, and their distribution pattern was accordant with T11 positive cells. Immunoglobulin synthesizing B cells were positively stained in 47 of 94 cases tested and IgG was the most predominant. In the mild and moderate lymphocytic infiltration groups, IgM was mostly stained at the mantle zone or in the lymphoid cluster of the interfollicular stroma, whereas IgM positive cells were present exclusively in the germinal center of the severely infiltrated group. The results of our study indicate that the major lymphocyte subpopulation in Graves' disease is B lymphocytes, and the degree of T lymphocytic infiltration correlated better with titers of antimicrosomal antibody than antithyroglobulin.     

Verification of the agreement of two dosimetric methods with radioiodine therapy in hyperthyroid patients

The aim of this study was to verify the capability of an MIRD formula-based dosimetric method to predict radioiodine kinetics (fraction of administered iodine transferred to the thyroid, U0, and effective clearance rate, lambda(eff)) and absorbed dose after oral therapeutic 131I administration. The method is based on 123I intravenous administration and five subsequent gamma camera measured uptake values determined separately on different structures within the thyroid. Another dosimetric method based on only the 123I 24-h uptake and a fixed lambda(eff) value was also considered. Eighty-nine hyperthyroid patients (10 with Graves' disease and 79 with autonomously functioning nodules) were studied and 132 thyroidal structures were evaluated. The mean time interval between dosimetry and therapy was 20 +/- 10d. Uptake values were measured at 2, 4, 24, 48, and 120 h during dosimetry and at 2, 4, 24, 48, 96, and 168 h during therapy. The value 0.125d(-1) was chosen in the fixed-lambda(eff) method. The planned doses to the target ranged from 120 to 250 Gy depending on the type and severity of hyperthyroidism. The following significant correlations between therapeutic and dosimetric parameters were found: U0(ther)=0.88U0(dos) (r=0.97,p<0.01), lambda(eff)ther = 1.01 lambda(eff)dos (r=0.85,p<0.01), and D(estimated)= 0.85D(planned) (r=0.88, p<0.01). The percent difference between U0(ther) and U0(dos) ranged from -44 to 32% and between lambda(eff)ther and lambda(eff)dos from -32 to 48%. U0(ther) was lower than U0(dos) in 74% of cases: this can be explained by the self-stunning effect of 131I therapeutic activity that produced a dose of about 20 Gy with a maximum dose rate of 0.6 Gy/h over the initial 24-48 h. The differences, deltaD, between the estimated and the planned doses ranged from -42% (-87 Gy) to 32% (59 Gy); in 73% of cases the difference was within +/- 35 Gy. Greater discrepancies were found with the fixed-lambda(eff) method, in which deltaD ranged from -69 to 95% (-202 to 88 Gy, respectively). In hyperthyroid patients, the five uptake value dosimetric method is able to predict with a good agreement the radioiodine kinetics and the dose after the therapeutic administration in about 73% of the analyzed thyroid structures. The fixed-lambda(eff) method is less reliable.     

Peripheral blood lymphocyte apoptosis and its relationship with thyroid function tests in adolescents with hyperthyroidism due to Graves' disease

Introduction

Failures in apoptotic pathways can contribute to various autoimmune diseases, including autoimmune hyperthyroidism due to Graves’ disease (GD). The aim of the present research was to assess changes in the degree of peripheral blood (PB) lymphocyte apoptosis during methimazole (MMI) treatment in the group of teenage children, and to describe its relationship with thyroid function tests.

Material and methods

The percentage of PB apoptotic lymphocytes, assessed by the decrease in mitochondrial transmembrane potential (CMXRos staining), was measured in 30 adolescents at the time of diagnosis and after obtaining normalization of the thyroid hormone levels.

Results

The percentage of apoptotic lymphocytes in previously untreated patients with GD (5.16 ±2.81%) was significantly lower (p = 0.000001) than the percentage of apoptotic cells in the same group of patients after obtaining methimazole-induced euthyroidism (10.72 ±4.66%). There was a correlation between the increase of the mean percentages of apoptotic lymphocytes and the reduction of FT4 levels (R = 0.63, p < 0.0001), as well as the reduction of TT3 levels (R = 0.95, p < 0.0001). The more signs and symptoms accompanying the diagnosis of GD, the higher was the increment of the degree of lymphocyte apoptosis observed during the MMI-treatment (R = 0.74, p < 0.0000001). The methimazole dosage correlated (R = 0.85, p < 0.0001) with the percentage of apoptotic cells.

Conclusions

The use of methimazole in treatment of hyperthyroidism due to GD leads to an increment of apoptotic cells in PB. Higher doses of methimazole cause a higher increase of apoptotic lymphocytes. Apoptosis induction of human PB lymphocytes seems to be one of the indicators of proper hyperthyroidism treatment.     

Identification of apoptotic proteins in thyroid gland from patients with Graves' disease and Hashimoto's thyroiditis

Apoptosis, i.e. natural programmed cell death, is a physiological phenomenon indispensable for normal functioning of the organism. The signal to apoptosis can be started practically in any cell. Disturbances in the apoptosis regulation determine the essential link of the pathogenesis of many diseases, including autoimmune thyroid disorders.

The aim of the study was to assess the expression of Fas/FasL and caspase eight in the tissues of the thyroid gland in patients with Graves' disease (GD), non-toxic nodular goiter (NTNG) and Hashimoto's thyroiditis (HT). The analysis of Fas/FasL expression was performed by western blot and immunohistochemical investigation with DAB-visualization and Mayer's hematoxylin staining. Caspase-8 expression in thyroid follicular cells was assayed by western blot method.

Identification of the proapoptotic proteins FasL and Fas exhibited their pronounced expression in the thyroid tissue in GD patients (++; ++) and HT (+++; +++) as compared to the NTNG group (0/+; 0/+). Among the study groups, the expression of caspase-8 was revealed in band 55 kDa from patients with autoimmune thyroid diseases.

In GD patients, the percentage of thyrocytes with FasL expression correlated positively with TRAb (R = 0.58, p < 0.02). However, no such correlations were noted in HT or non-toxic multinodular goiter. There were no significant correlations between thyroid hormones and the percentage of thyrocytes with Fas and FasL expression.

In conclusion, our findings suggest that the changes in the expression of apoptotic molecules on the surface of T lymphocytes and thyroid follicular cells in patients with autoimmune thyroid disorders reflect their substantial involvement in the pathogenesis of GD and HT. In addition, analysis of Fas/FasL and caspase-8 expression in thyroid tissue may indicate the disease activity and immunological phenotype.     

Identification of apoptotic proteins in thyroid gland from patients with Graves' disease and Hashimoto's thyroiditis

Apoptosis, i.e. natural programmed cell death, is a physiological phenomenon indispensable for normal functioning of the organism. The signal to apoptosis can be started practically in any cell. Disturbances in the apoptosis regulation determine the essential link of the pathogenesis of many diseases, including autoimmune thyroid disorders. The aim of the study was to assess the expression of Fas/FasL and caspase eight in the tissues of the thyroid gland in patients with Graves' disease (GD), non-toxic nodular goiter (NTNG) and Hashimoto's thyroiditis (HT). The analysis of Fas/FasL expression was performed by western blot and immunohistochemical investigation with DAB-visualization and Mayer's hematoxylin staining. Caspase-8 expression in thyroid follicular cells was assayed by western blot method. Identification of the proapoptotic proteins FasL and Fas exhibited their pronounced expression in the thyroid tissue in GD patients (++; ++) and HT (+++; +++) as compared to the NTNG group (0/+; 0/+). Among the study groups, the expression of caspase-8 was revealed in band 55 kDa from patients with autoimmune thyroid diseases. In GD patients, the percentage of thyrocytes with FasL expression correlated positively with TRAb (R = 0.58, p < 0.02). However, no such correlations were noted in HT or non-toxic multinodular goiter. There were no significant correlations between thyroid hormones and the percentage of thyrocytes with Fas and FasL expression. In conclusion, our findings suggest that the changes in the expression of apoptotic molecules on the surface of T lymphocytes and thyroid follicular cells in patients with autoimmune thyroid disorders reflect their substantial involvement in the pathogenesis of GD and HT. In addition, analysis of Fas/FasL and caspase-8 expression in thyroid tissue may indicate the disease activity and immunological phenotype.     

Separation and analysis of mononuclear cells infiltrating the thyroid of patients with Graves' disease

A simple method was established for separating lymphocytes infiltrating the thyroid from thyroid epithelial cells. Namely, suspensions of minced thyroid from patients with Graves' disease were layered on a Percoll two-step density gradient (p = 1.050 and 1.077 g/ml) and centrifuged (400g, 30 min, 4 degrees C). In this way 0.1-18 X 10(5) lymphocytes/g of thyroid tissue with a purity of 65-95% were obtained. Thyroid lymphocytes were analyzed quantitatively with monoclonal antibodies by laser flow cytometry and compared with peripheral lymphocytes. The proportion of OKT3+ cells was decreased with increase in OKIa+ cells. The percentage of OKIa+ cells was significantly correlated with that of Leu12+ cells. The percentages of OKT4+ cells and OKIa+ cells were higher when analyzed with an extended gate window, which was arranged for detection of activated, large-sized lymphocytes. The percentages of OKT8+ and Leu7+ cells were not significantly different from those in peripheral blood. From these results it was concluded that the proportion of B lymphocytes is increased and that of T lymphocytes is decreased, the proportion of activated B lymphocytes is increased, some helper/inducer T cells are activated in the thyroid gland in Graves' disease, and these activated lymphocytes may be important in local production of antithyroid autoantibodies.     

Treatment of Graves' disease with rituximab specifically reduces the production of thyroid stimulating autoantibodies

Treatment of Graves' disease (GD) with the B-lymphocyte depleting agent rituximab in addition to standard methimazole-therapy prolongs remission. Paradoxically, it does not mediate a reduction in thyrotropin receptor antibody (TRAb) levels over that of methimazole monotherapy. Using a bioassay involving Chinese hamster ovary cells transfected with the human thyrotropin receptor, we found that the stimulatory capacity of TRAbs was reduced markedly, by 66 ± 22%, upon treatment with rituximab and methimazole for 21 days (p < 0.0001), compared to an increase by 33% on average (NS) in patients receiving methimazole alone (p = 0.04 between groups). The overall levels of TRAbs decreased by around 15% in both groups. Within one year of follow-up, rituximab therapy mediated specific decreases in thyroid-peroxidase antibody- and IgM levels, whereas IgG levels were unaffected. The data indicate that rituximab therapy has differential effects on pathogenic and non-pathogenic autoantibodies, even when directed against the same antigen. The possible mechanisms underlying this hitherto unappreciated phenomenon are discussed.