TTF-1 Expression Research Articles

OA19.02 Sex Differences Are Detected in the Profile of Tumor Associated Inflammatory Cells (TAICs) Are Lung Adenocarcinoma

A number of studies have characterized TAICs in lung cancer and associated their levels of infiltration with patients’ outcome. There is limited information about the correlation of TAICs infiltration with clinical and pathological features of lung cancer. We investigated the association between patterns of tumor infiltrating lymphocytes and macrophages with detailed clinical and pathological features in lung adenocarcinoma. We studied archival tumor tissue from 93 surgically resected lung adenocarcinomas, stages I to III. Density of TAICs expressing CD3, CD4, CD8, and CD68 was evaluated using immunohistochemistry (IHC) and image analysis. TAICs density was correlated with tumor’s histological characteristics and patients’ characteristics. We found significant differences in the TAICs infiltrate density of lung adenocarcinomas based on patients’ characteristics. Overall: a) females showed higher levels of CD8+ (P=0.01) and CD3+ (P=0.03) cell density than males; b) smaller tumors (<3cms) showed more CD4+ (P=0.01) and CD3+ (P=0.03) cells than larger tumors; and, c) tumors with solid histology pattern showed higher levels of CD8+ (P=0.03) cells than non-solid pattern. No overall significant differences on TAICs infiltrates were detected by age, tobacco exposure by pack-years and TTF-1 IHC expression score. However when TAICs density of tumors was examined by sex we found the following: a) in larger tumor (>3cms), females demonstrated higher levels of CD8+ cells (P=0.0007) than males; b) tumors from females older than the median age (63 years) showed more CD4+ (P=0.04), CD8+ (P=0.009) and CD3+ (P=0.042) cells than males; c) tumors from females with <40 pack-years of tobacco history showed significantly higher levels of CD3+ (P=0.004) and CD68+ (P=0.004) cells than males; d) tumors from females with high levels of TTF1 expression (score >150) showed higher levels of CD8+ cells(P=0.03) than males; e) females with tumors having non-solid histology pattern showed higher CD8+ (P=0.02) and CD3+ (P=0.02) cells than males. Finally, tumors expressing low levels of TTF-1 and lower CD4+ cells correlated significantly with worse overall recurrence free survival and overall survival in both males (P<0.0001) and females (P=0.0072). In lung adenocarcinoma, TAICs infiltration correlates with clinical characteristics of patients and pathological features of tumors, particularly, sex, age, size and TTF-1 expression. Compared with men, lung adenocarcinomas from females showed higher levels of TAICs, particularly at older age, larger tumor size, less exposure to tobacco, and more differentiated histological patterns. (UT Lung SPORE and MD Anderson Moon Shot Program).

MA17.02 Genome-Wide Copy Number and Mutational Analysis in Longitudinal Biopsies of Matched Primary and Metastatic Pulmonary Adenocarcinomas

There are still limited data on the extent of intratumoral heterogeneity of cancer gene mutations and genome-wide copy number aberrations between primary tumors and metastases in non-small cell lung cancers (NSCLC). Deconvolution of the intermixture of tumor and stromal components remains a major challenge for such analysis. To overcome these limitations, we applied a refined nuclei flow sorting approach on matched longitudinal biopsies (primary/metastasis) from pulmonary adenocarcinomas. Multiparameter Ploidy Profiling (MPP) comprises the isolation of nuclei from frozen or formalin-fixed and paraffin embedded (FFPE) tissues, followed by multiparameter flow sorting by DAPI for DNA content (ploidy) and TTF1 as a lineage marker to enrich for tumor cell nuclei. Homogenous TTF1 expression was ascertained by immunohistochemistry. Sorted populations were subjected to genomic profiling by high resolution aCGH and NGS with the Ion Torrent™ Comprehensive Cancer Panel. This approach allows for the detection of genome-wide copy number aberrations and provides all exon-coverage of 409 well-known cancer genes. Sequencing was performed with a mean depth of 965x. MPP was successfully applied on 44 frozen or FFPE tissue specimens from 19 patients. Clonally unrelated secondary primaries were found in three patients, defined by the absence of both shared copy number (CN) transition and somatic mutations. The concordance rate between primary tumor and corresponding metastases was 65.2% and reached 85.5% for mutations and copy number amplifications/deletions in the top 12 affected genes (including CDKN2A, KRAS, ATM, KEAP1, EGFR and STK11). The correlation of the allele frequencies between primary tumors and metastases was linear (r=0.87, p<0.001), irrespective of the time interval between the tissue resections. Overall, ploidy was not different between primary tumors and metastases. Additionally, the metastases did not bear a higher burden of private events (CN transitions and somatic mutations) than the primary tumors. MPP is a powerful method to increase the precision of downstream analysis due to unprecedented purity of tumor DNA. Our data argue for a high concordance rate of mutations and CN transitions between primary tumors and their corresponding metastases. Intriguingly, the ploidy remains remarkably stable during progression even after long time-periods, which suggests chromosomal stability with a limited degree of macroevolutionary shifts over time and space. Taken together, our data suggest the presence of at least two evolutionary patterns: 1) early/branched and 2) late/linear progression, with a continuum from high to low genetic divergence of the primary tumor and metastases to their most recent common ancestor.

P2.03a-060 Favorable Survival of TTF-1 Expression in Pemetrexed Based Treated NSCLC Patients: Topic: Clinical Trials

The translational research analysis of the Pointbreak study could demonstrate superior overall survival (OS) for TTF-1 expressing NSCLC in comparison to TTF1-1 negatives treated with Pemetrexed based chemotherapy. The aim of this study was to prove retrospectively whether this finding can be reproduced in patients from the daily clinical practice in three different German hospitals. 323 patients (pts), 182m, 141f, median age 64 years (34-84) with non-squamous NSCLC stage UICC IIIB/IV undergoing pemetrexed based palliative chemotherapy were analyzed for TTF-1 expression by immunohistochemistry and outcome. 246 pts (76%) were TTF-1 positive, well balanced for age and gender. Median number of administered chemotherapy cycles was 4 in both groups; 19 pts (5.9%) received maintenance therapy. Response was superior in TTF-1 expressing (vs non-expressing) patients (CR 2% vs. 6%, PR 58% vs. 39%, NC 21% vs. 16%, PD 19% vs. 39%, X2=14.4, p<0.002) as well as OS (MST 14.5m vs. 8.3m, HR=0.44, CI 0.33-0.58, p<0.0001) and progression free survival (PFS: MST 7.7m vs. 4.8m, HR 0.51, CI 0.35-0.74, p<0.001). 1-y and 2-y OS were 59% and 23% for TTF-1 expressing pts compared to 34% and 0%, respectively In multivariate analysis TTF-1 expression (HR= 0.43, CI 0.30 - 0.63, p<0.0001) and 4 vs. 6 cycles of chemotherapy (HR=0.81, CI 0.70 - 0.95, p=0.009) were the only independent factors for OS. For PFS TTF-1 expression was the only independent predictive factor (HR=0.49, CI 0.31-0.77, p=0.002). These results confirm the relevance of TTF-1 expression on outcome in pemetrexed based chemotherapy with TTF-1 being of significant independent prognostic relevance. Further analyses with TTF-1 in non pemetrexed treated patients are ongoing to evaluate its predictive value.

P3.02a-003 ALK and ROS1 Rearrangements, Coexistence and Treatment in EGFR-Wild Type Lung Adenocarcinoma - A Multicenter Study of 732 Cases: Topic: ALK

Anaplastic lymphoma kinase (ALK) and c-ros oncogene 1 (ROS1) rearrangements represent two of the most frequency fusion targets in lung adenocarcinoma. The aim of this study was to investigate the clinicopathological characteristics, coexistence and treatment of ALK and ROS1- rearrangement patients in lung adenocarcinoma without EGFR mutations. Patients who harbored EGFR wild-type gene were screened for ALK and ROS1 gene in four Hospitals in China. ALK and ROS1 rearrangements were detected using RT-PCR. Progression free survival curves were plotted using the Kaplane-Meier method. Seven hundred and thirty-two patients enrolled in current study. Of the 732 patients, the median age was 59 years (range: 28–81). ALK and ROS1 rearrangements were detected in 89 (12.2%) and 32 (4.4%) patients, respectively. One patient was identified with ALK/ROS1 coexistence. Both of ALK and ROS1 positive were predominantly found in younger and non-smokers. More patients with ALK/ROS1 rearrangements were associated with TTF1 expression，napsin A expression and solid predominant adenocarcinoma subtype (Figure 1 ). ALK and ROS1 rearrangements frequency was enriched in EGFR wild-type patients and ALK/ROS1 coexistence was rare. The ALK and ROS1 arrangements were associated with age, smoking status, TTF1 expression, napsin A expression and solid predominant adnocarcinoma subtype.

P3.02b-019 TTF-1 Expression as a Predictor of Response to EGFR-TKIs in Patients with Lung Adenocarcinoma: Topic: EGFR Biomarkers

Several studies have shown that overexpression of thyroid transcription factor (TTF-1) may be associated with the presence of epidermal growth factor receptor (EGFR) gene mutations and predict survival in patients with non-small cell lung cancer (NSCLC). Nevertheless, the potential significance of TTF-1 immunostaining as a predictor of response to EGFR tyrosine kinase inhibitors (TKIs) has received limited research attention thus far. The aim of this study was to further explore the potential association between TTF-1 immunohistochemical expression and response to EGFR TKIs in patients with lung adenocarcinoma. The medical records of 129 patients with stage IV lung adenocarcinoma, treated at the Oncology Unit of “Sotiria” Athens General Hospital between January 2011 and December 2014, were retrospectively reviewed. All patients had received treatment with EGFR TKIs (erlotinib or gefitinib) and had a known TTF-1 immunohistochemical expression status in formalin-fixed, paraffin-embedded tumor tissue. Demographic and clinicopathological features (age, gender, smoking status and performance status), TTF-1 immunohistochemistry and EGFR mutation status results were correlated to each other as well as with the response rate (RR) to EGFR TKIs, using univariate and multivariate regression analysis. Median age of our study population was 68 years (range 26-88 years), while the majority were male (79/129 cases, 61.2%). Patients with EGFR mutant tumors had significantly higher response rates to EGFR TKIs compared to patients with wild-type EGFR tumors (p=0.001). TTF-1 positive staining was weakly associated with the presence of EGFR mutations, without reaching the level of statistical significance (p=0.053). Most importantly, TTF-1 positive staining was not significantly correlated with RR to EGFR TKIs, when gender, age, smoking status, EGFR mutation status and PS were included in the multivariate model. The present results failed to demonstrate any independent association between TTF-1 overexpression and the RR to EGFR TKIs in patients with advanced-stage lung adenocarcinoma. Prospective data from larger cohorts of patients are needed to clarify the exact predictive value of TTF-1 immunostaining in this setting.

PUB094 Correlation of Clinicopathologic Features and Lung Squamous Cell Carcinoma Subtypes According to the 2015 WHO Classification

This study aimed to determine the relationship between clinicopathologic features and lung squamous cell carcinoma subtypes according to the 2015 WHO classification. We identified 368 operable lung squamous cell carcinoma patients who had undergone a complete surgical resection at Shanghai Chest Hospital between April 2015 and March 2016. Among all patients, the percentages of lung squamous cell carcinoma subtypes were: 68.5% (252/368), 26.2% (97/368), and 5.1% (19/368) for keratinizing squamous cell carcinoma (KSCC), nonkeratinizing squamous cell carcinoma (NKSCC), and basaloid squamous cell carcinoma (BSCC), respectively. There were more smokers in patients with KSCC than in patients with other subtypes (p=0.004). There were no significant relationships between pathological subtypes and other clinicopathologic features, such as gender, age, location type, type of resection, stage, visceral pleural involvement, lymphovascular invasion, and lymph node involvement. The expressions of P40, CK5/6, TTF-1, Napsin A, and CK7 were also not significantly different in three subtypes. Our study revealed that there were no significant relationships between clinicopathologic features and lung squamous cell carcinoma subtypes.

Lung Adenocarcinoma With MUC4 Expression Is Associated With Smoking Status, HER2 Protein Expression, and Poor Prognosis: Clinicopathologic Analysis of 338 Cases

BackgroundMUC4 is a transmembrane glycoprotein that plays a role in the cell growth signaling pathway and has been studied in various organ malignancies. This study aimed to analyze MUC4 expression in resected lung adenocarcinomas (ADCs) to define the clinicopathologic characteristics of MUC4-positive cancers. Patients and MethodsImmunohistochemical MUC4 analysis was performed using tissue microarray slides containing 338 lung ADCs. Associations between MUC4 expression and the following clinicopathologic parameters were evaluated: sex; age; smoking status; tumor stage; tumor grade; lymphovascular invasion; pleural invasion; TTF-1 and HNF4α expression; EGFR, KRAS, BRAF, and HER2 mutation status; and ALK and ROS1 fusion status. ResultsNinety-four tumors (27.8%) were MUC4 positive. Most patients with MUC4-positive tumors were male (P < .001) and smokers (P = .006). Moreover, MUC4 expression was significantly associated with solid ADCs (P < .001) and vascular invasion (P = .001). MUC4 expression inversely correlated with TTF-1 expression (P = .020) and EGFR mutations (P = .004). Interestingly, MUC4 expression correlated with HER2 protein expression (P = .042), although MUC4 expression did not correlate with HER2 DNA amplification or HER2 gene mutations. Patients with MUC4-positive tumors had significantly worse prognoses compared to patients with MUC4-negative tumors (P = .025). ConclusionThe present study showed that MUC4-positive lung ADCs correlated with male smokers, solid ADCs, negative TTF-1 expression, the EGFR wild-type gene, HER2 protein expression, and poorer prognoses. These results suggest that MUC4-positive lung ADC may be a distinct subtype found in patients with smoking-related poor outcomes, mediated by HER2 signaling pathway.

High-level expression of divergent endodermal lineage markers in gonadal and extra-gonadal yolk sac tumors

AbstractYolk sac tumors occur at both gonadal and extra-gonadal sites. A recent case of ovarian endometrioid-pattern yolk sac tumor with strong diffuse expression of TTF-1 illustrated the potential for misdiagnosis due to divergent expression of endodermal lineage markers. The aim of this study was to investigate the expression of four divergent endodermal lineage markers, TTF-1, CDX2, Hep Par 1, and Napsin A, in gonadal and extra-gonadal yolk sac tumors of differing age, sex, and location (excluding foci of overt hepatoid differentiation). We identified 26 cases (5 ovarian, 15 testicular, and 6 extra-gonadal) containing yolk sac tumor as identified by typical histology and confirmed by positive immunohistochemical staining for alpha-fetoprotein and glypican-3. Mixed or ambiguous foci were confirmed by immunohistochemistry (SALL4 positive and Oct-4 negative). The relative proportion of three histologic patterns: reticular/cystic, solid/myxoid, and glandular was estimated. Percent positivity for the four divergent endodermal lineage markers was compared within yolk sac tumor areas according to site, age group, and histologic pattern. High-level (>25%) staining for one or more divergent endodermal lineage markers was seen in eleven cases: Hep Par 1 in seven cases, all post-pubertal, TTF-1 in four cases, two ovarian and two extra-gonadal, and CDX2 in three cases, with no age or site predilection. No case highly expressed all three divergent endodermal lineage markers, but four co-expressed high levels of two markers: two ovarian yolk sac tumors with TTF-1 and Hep Par 1, one testicular yolk sac tumor with CDX2 and Hep Par 1, and one extra-gonadal yolk sac tumors with TTF-1 and CDX2. While no absolute correlation of high-level divergent endodermal lineage marker expression with histologic subtype was observed, TTF-1 and CDX2 expression was predominantly seen in reticular/cystic and glandular areas while Hep Par 1 was most frequent in myxoid/solid and glandular areas.

Clinicopathologic features and epidermal growth factor receptor gene mutation of primary pulmonary mucoepidermoid carcinoma

To investigate the clinicopathological features and epidermal growth factor receptor (EGFR) gene mutation of primary pulmonary mucoepidermoid carcinoma (PMC). Fifth-five cases of PMC were included in the study; their clinicopathological, immunohistochemical features were evaluated, and in 31 cases, paraffin embedded specimens were subjected to mutation analysis of exons 18, 19, 20 and 21 of the EGFR gene by ARMS method. There were 32 females and 23 males. The patients' age ranged from 11 to 68 years(mean 36 years). The tumor size ranged from 0.7 to 5.5 cm(mean 2.4 cm). The tumors were located in the segmental bronchus and upper segmental bronchus. The tumors were well-demarcated, had no obvious capsules, and protruded into the bronchial lumen. Microscopically, there were three types of tumor cells including squamous cells, mucin-producing cells and intermediate cells. Fifty-three cases were low grade, two were high grade and both showed lymph node metastases. Immunohistochemically the tumor cells were uniformly negative for TTF1, Napsin A and ALK (Ventana D5F3) in all 55 cases. The tumor cells were positive for CK7, and some squamous and intermediate cells were positive for p63 and CK5/6 in all 55 cases; whereas they were positive for p40 in 53 cases. The mucin-producing cells were negative for CK5/6, p63 and p40, but were positive for PAS in all 55 cases. The Ki-67 positive rate was <10% in the low grade PMC, and was about 80%-90% in the high grade cases. Follow-up information was available in 37 patients, with no recurrence or death. There were no EGFR gene mutations in all 31 patients of PMC. PMC is a rare malignant salivary gland-type tumor occurring mainly in the central trachea of young patients.Most PMCs are low grade with good prognosis. The rate of lymph node metastasis of high grade PMC is high. Diagnosing PMC in small biopsies could be problematic. The lack of TTF1 expression is helpful to differentiate from other primary lung cancers. There is no EGFR gene mutation in PMC.

Dietary flavanones and citrus fruits influence cytokines and thyroid transcription factor-1 in an HDM-induced chronic asthma murine model

The effects of hesperetin plus naringenin, orange juice and grapefruit juice on mRNA expression of TNF-α, TGF-β1 and TTF-1 in a house dust mite (HDM)-induced asthma animal model were investigated. Interventional mice exposed to HDM for 6 weeks received hesperetin plus naringenin, orange plus grapefruit juice, orange juice, grapefruit juice or water during the last 4 weeks, whereas non-asthmatic control mice consumed water. Lung tissue TNF-α and TGF-β1 expressions in supplemented groups were significantly reduced (p < 0.0001 and p < 0.001, respectively) compared with asthmatic controls. Hesperetin plus naringenin and orange plus grapefruit juice intake reduced expression of TGF-β1 more than grapefruit juice (p ≤ 0.02) or orange juice compared with the asthmatic control group. All supplemented groups had non-significantly higher TTF-1 expression compared with water intake groups. Orange plus grapefruit juice or hesperetin plus naringenin with down-regulation of TNF-α and TGF-β1 may ameliorate lung damage in the lung tissue of asthmatic subjects.

Expression of SOX11, PAX5, TTF-1 and ISL-1 in medulloblastoma

The aim of our study was to evaluate the immunohistochemical expression of SOX11, PAX5, TTF-1 and ISL-1 in medulloblastoma (MB) to investigate their diagnostic usefulness. MethodsImmunohistochemical expression of PAX5 (two antibodies: Dako, DAK-Pax5; and BD, clone 24), TTF-1 (Dako, 8G7G3/1), SOX11 (CL0142; Abcam) and ISL-1 (1 H9, Abcam) was analyzed using the h-score and Remmele score in 25 cases of MB. ResultsThere were 18 MBs of classic and 7 of desmoplastic type. SOX11 was strongly expressed in all tumors. The expression of PAX5 was higher and more frequent in a case of DAK-Pax5 clone (25/25) than clone 24 (6/25). ISL-1 was positive in 11 (44%) and TTF-1 in 3 (12%) cases. ISL-1 expression correlated positively (p<0.001), while TTF-1 correlated negatively with the age of patients (p=0.039). PAX5 expression correlated with ISL-1 (p=0.039) and showed a trend toward higher expression in the desmoplastic subtype (p=0.069). ConclusionsSOX11 is strongly and robustly expressed in MBs. PAX5 expression pattern differs substantially among two antibody clones. TTF-1 and ISL-1 is associated with the age of patients.

Abstract 3887: Proteomic profiling identifies cMyc and TTF1 as biomarkers of response to the aurora kinase inhibitor alisertib in small cell lung cancer (SCLC)

Abstract Background: Small cell lung cancer (SCLC) is the most aggressive form of lung cancer, with a 5-year survival rate of only 6%. The treatment of SCLC has not changed in over 25 years, hence the development of new drugs for SCLC represents a major unmet need. A recent phase II trial of the aurora kinase inhibitor alisertib showed single agent activity in a subset (21%) of SCLC (Melichar et al, 2015) and a second study is underway testing alisertib in combination with chemotherapy (NCT02038647). However, there are currently no established biomarkers to identify patients likely to have the greatest benefit from alisertib. We have previously demonstrated the utility of proteomic profiling to identify targets and markers in SCLC. Here we use proteomics to identify markers of response to alisertib in SCLC. Methods: The sensitivity (IC50) of 51 SCLC cell lines to alisertib was assayed in 5-day proliferation assays. Expression levels of 171 total and/or phosphorylated proteins were measured by reverse phase protein array (RPPA) and correlated with IC50’s. Using two approaches – 1) Spearman correlation of IC50 to protein data for all cell lines and 2) t-test comparing protein data between the most and least sensitive cell lines – we generated consensus markers of response. Results: Proliferation assays showed sensitivity to alisertib at clinically achievable doses in 14/51 (27%) cell lines (based on Cmax = 1.8uM, Phase I single agent trial). High cMyc protein was the top marker of sensitivity to alisertib (R = -0.47, p = 0.0006 as continuous variables; fold difference = 3.52, p = 0.008 by t-test comparing extremes). Further analysis revealed a bi-modal distribution of cMyc protein, defined as high and low using a bimodality index. The cMyc protein high group (25%) included cell lines that were both cMyc amplified and non-amplified. While cMyc amplified cell lines (11%) are more sensitive to alisertib than non-amplified (p = 0.002), high cMyc protein captures a larger population of SCLC that is sensitive to alisertib. In contrast, expression of thyroid transcription factor 1 (TTF1, a standard IHC marker used in the diagnosis of lung cancer) was the top marker of alisertib resistance (R = 0.38, p = 0.006; fold difference = -4.22, p = 0.003). TTF1 protein expression was also bi-modal, with 32% of cell lines falling into a distinct TTF1-low (more sensitive to alisertib) group. Conclusions: High cMyc and low TTF1 protein expression identify a subset of SCLC cell lines (27%) that are sensitive to single agent alisertib. cMyc protein as a marker of response is consistent with other preclinical findings suggesting that cMyc amplified SCLC may be more sensitive to aurora kinase inhibition. The association of low TTF1 expression with alisertib sensitivity may prove to be of particular value in selecting patients for treatment given that immunohistochemical assessment of TTF1 is commonly used in the diagnosis of SCLC. Citation Format: Robert J. Cardnell, Lerong Li, Fatemeh Masrorpour, Huifeng Niu, Jeffrey Ecsedy, Jing Wang, Lauren A. Byers. Proteomic profiling identifies cMyc and TTF1 as biomarkers of response to the aurora kinase inhibitor alisertib in small cell lung cancer (SCLC). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3887.

Induction of TTF-1 or PAX-8 expression on proliferation and tumorigenicity in thyroid carcinomas.

TTF-1 and PAX-8 are responsible for thyroid organogenesis and for maintenance of differentiation in thyrocytes. Thus, we hypothesized that the induction of these two transcription factors could affect proliferation and tumorigenicity. Moreover, the ability of various pharmacological agents to modulate expression of the TTF-1 and PAX-8 and their effects on apoptosis were also analysed. For this purpose, cell lines derived from papillary (TPC-1 and BHP 10-3) and anaplastic (ARO) thyroid carcinomas were stably transfected with expression vectors containing TTF-1 or PAX-8 genes. Subsequently, the effects on expression at gene and protein levels, as well as on cell growth, cell cycle, migration and invivo tumorigenicity were studied. Our results showed that: i) TTF-1 reciprocally induces PAX-8 expression; ii) the basal state of TTF-1 or PAX-8 influences proliferation, migration and tumorigenicity; iii) the induction of TTF-1 acts on cell proliferation more than PAX-8 and mainly affects tumorigenicity; and iv) TTF-1 was found to be more sensitive to epigenetic modulators than PAX-8. Therefore, we postulated that both TTF-1 and PAX-8 when co-expressed have anti-proliferative and anti-tumorigenic properties up to a threshold expression level and beyond that, are able to induce pro-tumorigenic effects. Hence in future, it will be quite interesting to systematically take into account the basal state of expression of TTF-1 and PAX-8. It will also be important to study the two thyroid transcription factors as part of a duo. This could open in the long-term, new therapeutic perspectives for thyroid carcinomas.

Surfactant dysfunction during overexpression of TGF-β1 precedes profibrotic lung remodeling in vivo.

Transforming growth factor-β1 (TGF-β1) is involved in regulation of cellular proliferation, differentiation, and fibrogenesis, inducing myofibroblast migration and increasing extracellular matrix synthesis. Here, TGF-β1 effects on pulmonary structure and function were analyzed. Adenovirus-mediated gene transfer of TGF-β1 in mice lungs was performed and evaluated by design-based stereology, invasive pulmonary function testing, and detailed analyses of the surfactant system 1 and 2 wk after gene transfer. After 1 wk decreased static compliance was linked with a dramatic alveolar derecruitment without edema formation or increase in the volume of septal wall tissue or collagen fibrils. Abnormally high surface tension correlated with downregulation of surfactant proteins B and C. TTF-1 expression was reduced, and, using PLA (proximity ligand assay) technology, we found Smad3 and TTF-1 forming complexes in vivo, which are normally translocated into the nucleus of the alveolar epithelial type II cells (AE2C) but in the presence of TGF-β1 remain in the cytoplasm. AE2C show altered morphology, resulting in loss of total apical surface area per lung and polarity. These changes of AE2C were progressive 2 wk after gene transfer and correlated with lung compliance. Although static lung compliance remained low, the volume of septal wall tissue and collagen fibrils increased 2 wk after gene transfer. In this animal model, the primary effect of TGF-β1 signaling in the lung is downregulation of surfactant proteins, high surface tension, alveolar derecruitment, and mechanical stress, which precede fibrotic tissue remodeling and progressive loss of AE2C polarity. Initial TTF-1 dysfunction is potentially linked to downregulation of surfactant proteins.

Contribution of ¹⁸Fluorodeoxyglucose positron emission tomography uptake and TTF-1 expression in the evaluation of the EGFR mutation in patients with lung adenocarcinoma.

The aim of this study is to evaluate the diagnostic value of PET-CT scan for the prediction of EGFR mutation status and the contribution of TTF-1 expression to PET-CT scan. We retrospectively studied 218 cases with a diagnosis of pulmonary adenocarcinoma between 2012-2014 which underwent EGFR analysis, TTF-1 and PET-CT before treatment. The EGFR mutation was present in 28.9% (n= 63) of cases. TTF-1 positivity was 66.9% (n= 105). Standardized uptake value (SUV max) was 16.7 ± 6.8 in EGFR mutant type, 13.8 ± 7.6 in cases having no EGFR mutations. According to our evaluations, high SUVmax is positively correlated with EGFR mutation status. TTF-1 expression in multivariate analysis strengthens the accuracy of detecting an EGFR mutation. PET-CT FDG uptake may, together with TTF-1 expression, help diagnosis in lung adenocarcinoma cases when evaluating for EGFR mutation status.

Cytokeratin 5/6 and P63 immunophenotype of thyroid lymphoepithelial complexes.

Thyroid lymphoepithelial complexes (LECos) are rare, being reported in lymphoma, Graves-Basedow disease, Hashimoto thyroiditis, pericarcinomatous thyroid or in the context of branchial cleft-like cysts. Here we report immunohistochemical expression of cytokeratin 5/6, P63 and TTF1 in 6 cases of thyroid LECos. Two cases had carbimazole treatment for hyperthyroidia and Graves disease. Anti-thyroglobulin, -thyroperoxidase or -TSH antibodies were detected in 4 cases. NSAID or poviodone iodine allergy were present in 2 cases. The treatment consisted in total thyroidectomy or lobectomy. Microscopy showed nodular goiter and focal lymphocytic thyroiditis. Basaloid LECos were seen in all thyroids while squamoid LECos in 2. Associated lesions were papillary thyroid microcarcinoma (2 cases), solid cell nest, thyroglosal duct remnant, lymphoepithelial cyst and thymus-parathyroid unit (one case each). Cytokeratin 5/6 was expressed in both squamoid and basaloid LECos along with P63. TTF1 expression was faint or absent. In conclusion LECos may occur in the context of autoimmune thyroiditis or of a specific immune susceptibility background. The expression of CK5/6 and of P63 suggests a squamous differentiation including in the basaloid LECos. The etiologic relevance of these immunostainings remains limited although rather suggestive of a metaplastic process than of migration-abnormalities.

Thyroid Transcription Factor 1 Reprograms Angiogenic Activities of Secretome.

Through both gain- and loss-of-TTF-1 expression strategies, we show that TTF-1 positively regulates vascular endothelial growth factor (VEGF) and that the VEGF promoter element contains multiple TTF-1-responsive sequences. The major signaling receptor for VEGF, i.e VEGFR2, also appears to be under a direct and positive regulation of TTF-1. The TTF-1-dependent upregulation of VEGF was moderately sensitive to rapamycin, implicating a partial involvement of mammalian target of rapamycin (mTOR). However, hypoxia did not further increase the secreted VEGF level of the TTF-1+ lung cancer cells. The TTF-1-induced VEGF upregulation occurs in both compartments (exosomes and exosome-depleted media (EDM)) of the conditioned media. Surprisingly, the EDM of TTF-1+ lung cancer cells (designated EDM-TTF-1+) displayed an anti-angiogenic activity in the endothelial cell tube formation assay. Mechanistic studies suggest that the increased granulocyte-macrophage colony-stimulating factor (GM-CSF) level in the EDM-TTF-1+ conferred the antiangiogenic activities. In human lung cancer, the expression of TTF-1 and GM-CSF exhibits a statistically significant and positive correlation. In summary, this study provides evidence that TTF-1 may reprogram lung cancer secreted proteome into an antiangiogenic state, offering a novel basis to account for the long-standing observation of favorable prognosis associated with TTF-1+ lung adenocarcinomas.

ROS-1 rearrangements in non-small cell lung cancer (NSCLC): a new target for a small subset of patients but a giant leap in therapeutics

ROS1 is a membrane tyrosine kinase receptor of which encoding gene has recently been found rearranged in non-small cell lung cancer (NSCLC), leading to constitutive activation of the ROS1 kinase activity. The ROS1 gene rearrangement has been described in roughly 1 % of patients with NSCLC and such rearrangement needs to be assessed by a FISH break-apart assay (CD74-ROS1) with ROS1 immunohistochemistry as a screening tool in tumor histological samples. The clinical and histological features of patients with ROS1-rearranged NCSCL include adenocarcinoma histology with TTF-1 expression, predominantly young women patients, and never-smokers. Targeting ROS1 fusion proteins with a kinase inhibitor of ROS1 kinase domain, such as crizotinib, seems to be promising. Indeed, this potential inhibitor of ROS1 kinase shows evidence of clinical efficiency either in patients treated off-label, in the crizotinib phase I trial, or in retrospective series. New kinase inhibitors such as ceritinib or lorlatinib also showed promising activities either in vitro, in mice xenograft models, or in patients treated off-label.

Clinical and biological significance of transcription termination factor, RNA polymerase I in human liver hepatocellular carcinoma.

Recent studies have indicated that increased ribosomal activity contributes to cancer progression. Transcription termination factor, RNA polymerase I (TTF1) acts as a transcription factor for RNA polymerase I. However, the role which TTF1 plays in cancer progression still remains unknown. The present study aimed to determine whether TTF1 plays a critical role in the progression of human liver hepatocellular carcinoma (HCC). In the present study, quantitative real-time reverse transcription polymerase chain reaction was conducted to evaluate TTF1 mRNA expression in 60 HCC tissue samples in order to determine the clinicopathological significance of TTF1. To investigate whether the expression levels of TTF1 were associated known gene signatures which represented ribosomal activity, we applied gene set enrichment analysis (GSEA) to HCC cases in The Cancer Genome Atlas (TCGA) a. We also performed in vitro proliferation assays using TTF1‑overexpressing HCC cells. TTF1 expression was significantly higher in HCC tumor tissues than in adjacent liver tissues (P<0.001). The overall survival (OS) of patients with high TTF1 expression levels was significantly shorter than that of patients with low TTF1 expression (P=0.027). Multivariate analysis indicated that TTF1 expression was an independent prognostic factor for OS (P=0.020). GSEA revealed significant associations between TTF1 expression and gene sets involved in ribosomal function. In vitro, cell proliferation and rRNA transcription were significantly promoted by overexpression of TTF1 in the HCC cell lines HuH-7 and HepG2. From these results, it was suggested that TTF1 participate in poor prognoses and play a role in tumor cell growth in HCC, possibly by upregulating ribosomal activity. In conclusion, we first propose that TTF1 may be a novel biomarker and therapeutic target in HCC. Increased expression of TTF1 was significantly associated with poor prognosis in two independent sets of HCC cases. Furthermore, in vitro experiments provided an explanation for clinical data showing that overexpression of TTF1 contributed to the proliferation of cancer cells.

Expression pattern of ISL-1, TTF-1 and PAX5 in olfactory neuroblastoma

Olfactory neuroblastoma (ONB) is a rare neoplasm of the sinonasal area with neuroendocrine differentiation. ISL-1, TTF-1 and PAX5 are transcription factors that are frequently upregulated in tumors showing neuroendocrine differentiation. The aim of our study was to evaluate these markers in a group of ONBs. We included 11 ONBs from 4 large university hospitals. Immunohistochemical expression of TTF-1, PAX5 and ISL-1 was evaluated. TTF-1, ISL-1 and PAX5 were expressed in 3/11 cases (27.27%, h-score: 3-45), 7/11 cases (63.64%, h-score: 23-200), and in 3/11 cases (27.77%, h-score 3-85), respectively. The patient with the strongest PAX5 reactivity exhibited an aggressive clinical course with rapid dissemination to the spine and death shortly after the diagnosis. No significant correlation in the expression of PAX5 and TTF-1 ( = 0.43; p = 0.18) was observed. ISL-1 is widely expressed in tumors with neuroendocrine differentiation and therefore of limited value in their differential diagnosis. TTF-1 positivity does not exclude the diagnosis of primary ONB, although usually only a small percentage of cells are positive. PAX5 expression is infrequent (27.27%) in ONB; however, if present it can be associated with a very aggressive clinical course.