Anti-thyroid Stimulating Hormone Receptor Research Articles

A Comprehensive Analysis of Clinical and Laboratory Characteristics of Thyroid Abnormalities in a Two-Year Outpatient Cross-Sectional Study

Background: The prevalence of thyroid disorders, particularly disseminated Graves’ disease and thyroid nodules, is escalating within the general population. Irregular activation of the immune system is a key factor in causing autoimmune thyroid diseases such as Graves’ disease and Hashimoto thyroiditis. However, the development of autoimmune disorders is influenced by a combination of environmental factors such as urbanization, pollution, and iodine intake. Methods and Results: The study involved 204 patients with thyroid disorders at the department of endocrinology in the outpatient clinic over the past 2 years. The data collection of demographical and clinical information included age, gender, clinical features, laboratory tests, and instrumental analysis. The mean age of participants was 44.35±14.29 years, and females comprised 91.2% of the cases. Among them, 46.1% had thyroid nodules, 36.8% presented with Grave’s disease, 13.2% with Hashimoto’s thyroiditis, and 3.9% with thyroid hyperplasia. The patients with Grave’s disease, the average levels of thyroid antibodies were as follows: anti-thyroid stimulating hormone receptor (Anti-TSHR ) at 21.91±14.02 IU/L, anti-thyroglobulin (Anti-TG) at 265.91±406.03 IU/ml, and anti-thyroid peroxidase (Anti-TPO) at 319.3±327.1 IU/ml, respectively. Additionally, the mean thyroid enlargement was 29.60±16.08 ml. The participants were categorized based on their geographical region, with the distribution as follows: central region 59.3%, northern region 20.1% western region 10.8%, and eastern region 9.8%. Conclusion: Thyroid disorders are prevalent in our population, with greater incidence in females than males. The majority of cases (46.1%) involved thyroid nodules, with a peak age of above 30 years among participants. The central region, particularly the capital city (Ulaanbaatar), exhibited the highest prevalence at 59.3%, as highlighted by the distribution of thyroid abnormalities.

Thyroid dysfunction and autoantibodies among patients infected with Covid 19 in Erbil city /IRAQ

Background and objectives: Autoimmune thyroid diseases or specifically Anti-thyroid peroxidase antibodies increasing after corona virus infection pandemic. This study investigating Anti-thyroperoxidase, anti-thyroid stimulating receptor antibodies in conjunction with traditional thyroid markers after corona virus infection hoping that this would aid in reduction of enduring morbidity and associated health concerns. Methods: This cross-sectional study was conducted in Rizgary Teaching Hospital and Erbil Teaching Hospital in Erbil, Iraq, within the period of February to September 2022. The study comprised hundred patients; 80 females and 20 males, these patients had clinical symptoms suspecting thyroid abnormalities and were suspected for having a sort of thyroid disease. They had abnormal thyroid function tests, with investigating thyroid autoantibodies and corona virus infection. Investigations include thyroid function test and antithyroid antibodies. Results: The level of Anti-thyroid peroxidase antibodies was positive among 75 hypothyroidism patients than among hyperthyroidism cases 2, p-value 0.012. Positive Anti-thyroperoxidase antibodies and hypothyroidism were significantly more common among corona virus infected cases, p-value 0.012 and 0.0109 respectively. However; anti-thyroid stimulating hormone receptor antibody was positive only among hyperthyroidism group (18%); p-value was significant 0.019. Conclusions: This study confirmed the correlation between thyroid function test, Anti-thyroperoxidase antibodies values and corona virus infection. Autoimmune thyroid diseases are more common among female cases, in addition; antithyroid antibodies especially Anti-thyroperoxidase antibodies were more common among female patients. There was high prevalence of both hypothyroidism and positive anti-TPO Ab in COVID-19 cases.

Autoimmune thyroid status in subclinical thyroid disorders in patients attending a tertiary care center in Nepal: a hospital-based cross-sectional study

BackgroundThyroid dysfunction is the leading endocrine disorder worldwide. Iodine deficiency disorders, which were once the major etiology of thyroid dysfunctions, now have been succeeded by autoimmune thyroid diseases with the rise in aberrant salt ionization protocols. This study endeavors to access the level of thyroid autoantibodies viz. anti-thyroid peroxidase (anti-TPO), anti-thyroglobulin (TGA), and anti-thyroid stimulating hormone receptor (TRAb) in individuals with subnormal thyroid profiles.MethodsThis hospital-based cross-sectional study was conducted at the Department of Clinical Biochemistry, Tribhuvan University for a period of six months. Using non-probability (purposive) sampling method, a total of 60 patients were enrolled with subnormal thyroid profiles to include the population who have not yet started medication. Thyroid hormones (free T3, free T4, TSH) and thyroid antibodies (anti-TPO, TGA, and TRAb) were measured. For non-parametric data, Chi-square test and Kruskal-Wallis test were used. Spearman’s correlation was done to determine the association between variables.ResultsOut of 60 participants, the majority of the population between 25 and 44 years were diagnosed with thyroid dysfunction with female preponderance. Among all, 40% (n = 24) had subclinical hyperthyroid states while, 60% (n = 36) had subclinical hypothyroid states, and 75% (n = 45) of the total exhibited positive thyroid antibodies. In subclinical hypothyroid patients with TSH above 10 µIU/ml, anti TPO (58.5%) and TGA (66.7%) positivity were highly prevalent. On the other hand, TRAb was exclusively positive in hyperthyroid condition (50% among the group) which is by far the first of its kind reported in Nepal.ConclusionThe rise in autoimmune thyroid disease among the Nepalese population infers that addressing iodine deficiency simply through salt iodinization may not be adequate to deal with the rising burden of thyroid disorders, especially in iodine-depleted areas. Also, the increasing prevalence of thyroid autoantibodies positivity in subclinical hypothyroidism in the Nepalese population accounts for the arduous screening and monitoring of autoimmune thyroid disorders in Nepal.

Significance of the autoantibody assay in predicting the development of immune-related adverse events in patients receiving atezolizumab plus bevacizumab combination therapy for unresectable hepatocellular carcinoma.

Atezolizumab plus bevacizumab (AB) combination therapy is the first-line treatment for unresectable hepatocellular carcinoma (u-HCC). The management of immune-related adverse events (irAEs) is an important issue associated with achieving a good therapeutic response in patients receiving AB therapy. However, few studies have reported irAE development in patients receiving AB therapy. This study focused on the association between irAE development and autoantibodies at baseline in patients receiving AB therapy. Sixty-one patients receiving AB therapy were enrolled. For autoantibodies, the following antibodies were tested before the start of AB therapy: antinuclear antibodies, rheumatoid factor (RF), anti-thyroglobulin antibodies, thyroid peroxidase antibodies, anti-thyroid stimulating hormone receptor antibodies, and acetylcholine receptor antibodies. A patient was considered to have pre-existing antibodies if any of the listed antibodies were present at baseline. Ten patients (16%) developed irAEs during the observation period. The irAEs included liver injury, hypothyroidism, adrenal insufficiency, adrenocorticotropic hormone deficiency, and rhabdomyolysis. Patients with irAE (n=10) were more likely to be positive for any autoantibody (hazard ratio [HR] 3.7, p=0.047) and RF at baseline (HR 5.4, p=0.035) and to achieve complete response (HR 5.8, p=0.027) than those without. The presence of autoantibodies at baseline was an independent factor associated with irAE development. In the real world, 16% of patients receiving AB therapy for u-HCC developed irAEs. Patients with autoantibodies at baseline are at high risk of developing irAEs and require cautious follow-up.

Thyroid Hormones and Autoantibodies in Iraqi Hyperthyroid Patients

Hyperthyroidism can be defined as the status of over production of thyroid hormones: tri-iodothyronine (T3) and thyroxine (T4) as a results of thyroid gland dysfunction. The aim of the current study was to investigate the association between the abnormalities of thyroid hormones (T3 and T4) and serum levels of thyroid auto-antibodies: anti-thyroid peroxidase (TPO-Ab) and anti-thyroid stimulating hormone receptor (TR-Ab) and the state of hyperthyroidism. Blood samples were collected from 75 Iraqi individualswere early diagnosed as hyperthyroidism patients in addition to 25 Iraqi individuals as a healthy control group. Enzyme Linked Fluorescent Assay (ELFA) used to determine the levels of thyroid hormones (T3 and T4) and the level of thyroid stimulating hormone (TSH), Enzyme Linked Immuno-Sorbent Assay (ELISA) was used to determine the levels of auto-antibodies (TPO-Ab and TR-Ab). The results show that T3 levels and T4 levels in hyperthyroidism patients (2.07 nmol/L and 14.02 μg/dl respectively) increased significantly (푃<0.01)compared to healthy control levels (1.33 nmol/L and 6.4 μg/dl respectively). While the levels of thyroid stimulating hormone (TSH) in hyperthyroidism patient (0.17 μIU/Ml) decreased significantly (푃<0.01)when compared with healthycontrol group (1.84 μIU/Ml). The results also showed that levels of TPO-Ab and TR-Ab in hyperthyroidism patients (235.29 IU/mL and 32.05 IU/mL respectively) increased significantly (푃<0.01)when compared to healthy control group (20.76 IU/mL and 0.3 IU/mL respectively). This study revealed that there is an association between thyroid hormones disorders and the positivity of thyroid auto-antibodies in hyperthyroidism patient

HLA typing of patients who developed subacute thyroiditis and Graves’ disease after SARS-CoV-2 vaccination: a case report

BackgroundCases of subacute thyroiditis (SAT) after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination have been reported. A human leukocyte antigen (HLA) allele, HLA-B*35, appears to be involved in the pathogenesis of SAT.Case presentationWe conducted HLA typing of one patient with SAT and another with both SAT and Graves’ disease (GD), which developed after SARS-CoV-2 vaccination. Patient 1, a 58-year-old Japanese man, was inoculated with a SARS-CoV-2 vaccine (BNT162b2; Pfizer, New York, NY, USA). He developed fever (38 °C), cervical pain, palpitations, and fatigue on day 10 after vaccination. Blood chemistry tests revealed thyrotoxicosis and elevated serum C-reactive protein (CRP) and slightly increased serum antithyroid-stimulating antibody (TSAb) levels. Thyroid ultrasonography revealed the characteristic findings of SAT. Patient 2, a 36-year-old Japanese woman, was inoculated twice with a SARS-CoV-2 vaccine (mRNA-1273; Moderna, Cambridge, MA, USA). She developed fever (37.8 °C) and thyroid gland pain on day 3 after the second vaccination. Blood chemistry tests revealed thyrotoxicosis and elevated serum CRP, TSAb, and antithyroid-stimulating hormone receptor antibody levels. Fever and thyroid gland pain persisted. Thyroid ultrasonography revealed the characteristic findings of SAT (i.e., slight swelling and a focal hypoechoic area with decreased blood flow). Prednisolone treatment was effective for SAT. However, thyrotoxicosis causing palpitations relapsed thereafter, for which thyroid scintigraphy with 99mtechnetium pertechnetate was conducted, and the patient was diagnosed with GD. Thiamazole treatment was then initiated, which led to improvement in symptoms.ConclusionHLA typing revealed that both patients had the HLA-B*35:01, -C*04:01, and -DPB1*05:01 alleles. Only patient 2 had the HLA-DRB1*11:01 and HLA-DQB1*03:01 alleles. The HLA-B*35:01 and HLA-C*04:01 alleles appeared to be involved in the pathogenesis of SAT after SARS-CoV-2 vaccination, and the HLA-DRB1*11:01 and HLA-DQB1*03:01 alleles were speculated to be involved in the postvaccination pathogenesis of GD.

Vitamin D Status in Sample of Iraqi Women with Autoimmune Thyroiditis

The most common cause of acquired thyroid dysfunction is autoimmune thyroid disease (AITD), which most commonly manifests as Hashimoto's thyroiditis (HT) or Graves' disease (GD). The importance of vitamin D (vit D) as an immune modulator has recently been emphasized in several types of disorders. However, its significance in thyroid illnesses is not fully understood. The purpose of this study is to investigate how vitamin D affects the pathophysiology of hyperthyroidism and hypothyroidism in Iraqi women. One hundred Iraqi women with age ranged from 18 to 60 years participate in this research, 50 of them were hypothyroidism patients, 30 were hyperthyroidism patients and the other 20 were euthyroidism served as controls. Blood samples were collected from the studied subjects to determine thyroid profile [free triiodothyronine (FT3), free tetraiodothyronine (FT4) and thyroid stimulating hormone (TSH)], thyroid antibodies [anti-thyroid peroxidase (anti-TPO), anti-thyroglobulin (anti-Tg), and anti-thyroid stimulating hormone receptor (anti-TSHR)], vit D, calcium (Ca), and phosphorus (P) using different analysis techniques. Levels of FT3 and FT4 revealed a significant (P<0.01) increase in hyperthyroidism patients and a significant (P<0.01) decrease in hypothyroidism patients compared with euthyroidism control. While level of TSH was significantly (P<0.01) decreased in hyperthyroidism patients and significantly (P<0.01) increased in hypothyroidism compared with euthyroidism control. When a comparison was made between hyperthyroidism group and hypothyroidism group, the results showed that levels of FT3 and FT4 were significantly (P<0.01) higher in hyerthyroidism than hyporthyroidism while level of TSH was significantly (P<0.01) lower in hyperthyroidism than hyporthyroidism. Levels of anti-TPO and anti-Tg significantly (P<0.01) increased in hyperthyroidism and hypothyroidism patients compared with euthyroidism individuals, while level of anti-TSHR was significantly (P<0.01) decreased in hyperthyroidism patients and significantly (P<0.01) increased in hypothyroidism patients compared with euthyroidism individuals. On the other hand, the findings revealed that level of anti-TPO was significantly (P<0.01) higher while levels of anti-Tg and anti-TSHR were significantly (P<0.01) higher in hypothyroidism than hyperthyroidism. The results revealed a significant (P<0.01) decrease in level of vit D in hyperthyroidism patients and hypothyroidism patients compared with euthyroidism control, a significant (P<0.01) increase in levels of Ca and P in hyperthyroidism patients compared with euthyroidism control, while non-significant (P>0.01) differences were found between hypothyroidism patients and euthyroidism control. Also, the findings revealed that levels of vit D, Ca, and P were significantly (P<0.01) higher in hyperthyroidism than hypothyroidism. Conclusion: It can be concluded that Vit D deficiency may be act as a factor in both hyperthyroidism and hypothyroidism, and the status of vit D impact on autoimmune thyroiditis. Keyword: Autoimmune thyroiditis, Vitamin D, Hyperthyroidism, Hypothyroidism, Euthyroidism

Stability of Anti-thyroid Stimulating Hormone Receptor Antibody in Whole Blood and Serum: Caution Required for Reflective and Batch Testing

Anti-thyroid stimulating hormone receptor antibody (TRAb) stability is stated as 7h at 20-25°C in the Roche Elecsys assay kit insert. The purpose of this study was to determine TRAb stability in whole blood and serum to assess the suitability of samples for reflective and weekly batch testing (with a single freeze-thaw cycle). In the first study, blood from n = 5 volunteers was used to assess: (1) stability in whole blood at room temperature up to 24h, and (2) stability in serum at 4-8°C up to 72h. In the second study, n = 21 patient samples were analysed in serum stored at 4-8°C for two and fivedays post-preliminary analysis. There was a statistically significant decrease in TRAb concentration caused by the single freeze-thaw cycle of -8.9% ± 5.2% (p = 0.03). TRAb concentration decreased in whole blood between 0 and 24h by -16.5% ±9.2%, and declined in serum over time by -11.6% ±6.6% (at 12h). In the patient samples, serum TRAb concentration decreased over time by -4.6% ± 2.5% at day two and -6.5% ± 4.0% at day five. TRAb concentration decreases over time in both whole blood at room temperature and serum samples stored at 4-8°C. A single freeze-thaw cycle also has a statistically significant effect on TRAb concentration.

Graves\u2019 disease coexisted with resistance to thyroid hormone: a case report

BackgroundResistance to thyroid hormone is a rare autosomal dominant disorder characterized by reduced responsiveness to thyroid hormone and can cause syndrome of inappropriate secretion of thyroid stimulating hormone. Although Graves’ disease is a common autoimmune thyroid disorder, the coexistence of these two diseases is extremely rare and makes the diagnosis and treatment complicated, leading to the delayed diagnosis of resistance to thyroid hormone. We describe the case of a Japanese man with resistance to thyroid hormone coexisting with Graves’ disease, in which the correct diagnosis of resistance to thyroid hormone was delayed by masking of the signs of syndrome of inappropriate secretion of thyroid stimulating hormone, with final diagnosis 30 years after the initial treatment for Graves’ disease.Case presentationA 30-year-old Japanese man presented with diffuse goiter and thyrotoxicosis. Anti-thyroid stimulating hormone receptor antibody was positive. He was diagnosed with Graves’ disease. Anti-thyroid medication was chosen as the initial treatment for Graves’ disease. However, this treatment failed to normalize the free triiodothyronine, free thyroxine, and thyroid stimulating hormone levels. His thyroid hormone levels indicated syndrome of inappropriate secretion of thyroid stimulating hormone. After cessation of methimazole treatment by remission of Graves’ disease, his state of syndrome of inappropriate secretion of thyroid stimulating hormone persisted. Magnetic resonance imaging revealed no pituitary tumor lesions. The results of thyroid stimulating hormone-releasing hormone stimulation test showed a normal response of thyroid stimulating hormone. He was suspected to have resistance to thyroid hormone. Direct sequencing analysis of the thyroid hormone receptor β gene identified a heterozygous missense mutation, R282S. Coexistence of resistance to thyroid hormone with Graves’ disease was confirmed. He has no signs of thyrotoxic symptoms, and is capable in activities of daily living at the present time.ConclusionWe described a rare case of resistance to thyroid hormone simultaneously existing with Graves’ disease. This case demonstrated that these diseases can coexist, and indicated some of the difficulties in diagnosis of resistance to thyroid hormone with coexisting Graves’ disease. The diagnosis of resistance to thyroid hormone did not become apparent until after anti-hyperthyroidism treatment. Although rare, careful follow-up after the initial treatment of Graves’ disease is necessary. The coexistence of these two diseases should be considered in patients showing occasional syndrome of inappropriate secretion of thyroid stimulating hormone.

Abstract LB145: CoupledCARTMtechnology for treating thyroid cancer

Abstract Chimeric antigen receptor modified T cells (CAR T) have demonstrated remarkable clinical efficacy in the treatment of B cell malignancies and multiple myeloma. Significant challenges restrict their application across solid tumors due to multiple obstacles, including the lack of robust in vivo CAR-T cell expansion and persistence, the immunosuppressive tumor microenvironment.To address these difficulties, we generated CAR T cells using a novel CoupledCAR® technology. Specifically, we engineered CoupledCAR T cells with lentiviral vectors encoding an anti-thyroid stimulating hormone receptor (TSHR) CAR molecule. Immunohistochemistry (IHC) results showed that TSHR was highly expressed in thyroid cancer cells making it an ideal tumor-specific target antigen. In vitro co-culture experiments showed that TSHR CAR T cells specifically recognized and subsequently killed TSHR-positive tumor cells. Animal model experiments showed that TSHR CAR T cells inhibited the proliferation of TSHR-positive tumor cells.To evaluate the clinical safety and efficacy of anti-TSHR CoupledCAR T cells on refractory or relapsed thyroid cancer, we treated refractory/relapsed post-thyroidectomy thyroid cancer patients according to an IRB approved protocol. We treated two patients using anti-TSHR CoupledCAR T cells and observed the rapid expansion of CAR T cells and enhanced the killing of tumor cells. One patient's best response was complete remission, and the other was near complete remission.Patient 1 Male, 64Y, Papillary Thyroid Carcinoma. In May 2017, Thyroid cancer was diagnosed, bilateral total thyroidectomy, and right cervical lymph node functional dissection were performed in Jun 2018, followed by iodine 131 isotope therapy. In December 2018, bilateral multiple cervical lymph nodes were enlarged, especially on the right side. In February 2019, right neck lymphadenectomy was performed.Patient 2 Female, 60Y, Thyroid Carcinoma. In Aug 2013, a "double lobectomy of the thyroid gland” was performed. From Oct 2013 to Jan 2014, she received iodine 131 isotope therapy. In Sep 2014, she was diagnosed with iodine - resistant thyroid cancer. From Sep to Jan 2016, 5 cycles of chemotherapy were performed. In Jun 2016, she enrolled in the Anlotinib experimental group. In Mar 2019, multiple metastases in both lungs and multiple enlarged lymph nodes in the mediastinum were observed.Patient 1: One month after infusion (M1), the patient was evaluated as PR. Three months after infusion (M3), the patient was evaluated as CR, and the patient's CR lasted from M3 to M12 after infused anti-TSHR CoupledCAR T cells , and we are still following up.Patient 2: M1, the patient was evaluated as PR (Partial Response): the tumor volume in the right lower lobe of the lung was reduced by approximately 67.51% (decreased from 65*55mm to 42*39mm). Three months after infusion (M3), compared with that before, the tumor volume was reduced by approximately 73.54% and SUV max value decreased from 14.9 to 2.8, therefore, the patient was evaluated as nCR (near complete remission).We show that TSHR is a good target for treating thyroid cancer, and our anti-TSHR CoupledCAR T cells are safe and effective for treating thyroid cancer. Recruitment is ongoing to evaluate the safety and efficacy of our CoupledCAR T cells. Further, since our CoupledCAR® technology is a platform technology, we are developing it to treat other solid tumors using different target tumor markers. Citation Format: Lei Xiao, Xingchen Liu, Keshu Zhou, Yu Liu, Yong Huang, Chengfei Pu, Zhiyuan Cao, Ruihong Zhu, Haiyang Tang, Zhipeng Huang, Hang Yang, Xi Huang, Yongping Song, Renbin Liu, Zhao Wu, Victor Lu. CoupledCARTMtechnology for treating thyroid cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB145.

Insights from a Prospective Follow-up of Thyroid Function and Autoimmunity among COVID-19 Survivors.

BackgroundThe occurrence of Graves’ disease and Hashimoto thyroiditis after coronavirus disease 2019 (COVID-19) raised concerns that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may trigger thyroid autoimmunity. We aimed to address the current uncertainties regarding incident thyroid dysfunction and autoimmunity among COVID-19 survivors.MethodsWe included consecutive adult COVID-19 patients without known thyroid disorders, who were admitted to Queen Mary Hospital from July 21 to September 21, 2020 and had serum levels of thyroid-stimulating hormone, free thyroxine, free triiodothyronine (fT3), and anti-thyroid antibodies measured both on admission and at 3 months.ResultsIn total, 122 patients were included. Among 20 patients with abnormal thyroid function tests (TFTs) on admission (mostly low fT3), 15 recovered. Among 102 patients with initial normal TFTs, two had new-onset abnormalities that could represent different phases of thyroiditis. Among 104 patients whose anti-thyroid antibody titers were reassessed, we observed increases in anti-thyroid peroxidase (TPO) (P<0.001) and anti-thyroglobulin (P<0.001), but not anti-thyroid stimulating hormone receptor titers (P=0.486). Of 82 patients with negative anti-TPO findings at baseline, 16 had a significant interval increase in anti-TPO titer by >12 U, and four became anti-TPO-positive. Worse baseline clinical severity (P=0.018), elevated C-reactive protein during hospitalization (P=0.033), and higher baseline anti-TPO titer (P=0.005) were associated with a significant increase in anti-TPO titer.ConclusionMost patients with thyroid dysfunction on admission recovered during convalescence. Abnormal TFTs suggestive of thyroiditis occurred during convalescence, but infrequently. Importantly, our novel observation of an increase in anti-thyroid antibody titers post-COVID-19 warrants further follow-up for incident thyroid dysfunction among COVID-19 survivors.

CoupledCAR technology for treating thyroid cancer.

e14507 Background: Chimeric antigen receptor modified T-cells (CAR-T) have demonstrated remarkable clinical efficacy in the treatment of B-cell malignancies. Significant challenges restrict their application across solid tumors due to multiple obstacles, including the lack of robust in vivo CAR-T cell expansion and persistence in the immunosuppressive tumor microenvironment. Methods: To address these difficulties, we generated CAR-T cells using a novel CoupledCAR technology. Specifically, we engineered CoupledCAR-T cells with lentiviral vectors encoding an anti-thyroid stimulating hormone receptor (TSHR) CAR molecule. In vitro co-culture experiments showed that TSHR CAR-T cells specifically recognized and subsequently killed TSHR-positive tumor cells. Animal model experiments showed that TSHR CAR-T cells inhibited the proliferation of TSHR-positive tumor cells. Results: Patient 1: Male, 64Y, Papillary Thyroid Carcinoma. In May 2017, his Thyroid cancer was diagnosed, bilateral total thyroidectomy, and right cervical lymph node functional dissection were performed in Jun 2018, followed by iodine 131 isotope therapy. In December 2018, bilateral multiple cervical lymph nodes were enlarged, especially on the right side. In February 2019, right neck lymphadenectomy was performed. One month after infusion (M1) of the anti-TSHR CoupledCAR-T cells, the patient was evaluated as PR. Three months after infusion (M3), the patient was evaluated as CR, and the patient's CR lasted from M3 to M12 after infusion of the CoupledCAR-T cells. We are still following the patient for long-term clinical effects. Patient 2: Female, 60Y, Thyroid Carcinoma: In Aug 2013, a "double lobectomy of the thyroid gland” was performed. From Oct 2013 to Jan 2014, she received iodine 131 isotope therapy. In Sep 2014, she was diagnosed with iodine-resistant thyroid cancer. From Sep to Jan 2016, 5 cycles of chemotherapy were performed. In Jun 2016, she enrolled in the Anlotinib experimental group. In Mar 2019, multiple metastases in both lungs and multiple enlarged lymph nodes in the mediastinum were observed. At month 1 (M1) post cell infusion, the patient was evaluated as PR (Partial Response): the tumor volume in the right lower lobe of the lung was reduced by approximately 67.51% (decreased from 65*55mm to 42*39mm). Three months after infusion (M3), the tumor volume was reduced by approximately 73.54% and SUV max value decreased from 14.9 to 2.8. Therefore, the patient was evaluated as nCR (near complete remission). Conclusions: In summary, we showed that TSHR is an attractive and specific target for treating thyroid cancer and our anti-TSHR CoupledCAR-T cells are safe and effective for treating thyroid cancer. Recruitment is ongoing to evaluate the safety and efficacy of our CoupledCAR-T cells. Further, since our CoupledCAR technology is a platform technology, we are developing additional CoulpledCAR-T cells to treat other solid tumors using different target tumor markers.

Decreased Treg Cell and TCR Expansion Are Involved in Long-Lasting Graves’ Disease

Graves’ disease (GD) is a T cell-mediated organ-specific autoimmune disorder. GD patients who have taken anti-thyroid drugs (ATDs) for more than 5 years with positive anti-thyroid stimulating hormone receptor autoantibodies value were defined as persistent GD (pGD). To develop novel immunotherapies for pGD, we investigated the role of T cells in the long-lasting phase of GD. Clinical characteristics were compared between the pGD and newly diagnosed GD (nGD) (N = 20 respectively). Flow cytometric analysis was utilized to determine the proportions of Treg and Th17 cells (pGD, N = 12; nGD, N = 14). T cell receptor sequencing (TCR-seq) and RNA sequencing (RNA-seq) were also performed (pGD, N = 13; nGD, N = 20). Flow cytometric analysis identified lower proportions of Th17 and Treg cells in pGD than in nGD (P = 0.0306 and P = 0.0223). TCR-seq analysis revealed a lower diversity (P = 0.0025) in pGD. Specifically, marked clonal expansion, represented by an increased percentage of top V-J recombination, was observed in pGD patients. Interestingly, pGD patients showed more public T cell clonotypes than nGD patients (2,741 versus 966). Meanwhile, RNA-seq analysis revealed upregulation of the inflammation and chemotaxis pathways in pGD. Specifically, the expression of pro-inflammatory and chemotactic genes (IL1B, IL13, IL8, and CCL4) was increased in pGD, whereas Th17 and Treg cells associated genes (RORC, CARD9, STAT5A, and SATB1) decreased in pGD. Additionally, TCR diversity was negatively correlated with the expression of pro-inflammatory or chemotactic genes (FASLG, IL18R1, CCL24, and CCL14). These results indicated that Treg dysregulation and the expansion of pathogenic T cell clones might be involved in the long-lasting phase of GD via upregulating chemotaxis or inflammation response. To improve the treatment of pGD patients, ATDs combined therapies, especially those aimed at improving Treg cell frequencies or targeting specific expanded pathogenic TCR clones, are worth exploring in the future.

Diagnostic Utility of Mean Peak Systolic Velocity of Superior Thyroid Artery in Differentiating Graves' Disease from Thyroiditis.

Background Differentiating Graves’ disease from thyroiditis can be at times clinically challenging. The gold standard test (thyroid nuclear imaging scan) is expensive, not routinely available, and has radiation exposure. Color Doppler ultrasonography of thyroid represents a suitable alternate which can be used for differentiating these conditions by studying thyroid blood flow parameters. Aim We aimed to investigate the use of thyroid blood flow parameters’ assessment of the superior thyroid artery (STA) and common carotid artery (CCA) with color Doppler ultrasonography for differentiating Graves’ disease from thyroiditis. Materials and Methods This is a cross-sectional study on 111 patients with newly diagnosed thyrotoxicosis (82 with Graves’ disease and 29 with thyroiditis) and 45 years of age and sex-matched healthy controls. All patients underwent detailed clinical and necessary investigations. Color Doppler ultrasonography of the thyroid gland and spectral flow analysis of both superior thyroid arteries was done using standard protocol. Sensitivity and specificity for mean peak systolic velocity of STA (STA-PSV) cut-offs were calculated using receiver operating characteristic curves. Results Patients with Graves’ disease have significantly higher free tri-iodothyronine (FT3) levels, free thyroxine (FT4) levels, antithyroid stimulating hormone receptor antibody (TRAb) levels, and thyroid volume as compared with those with thyroiditis. The mean STA-PSV of patients with Graves’ disease was significantly higher than thyroiditis and control group. Mean STA-PSV greater than 54.3 cm/s had 82.9% sensitivity and 86.2% specificity in diagnosing Graves’ disease. Mean PSV-STA/PSV-CCA ratio of 0.40 was 80.5% sensitive and 86.2% specific for Graves’ disease. Conclusion Mean STA-PSV has high sensitivity and specificity in differentiating Graves’ disease from thyroiditis and can be used routinely in clinical practice as a cheap and invaluable diagnostic tool.

The role of oxidative stress in the pathogenesis of Graves’ orbitopathy

Graves' disease (GD) is a chronic autoimmune condition, in which the anti-thyroid stimulating hormone receptor antibodies (TRAb) activate the thyrotropin receptor (TSHR) located on thyrocytes, leading to excessive thyroid hormone production. TSHR is also expressed in extrathyroidal tissues, in particular, within the orbit. The serum levels of TRAb corelate with severity and activity of thyroid orbitopathy (TO). TO is the most common extrathyroidal manifestation of GD. It is an autoimmune inflammation of orbital tissues, that is, extraocular muscles, orbital adipose tissue or a lacrimal gland. Increased orbital fibroblast and adipocyte proliferation, overproduction of glycosaminoglycans, as well as extraocular muscle oedema result in an increased orbital tissue volume and trigger the onset of TO symptoms. The pathophysiology of TO is complex and has not been fully unexplained to date. Orbital fibroblasts show expression of the TSHR, which is the main target of autoimmunity. It has been hypothesised that T-cell activation induced by orbital receptor stimulation by the target antibody results in orbital tissue infiltration, triggering a cascade of events which leads to the production of cytokines, growth factors and reactive oxygen species (ROS). ROS cause damage to many components of the cell: the cell membrane through the peroxidation of lipids and proteins leading to a loss of their function and enzymatic activity. Oxidative stress leads to activation of the antioxidant system which operates through two mechanisms: enzymatic and non-enzymatic. Assessment of the concentration of oxidative stress markers and the concentration or activity of antioxidative system parameters enables evaluation of oxidative stress severity, which in the future may be utilized for assessment of treatment efficacy and prognosis in patients with active OT.

Rola stresu oksydacyjnego w patogenezie orbitopatii Gravesa

Streszczenie Choroba Gravesa-Basedowa (chGB) jest przewlekłą chorobą autoimmunologiczną, której auto-antygenem jest receptor TSH (TSHR) umiejscowiony na tyreocytach, a jego pobudzenie przez przeciwciała przeciwko TSHR (TRAb) powoduje nadmierne wytwarzanie hormonów tarczycy. TSHR wykazuje także ekspresję w tkankach pozatarczycowych, przede wszystkim w tkankach oczodołu, a stężenie TRAb w surowicy krwi dodatnio koreluje z ciężkością oraz aktywnością orbitopatii tarczycowej (OT). OT jest najczęstszym pozatarczycowym objawem chGB. Jest to choroba autoimmunologiczna, w której dochodzi do zmian zapalnych w obrębie tkanek oczodołów, tj.: mięśniach okoruchowych, tkance tłuszczowej oczodołu czy gruczole łzowym. Wzrost proliferacji fibroblastów oczodołowych i adipocytów oraz nadmierne wytwarzanie glikozaminoglikanów, obrzęk mięśni okoruchowych zwiększają objętość tkanek oczodołu i powodują powstanie objawów klinicznych choroby. Patogeneza OT jest złożona i nadal pozostaje niewyjaśniona. Fibroblasty oczodołów wykazują ekspresję TSHR, który jest głównym miejscem ataku autoimmunologicznego. Zgodnie z szeroko akceptowaną hipotezą po pobudzeniu receptorów dochodzi do aktywacji limfocytów T, które naciekając tkanki oczodołu stymulują je do wytwarzania cytokin, czynników wzrostu oraz wolnych rodników tlenowych (WRT). WRT powodują uszkodzenie wielu składowych komórki m.in. błony komórkowej przez peroksydację lipidów oraz białek doprowadzając do utraty ich funkcji i aktywności enzymatycznej. Stres oksydacyjny uaktywnia układ antyoksydacyjny działający poprzez dwa mechanizmy: enzymatyczny oraz nieenzymatyczny. Ocena stężenia markerów stresu oksydacyjnego oraz stężenia lub aktywności parametrów układu antyoksydacyjnego umożliwia ocenę nasilenia stresu oksydacyjnego, co w przyszłości może być wykorzystywane do oceny skuteczności leczenia i rokowania u chorych z aktywną OT.

The profile of autoimmunity in Type 1 diabetes patients.

Background:Type 1 diabetes mellitus (T1DM) is an autoimmune disorder caused by pancreatic β-cells destruction. Anti-pancreatic antibodies are the witness of β-cell destruction and their dosage is mainly used for etiological diagnosis. Patients with T1DM are at increased risk of developing other autoimmune reactions, which may involve other organs, resulting in organ specific autoimmune disease. The most frequently encountered are autoimmune thyroid disease, followed by celiac and gastric disease and other rare autoimmune diseases.Objectives:The purpose of this study is to investigate the prevalence of autoimmune markers in patients with T1DM.Methods:The study was conducted at the Department of Endocrinology of the Military Hospital Moulay Ismail in Meknes Morocco, from January 2016 to December 2018. All Type 1 diabetes patients consulting during the study period were included in the study. Their clinical and biochemical data were collected at their first presentation, made up of anti-pancreatic antibodies (glutamic acid decarboxylase [GAD] antibody, tyrosine phosphatase antibody, and islet cell antibody) and other organ-specific antibodies: the thyroid (antithyroid peroxidase antibody, antithyroglobulin antibody, and antithyroid-stimulating hormone receptor antibody), the intestine (IgA antitissue transglutaminase antibody), the adrenal gland (anti-21 hydroxylase antibody), and the stomach (antigastric parietal cell antibody and anti-intrinsic factor antibody).Results:Fifty-four patients were included, with an average age of 26 years. GAD, tyrosine phosphatase, and islet cell antibodies were detected in 74%, 22%, and 3.7%, respectively, of the 54 patients examined. The prevalence of extrapancreatic autoimmunity was 45% with a large preponderance among different immunities of those from thyroid and celiac diseases (CDs).Conclusion:Our results confirm that patients with Type 1 diabetes should be investigated for the presence of autoimmune diseases mainly from thyroid and CDs.

CoupledCARTM Technology for Treating Thyroid Cancer

CoupledCAR TM Technology for Treating Thyroid Cancer Chimeric antigen receptor modified T cells (CAR T) have demonstrated remarkable clinical efficacy in the treatment of B cell malignancies and multiple myeloma. Significant challenges restrict their application across solid tumors due to multiple obstacles, including the lack of robust in vivo CAR-T cell expansion and persistence, the immunosuppressive tumor microenvironment, and tumor escape due to heterogeneous tumor cell composition with a potential loss of the targeted tumor antigen. To address these difficulties, we generated CAR T cells using a novel CoupledCARTM technology. Specifically, we engineered CoupledCAR T cells with lentiviral vectors encoding an anti-thyroid stimulating hormone receptor (TSHR) CAR molecule. Immunohistochemistry (IHC) results showed that TSHR was highly expressed in thyroid cancer cells making it an ideal tumor-specific target antigen. In vitro co-culture experiments showed that TSHR CAR T cells specifically recognized and consequently killed TSHR-positive tumor cells. Animal experiments showed that TSHR CAR T cells inhibited the proliferation of TSHR-positive tumor cells. To evaluate the clinical safety and efficacy of anti-TSHR CoupledCAR T cells on refractory or relapsed thyroid cancer, we treated refractory/relapsed post-thyroidectomy thyroid cancer patients according to an IRB approved protocol. We treated two patients using anti-TSHR CoupledCAR T cells and observed the rapid expansion of CAR T cells and enhanced the killing of tumor cells. One patient's best response was complete remission, and the other was near complete remission. Patient Profile: Patient 1 Male, 64Y, Papillary Thyroid Carcinoma. In May 2017, Thyroid cancer was diagnosed, bilateral total thyroidectomy, and right cervical lymph node functional dissection were performed in June, followed by iodine 131 isotope therapy. In December 2018, bilateral multiple cervical lymph nodes were enlarged, especially on the right side. In February 2019, right neck lymphadenectomy was performed. Patient 2 Female, 60Y, Thyroid Carcinoma. In Aug 2013, a "double lobectomy of the thyroid gland" was performed. From Oct 2013 to Jan 2014, she received iodine 131 isotope therapy. In Sep 2014, she was diagnosed with iodine - resistant thyroid cancer. From Sep to Jan 2016, 5 cycles of chemotherapy were performed. In Jun 2016, she enrolled in the Anlotinib experimental group. In Mar 2019, multiple metastases in both lungs and multiple enlarged lymph nodes in the mediastinum were observed. Observations and Results: Patient 1: One month after infusion (M1), the patient was evaluated as PR: lymph node metastasis became undetectable and the size of the thoracic paratracheal tumor nodules decreased significantly. Three months after infusion (M3), the patient was evaluated as CR, and the tumor tissue was substantially smaller than M1. Patient 2: M1, the patient was evaluated as PR (Partial Response): the tumor volume in the right lower lobe of the lung was reduced by approximately 67.51% (decreased from 65*55mm to 42*39mm). Three months after infusion (M3), compared with that before, the tumor volume was reduced by approximately 73.54% and SUV max value decreased from 14.9 to 2.8, therefore, the patient was evaluated as nCR (near complete remission). We show that TSHR is a good target for treating thyroid cancer, and our anti-TSHR CoupledCAR T cells are safe and effective for treating thyroid cancer. Recruitment is ongoing to evaluate the safety and efficacy of our CoupledCAR T cells. Further, since our CoupledCARTM technology is a platform technology, we are developing it to treat other solid tumors using different target markers. Disclosures Xiao: Innovative Cellular Therapeutics: Other: stockholder.

Short-Term Versus 6-Week Prednisone In The Treatment Of Subacute Thyroiditis: A Randomized Controlled Trial

Moderate-to-severe subacute thyroiditis is clinically managed with 6 to 8 weeks of glucocorticoid therapy. However, no studies have evaluated short-term prednisone treatment for subacute thyroiditis. This 24-week, prospective, single-blind, randomized controlled study enrolled patients (aged 18 to 70 years) with subacute thyroiditis who were hospitalized between August, 2013, and December, 2014. Patients with moderate-to-severe symptoms were randomly assigned to receive either 30 mg/day prednisone for 1 week, followed by 1 week of nonsteroidal anti-inflammatory drugs, or the conventional 6-week prednisone therapy. The primary endpoint was intergroup differences in treatment efficacy at the end of the treatment course. Secondary endpoints included between-group differences in post-withdrawal adverse effect parameters and thyroid function at weeks 6, 12, and 24. We screened 96 patients, randomized 52 participants, and 50 participants completed the study. Efficacy and recurrence rates were not significantly different at withdrawal in both groups (P = .65). At treatment discontinuation, parathyroid hormone (28.8 versus 38.9 pg/mL; P = .011) and systolic blood pressure (113.9 versus 122.4mm Hg; P = .023) were significantly lower in the experimental group than in the control group. There were no significant intergroup differences in other secondary endpoints at withdrawal and in thyroid function at weeks 6, 12, and 24. Fewer side effects of glucocorticoids and similar efficacy and recurrence rates were observed with short-term prednisone compared with the 6-week treatment for subacute thyroiditis. Short-term prednisone, with a better safety profile, may be an alternative strategy for ameliorating moderate-to-severe symptoms of subacute thyroiditis. BPPG = breakfast postprandial plasma glucose; CRP = C-reactive protein; ESR = erythrocyte sedimentation rate; FPG = fasting plasma glucose; FT3 = free triiodothyronine; FT4 = free thyroxine; GA = glycated albumin; NSAIDs = nonsteroidal anti-inflammatory drugs; OC = osteocalcin; PINP = type I procollagen amino-terminal peptide; PTH = parathyroid hormone; RCT = randomized controlled trial; SAT = subacute thyroiditis; SBP = systolic blood pressure; TC = total cholesterol; TSH = thyroid-stimulating hormone; TBG = thyroid-binding globulin; TG = triglyceride; TGAb = antithyroglobulin antibody; TPOAb = antithyroid peroxidase antibody; TRAb = antithyroid-stimulating hormone receptor antibody.

CoupledCAR technology for treating thyroid cancer.

e15027 Background: Significant challenges restrict CAR-T cell therapy to treat solid tumors. Methods: Here, we generated CAR-T cells using a novel CoupledCAR technology. Specifically, we engineered CoupledCAR T cells with lentiviral vectors encoding an anti-thyroid stimulating hormone receptor (TSHR) CAR molecule, and anti-TSHR CAR-T cells showed anti-tumor activities in vitro and in vivo experiments. Further, we treated refractory/relapsed post-thyroidectomy thyroid cancer patients using anti-TSHR CoupledCAR T cells, and observed the rapid expansion of CAR-T cells and the enhanced killing of tumor cells. Results: Both patients achieved PR (Partial Response). Patient Profile: Patient 1 Male, 64Y, Papillary Thyroid Carcinoma. In May 2017, thyroid cancer was diagnosed, bilateral total thyroidectomy, and right cervical lymph node functional dissection were performed in June, followed by iodine 131 isotope therapy. In December 2018, bilateral multiple cervical lymph nodes were enlarged. In February 2019, right neck lymphadenectomy was performed. Patient 2 Female, 60Y, Thyroid Carcinoma. In Aug 2013, a "double lobectomy of the thyroid gland” was performed. From Oct 2013 to Jan 2014, she received iodine 131 isotope therapy. In Sep 2014, she was diagnosed with iodine-resistant thyroid cancer. In 2016, 5 cycles of chemotherapy were performed. In Mar 2019, multiple metastases in both lungs and multiple enlarged lymph nodes were observed. Observations and Results: Patient 1: One month after infusion (M1), the patient was evaluated as PR: lymph node metastasis became undetectable and the size of the thoracic paratracheal tumor nodules decreased significantly. Three months after infusion (M3), the patient was evaluated as having a durable response, and the tumor tissue was substantially smaller than M1. Patient 2: M1, the patient was evaluated as PR (Partial Response): the tumor volume in the right lower lobe of the lung was reduced by approximately 67.51% (decreased from 65*55mm to 42*39mm). Three months after infusion (M3), compared with that before, the tumor volume was reduced by approximately 73.54% and SUV max value decreased from 14.9 to 2.8, therefore, the patient was evaluated as nCR (near complete remission). Conclusions: We show that TSHR is a good target for treating thyroid cancer, and our anti-TSHR CoupledCAR T cells are safe and effective for treating thyroid cancer. Recruitment is ongoing to evaluate the safety and efficacy of our CoupledCAR T cells. Further, since our CoupledCAR technology is a platform technology, we are developing it to treat other solid tumors using different target markers.