Malignant Thyroid Tissues Research Articles

Zirconium- 89 Labeled Antibody K1-70 for PET Imaging of Thyroid-stimulating Hormone Receptor Expression in Thyroid Cancer.

Thyroid-stimulating hormone receptor (TSHR) is a G-protein coupled receptor that is highly expressed on benign and malignant thyroid tissues. TSHR binding and activation has long been a component of thyroid cancer molecular imaging and radiotherapy, by promoting expression of the sodium-iodide symporter (NIS) and incorporation of I-131 into thyroid hormones. Here, we report the radiosynthesis and preclinical evaluation of a Zirconium-89 (89Zr) labeled TSHR antibody to serve as a positron emission tomography (PET) diagnostic correlate for therapeutic agents targeting TSHR without reliance on NIS. TSHR human monoclonal antibody K1-70 was conjugated to chelator desferrioxamine-p-benzyl-isothiocyanate, followed by labeling with Zr-89, yielding the radiotracer 89Zr-DFO-TSHR-Ab. The in vitro cellar uptake and binding affinity of 89Zr-DFO-TSHR-Ab were analyzed in three new TSHR stable overexpressing tumor cell lines and their corresponding wild types (WT) with low or no TSHR expression. 89Zr-DFO-TSHR-Ab PET/CT imaging of TSHR expression was evaluated in tumor mouse models bearing one TSHR-positive tumor and other negative control with or without the coinjection of antibody K1-70, and then verified by radiotracer biodistribution study and tumor immunohistochemistry (IHC). The conjugate DFO-TSHR-Ab was labeled with Zr-89 at 37°C for 60min and purified by PD-10 column in radiochemical yields of 68.8 ± 9.9%, radiochemical purities of 98.7 ± 0.8%, and specific activities of 19.1 ± 2.7mCi/mg (n = 5). In vitro cell studies showed 89Zr-DFO-TSHR-Ab had significantly high uptake on TSHR expressing tumor cells with nanomolar affinity and high potency. Preclinical PET/CT imaging revealed that 89Zr-DFO-TSHR-Ab selectively detected TSHR expressing thyroid tumors and displayed improved in vivo performance with the coinjection of unlabeled TSHR antibody K1-70 leading to higher uptake in TSHR expressing tumors than parental WT tumors and physiologic tissues; this observation was confirmed by the biodistribution and immunostaining analyses. We synthesized 89Zr-labeled antibody K1-70 as a new radiopharmaceutical for PET imaging of TSHR. 89Zr-DFO-TSHR-Ab has high radioactive uptake and retention in TSHR expressing tumors and cleared quickly from most background tissues in mouse models. Our study demonstrated that 89Zr-DFO-TSHR-Ab has the potential for PET imaging of TSHR-positive thyroid cancer and monitoring TSHR-targeted therapy.

The expression of c-MYC, Cyclin D1 and Ki-67/MIB-1 in benign and malignant thyroid tissues: is there any diagnostic value?

To investigate the immunohistochemical (IHC) expression and the diagnostic value of c-MYC, Cyclin D1, and Ki-67∕MIB-1 in follicular adenomas (FAs), follicular carcinomas (FCs), and anaplastic carcinomas (ACs) of the thyroid gland, as well as in their corresponding adjacent, non-neoplastic thyroid tissue (NNTT). We conducted a retrospective study of patients who were pathologically diagnosed with FA, FC, or AC after total thyroidectomy. Tissue microarrays with cores taken from neoplastic and adjacent NNTT were constructed. Immunohistochemistry for anti-c-MYC, anti-Cyclin D1, and anti-Ki-67∕MIB-1 antibodies was performed, and the positivity was evaluated. Twenty-eight specimens were included. Nuclear c-MYC positivity was observed in 4∕11 FCs, and 3∕4 ACs, whereas cytoplasmic c-MYC positivity was found in 16∕24 NNTTs. Globally, there were statistically significant differences between neoplasms and NNTTs, with higher nuclear c-MYC and Cyclin D1 expression observed in neoplasms (p=0.017 and p=0.001, respectively). In contrast, cytoplasmic positivity was seen solely in NNTTs (p=0.001). Cyclin D1 positivity was noted in 11∕13 FAs, 7∕11 FCs, 2∕4 ATCs, and only in one NNTT. A statistically significant correlation was found between MIB1 and c-MYC nuclear positivity (p=0.040). Our findings exhibit a clear difference in the IHC expression of c-MYC and Cyclin D1 between different types of thyroid tumors, as well as between the neoplastic and NNTT. Nuclear c-MYC positivity excludes the benign nature of a thyroid lesion, in contrast to cytoplasmic positivity, which demonstrates normal or hyperplastic nature.

Identification of Germline FOXE1 and Somatic MAPK Pathway Gene Alterations in Patients with Malignant Struma Ovarii, Cleft Palate and Thyroid Cancer.

Germline variants in the FOXE1 transcription factor have been associated with thyroid ectopy, cleft palate (CP) and thyroid cancer (TC). Here, we aimed to clarify the role of FOXE1 in Portuguese families (F1 and F2) with members diagnosed with malignant struma ovarii (MSO), an ovarian teratoma with ectopic malignant thyroid tissue, papillary TC (PTC) and CP. Two rare germline heterozygous variants in the FOXE1 promoter were identified: F1) c.-522G>C, in the proband (MSO) and her mother (asymptomatic); F2) c.9C>T, in the proband (PTC), her sister and her mother (CP). Functional studies using rat normal thyroid (PCCL3) and human PTC (TPC-1) cells revealed that c.9C>T decreased FOXE1 promoter transcriptional activity in both cell models, while c.-522G>C led to opposing activities in the two models, when compared to the wild type. Immunohistochemistry and RT-qPCR analyses of patients' thyroid tumours revealed lower FOXE1 expression compared to adjacent normal and hyperplastic thyroid tissues. The patient with MSO also harboured a novel germline AXIN1 variant, presenting a loss of heterozygosity in its benign and malignant teratoma tissues and observable β-catenin cytoplasmic accumulation. The sequencing of the F1 (MSO) and F2 (PTC) probands' tumours unveiled somatic BRAF and HRAS variants, respectively. Germline FOXE1 and AXIN1 variants might have a role in thyroid ectopy and cleft palate, which, together with MAPK pathway activation, may contribute to tumours' malignant transformation.

Evaluation of Pendrin Expression Using Nuclear Imaging Modalities and Immunohistochemistry in Animal Thyroid Cancer Model.

The impaired ability of thyroid cancer (TC) cells to uptake and concentrate iodine represents a major therapeutic challenge in malignant TC management. This has been reported probably due to reduced or loss of expression of pendrin in thyroid tumors. In view of this, we evaluated the pendrin expression in the chemically induced (using N-bis[2-hydroxypropyl] nitrosamine [DHPN]) TC model in Wistar rats. Uptake in the thyroid gland was evaluated by positron emission tomography with computed tomography (PET-CT) and scintigraphy imaging. Further histopathology (HP) and immunohistochemistry (IHC) were performed for confirming malignancy. The altered uptake in the thyroid gland was observed by PET-CT and scintigraphy imaging. Significant pathological changes in the thyroid were observed using 2-deoxy-2-(fluorine-18) fluoro-D-glucose PET-CT, technetium-99m pertechnetate imaging, and reduced iodine-131 uptake (n = 4) in DHPN-induced animals compared to control indicative of thyroid cell proliferation. In treated groups, tissue HP revealed hyperplastic follicular to papillary cell proliferation with variable mitotic activity. The malignant nature of the tissue and variable uptake of the tracer were further reconfirmed by IHC. IHC revealed reduced pendrin expression in malignant thyroid tissue. Hence, nuclear imaging techniques can be of aid in the early identification and evaluation of cellular changes during the early development of tumor models in laboratory animals. In conclusion, our study reveals that pendrin expression plays a vital role in thyroid uptake, and its reduction was observed in TC in a chemically induced TC model.

Proteomics Reveals mRNA Regulation and the Action of Annexins in Thyroid Cancer.

Differentiated thyroid cancer is the most common malignancy of the endocrine system. Although most thyroid nodules are benign, given the high incidence of thyroid nodules in the population, it is important to understand the differences between benign and malignant thyroid cancer and the molecular alterations associated with malignancy to improve detection and signal potential diagnostic, prognostic, and therapeutic targets. Proteomics analysis of benign and malignant human thyroid tissue largely revealed changes indicating modifications in RNA regulation, a common cancer characteristic. In addition, changes in the immune system and cell membrane/endocytic processes were also suggested to be involved. Annexin A1 was considered a potential malignancy biomarker and, similarly to other annexins, it was found to increase in the malignant group. Furthermore, a bioinformatics approach points to the transcription factor Sp1 as being potentially involved in most of the alterations seen in the malignant thyroid nodules.

Changes in antioxidant status and DNA repair capacity are corroborated with molecular alterations in malignant thyroid tissue of patients with papillary thyroid cancer.

Introduction: Papillary thyroid cancer (PTC) accounts for approximately 80% of all thyroid cancer cases. The mechanism of PTC tumourigenesis is not fully understood, but oxidative imbalance is thought to play a role. To gain further insight, this study evaluated antioxidant status, DNA repair capacity and genetic alterations in individuals diagnosed with benign thyroid lesion in one lobe (BTG) and PTC lesion in another. Methods: Individuals with coexisting BTG and PTC lesions in their thyroid lobes were included in this study. Reactive oxygen species (ROS) level, ABTS radical scavenging activity, ferric reducing antioxidant capacity, glutathione peroxidase and superoxide dismutase activities were measured in the thyroid tissue lysate. The expression of selected genes and proteins associated with oxidative stress defence and DNA repair were analysed through quantitative real-time PCR and Western blotting. Molecular alterations in genomic DNA were analysed through whole-exome sequencing and the potentially pathogenic driver genes filtered through Cancer-Related Analysis of Variants Toolkit (CRAVAT) analysis were subjected to pathway enrichment analysis using Metascape. Results: Significantly higher ROS level was detected in the PTC compared to the BTG lesions. The PTC lesions had significantly higher expression of GPX1, SOD2 and OGG1 but significantly lower expression of CAT and PRDX1 genes than the BTG lesions. Pathway enrichment analysis identified "regulation of MAPK cascade," "positive regulation of ERK1 and ERK2 cascade" and "negative regulation of reactive oxygen species metabolic process" to be significantly enriched in the PTC lesions only. Four pathogenic genetic variants were identified in the PTC lesions; BRAF V600E, MAP2K7-rs2145142862, BCR-rs372013175 and CD24 NM_001291737.1:p.Gln23fs while MAP3K9 and G6PD were among 11 genes that were mutated in both BTG and PTC lesions. Conclusion: Our findings provided further insight into the connection between oxidative stress, DNA damage, and genetic changes associated with BTG-to-PTC transformation. The increased oxidative DNA damage due to the heightened ROS levels could have heralded the BTG-to-PTC transformation, potentially through mutations in the genes involved in the MAPK signalling pathway and stress-activated MAPK/JNK cascade. Further in-vitro functional analyses and studies involving a larger sample size would need to be carried out to validate the findings from this pilot study.

Expression of CK19 in papillary thyroid carcinoma in comparison with benign thyroid lesions

Background: The Papillary variant of thyroid carcinoma is one of the common types of thyroid malignancies , the commonest. Its diagnosis usually is microscopic which based on nuclear features. Objective: To assess the immunohistochemical expression of cytokeratin 19 (CK19) in PTC and compare its reactivity with other thyroid conditions to approve its importance as a useful test for diagnosing PTC. Patients and Methods: Twenty- cases of papillary thyroid carcinoma (with different variants), five follicular adenomas and ten multinodular goiters (MNG) were collected over three years (2018-2021).10% formalin-fixed sections were collected from thyroidectomy specimens. Stained with Hematoxylin and eosin then immunohistochemical staining for immune-marker CK 19 was done using standard techniques. The results were quantitatively scored as the following: score 0 (negative ), score 1+ (<5%) positive cells), score 2+ (5- 25%), score 3+ (25-75%) ,finally score 4+ (>75%) positive cells), and then calculated. Results: An 18 (90%) papillary carcinomas show diffuse and strong (3+ and 4+) expression of CK19 . ,three out of ten (30%) MNG were weakly positive(1+ and 2+) and focal in distribution while in follicular adenoma all cases were negative for CK19 . Conclusion: Ck 19 can stain benign and malignant thyroid tissue, but strong membranous staining applied in cases of papillary ca , thus can be used as confirmatorytool for PTC diagnosis.

Expression of CK19 in papillary thyroid carcinoma in comparison with benign thyroid lesions

Background: The Papillary variant of thyroid carcinoma is one of the common types of thyroid malignancies , the commonest. Its diagnosis usually is microscopic which based on nuclear features.
 Objective: To assess the immunohistochemical expression of cytokeratin 19 (CK19) in PTC and compare its reactivity with other thyroid conditions to approve its importance as a useful test for diagnosing PTC.
 Patients and Methods: Twenty- cases of papillary thyroid carcinoma (with different variants), five follicular adenomas and ten multinodular goiters (MNG) were collected over three years (2018-2021).10% formalin-fixed sections were collected from thyroidectomy specimens. Stained with Hematoxylin and eosin then immunohistochemical staining for immune-marker CK 19 was done using standard techniques. The results were quantitatively scored as the following: score 0 (negative ), score 1+ (<5%) positive cells), score 2+ (5- 25%), score 3+ (25-75%) ,finally score 4+ (>75%) positive cells), and then calculated.
 Results: An 18 (90%) papillary carcinomas show diffuse and strong (3+ and 4+) expression of CK19 . ,three out of ten (30%) MNG were weakly positive(1+ and 2+) and focal in distribution while in follicular adenoma all cases were negative for CK19 .
 Conclusion: Ck 19 can stain benign and malignant thyroid tissue, but strong membranous staining applied in cases of papillary ca , thus can be used as confirmatorytool for PTC diagnosis.

The Role of Thyroglobulin in Preoperative and Postoperative Evaluation of Patients With Differentiated Thyroid Cancer.

Thyroglobulin (Tg) is secreted by thyroid follicular cells and stored in the thyroid follicular lumen as a component of thyroid hormone. It is known that both benign and well-differentiated malignant thyroid tissue can secrete Tg. In recent years, growing lines of evidence have shown that Tg plays an important role in the diagnosis and metastasis of preoperative differentiated thyroid carcinoma (DTC). The levels of Tg, whether in the serum or in a fine-needle aspiration washout fluid, are usually viewed as an excellent indicator in the monitoring of postoperative DTC, including the guidance and evaluation of radioactive iodine ablation. Nevertheless, some factors limit the application of Tg, such as the method used to measure Tg and the presence of Tg antibodies. This review aimed to summarize the role of Tg in the preoperative and postoperative evaluation of patients with DTC, and the factors influencing Tg. This review could provide a reference for a more accurate application of Tg in patients with DTC.

Knockdown of KRAB domain-associated protein 1 suppresses the proliferation, migration and invasion of thyroid cancer cells by regulating P68/DEAD box protein 5

ABSTRACT KRAB domain-associated protein 1 (KAP-1) has been reported to be an oncogene in diverse tumors. KAP-1 was found to have abundant existence in malignant thyroid tissues, but its role in thyroid cancer hasn’t been elucidated clearly. This study was carried out to explore the role of KAP-1 in thyroid cancer, and to clarify its molecular mechanism. The expressions of KAP-1 and P68/DEAD box protein 5 (DDX5) were assessed under the help of qRT-PCR and western blot. Then, we downregulated KAP-1 or upregulated DDX5 by cell transfection in TPC-1 cells. A series of cellular experiments on proliferation, apoptosis, migration and invasion were conducted with CCK-8, EdU, TUNEL, wound-healing and Transwell assays. Besides, the relationship between KAP-1 and DDX5 was verified by co-immunoprecipitation (Co-IP). The results showed that both of KAP-1 and DDX5 were upregulated in thyroid cancer cells. Loss-of-function experiments revealed that KAP-1 knockdown imparted suppressive effects on cell proliferation, migration and invasion, but promoted cell apoptosis. Additionally, KAP-1 was demonstrated to interact with DDX5 and positively regulate DDX5 expression. The following rescued experiments exhibited that the inhibitory effects of KAP-1 knockdown on cellular activities of thyroid cancer and Wnt/β-catenin signaling were also partly reversed by DDX5 overexpression. Moreover, activation of Wnt/β-catenin signaling retarded the anti-tumor activity of KAP-1 knockdown. In conclusion, the data in this study disclosed that KAP-1 silence helped to repress the cell proliferation, migration and invasion by degrading DDK5, so as to hinder the development of thyroid cancer.

Content of Copper, Iron, Iodine, Rubidium, Strontium and Zinc in Thyroid Malignant Nodules and Thyroid Tissue adjacent to Nodules

Thyroid malignant nodules (TMNs) are the most common endocrine cancer. The etiology and pathogenesis of TMNs must be considered as multifactorial. Diagnostic evaluation of TMNs represents a challenge, since there are numerous benign and malignant thyroid disorders that need to be exactly attributed. The present study was performed to clarify the possible role of some trace elements (TEs) as cancer biomarker. For this aim thyroid tissue levels of copper (Cu), iron (Fe), iodine (I), rubidium (Rb), strontium (Sr), and zinc (Zn) were prospectively evaluated in malignant tumor and thyroid tissue adjacent to tumor of 41 patients with TMNs. Measurements were performed using energy-dispersive X-ray fluorescent analysis. Results of the study were additionally compared with previously obtained data for the same TEs in “normal” thyroid tissue. From results obtained, it was possible to conclude that the common characteristics of TMNs in comparison with “normal” thyroid and visually “intact” thyroid tissue adjacent to tumor were drastically reduced level of I. It was supposed that the drastically reduced level of I content in cancerous tissue could possibly be explored for differential diagnosis of benign and malignant thyroid nodules.

Content of 31 trace elements in thyroid malignant nodules and thyroid tissue adjacent to nodules investigated using neutron activation analysis and inductively coupled plasma mass spectrometry

Thyroid malignant nodules (TMNs) are the most common endocrine cancer and the fifth most frequently occurring type of malignancies. Women are at particular risk for this thyroid disease The etiology and pathogenesis of TMNs must be considered as multifactorial. The present study was performed to clarify the role of some trace elements (TEs) in the etiology of these thyroid disorders. Thyroid tissue levels of silver (Ag), aluminum (Al), boron (B),, beryllium (Be), bismuth (Bi), cadmium (Cd), cerium (Ce), cobalt (Co), chromium (Cr), cesium (Cs), iron (Fe), gallium (Ga), mercury (Hg), lanthanum (La), lithium (Li), manganese (Mn), molybdenum (Mo), neodymium (Nd), nickel (Ni), lead (Pb), praseodymium (Pr), rubidium (Rb), antimony (Sb), scandium (Sc), selenium (Se), samarium (Sm), tin (Sn), thallium (Tl), uranium (U), yttrium (Y), and zinc (Zn) were prospectively evaluated in malignant tumor and thyroid tissue adjacent to tumor of 41 patients with TMNs. Measurements were performed using a combination of non-destructive instrumental neutron activation analysis and destructive method such as inductively coupled plasma mass spectrometry. Results of the study were additionally compared with previously obtained data for the same TEs in “normal” thyroid tissue. It was observed that main characteristics of TMNs in comparison with “normal” thyroid were significantly elevated levels of Ag, Al, B, Bi, Ce, Cr, Hg, Mn, Mo, Ni, Pb, Rb, Sm, and Tl. Furthermore, the TEs composition of thyroid tissue adjacent to tumor did not equal TEs contents of “normal” thyroid. Moreover, contents of such elements as Ag, Hg, and Se in adjacent tissue were higher than in tumor. Thus, from results obtained, it was possible to conclude that at least the excessive accumulation of Ag, Hg, and Se by thyroid tissue is likely to precede the TMNs origination and development. Elevated levels of Ag, Al, B, Bi, Ce, Cr, Hg, Mn, Mo, Ni, Pb, Rb, Sm, and Tl in nodular tissue could possibly be explored for differential diagnosis of benign and malignant thyroid nodules.

Content of 31 trace elements in thyroid malignant nodules and thyroid tissue adjacent to nodules investigated using neutron activation analysis and inductively coupled plasma mass spectrometry

Content of 31 trace elements in thyroid malignant nodules and thyroid tissue adjacent to nodules investigated using neutron activation analysis and inductively coupled plasma mass spectrometry

Measurement of Nineteen Chemical Elements in Thyroid Malignant Nodules and Thyroid Tissue Adjacent to Nodules using Neutron Activation Analysis and Inductively Coupled Plasma Atomic Emission Spectrometry

Thyroid malignant nodules (TMNs) are the most common endocrine cancer and the fifth most frequently occurring type of malignancies. The etiology and pathogenesis of TMNs must be considered as multifactorial. The present study was performed to clarify the role of some chemical elements (ChEs) in the etiology of these thyroid disorders. Thyroid tissue levels of nineteen ChEs were prospectively evaluated in malignant tumor and tissue adjacent to tumor of 41 patients with TMNs. Measurements were performed using a combination of non-destructive and destructive methods: instrumental neutron activation analysis with high resolution spectrometry of short-lived radionuclides and inductively coupled plasma atomic emission spectrometry. Results of the study were additionally compared with previously obtained data for the same ChEs in “normal” thyroid tissue. It was observed that in malignant tissue the mass fractions of Al, B, Ca, Cl, Cu, K, Mg, Mn, Na, P, S, and Si were 3.14, 4.64, 1.79, 2.26, 3.55, 1.57, 1.61, 1.57, 1.24, 2.45, 1.14, and 2.81 times, respectively, higher whereas mass fraction of I was 25.6 times lower than in the normal thyroid. In the “adjacent” group mean mass fractions of Cl and I were approximately 1.6 and 1.7 times, respectively, higher, while mean values of Ba, Ca and Sr content were 55%, 45%, and 70%, respectively, lower than in the “normal” group. In tumor Ca, Cl, Cu, K, and Sr contents were approximately 2.8, 1.4, 1.8, 1.7, and 5.4 times, respectively, higher, while I content was 43 times lower than in “adjacent” group of tissue samples. It was concluded that thyroid tissue adjacent to malignant nodules kept the main function of thyroid gland, while malignantly transformed thyroid cells lost its capacity to accumulate I. It was supposed that the excessive accumulation of Cl and I by thyroid tissue probably precede the TMNs origination and development. It was also supposed that elevated levels of Al, B, Ca, Cl, Cu, K, Mg, Mn, Na, P, S, and Si, as well as drastically reduced level of me in cancerous tissue could possibly be explored for differential diagnosis of benign and malignant thyroid nodules.

Content of Eleven Trace Elements in Thyroid Malignant Nodules and Thyroid Tissue adjacent to Nodules

Content of Eleven Trace Elements in Thyroid Malignant Nodules and Thyroid Tissue adjacent to Nodules

Analysis of NIS Plasma Membrane Interactors Discloses Key Regulation by a SRC/RAC1/PAK1/PIP5K/EZRIN Pathway with Potential Implications for Radioiodine Re-Sensitization Therapy in Thyroid Cancer.

Simple SummaryThe presence of the sodium–iodide symporter (NIS) at the plasma membrane (PM) of differentiated thyroid cancer (DTC) cells is required for the successful use of radioiodine (RAI) therapy in these malignancies. However, NIS is frequently downregulated in malignant thyroid tissue, and 30% to 50% of metastatic DTCs become refractory to RAI treatment. Notably, many RAI-refractory DTCs still show relatively abundant NIS protein levels despite failing to uptake sufficient iodide to enable RAI therapy. This suggests an impairment in the NIS delivery and retention at the PM. Because little is known about these mechanisms, we employed a new experimental strategy to purify and characterize proteins which are selectively associated with NIS at the PM. We found that, when at the PM, NIS is particularly associated with proteins that regulate the cell’s actin cytoskeleton. Further investigation revealed that several of these proteins integrate an intracellular signaling pathway that modulates the functional localization of NIS at the PM, in normal and DTC cells. Our findings open new venues for therapeutic intervention in refractory DTC, envisioning the re-sensitization of these tumors to RAI therapy.The functional expression of the sodium–iodide symporter (NIS) at the membrane of differentiated thyroid cancer (DTC) cells is the cornerstone for the use of radioiodine (RAI) therapy in these malignancies. However, NIS gene expression is frequently downregulated in malignant thyroid tissue, and 30% to 50% of metastatic DTCs become refractory to RAI treatment, which dramatically decreases patient survival. Several strategies have been attempted to increase the NIS mRNA levels in refractory DTC cells, so as to re-sensitize refractory tumors to RAI. However, there are many RAI-refractory DTCs in which the NIS mRNA and protein levels are relatively abundant but only reduced levels of iodide uptake are detected, suggesting a posttranslational failure in the delivery of NIS to the plasma membrane (PM), or an impaired residency at the PM. Because little is known about the molecules and pathways regulating NIS delivery to, and residency at, the PM of thyroid cells, we here employed an intact-cell labeling/immunoprecipitation methodology to selectively purify NIS-containing macromolecular complexes from the PM. Using mass spectrometry, we characterized and compared the composition of NIS PM complexes to that of NIS complexes isolated from whole cell (WC) lysates. Applying gene ontology analysis to the obtained MS data, we found that while both the PM-NIS and WC-NIS datasets had in common a considerable number of proteins involved in vesicle transport and protein trafficking, the NIS PM complexes were particularly enriched in proteins associated with the regulation of the actin cytoskeleton. Through a systematic validation of the detected interactions by co-immunoprecipitation and Western blot, followed by the biochemical and functional characterization of the contribution of each interactor to NIS PM residency and iodide uptake, we were able to identify a pathway by which the PM localization and function of NIS depends on its binding to SRC kinase, which leads to the recruitment and activation of the small GTPase RAC1. RAC1 signals through PAK1 and PIP5K to promote ARP2/3-mediated actin polymerization, and the recruitment and binding of the actin anchoring protein EZRIN to NIS, promoting its residency and function at the PM of normal and TC cells. Besides providing novel insights into the regulation of NIS localization and function at the PM of TC cells, our results open new venues for therapeutic intervention in TC, namely the possibility of modulating abnormal SRC signaling in refractory TC from a proliferative/invasive effect to the re-sensitization of these tumors to RAI therapy by inducing NIS retention at the PM.

Feasibility of Quantitative Tissue Characterization Using Novel Parameters Extracted From Photoacoustic Power Spectrum Considering Multiple Absorbers

Frequency domain analysis of radio frequency signal is performed to differentiate between different tissue categories in terms of spectral parameters. However, due to complex relationship between the absorber size and spectral parameters, they cannot be used for quantitative tissue characterization. In an earlier study, we showed that using linear relationship between absorber size and two new spectral parameters namely number of lobes and average lobe width, absorber size can be successfully recovered from photoacoustic signal generated by single absorber. As actual biological tissue contains multiple absorbers, in this study we extended the application of these two new spectral parameters for computing absorber size from signals generated by multiple PA absorbers. We revisited our analytical model to establish two new linear relationships between the absorber radius and number of lobes as well as average lobe width considering multiple absorbers with bandlimited acquisition. A simulation study was performed to validate these linear relationships. A retrospective ex vivo study, in which the spectral parameters were computed using multiwavelength photoacoustic signals, was performed with freshly exercised thyroid specimens from 38 actual human patients undergoing thyroidectomy after having a diagnosis of suspected thyroid lesions. From statistical analysis it is shown that both the parameters were significantly different between malignant and non-malignant thyroid and malignant and normal thyroid tissue. Performance of the supervised classification with the computed spectral parameters showed that the extracted parameters could be successfully used to differentiate malignant thyroid tissue from normal thyroid tissue with reasonable degree of accuracy.

Completion Total Thyroidectomy in Occult Thyroid Cancer Patients after Partial Thyroidectomy

Thyroid cancer is one of the commonest cancer while the incidence of occult thyroid cancer is only 0,05%. Completion total thyroidectomy as one of the treatment choice for thyroid cancer which initially diagnosed preoperatively as benign thyroid mass, remains controversial. The aim of this study is to understand the effectiveness of completion total thyroidectomy after partial thyroidectomy, by analyzing from: proportion of malignant contralateral thyroid tissue, post operative complications, and recurrence of cancer. This retrospective study collected medical record datan between 2011 and 2016. Total sample were 16 patients (15 females, 1 male). There was no significant difference on contralateral thyroid tissue malignancy proportion between completion total thyroidectomy and without total thyroidectomy (p=0,375). It was found 3 surgical complications (50%) from 6 patients with completion total thyroidectomy and 3 complications (30%) from 10 patients without completion total thyroidectomy. The complications were temporary injury of recurrent laryngeal nerve (p=0,511), seroma (p=0,375), and hypoparathyroidism (p=0,375). No recurrence event after 4 years follow up amount both groups. In conclusion, there is no advantage in completion total thyroidectomy analized from proportion of malignant contralateral thyroid tissue, post operative complication and recurrence in 4 years-follow up.

Effective rational humanization of a PASylated anti-galectin-3 Fab for the sensitive PET imaging of thyroid cancer in vivo

The lack of a non-invasive test for malignant thyroid nodules makes the diagnosis of thyroid cancer (TC) challenging. Human galectin-3 (hGal3) has emerged as a promising target for medical TC imaging and diagnosis because of its exclusive overexpression in malignant thyroid tissues. We previously developed a human-chimeric αhGal3 Fab fragment derived from the rat monoclonal antibody (mAb) M3/38 with optimized clearance characteristics using PASylation technology. Here, we describe the elucidation of the hGal3 epitope recognized by mAb M3/38, X-ray crystallographic analysis of its complex with the chimeric Fab and, based on the three-dimensional structure, the rational humanization of the Fab by CDR grafting. Four CDR-grafted versions were designed using structurally most closely related fully human immunoglobulin VH/VL regions of which one—employing the acceptor framework regions of the HIV-1 neutralizing human antibody m66—showed the highest antigen affinity. By introducing two additional back-mutations to the rodent donor sequence, an affinity toward hGal3 indistinguishable from the chimeric Fab was achieved (KD = 0.34 ± 0.02 nM in SPR). The PASylated humanized Fab was site-specifically labelled with the fluorescent dye Cy7 and applied for the immuno-histochemical staining of human tissue sections representative for different TCs. The same protein was conjugated with the metal chelator Dfo, followed by radiolabelling with 89Zr(IV). The resulting protein tracer allowed the highly sensitive and specific PET/CT imaging of orthotopic tumors in mice, which was confirmed by quantitative analysis of radiotracer accumulation. Thus, the PASylated humanized αhGal3 Fab offers clinical potential for the diagnostic imaging of TC.

Content of Eleven Trace Elements in Thyroid Malignant Nodules and Thyroid Tissue adjacent to Nodules

Content of Eleven Trace Elements in Thyroid Malignant Nodules and Thyroid Tissue adjacent to Nodules