Undifferentiated Thyroid Cancer Research Articles

KLHL14 is a tumor suppressor downregulated in undifferentiated thyroid cancer

KLHL14 is a substrate-binding subunit of Cullin-RING ligase 3 ubiquitin ligase complex, highly enriched in thyroid since early embryonic development, together with its antisense RNA KLHL14-AS. We have previously demonstrated that Klhl14-AS is a competing endogenous RNA regulating several differentiation and survival factors in thyroid cancer, acting as tumor suppressor. Recently, also KLHL14 has been shown to function as tumor suppressor in diffuse large B-cell lymphoma and in malignant mesothelioma. Here we show that KLHL14 expression is strongly reduced in anaplastic thyroid cancer, the less differentiated and most aggressive type of thyroid neoplasia. Such reduction is reproduced in different in vivo and in vitro models of thyroid cancer, being invariably associated with loss of differentiation. When Klhl14 expression is rescued in thyroid transformed cells, it reduces the cell proliferation rate and increase the number of apoptotic cells. On the other side, Klhl14 loss of function in normal thyroid cells affects the expression of several regulatory as well as functional thyroid markers. All these findings suggest that KLHL14 could be considered as a novel tumor suppressor in thyroid cancer, by also revealing its physiological role in the maintenance of a fully differentiated and functional thyroid phenotype.

Neutrophil diversity is associated with T-cell immunity and clinical relevance in patients with thyroid cancer

Neutrophil heterogeneity is involved in autoimmune diseases, sepsis, and several cancers. However, the link between neutrophil heterogeneity and T-cell immunity in thyroid cancer is incompletely understood. We investigated the circulating neutrophil heterogeneity in 3 undifferentiated thyroid cancer (UTC), 14 differentiated thyroid cancer (DTC) (4 Stage IV, 10 Stage I–II), and healthy controls (n = 10) by transcriptomic data and cytometry. Participants with UTC had a significantly higher proportion of immature high-density neutrophils (HDN) and lower proportion of mature HDN in peripheral blood compared to DTC. The proportion of circulating PD-L1+ immature neutrophils were significantly increased in advanced cancer patients. Unsupervised analysis of transcriptomics data from circulating HDN revealed downregulation of innate immune response and T-cell receptor signaling pathway in cancer patients. Moreover, UTC patients revealed the upregulation of glycolytic process and glutamate receptor signaling pathway. Comparative analysis across tumor types and stages revealed the downregulation of various T-cell-related pathways, such as T-cell receptor signaling pathway and T-cell proliferation in advanced cancer patients. Moreover, the proportions of CD8+ and CD4+ T effector memory CD45RA+ (TEMRA) cells from peripheral blood were significantly decreased in UTC patients compared to DTC patients. Finally, we demonstrated that proportions of tumor-infiltrated neutrophils were increased and related with poor prognosis in advanced thyroid cancer using data from our RNA-seq and TCGA (The Cancer Genome Atlas) data. In conclusion, observed prevalence of circulating immature high-density neutrophils and their immunosuppressive features in undifferentiated thyroid cancers underscore the importance of understanding neutrophil dynamics in the context of tumor progression in thyroid cancer.

Long Non-Coding RNAs as Determinants of Thyroid Cancer Phenotypes: Investigating Differential Gene Expression Patterns and Novel Biomarker Discovery.

Thyroid Cancer (TC) is the most common endocrine malignancy, with increasing incidence globally. Papillary thyroid cancer (PTC), a differentiated form of TC, accounts for approximately 90% of TC and occurs predominantly in women of childbearing age. Although responsive to current treatments, recurrence of PTC by middle age is common and is much more refractive to treatment. Undifferentiated TC, particularly anaplastic thyroid cancer (ATC), is the most aggressive TC subtype, characterized by it being resistant and unresponsive to all therapeutic and surgical interventions. Further, ATC is one of the most aggressive and lethal malignancies across all cancer types. Despite the differences in therapeutic needs in differentiated vs. undifferentiated TC subtypes, there is a critical unmet need for the identification of molecular biomarkers that can aid in early diagnosis, prognosis, and actionable therapeutic targets for intervention. Advances in the field of cancer genomics have enabled for the elucidation of differential gene expression patterns between tumors and healthy tissue. A novel category of molecules, known as non-coding RNAs, can themselves be differentially expressed, and extensively contribute to the up- and downregulation of protein coding genes, serving as master orchestrators of regulated and dysregulated gene expression patterns. These non-coding RNAs have been identified for their roles in driving carcinogenic patterns at various stages of tumor development and have become attractive targets for study. The identification of specific genes that are differentially expressed can give insight into mechanisms that drive carcinogenic patterns, filling the gaps of deciphering molecular and cellular processes that modulate TC subtypes, outside of well-known driver mutations.

Prognostic analysis of 23 cases of undifferentiated thyroid carcinoma

Objective: To investigate the relationship between clinical features, treatment and prognosis of undifferentiated thyroid carcinoma. Methods: The clinical and follow-up data of 23 patients with undifferentiated thyroid carcinoma admitted to Chongqing University Cancer Hospital from March 2012 to September 2019 were retrospectively analyzed. Results: The median survival time of 23 patients with undifferentiated thyroid carcinoma was 15 weeks, 3-month survival time was 56.5%, 6-month survival rate was 39.1%, and 1-year survival rate was 30.4%. Univariate analysis showed that the size of primary tumor lesion, normal albumin, normal platelets, activated partial thrombin time, fibrinogen, thyroglobulin antibody and radiotherapy were the factors influencing the prognosis of undifferentiated thyroid cancer. The analysis of the effect of treatment on the prognosis showed that surgery, radiotherapy and combined treatment can obviously improve the prognosis. Multivariate analysis showed that the independent factors affecting prognosis were the diameter of the primary lesion, whether radiotherapy was given or not, the dose of radiotherapy, and whether radiotherapy was combined with surgery. Conclusion: The malignant degree of undifferentiated thyroid cancer is very high, so R0 or R1 resection of neck lesions plus radiotherapy should be taken as far as possible for patients with surgical opportunity, and the same effect may be achieved for patients who have lost the surgical opportunity. Anticoagulant and antiplatelet therapy may benefit patients.

Unraveling the role of the mitochondrial one-carbon pathway in undifferentiated thyroid cancer by multi-omics analyses

The role of the serine/glycine metabolic pathway (SGP) has recently been demonstrated in tumors; however, the pathological relevance of the SGP in thyroid cancer remains unexplored. Here, we perform metabolomic profiling of 17 tumor-normal pairs; bulk transcriptomics of 263 normal thyroid, 348 papillary, and 21 undifferentiated thyroid cancer samples; and single-cell transcriptomes from 15 cases, showing the impact of mitochondrial one-carbon metabolism in thyroid tumors. High expression of serine hydroxymethyltransferase-2 (SHMT2) and methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) is associated with low thyroid differentiation scores and poor clinical features. A subpopulation of tumor cells with high mitochondrial one-carbon pathway activity is observed in the single-cell dataset. SHMT2 inhibition significantly compromises mitochondrial respiration and decreases cell proliferation and tumor size in vitro and in vivo. Collectively, our results highlight the importance of the mitochondrial one-carbon pathway in undifferentiated thyroid cancer and suggest that SHMT2 is a potent therapeutic target.

SAT525 Long-Term Outcomes in Stage I Thyroid Cancer Without Radioiodine Remnant Ablation

Abstract Disclosure: F. Mubeen: None. M. Zahid: None. T.D. Cubb: None. J. Shakil: None. J. Xu: None. R.J. Robbins: None. Introduction: Although differentiated thyroid cancer (DTC) has low mortality, the risk of recurrence after thyroidectomy is not negligible. In the past radioiodine was routinely given to almost all patients following surgery to achieve radioiodine remnant ablation (RRA). We hypothesized, omission of RRA in Stage I (American Joint Committee on Cancer [AJCC] 8th edition) DTC survivors would not be inferior to RRA. Methods: We did a retrospective analysis of thyroid cancer survivors at a single academic medical center from 2000 to 2021. Patients with under six months follow-up, medullary or undifferentiated thyroid cancer, Stage II, III, or IV cancer or those who did not have total thyroidectomy were excluded. 208 Stage I (AJCC) DTC patients met inclusion criteria and were divided into two groups: those who had RRA (RRA+; n=121) and those who did not (RRA-; n=87). Primary outcome assessed was persistent/recurrent disease (P/RD). Results: P/RD was seen in 6.25% of patients at closing the study. RRA+ patients were significantly younger (42.88±13.02 vs. 46.84±13.98, p = 0.037), had a longer follow-up duration (111.23 (IQR: 51.80-161.27) vs. 78.87 (53.13-111.93) months, p=0.003), but there was no difference in sex distribution (p = 1.0). The RRA+ group had significantly higher multicentric tumors (66 (54.55%) vs. 32 (36.78%), p<0.001), larger tumors ( T3 and T4 , 25(20.67%) vs 3 (3.45%), p<0.001) and spread to loco-regional LN (48 (39.67%) vs. 8 (9.20%), p=0.001). P/RD was present in 13 survivors with RRA+ group having a slightly, but not significantly higher incidence of P/RD (11 (9%) in RRA+ vs. 2 (2.3%) in RRA-, P=0.078). In the 98 patients with presence of multicentricity, P/RD was seen in 1 (3.1%) of 32 RRA- and 8 (12.1%) of 66 RRA+ patients. The percentage of RRA+ patients who had P/RD increased as the size of primary tumor increased with PR/D seen in 28.5% (2/7) of T4 tumors. Sub-analysis of P/RD in patients who had LN metastases (N1) and in those without known LN metastases (N0 and Nx) did not show significant difference in P/RD. When disease-free survival was compared, there was no statistically significant difference in the two groups. In the 56 patients with pathological LN involvement (N1), all P/RD patients were in RRA+ group (n=48) with no statistical significance (7 (14.58%) vs. 0 (0), p=0.58). On comparing the amount of radioiodine administered to RRA+ patients, there was no trend of higher P/RD with increased doses (p=0.88). Conclusion:Our study found that P/RD was not higher in Stage I DTC patients who did not undergo RRA compared to those who received RRA at a median follow-up of 7.4 years. We confirmed our hypothesis that omission of RRA in Stage I patients was not inferior to RRA regarding disease-free survival and long-term clinical outcome proving it to be a safe and reasonable clinical decision to omit RRA. Presentation Date: Saturday, June 17, 2023

SAT566 A Rare Case Of Aggressive Anaplastic Thyroid Presenting As Thyrotoxicosis

Abstract Disclosure: A. Al-Juboori: None. Y. Al-Yousif: None. S. Fomin: None. M. Sundaram: None. Background:Anaplastic Thyroid cancer is a rare neoplasm with a poor prognosis and high mortality rate. Usually, it presents with symptoms like dysphagia and hoarseness. However, it is rarely presented with thyrotoxicosis from the rapid destruction of thyroid follicle cells, causing excessive leakage of preformed thyroid hormones into the circulation. Clinical presentation can vary from a subclinical state to a life-threatening thyroid storm. Management requires an inter-professional team approach. Clinical Case:77-year-old female patient, who visited the urgent care on August 12 complaining of sore throat, Amoxicillin was prescribed. Presented to ER 10 days later complaining of a headache, gum discomfort, shortness of breath, worsening sore throat, neck pain, and gradual worsening dysphagia to liquids and solids. Physical exam revealed anterior painful and firm neck mass upon palpation. Stridor was noted upon auscultation and was found to be in arterial fibrillation with a rapid ventricular rate for which Amiodarone drip and heparin drip were initiated. IV hydrocortisone 50 mg TID was added to the treatment right away for symptomatic relief. The results of the thyroid function test revealed an increased serum FT4 and FT3, and low serum TSH. Antibodies were normal. Clinical presentation increases the suspicion of undifferentiated thyroid cancer versus lymphoma. CT revealed leftward tracheal deviation along with a big undetermined right thyroid mass with accompanying cervical and supraclavicular adenopathy. IR was consulted urgently for a thyroid biopsy. High-grade anaplastic cancer was discovered after a biopsy with metastasis to the surrounding lymph. The decision was made to transfer the patient to a tertiary care facility for invasive interventions. Anaplastic thyroid cancer accounts for 2-3% of all thyroid gland neoplasm, it often affects persons over the age of 60, and the mean age at the time of diagnosis is 65 years, most common in female population compared to males with a ratio of 2:1. There have been only around 9 other cases reported in the literature in which thyrotoxicosis is the initial presentation of anaplastic thyroid cancer. Diagnosis is usually made via core needle biopsy. 70% of individuals with anaplastic thyroid carcinoma will have contiguous spread and distant metastasis at the time of diagnosis. It most commonly compresses the trachea and esophagus. Given the rapid progression of illness, patients can die from local airway obstruction. Average survival is just 6 months with only 1 in 5 people surviving after 12 months. Conclusion:Anaplastic thyroid cancer-induced thyrotoxicosis is an uncommon and rare presentation of thyroid neoplasm. Debulking surgery is the cornerstone for removing tumors and securing the patient’s airway along with controlling thyrotoxicosis. A low survival rate was reported in these patients. Presentation: Saturday, June 17, 2023

THU672 Fungating And Inflammatory Anaplastic Thyroid Carcinoma In A 64-Year Old Filipino

Abstract Disclosure: R.M. Tarrazona - Dominisac: None. M.C. Isidro: None. Background: Anaplastic Thyroid Carcinoma is a rare and lethal undifferentiated thyroid cancer that occurs in less than 2% of all thyroid malignancies. In the Philippines, this rare entity has an estimated incidence of 0.7%. Its aggressive nature is strikingly significant because it carries a poor prognosis with a median survival of 5-6 months. An exceptionally rare presentation of an Anaplastic Thyroid Carcinoma is a fungating, exophytic and inflammatory type. Case Summary: This is a case of a 64-year old female with a 2 x 2 cm erythematous patch at her anterior neck. There is progressive enlargement of the reddish neck lesion associated by burning sensation with subsequent development whitish nodules with purulent exudate. The nodules were fragile with noted bleeding on minimal manipulation. Initial impression was anterior neck cellulitis and she was given 7 days of IV antibiotics with no resolution of the lesion. Neck CT scan showed a multinodular heterogeneously enhancing mass in the left neck region measuring 6.3 x 5.2 x 5.5 cm. A core needle biopsy of the mass revealed malignant cells present: anaplastic carcinoma vs squamous cell carcinoma, with immunohistochemical stains: PAX 8 positive, p63 negative, CK 5/6 negative, BRAF negative, TTF-1 focally positive. The patient underwent tumor debulking surgery with tracheostomy. The gross specimen was sent for histopathologic examination and finally revealed Anaplastic Thyroid Carcinoma. Conclusion: The case reported was a diagnostic dilemma early during the clinical presentation due to presence of rapidly enlarging, inflammatory and fungating anterior neck mass with presence of whitish nodules with purulent exudates, bleeding and pain. She was initially treated as an abscess only to discover presence of malignancy upon further work-up. Anaplastic thyroid cancer is the most advanced and aggressive thyroid malignancy. It is very rare and the atypical fungating presentation of this ATC makes it an exceptionally rare entity. Fungating wounds are the result of cancer cells infiltrating and invading the cutaneous structure. This case report highlights the importance of harnessing a high level of clinical suspicion for anaplastic thyroid cancer even in atypical looking anterior neck mass to ensure prompt definitive diagnosis and improve overall prognosis. Presentation: Thursday, June 15, 2023

Correlation between preoperative platelet parameters and clinicopathological features of differentiated thyroid cancer

Objective:To investigate the correlation between preoperative platelet parameters and the clinicopathological features of differentiated thyroid cancer. Methods:We retrospectively analyzed the medical records of patients with thyroid tumors admitted to Zhongda Hospital affiliated to Southeast University and healthy adults with normal physical examination results in our hospital from January 2019 to December 2020, and collected their general information and preoperative blood routine data. Patients with undifferentiated thyroid cancer, diabetes, coronary heart disease, hematological diseases, kidney diseases, autoimmune diseases, genetic diseases, infectious diseases, other systemic tumors, hepatitis or cirrhosis, or those taking anticoagulants were excluded. The exclusion criteria for healthy adults were the absence of the above diseases and normal physical examination results. Differences in platelet parameters among the three groups were compared, and the correlation between clinicopathological characteristics of thyroid cancer, accompanying cervical lymph node metastasis, and platelet parameters of patients was analyzed. A multivariate logistic regression model was used to analyze the risk factors of thyroid cancer with cervical lymph node metastasis. Results:A total of 117 cases of differentiated thyroid cancer were collected, including 33 males and 84 females, with an average age of （41.64±12.25） years; 46 patients had benign thyroid tumors, including 15 males and 31 females, with an average age of （41.35±12.52） years; 50 healthy adults with normal physical examination results in our hospital during the same period were also included, including 18 males and 32 females, with an average age of（42.02±9.62） years, without underlying diseases. The platelet count of the differentiated thyroid cancer group was higher than that of the benign thyroid tumor group（t=-2.219, P=0.028） and the normal control group（t=2.069, P=0.04）, while the platelet distribution width of the differentiated thyroid cancer group was lower than that of the benign thyroid tumor group（t=2.238, P=0.027） and the normal control group（t=-2.618, P=0.002）. These differences were statistically significant. Preoperative age ≤45 years（χ²=4.225, P=0.04）, tumor diameter>1 cm（χ²=4.415, P=0.036）, PLT（t=-4.018, P<0.01） increase, and PDW（t=4.568, P<0.01） decrease were significantly correlated with cervical lymph node metastasis of thyroid cancer and had statistical significance. Univariate analysis showed that age ≤45 years（OR=0.447, 95%CI 0.206-0.970, P=0.042）, tumor diameter>1 cm（OR=2.3, 95%CI 1.050-5.039, P=0.037）, PLT（OR=1.012, 95%CI 1.005-1.019, P=0.001）, and PDW（OR=0.693, 95%CI 0.518-0.827, P<0.01） were risk factors for cervical lymph node metastasis of thyroid cancer. The results of multifactorial logistic regression analysis showed that PLT（OR=1.008, 95%CI 1.001-1.016, P=0.026） and PDW（OR=0.692, 95%CI 0.564-0.848, P<0.01） were independent risk factors for thyroid cancer with cervical lymph node metastasis. Conclusion:PLT and PDW may be useful predictive factors for the differentiation of thyroid cancer malignancy and central lymph node metastasis.

The lncRNA RMST is drastically downregulated in anaplastic thyroid carcinomas where exerts a tumor suppressor activity impairing epithelial-mesenchymal transition and stemness

Thyroid cancer is the most prevalent endocrine malignancy and comprises a wide range of lesions subdivided into differentiated (DTC) and undifferentiated thyroid cancer (UTC), mainly represented by the anaplastic thyroid carcinoma (ATC). This is one of the most lethal malignancies in humankind leading invariably to patient death in few months. Then, a better comprehension of the mechanisms underlying the development of ATC is required to set up new therapeutic approaches. Long non-coding RNAs (lncRNAs) are transcripts over 200 nucleotides in length that do not code for proteins. They show a strong regulatory function at both transcriptional and post-transcriptional level and are emerging as key players in regulating developmental processes. Their aberrant expression has been linked to several biological processes, including cancer, making them potential diagnostic and prognostic markers. We have recently analyzed the lncRNA expression profile in ATC through a microarray technique and have identified rhabdomyosarcoma 2-associated transcript (RMST) as one of the most downregulated lncRNA in ATC. RMST has been reported to be deregulated in a series of human cancers, to play an anti-oncogenic role in triple-negative breast cancer, and to modulate neurogenesis by interacting with SOX2. Therefore, these findings prompted us to investigate the role of RMST in ATC development. In this study we show that RMST levels are strongly decreased in ATC, but only slightly in DTC, indicating that the loss of this lncRNA could be related to the loss of the differentiation and high aggressiveness. We also report a concomitant increase of SOX2 levels in the same subset of ATC, that inversely correlated with RMST levels, further supporting the RMST/SOX2 relationship. Finally, functional studies demonstrate that the restoration of RMST in ATC cells reduces cell growth, migration and the stemness properties of ATC stem cells. In conclusion, these findings support a critical role of RMST downregulation in ATC development.

TIM3 Expression in Anaplastic-Thyroid-Cancer-Infiltrating Macrophages: An Emerging Immunotherapeutic Target.

Anaplastic thyroid cancer (ATC) is a clinically aggressive form of undifferentiated thyroid cancer with limited treatment options. Immunotherapy for patients with ATC remains challenging. Tumor-associated macrophages (TAMs) constitute over 50% of ATC-infiltrating cells, and their presence is associated with a poor prognosis. Consequently, the development of new therapies targeting immune checkpoints in TAMs is considered a promising therapeutic approach for ATC. We have previously shown that soluble factors secreted by ATC cells induced pro-tumor M2-like polarization of human monocytes by upregulating the levels of the inhibitory receptor TIM3. Here, we extended our observations on ATC-cell-induced xenograft tumors. We observed a large number of immune cells infiltrating the ATC xenograft tumors. Significantly, 24-28% of CD45+ immune cells were macrophages (CD11b+ F4/80+). We further showed that 40% of macrophages were polarized toward a M2-like phenotype, as assessed by CD206 expression and by a significant increase in the Arg1/iNOS (M2/M1) ratio. Additionally, we found that ATC xenograft tumors had levels of TIM3 expression when determined by RT-PCR and immunofluorescence assays. Interestingly, we detected the expression of TIM3 in macrophages in ATC tumors by flow cytometry assays. Furthermore, TIM3 expression correlated with macrophage marker expression in human ATC. Our studies show that TIM3 is a newly identified immune checkpoint in macrophages. Since TIM3 is known as a negative immune regulator, it should be considered as a promising immunotherapeutic target for ATC.

ODP489 Insular Thyroid Cancer: A Rare Case of Undifferentiated Thyroid Carcinoma

Abstract Introduction Insular thyroid cancer (ITC) is a rare form of thyroid cancer derived from follicular cells. ITC requires a histopathologic diagnosis and has an intermediate morphology between well differentiated thyroid cancers (follicular and papillary) and undifferentiated thyroid cancer (anaplastic). The rate of metastasis and mortality are higher compared with well-differentiated thyroid carcinomas. Case presentation A 74-year-old woman presented for a 1 year history of rapidly progressive neck mass associated with neck pain, limited neck motion, loss of appetite, weight loss, and dysphagia. Past medical history included hypertension, type 2 diabetes, dyslipidemia, and vitamin D deficiency. There was no history of neck radiation or family history of cancer. Physical examination revealed a 10×7 cm neck mass with cervical adenopathy. Vital signs were unremarkable. Laboratory studies showed TSH of 0.71 uIU/mL (0.270-4.200), free T4 of 0.57 ng/dL (0.93-1.70) and thyroglobulins of 213.616 ng/dL (1.6-59.9). Neck CT demonstrated a multilobulated mass in the left thyroid lobe measuring 12×5.2×5.6 cm with tracheal deviation, multiple nodules surrounding the carotids and bone metastasis. Fine needle aspiration showed a mixed pattern of thyroid follicles. An excisional biopsy showed nests and sheets of small uniform cells with scattered abortive follicles and mitosis consistent with ITC. There was no necrosis. The Ki67 index was 10-20%. On immunohistochemical analysis TTF1 and thyroglobulin were positive and P63, calcitonin, and PTH were negative. Tracheostomy and PEG tube placement were required due to mass obstructive effect. Radiation and steroids were initiated before starting lenvatinib. The patient was not a surgical candidate due to tumor extension to the carotids. She ultimately succumbed to the disease after a 45-day hospital course complicated by pneumonia. Discussion ITC is a rare type of thyroid tumor with a mortality rate of 12-75%. The diagnosis is based on histopathology, demonstrating solid nests (insulae) of small, uniform carcinoma cells, small follicles containing thyroglobulin, frequent necrotic foci and variable mitotic activity. The tumor cells in ITC are derived from poorly differentiated follicular cells expressing thyroglobulin. The immunostaining of thyroglobulin differentiates ITC from other poorly differentiated thyroid tumors. In this case the immunohistochemical analysis was positive for thyroglobulin and negative for calcitonin, which confirmed that the tumor cells were follicular in origin 1 . ITC behaves differently compared with other thyroid carcinomas, with more frequent metastasis and lower average 20-year post-diagnosis survival. Our patient had other characteristics that conferred a poor prognosis, including advanced age, high rate of mitosis, and multiple metastases. Although surgery and radioactive iodine are first line, this patient's tumor extension required alternative therapies. Radiotherapy and lenvatinib, a tyrosine kinase receptor inhibitor, were employed unsuccessfully. 1. Soza, J., et al. Insular Thyroid Cancer. Cancer, 3260-3267. Presentation: No date and time listed

Management of anaplastic thyroid cancer and proposed treatment guidelines-A 5-year case series study.

Anaplastic thyroid cancer is a rare and rapidly progressive cancer with an extremely poor prognosis. Besides surgical control, no clear treatment has been found, mainly due to the small population affected and high mortality rate. To propose evidence-based treatment guidelines based on a 5-year retrospective study of patients with anaplastic thyroid cancer treated at our facility. There have been no clearly defined guidelines for treatment plan for undifferentiated thyroid cancer. Our paper presents a retrospective analysis on the treatment of patients with undifferentiated thyroid cancer at our hospital. We retrospectively evaluated the data of patients diagnosed with anaplastic thyroid cancer from April 2017 to March 2022. The total number of patients diagnosed and treated was seven. Two of these patients had operable cancer; five were inoperable and treated with lenvatinib or paclitaxel maintenance therapy. The median time from the first visit to death was 3.84 months, and six of the seven patients died before this study started. Three of them had Stage IVB cancer and died due to deterioration of their general condition, including lung metastasis; the other three had Stage IVC cancer and died of suffocation. The survivor had Stage IVB cancer, was treated by surgery combined with chemical radiotherapy, and survived >240 days. Considering the above findings, personalized surgical treatment should be prioritized to prevent suffocation. Especially in Stage IVB cancer, local control can be achieved by surgical and anticancer drug treatment to avoid death from suffocation.

Aberrant Expression of Thymosin Beta-4 Correlates With Advanced Disease and BRAF V600E Mutation in Thyroid Cancer.

Thymosin beta-4 (TMSB4X) was recently identified as a differentially expressed gene between malignant and non-malignant thyroid cells via single-cell RNA sequencing. In the present study, we aimed to study the immunostaining pattern of TMSB4X in benign and malignant thyroid neoplasms. Immunohistochemical analysis revealed that normal thyroid tissue or benign thyroid disorders exhibited undetectable immunoreactivity against TMSB4X except for positive staining of inflammatory infiltrates and stromal cells associated with autoimmune thyroid disease. By contrast, overexpression of TMSB4X was observed in a variety of thyroid malignancies, including papillary, follicular, poorly differentiated, and undifferentiated thyroid cancer. Among 141 patients with differentiated thyroid cancer, higher TMSB4X expression was associated with papillary tumor type, extrathyroidal extension, lymph node metastasis, and BRAF V600E mutation. The results were consistent with those from the public transcriptomic datasets. In summary, TMSB4X expression was aberrantly increased in various types of thyroid cancer, and higher TMSB4X expression was correlated with advanced disease characteristics. Thymosin beta-4 may be a novel downstream effector of the BRAF V600E mutation.

Anaplastik tiroid kanseri hücre hattının morfolojik analizi

Aim: Thyroid follicular cell derived cancers are classified into three groups such as papillary thyroid cancer (85%), follicular thyroid cancer (12%) and anaplastic (undifferentiated) thyroid cancer (ATC) (3%). ATCs have very rapid course, poor treatment outcomes and they are very aggressive. The aim of current study was to assess the analysis of the morphological differences of ATC cell line with the normal thyroid cell line (NTC).&#x0D; &#x0D; Materials and Methods: NTH and ATC cells were examined with haematoxylin and eosin, the nucleus: cytoplasm (N:C) ratios were detected, and cell cycles were investigated. These cell lines were compared according to their N:C ratio and their abundance in cell cycle phases.&#x0D; &#x0D; Results: The N:C ratio was higher in ATC than NTC. Both cell groups were mostly found in G0/G1 phase (68.4; 82.8) and have statistical difference in both G0/G1 and S phases.&#x0D; &#x0D; Conclusion: The rapid course and the rarity of ATC are significant barriers for clinical trials. Cultured cell lines are very important to explore the behaviour in the biology of ATC cells (such as the cell cycle), to understand the course of the disease, and to find an effective target for treatment.

RREB1 promotes the development of parafollicular carcinogenesis through the Ras-Raf-1-ELK3 signaling pathway

Thyroid cancer (TC) is the most common endocrine malignancy. Medullary thyroid carcinoma (MTC) is derived from parathyroid follicle cells (C cells) secrete calcitonin, accounting for approximately 5-10% of all thyroid cancers. The malignancy is between differentiated and undifferentiated thyroid cancer and undifferentiated thyroid cancer and has a relatively poor prognosis. In MTC tumor cells, RREB1 regulates the differentiation of parathyroid cells via RAS-Raf-1-ELK3 signaling and induce calcitonin secretion. Therefore, it is easy to induce parathyroid parafollicular cells canceration and medullary thyroid carcinoma. Here, we investigated the correlation between RREB1, RAS-Raf-1-ELK3 signaling pathway and medullary thyroid carcinoma with various phases. Our results suggest that RREB1 promotes parafollicular carcinoma through the Ras-Raf1-elk3 signaling pathway, providing a rationale to further investigate the role of RREB1 in parafollicular carcinoma. It provides theoretical guidance for the clinical treatment of medullary thyroid cancer.

Eugenol: A New Option in Combination Therapy with Sorafenib for the Treatment of Undifferentiated Thyroid Cancer.

Thyroid cancer (TC) is the most common endocrine malignancy. Thyroidectomy and radiotherapy are common treatment modalities for patients with undifferentiated TC (UTC), and sorafenib is usually recommended to prevent a recurrence. However, malignant cells may evade chemotherapy-induced apoptosis, and combination therapy was developed to achieve better outcomes. This study investigated whether eugenol in combination with sorafenib was more effective than either substance individually in triggering apoptosis in the UTC. The IC50 of sorafenib and eugenol was determined in a UTC cell line (8305C) by MTT assay, and their synergistic effect in combination therapy was investigated. Flow cytometry was used to evaluate the rate of apoptosis in treated cells. To confirm that cell death occurred through apoptosis, immunoblotting was used to determine the relative cleavage of caspase-8 and caspase-9. The IC50 of sorafenib was 20 µM, and that of eugenol was 2100 µM. The sorafenib-eugenol combination (1:105) showed synergistic effects at concentrations equal to or less than their IC50. The rate of apoptosis induction was higher in cells treated with eugenol or the eugenol-sorafenib combination compared to sorafenib-treated cells. The relative intensity of cleaved/un cleaved forms of caspase-8 increased in eugenol-treated cells compared to sorafenib-treated cells.Sorafenib and eugenol at concentrations equal to or less than their IC50 had a synergistic effect in 8305C cells. The most potent apoptotic effect was achieved with sorafenib and eugenol at their IC50. Lower doses of sorafenib could be used with eugenol to improve its efficacy while reducing its side effects.

Abstract 1410: Metabolic modulation of anaplastic thyroid cancer by Berberine: A targeted intervention

Abstract Anaplastic thyroid cancer (ATC) is an undifferentiated thyroid cancer. A hallmark of ATC is increased inflammatory cytokine expression and inflammatory cell infiltration in the tumor microenvironment that is responsible for its aggressive nature. In contrast, papillary thyroid cancer (PTC) is well-differentiated and subsequently significantly less aggressive and more responsive to available therapies. The metabolic differences between ATC and PTC that correlate with differentiation status, and hence metastatic propensity, are poorly understood. Owing to its lack of differentiation, ATC is radioiodine therapy resistant, whereas differentiated thyroid cancers are susceptible. AMPK in the thyroid gland is responsible for glucose and iodine uptake. The AMPK pathway is a potential target for modulation due to the higher glucose uptake and metabolism associated with treatment resistance seen in ATC. Berberine, a naturally occurring plant alkaloid, regulates glycometabolism and lipid metabolism, improves energy expenditure, and reduces body weight. Exploration of a possible connection between the metabolic regulation displayed by BBR and AMPKα expression was warranted. Increased AMPKα signaling, observed through increased AMPKα phosphorylation, has an anti-tumor effect. When comparing PTC (TPC1) and ATC (T238) cell lines there was a 4.75- and 2.34-fold change, respectively, in phosphorylation of AMPKα upon BBR treatment. This demonstrates that BBR increases metabolic regulation in thyroid cancer cell lines, importantly, even when poorly differentiated. Further analysis of the correlation between differentiation status and metabolic characteristics between ATC and PTC was investigated using RNASeq. Differential gene expression was conducted to compare ATC and PTC using human thyroid cancer cell lines T238 and K1, respectively. These thyroid cancer cell lines were treated with 100 μM BBR or the same concentration of DMSO as a vehicle control for 24 hours. RNA was isolated from T238 and K1 cells and RNASeq was conducted by GeneWiz. This differential gene expression will help identify specific genes that are regulating metabolic patterns and is currently ongoing. Berberine (BBR) is a natural compound used widely in Traditional Chinese Medicine for its many pharmacological properties including anti-inflammatory, metabolic, antioxidant, and anti-cancer. BBR may be able to mitigate the metabolic pattern of ATC to potentially affect differentiation status as an intervening product. Intervention of the differentiation status of ATC will impact its progression, specifically metastasis. Therefore, identifying gene patterns that correlate with differentiation status will identify novel intervention targets for BBR. Citation Format: Kaci Kopec, Tara Jarboe, Mordechai Sternman, Nicole R. DeSouza, Sarnath Singh, Augustine Moscatello, Jan Geliebter, Raj K. Tiwari, Xiu-Min Li. Metabolic modulation of anaplastic thyroid cancer by Berberine: A targeted intervention [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1410.

Introduction and expression of PIK3CAE545K in a papillary thyroid cancer BRAFV600E cell line leads to a dedifferentiated aggressive phenotype

Anaplastic thyroid cancer (ATC) is a rare, aggressive form of undifferentiated thyroid cancer, which exhibits rapid progression and is almost universally fatal. At least a subset of ATC is thought to arise from pre-existing well-differentiated thyroid cancer, most frequently papillary thyroid cancer (PTC). While PIK3CA mutations are rare in PTC, they are common in ATC and tend to co-occur with BRAF mutations. This provided the rationale for our study to identify the potential role of PIK3CA mutations in the progression from well-differentiated to undifferentiated thyroid cancer. We introduced PIK3CAE545K into the LAM1 PTC cell line, which carries a BRAFV600E mutation. In culture, the engineered cell line (LAM1:PIK3CAE545K) proliferated faster and demonstrated increased clonogenic potential relative to the parental line carrying an empty vector (LAM1EV). Both the LAM1EV and LAM1:PIK3CAE545K edited lines were implanted into hind flanks of athymic nude mice for in vivo determination of disease progression. While tumour weights and volumes were not significantly higher in LAM1:PIK3CAE545K mice, there was a decrease in expression of thyroid differentiation markers TTF-1, thyroglobulin, PAX8 and B-catenin, suggesting that introduction of PIK3CAE545K led to dedifferentiation in vivo. Collectively, this study provides evidence of a role for PIK3CAE545K in driving disease progression from a well-differentiated to an undifferentiated thyroid cancer; however, over-expression was not a determinant of an accelerated growth phenotype in ATC.Graphical

Effects of Anti-Cancer Drug Sensitivity-Related Genetic Differences on Therapeutic Approaches in Refractory Papillary Thyroid Cancer.

Thyroid cancer (TC) includes tumors of follicular cells; it ranges from well differentiated TC (WDTC) with generally favorable prognosis to clinically aggressive poorly differentiated TC (PDTC) and undifferentiated TC (UTC). Papillary thyroid cancer (PTC) is a WDTC and the most common type of thyroid cancer that comprises almost 70–80% of all TC. PTC can present as a solid, cystic, or uneven mass that originates from normal thyroid tissue. Prognosis of PTC is excellent, with an overall 10-year survival rate >90%. However, more than 30% of patients with PTC advance to recurrence or metastasis despite anti-cancer therapy; consequently, systemic therapy is limited, which necessitates expansion of improved clinical approaches. We strived to elucidate genetic distinctions due to patient-derived anti-cancer drug-sensitive or -resistant PTC, which can support in progress novel therapies. Patients with histologically proven PTC were evaluated. PTC cells were gained from drug-sensitive and -resistant patients and were compared using mRNA-Seq. We aimed to assess the in vitro and in vivo synergistic anti-cancer effects of a novel combination therapy in patient-derived refractory PTC. This combination therapy acts synergistically to promote tumor suppression compared with either agent alone. Therefore, genetically altered combination therapy might be a novel therapeutic approach for refractory PTC.