Brain metastasis from follicular thyroid carcinoma: treatment with sorafenib

Background: Sorafenib has shown promise in the treatment of patients with advanced or metastatic thyroid carcinoma. However, its therapeutic effect has not been assessed in patients with brain metastases from follicular thyroid carcinoma (FTC). Here, we report a patient in whom this treatment was employed with a relatively favorable response. Patient and Methods: A 56-year-old woman had a thyroidectomy 8 years previously for FTC. She subsequently developed lung metastases, for which she received seven courses of radioiodine ((131)I) therapy. She developed right hemiplegia and other symptoms and was found to have a ～5-cm lesion in the left parietal lobe. Radiosurgery with a total dose of 28?Gy (7?Gy/day, for 4 days) to treat her brain metastatic lesion was ineffective, and she was referred to us. We treated her with sorafenib, 200?mg orally, on a twice-daily basis. The effect of this intervention was assessed clinically and radiographically using Response Evaluation Criteria in Solid Tumors (RECIST). Summary: Symptoms and signs improved dramatically and continuously after initiation of sorafenib treatment. Partial response (PR) in the brain metastasis and stable disease (SD) in lung metastatic lesions were verified by consecutive imaging findings for more than one year. Despite alopecia, other treatment-related adverse events did not occur. Conclusions: Targeted therapy such as with sorafenib could be an effective alternative therapeutic strategy in the treatment of progressive brain metastasis from differentiated thyroid carcinoma (DTC) when surgery, external beam radiation, and (131)I are not suitable or give poor outcomes. A paradigm of sustained low dose of sorafenib (200?mg,twice a day) may be well-tolerated without compromising maintenance of the therapeutic effect.     

Solitary adrenal gland metastasis of a follicular thyroid carcinoma presenting with hyperthyroidism.

Follicular thyroid carcinoma typically manifests under euthyroid conditions, and diagnostic scintigraphy usually identifies a cold nodule. Sometimes, such tumors can appear in the context of hyperthyroidism, which can be caused by a toxic multinodular goitre, a toxic adenoma, or even carcinoma. We report a case of follicular thyroid carcinoma discovered after surgical treatment of a toxic multinodular goiter, in which solitary adrenal gland metastasis was detected five years later. A (131)I whole body scan is the diagnostic method of choice for functioning thyroid metastasis.     

Isolated metastasis of malignant melanoma into follicular carcinoma of the thyroid gland

A 38-yr-old woman with a history of malignant melanoma (MM) presented with a thyroidal nodule. Fine needle aspiration biopsy of the thyroid was consistent with metastatic MM. The patient underwent thyroidectomy: microscopic examination revealed a follicular carcinoma nodule harboring a focus of metastatic melanoma. On review of the fine needle aspiration biopsy specimen, the population of cells with more uniform nuclei with focal follicle formation, which initially was interpreted as cells originating from normal thyroid tissue, was seen to actually represent the follicular carcinoma component. Tumor-to-tumor metastasis is an interesting phenomenon and there are only few cases of MM metastasis to other tumors. MM metastasis into a neoplastic thyroid nodule is a very rare combination and may be explained because the nodule in question represents the most highly vascularized component of the thyroid.     

Papillary and follicular thyroid carcinoma

Papillary and follicular thyroid carcinomas are among the most curable of all cancers. However, some patients are at high risk of recurrence or even death from their cancer. Most of these patients can be identified at the time of diagnosis using well-established prognostic indicators. The extent of initial treatment and follow-up should therefore be individualized. The early discovery of persistent and recurrent disease is based on the combined use of serum thyroglobulin determination and of total body scanning with 131I. The recent availability of recombinant human thyroid stimulating hormone has greatly improved the quality of the patient's life during follow-up. Treatment of recurrences is based mainly on surgery and 131I treatment.     

Papillary and follicular thyroid carcinoma

Papillary and follicular carcinomas of the thyroid are differentiated carcinomas developed from the follicular epithelium, that keep some of its morphological and functional characteristics. Their increased incidence is related to an improved screening. Thyroid carcinoma usually presents as a thyroid nodule. Only 5% of nodules are malignant and fine needle biopsy is the most accurate tool for their diagnosis. Initial treatment is standardized and includes a total thyroidectomy with central lymph node dissection in case of papillary carcinoma, that is followed by the administration of a large activity of radioiodine in case of incomplete surgery, distant metastases or high risk factors. Papillary carcinomas of less than 1 cm in diameter, when unifocal and intra-thyroid are treated with surgery only, and radioiodine is not indicated. Thyroxine treatment is given to all patients. The majority of patients are cured, as demonstrated by the work-up performed at 1 year (undetectable serum thyroglobulin following stimulation with recombinant human TSH and normal neck ultrasonography). Subsequent follow-up is yearly with serum Tg and TSH determinations that is maintained within the normal range. In the other patients, other tests may be indicated, starting with the administration of a large activity of radioiodine. In these patients, serum TSH should be decreased to a low level.     

Gelsolin: a novel thyroid hormone receptor-beta interacting protein that modulates tumor progression in a mouse model of follicular thyroid cancer

Follicular thyroid cancer (FTC) is known to metastasize to distant sites via hematogenous spread; however, the underlying pathways that contribute to metastasis remain unknown. Recent creation of a knockin mutant mouse that expresses a mutant thyroid hormone receptor-beta (TRbeta(PV/PV) mouse) that spontaneously develops thyroid cancer with metastasis similar to humans has provided new opportunities to study contributors to FTC metastasis. This study evaluates the role of gelsolin, an actin-regulatory protein, in modulating the metastatic potential of FTC. Gelsolin was previously found by cDNA microarray analysis to be down-regulated in TRbeta(PV/PV) mice as compared with wild-type mice. This study found an age-dependent reduction of gelsolin protein abundance in TRbeta(PV/PV) mice as tumorigenesis progressed. Knockdown of gelsolin by small interfering RNA resulted in increased tumor cell motility and increased gelsolin expression by histone deacetylase inhibitor (trichostatin A) led to decreased cell motility. Additional biochemical analyses demonstrated that gelsolin physically interacted with TRbeta1 or PV in vivo and in vitro. The interaction regions were mapped to the C terminus of gelsolin and the DNA binding domain of TR. The physical interaction of gelsolin with PV reduced its binding to actin, leading to disarrayed cytoskeletal architectures. These results suggest that PV-induced alteration of the actin/gelsolin cytoskeleton contributes to increased cell motility. Thus, the present study uncovered a novel PV-mediated oncogenic pathway that could contribute to the local tumor progression and metastatic potential of thyroid carcinogenesis.     

Establishment of a DEN-induced mouse model of esophageal squamous cell carcinoma metastasis

Background

Syngeneic transplantation mouse models have been used to evaluate the efficacy of immunotherapy in addition to radiotherapy and chemotherapy for treating cancer. However, the mouse models of esophageal squamous cell carcinoma have yet to be established. Therefore, we aimed to develop a mouse model of esophageal squamous cell carcinoma.

Methods

Male and female Balb/c, C3H, and C57Bl mice received diethyl nitrosamine continuously for 4 months. On completion of the 200-day treatment period, animals were killed, and esophageal, forestomach, lung, and liver samples were examined macroscopically and by the histopathological analysis. Induced tumors from C3H female mice were mechanically dissociated into small pieces and were mixed with the brain homogenates, and injected into interscapular region subcutaneously into syngeneic C3H female mice to evaluate tumor growth and/or metastatic potential.

Results

The incidence of esophageal/forestomach squamous cell carcinoma varied according to mouse strain and gender, and the C3H mouse was found to be most susceptible. Pathologically, tumors were predominantly well-differentiated squamous cell carcinoma, with a proportion of tumors developing distant metastases. Transplanted esophageal squamous cell carcinoma cells developed subcutaneous tumors in syngeneic mice, with distant metastases into the lung. Metastatic tumors had poorly differentiated components histologically with Ki-67 and p53 expression. Metastatic tumors had lower p21 expression than transplanted tumors.

Conclusion

In the present study, we demonstrate the establishment of a mouse model esophageal squamous cell carcinoma allowing transplantation into syngeneic mice with distant metastatic potential. We believe that the present syngeneic mouse model will have utility in various preclinical research fields, including cancer immunotherapy.

     

Retroviral-mediated transmission of a mouse VL30 RNA to human melanoma cells promotes metastasis in an immunodeficient mouse model

Infection of a human melanoma cell line by a retroviral vector resulted in transmission of a mouse VL30 (mVL30-1) retroelement RNA to some of the cells infected by the retrovirus, followed by synthesis, integration, and expression of the mVL30-1 cDNA. One vector carried a tissue factor (TF) transgene that generated high TF melanoma clones, and another vector was a control without the TF transgene that generated low TF clones. Some high TF melanoma clones contained the mVL30-1 retroelement and others did not, and some low TF melanoma clones contained the mVL30-1 retroelement and others did not. Each type of melanoma clone was tested for its metastatic potential in severe combined immunodeficient (SCID) mice, by i.v. injection of the cells to generate lung tumors. None of the low TF clones that either contained or lacked the mVL30-1 retroelement generated lung tumors, consistent with earlier results showing that high TF expression promoted metastasis. The high TF clones containing the mVL30-1 retroelement were strongly metastatic, in contrast to the high TF clones lacking the mVL30-1 retroelement, which were weakly metastatic. Southern hybridization analyses showed that the mVL30-1 cDNA integrated into different genomic sites in different melanoma clones, suggesting that the effect of the mVL30-1 retroelement on metastasis depends not on integration per se but instead on expression of the mVL30-1 RNA. A role for the mVL30-1 RNA in metastasis and possibly other cell functions is an unexpected finding, because the RNA appears to lack significant coding potential for a functional protein. The metastatic effect might be mediated directly by a noncoding mVL30-1 RNA or by a peptide or small protein encoded by one of the short ORFs in the mVL30-1 RNA.     

Invasion of human follicular thyroid carcinoma cells in an in vivo invasion model.

Models that demonstrate histological invasion of extracellular matrix barriers by tumor cell lines are useful for assessing new methods to treat or prevent tumor metastasis. An in vivo invasion model using acellular human dermal matrix has been described in a murine squamous cell carcinoma line. The present study examined the application of this tumor invasion model to another epithelial cell line derived from a different species. A human follicular thyroid carcinoma cell line, known to be invasive by other assays, was grown on the dermal-epidermal basement membrane surface of human acellular dermal matrix in culture and then grafted in athymic mice. Immunohistochemical staining of type IV collagen was used to identify the basement membrane and invasion was determined as penetration of the basement membrane by tumor cells. Identification of the human tumor cells in the in vivo grafts was done by in situ hybridization with species specific probes. FTC-133 tumor cells did not invade the matrix after 4 weeks of growth in in vitro culture, but there was extensive loss of the basement membrane and infiltration of the tumor cells into the dermis after 2 weeks growth in vivo. This study suggests that the in vivo dermal matrix model of invasion is applicable to a broad range of epithelial carcinoma cell lines to study their capability to penetrate basement membrane. A model such as this may be useful for studying the local effects of genetic manipulations of implanted tumor cell populations, leading to the development of therapeutic agents that block invasion.     

p53 constrains progression to anaplastic thyroid carcinoma in a Braf-mutant mouse model of papillary thyroid cancer

Anaplastic thyroid carcinoma (ATC) has among the worst prognoses of any solid malignancy. The low incidence of the disease has in part precluded systematic clinical trials and tissue collection, and there has been little progress in developing effective therapies. v-raf murine sarcoma viral oncogene homolog B (BRAF) and tumor protein p53 (TP53) mutations cooccur in a high proportion of ATCs, particularly those associated with a precursor papillary thyroid carcinoma (PTC). To develop an adult-onset model of BRAF-mutant ATC, we generated a thyroid-specific CreER transgenic mouse. We used a Cre-regulated Braf^V600E mouse and a conditional Trp53 allelic series to demonstrate that p53 constrains progression from PTC to ATC. Gene expression and immunohistochemical analyses of murine tumors identified the cardinal features of human ATC including loss of differentiation, local invasion, distant metastasis, and rapid lethality. We used small-animal ultrasound imaging to monitor autochthonous tumors and showed that treatment with the selective BRAF inhibitor PLX4720 improved survival but did not lead to tumor regression or suppress signaling through the MAPK pathway. The combination of PLX4720 and the mapk/Erk kinase (MEK) inhibitor PD0325901 more completely suppressed MAPK pathway activation in mouse and human ATC cell lines and improved the structural response and survival of ATC-bearing animals. This model expands the limited repertoire of autochthonous models of clinically aggressive thyroid cancer, and these data suggest that small-molecule MAPK pathway inhibitors hold clinical promise in the treatment of advanced thyroid carcinoma.Mutations in the v-raf murine sarcoma viral oncogene homolog B (BRAF) kinase occur in ∼60% of papillary thyroid carcinomas (PTCs) (www.cbioportal.org/public-portal/data_sets.jsp). PTC generally exhibits an excellent prognosis with conventional therapy, including surgery and selective use of radioiodine (1). PTC may progress to clinically aggressive forms of thyroid cancer, including poorly differentiated thyroid carcinoma (PDTC), which exhibits more rapid growth and poorer clinical outcome. Less commonly, PTC progresses to undifferentiated (anaplastic) thyroid carcinoma (ATC) that is associated with a grim prognosis with a median survival of 5 mo and a 1-y survival of only 20% (2).Focused sequencing of clinically aggressive subsets of thyroid cancers including PDTC and ATC suggests acquired cooperating mutations drive thyroid cancer progression (3, 4). Mutations in tumor protein p53 (TP53) occur with increasing frequency in more aggressive forms of thyroid cancer, culminating in ATC, which harbors the highest frequency of TP53 mutations (5–7). ATC may progress from well-differentiated thyroid carcinomas and is also believed to arise spontaneously, possibly from clinically undetectable microscopic well-differentiated thyroid tumors. In the former scenario, ATCs frequently harbor mutations in BRAF, and these mutations are concordant between the anaplastic and papillary components. This implicates BRAF mutation as an initiating somatic genetic event and supports the hypothesis that loss of p53 function is important for progression to ATC (3, 8).Mouse models of thyroid cancer have supported the model of acquired mutations driving tumor progression. Although each study has technical limitations, including embryonic oncogene expression and/or elevated circulating thyroid-stimulating hormone (TSH) levels, this work generally supports the notion that BRAF^T1799A is sufficient to initiate PTC (9–12). In addition, deletion of p53 enabled tumor progression to high-grade thyroid carcinomas in a transgenic mouse model of translocations targeting the ret proto-oncogene (RET/PTC) driven PTC, and a model of follicular thyroid carcinoma initiated by tissue-specific phosphatase and tensin homolog (Pten) deletion (13, 14). These studies provide functional evidence of an important tumor suppressive role for p53 during thyroid carcinoma progression, although to date this has not been tested in models of BRAF-mutant PTC.Given the high frequency of BRAF and RAS mutations in thyroid carcinomas and the success of targeted therapy trials for advanced thyroid cancers, the potential utility of small-molecule inhibitors of the MAPK pathway has garnered much recent attention (15). These drugs have also been studied in models of BRAF-mutant thyroid carcinoma. Initial observations using a thyroid-specific doxycycline-inducible BRAF^T1799A allele suggested that BRAF or mapk/Erk kinase (MEK) inhibition induced thyroid carcinoma regression and differentiation (9). However, a recent study from the same laboratory showed a mitigated response to BRAF (PLX4032, vemurafenib) inhibition in human papillary and ATC cell lines and in an endogenous Braf^V600E-driven PTC mouse model. In response to PLX4032/vemurafenib, feedback inhibition of the human epidermal growth factor receptor 3 (HER3) receptor tyrosine kinase was abrogated, leading to reactivation of MAPK signaling (16). In addition, responses in patients treated with the BRAF inhibitor vemurafenib have exhibited modest activity (17).To develop an adult-onset autochthonous model of clinically aggressive thyroid carcinoma, we generate a thyroid-specific CreER transgenic mouse and use conditional Braf^T1799A and Trp53 alleles. We demonstrate that expression of BRAF^V600E is sufficient to initiate tumorigenesis in adult animals, and p53 loss enables progression to bona fide ATC recapitulating the cardinal features of the human disease including intrinsic resistance to BRAF inhibitors.     

Tissue factor promotes activation of coagulation and inflammation in a mouse model of sickle cell disease

Sickle cell disease (SCD) is associated with a complex vascular pathophysiology that includes activation of coagulation and inflammation. However, the crosstalk between these 2 systems in SCD has not been investigated. Here, we examined the role of tissue factor (TF) in the activation of coagulation and inflammation in 2 different mouse models of SCD (BERK and Townes). Leukocytes isolated from BERK mice expressed TF protein and had increased TF activity compared with control mice. We found that an inhibitory anti-TF antibody abrogated the activation of coagulation but had no effect on hemolysis or anemia. Importantly, inhibition of TF also attenuated inflammation and endothelial cell injury as demonstrated by reduced plasma levels of IL-6, serum amyloid P, and soluble vascular cell adhesion molecule-1. In addition, we found decreased levels of the chemokines MCP-1 and KC, as well as myeloperoxidase in the lungs of sickle cell mice treated with the anti-TF antibody. Finally, we found that endothelial cell-specific deletion of TF had no effect on coagulation but selectively attenuated plasma levels of IL-6. Our data indicate that different cellular sources of TF contribute to activation of coagulation, vascular inflammation, and endothelial cell injury. Furthermore, it appears that TF contributes to these processes without affecting intravascular hemolysis.     

Gene therapy of a rat follicular thyroid carcinoma model with adenoviral vectors transducing murine interleukin-12

Interleukin-12 (IL12) is a heterodimeric cytokine that plays an important role in the development of cellular immunity. We previously reported the antitumor activity of mouse IL12 (mIL12) transduced by adenovirus in a medullary thyroid carcinoma model. In this study, a rat thyroid follicular cancer cell line (RTC-R2) was employed to develop tumors after sc injection in Wistar rats. In five of five animals, RTC-R2 cells infected with mIL12 transducing adenovirus (AdCMVmIL12) in vitro failed to be tumorigenic in vivo in syngenic rats, whereas four of five animals developed tumors after injection of luciferase transducing adenovirus (AdCMVLuc)-infected cells. After intratumoral treatment with AdCMVmIL12 at 1 x 10(9) plaque-forming units per rat, 90% (26/29) of animals bearing small (<100 mm(3)) tumors were apparently cured. Larger tumors treated by injection of AdCMVmIL12 became significantly smaller than AdCMVLuc-treated animals, and growth stabilized. Challenge studies showed that only 3 of 28 animals previously treated and cured with AdCMVmIL12 developed a tumor after sc reinjection of RTC-R2 cells, whereas all animals developed tumors in na?ve animals. Thus, AdCMVmIL12-treated animals developed long-term antitumor immunity. We also studied animals with two tumors, injecting virus in one. Tumors regressed at both sites in five of six animals after treatment of one tumor with AdCMVmIL12, and in the sixth animal one site tumor regressed and another tumor continued to grow. In AdCMVLuc-treated animals, both tumors regressed in only one animal, and the reminder continued to grow. To detect toxicity to liver and other tissues after administration of AdCMVmIL2, the vector was administrated intratumorally or iv at the dose of 2 x 10(9) plaque-forming units per rat. No change in behavior was observed in any of the treated animals. Rats were killed at different time after virus administration. An overt increase of spleen size was observed 7 d after infection in all animals treated with AdCMVmIL12. All animals given virus IV had some lymphocyte infiltration in the sinusoids and triads of the liver, whereas AdCMVmIL12 injected intratumorally did not cause this effect. Spleens of some virus-treated animals showed decreased white pulp, with apparently increased hematopoiesis. No specific changes were found in lungs and kidneys. Iv administration of AdCMVmIL12 induced a moderate increase of glutamic-oxalacetic transaminase and glutamic-pyruvic transaminase, whereas AdCMVmIL12 injected intratumorally did not. This study confirms the efficient antitumor activity of an adenovirus expressing mIL12 after in vivo delivery in an animal model and indicates the possibility of application to patients because of the low toxicity.     

Receptor for hyaluronan-mediated motility isoform B promotes liver metastasis in a mouse model of multistep tumorigenesis and a tail vein assay for metastasis

The gene encoding the receptor for hyaluronan-mediated motility (RHAMM) is overexpressed in many human cancers. However, it is unclear whether RHAMM plays a causal role in tumor initiation or progression. Using somatic gene transfer in a mouse model of islet cell tumorigenesis, we demonstrate that RHAMM isoform B (RHAMM(B)) promotes tumor growth and metastases to lymph nodes and the liver. The propensity of RHAMM(B)-expressing cells to metastasize to the liver was confirmed using an experimental metastasis assay in which cells were injected into the tail vein of immunodeficient mice. However, RHAMM(B) did not increase cell migration or proliferation in culture. In initial efforts to identify signaling pathways activated by RHAMM(B), we found that RHAMM(B) induced phosphorylation of epidermal growth factor receptor (EGFR), Erk1/2, and STAT3 and conferred susceptibility to apoptosis after treatment with an EGFR inhibitor, gefitinib. Taken together, the results indicate that RHAMM(B) promotes hepatic metastasis by islet tumor cells, perhaps through growth factor receptor-mediated signaling.     

Cardiac metastasis from carcinoma of thyroid

A case of primary carcinoma of the thyroid with myocardial metastasis is reported.     

人肝癌肺和淋巴结靶向转移细胞及裸鼠移植模型的建立

目的 克隆具有肺和淋巴结转移亲嗜性的人肝癌细胞并建立相应的裸鼠移植模型.方法 将荧光人肝癌细胞HCCLM3-R接种于4周龄的裸鼠肝脏,6周后在荧光解剖下观察并获取肺及腹腔淋巴结转移灶组织,经体外克隆培养,所获细胞分别标记为HCCLM3-R-LM1和HCCLM3-R-LnM1.接种上述两种细胞于4周龄裸鼠肝脏,观察各自肺和腹腔淋巴结转移灶荧光数量,并与肺组织连续切片中的转移灶数目进行比较.计量资料采用Wilcoxon秩和检验和Kruskal-Wallis秩和检验进行统计学分析.结果 HCCLM3-R-LM1、HCCLM3-R和HCCLM3-R-LnM1细胞接种于裸鼠肝脏后第6周,在肺和腹腔淋巴结中均能发现肿瘤转移.3株细胞接种后,裸鼠肺和腹腔淋巴结转移灶荧光面积分别为8687.00±1844.63和2570.00±318.20(P＜0.001),6457.67±832.62和10 994.33±2212.31(P＜0.001),2968.67±2571.00和24 416.00±7186.13(P＜0.001),每只裸鼠光镜下肺转移灶中位数分别为755、430、310个(P＜0.001),与荧光定量结果相吻合.结论 成功建成人肝癌肺和淋巴结亲嗜性转移细胞和裸鼠移植模型,其中HCCLM3-R-LM1细胞具有明显的肺转移特性,而HCCLM3-R-LnM1细胞具有明显的淋巴结转移特性,为肝癌器官靶向转移研究提供了理想的体内外模型.

Abstract：

Objective To establish a systematic site-specific metastatsis model of human hepatocellular carcinoma (HCC) in nude mouse.Methods HCCLM3-R cells were seeded into mice liver to establish xenograft mouse models.With the help of RFP,metastasis foci in lungs and lymph nodes in mice were detected using fluorescent stereomicroscopy and were removed.Cells derived from the metastasis foci were named HCCLM3-R-LM1 and HCCLM3-R-LnM1 respectively.HCCLM3-R-LM1 and HCCLM3-R-LnM1 cells were seeded into mice livers to analyze the lung and lymph node metastasis.Lungs of all tested mice were collected,examined by pathological evaluation and counted lung metastasis.Results Both lung and lymph node metastasis were found in HCCLM3-R-LM1,HCCLM3-R and HCCLM3-R-LnM1 cells and a significant difference was found between the lung and the lymph node metastasis levels in the three cells.The fluorescent areas (pixels) of lung and lymph node metastasis were 8687.00 ± 1844.63 versus 2570.00 ±318.20 (P = 0.0031) in HCCLM3-R-LM1 cells,6457.67±832.62 versus 10 994.33±2 212.31(P=0.0036) in HCCLM3-R cells,and 2968.67 ± 2571.00 versus 24 416.00 ± 7 186.13 (P = 0.0094) in HCCLM3-R-LnM 1 cells,respectively.The middle numbers of microscopic lung metastatic foci were 775,430 and 310in HCCLM3-R-LM1,HCCLM3-R and HCCLM3-R-LnM1 cells (P＜0.001),respectively,consist with the results quantified by RFP.Conclusion We established the systematic site-specific metastasis models which demonstrates lung- and lymph node-specific metastasis potential in nude mice and can be used as a model for researches on site-specific metastasis of HCC.