Immunoreactive Score Research Articles

Mesonephric-like Adenocarcinoma of the Uterine Corpus: Comprehensive Immunohistochemical Analyses Using Markers for Mesonephric, Endometrioid and Serous Tumors.

Mesonephric-like adenocarcinoma (MLA) of the uterine corpus is a rare but distinct malignant tumor of the female genital tract, demonstrating a characteristic morphology and unique immunohistochemical profiles and molecular alterations. We conducted immunohistochemical staining (IHC) to make precise differential diagnoses of uterine MLAs from common histological subtypes of endometrial carcinomas. We collected 25 uterine MLAs and performed IHC for GATA3, TTF1, CD10, ER, PR, p16, p53, and HER2. Seventeen cases (68.0%) showed at least moderate nuclear GATA3 immunoreactivity in ≥25% of tumor cells. Most cases expressed TTF1 (17/21, 81.0%) and CD10 (luminal; 17/21, 81.0%). Heterogeneous TTF1 expression was noted in 12 cases. An inverse pattern of GATA3 and TTF1 staining was observed in eight cases (32.0%). Three cases (12.0%) showed moderate-to-strong ER expression in ≥25% of tumor cells, and two cases (8.0%) showed moderate-to-strong PR expression in ≥5% of tumor cells. These hormone receptor-positive MLAs varied in intensity and proportion of GATA3 staining. None of the 25 cases exhibited either diffuse and strong p16 expression or aberrant p53 expression. Five cases (20.0%) showed equivocal HER2 immunoreactivity (score 2+), but HER2 FISH confirmed that none of them exhibited HER2 gene amplification. In summary, a small subset of uterine MLAs displayed atypical IHC results: focal but strong expression of ER or PR, the complete absence of GATA3 immunoreactivity, the concurrent expression of mesonephric and hormone receptors, and the inverse pattern of GATA3 and TTF1 staining. These unusual immunophenotypes may complicate the differential diagnosis of MLA. Moreover, pathologists should be encouraged to interpret the IHC results cautiously.

Correlation between CD44+ cancer stem cell expressionand histopathologicaltypesof nasopharyngeal carcinoma.

Background:Nasopharyngeal carcinoma (NPC) recurrency rate is still high despite patients receiving complete treatment. The response to treatment may vary depending on the type of histopathology and Epstein-Barr virus,howeverthe mechanism remains unclear. Recent studies have found that there is a relationship between response to treatment and the presence of cancer stem cells (CSCs). CD44+ cancer stem cells may cause cancer cells to be resistant to treatment. Therefore, this cross-sectional study aims to determine the correlation between CD44 + cancer stem cell expression and the histopathological types of NPC. Method:Samples were obtained from NPC biopsies of type I, II, III patients (based on WHO histopathology criteria), who had not received prior treatment.CD44+ expression was examined using immunohistochemistry methods by staining CD44+ monoclonal antibodies. The degree of CD44+ cell membrane expression was based on the immunoreactive score scale or theRemmeleindex scale. Results: Most histopathological types wereWHOtype III (21 patients, 50%), followed by type II (18 patients, 42.86%), and type I (3 patients, 7.14%). CD44 + expression on type I showed one patient had moderate positive and two patients had a high-positive expression. In type II, 10 were moderatepositive and eight were high-positive. In type III, one patient was low-positive, 11 were moderate positive and nine patients were high-positive. Statistical analysis showed that the CD44+ expression difference between the three histopathology types were not statistically significant. Conclusion: There were no correlations between CD44 + expression and histopathological type of NPC.

Expressions of Nuclear Factor-kappa B and Peroxisome Proliferator-activated Receptor-Gamma Proportional with Clinical Staging of Nasopharyngeal Carcinoma

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a malignancy induced by the mutation of the transcription factors nuclear factor-kappa B (NF-kB) and peroxisome proliferator-activated receptor-gamma (PPAR-gamma). There was no known of the study about the association and targeted therapy of NF-kB and PPAR-gamma-induced NPC. AIM: This study analyzed and compared the proportion of NF-kB and PPAR-gamma and its association with the clinical characteristic of various NPC patients. METHODS: This was a cross-sectional study and conducted in Adam Malik General Hospital. The samples were paraffin block tissue obtained from 58 NPC patients and underwent immunohistochemistry staining for NF-kB or PPAR-gamma overexpression in March–November 2018. Determination of overexpression was based on the immunoreactive score. The association of NF-kB or PPAR-gamma overexpression with the clinical characteristics of the patients was analyzed using Fisher’s exact test. RESULTS: This study showed a significant increase of NF-kB and PPAR-gamma (p < 0.05). Male was found common than women (3.46:1) with non-keratinizing squamous cell carcinoma as the most common form of NPC (75.9%) and the 41–60 years old is the most common age (56.9%). Overexpression of NF-kB and PPAR-gamma was found mostly in T3-T4 (66.0%; 69.6%), N+ (92.5%; 91.3%), and clinical Stage IV (67.9%; 73.9%), respectively. CONCLUSION: The number of samples overexpressed was proportional to the clinical stage of NPC. This study provides an insight into the relationship of NF-kB and PPAR-gamma to NPC, suggesting their role in the development of malignancy.

Expression of Cytokeratin 6 and 16 in Middle Ear Cholesteatoma

BACKGROUND: Cholesteatoma is hyperproliferative because of the response of direct biomechanical trauma, and inflammation processes then lead to temporal bone destruction with some clinical manifestations of complications. The hyperproliferation mechanism occurred because of the activation of intermediate filament protein type I and type II known as cytokeratin (CK). AIM: This study aimed to examine the expression CK 6 and CK 16 in cholesteatoma. METHODS: This is a cross-sectional comparative study. Cholesteatoma specimens obtained from 15 patients who underwent surgery were considered as the case, and 15 normal retro-auricular skins were considered as the control. All samples were examined for expression through immunohistochemistry and scored using the immunoreactivity score. Data were analyzed using SPSS via χ2 test, and the difference was significant (p < 0.05). RESULTS: The expression of CK 6 was high in cholesteatoma (33.3%) and low in retro-auricular skin. The expression of CK 16 was high in all samples of cholesteatoma and mostly high in the retro-auricular skin; both expressions were statistically significant (p < 0.05). CONCLUSION: The expression of CK 6 and CK 16 in cholesteatoma was higher than in normal retro-auricular skin.

Immunohistochemical expression of chemokine receptor in neuroendocrine neoplasms (CXCR4) of the gastrointestinal tract: a retrospective study of 71 cases.

Aim: C-X-C motif chemokine receptor 4 (CXCR4) is expressed in many tumor entities, including gastrointestinal neuroendocrine neoplasms (GI-NENs). However, the role of CXCR4 expression in GI-NENs has been less studied. Our objective was to investigate the expression of CXCR4 in a series of GI-NENs with various clinical and pathological features. Methods: The immunohistochemical (IHC) expression of CXCR4 (clone UMB2) was examined in 71 GI-NENs and a semiquantitative immunoreactivity score (IRS) was calculated taking into consideration the intensity of the IHC reaction and the percentage of the tumor cells which showed positive expression. Results were compared with several clinical and pathological prognostic factors. Results: High CXCR4 expression was noted in 31 (43.7%) cases. Low IRS values were more frequent in NENs from the small intestine (66.7%) and stomach (60%). Also, all appendix tumors had IRS value of zero. High CXCR4 expression was noticed in 52.5% of liver metastases, compared to 40.4% primary tumors. A significant relationship was observed between the CXCR4 expression and the tumor grade (p=0.0216), and high IRS value was correlated with clinical stages III and IV (p=0.0142) and lympho-vascular invasion (p=0.0129). 74.1% of G1 neuroendocrine tumors (NETs) had a low IRS, G3 NETs showed minor differences between low (42.9%) and high (57.1%) expression and 66.7% of neuroendocrine carcinomas (NECs) presented high expression of CXCR4. Conclusions: The present study highlighted that high CXCR4 expression is associated with high grade and advanced stage GI-NENs, as well as with metastatic cases. In these cases, high CXCR4 expression could serve as an important target for CXCR4 antagonists.

Immunohistochemical evaluation of D2-40, Galectin-3, Maspin and MCM7 expression in palate squamous cell carcinomas.

Squamous cell carcinoma (SCC) is the most frequent cancer in oral cavity and its prognosis has exhibited little improvement in the last decades. Although much less common palate SCCs manifests a higher local aggression invading very quickly the adjacent muscles and jawbones, thus being able frequently to lead to dysfunctions in chewing, swallowing, and speech. To elucidate what underlies such local aggression, we investigated the immunohistochemical expression in palate SCCs of Podoplanin (D2-40), Galectin-3 (Gal-3), mammary serine protease inhibitor (Maspin) and minichromosome maintenance complex component 7 (MCM7), markers that are known to be involved in tumor invasiveness. We found a progressive increase in reactivity for D2-40 and MCM7 from the normal epithelium toward dysplastic epithelium and respectively to SCC, which suggests the intervention of these markers in the early stages of squamous cell carcinogenesis in the palate. The highest D2-40, Gal-3 and MCM7 reactivity was observed in basaloid and in poorly differentiated (G3) palate SCCs, while for Maspin the well-differentiated (G1) palate SCCs were the most reactive. The first three markers mentioned above were most intensely expressed at the invasion front, while the Maspin reactivity was low or absent at this level. Statistically, we found significant stratification on localization, grading, muscle invasion, and survival for all investigated markers, but with very high direct correlations between D2-40, Gal-3, and MCM7 immunoreactive score (IRS) values, while between the Maspin and each of the previous markers there were very high inverse correlations. Overall, all these investigate markers proved to be responsible for the local invasiveness and regional lymph node metastasis, thus allowing a prognostic and therapeutic stratification of patients with palate SCCs.

The predictive potential of Neuronatin for neoadjuvant chemotherapy of breast cancer.

Neuronatin (NNAT) determined by immunohistochemistry is a negative prognostic biomarker for breast cancer, independent of the major clinicopathological markers. Here, we investigated whether NNAT is also a predictive biomarker for pathological remission after neoadjuvant chemotherapy. One hundred and four breast cancer patients, treated with systemic neoadjuvant chemotherapy were included in this retrospective study. NNAT was detected in formaldehyde fixed, paraffin embedded primary cancer tissue by immunohistochemistry and an immuno-reactive score (IRS) determined. Pathological remission was scored according to Sinn and by evaluation of cytopathic effects. NNAT-IRS was correlated with clinicopathological parameters as well as relapse free and overall survival and for pathological remission after neoadjuvant therapy. NNAT IRS was an independent prognostic marker for relapse free and overall survival and the time from diagnosis to the "tumor-free" state. NNAT IRS was associated with Luminal-A tumors and correlated slightly negative with age and lymph-node metastasis. There was no significant correlation of NNAT-IRS with Sinn's remission score, but with cytopathic effects of chemotherapy. We confirmed the prognostic impact of NNAT-IRS in an independent cohort of neoadjuvantly treated patients. Additionally, a correlation with a score for pathological remission under systemic neoadjuvant chemotherapy for breast cancer was found.

β2-adrenergic receptor expression and the effects of norepinephrine and propranolol on various head and neck cancer subtypes.

The present study aimed to investigate expression of β2-adrenergic receptor (AR), the effect of the stress-related neurotransmitter norepinephrine (NE) on cell viability, proliferation and the therapeutic effect of propranolol, which is a typical β-blocker in various type of head and neck cancers for the first time. The β2-AR expression was investigated using immunohistochemistry and an immunoreactive scoring (IRS) system in 57 different head and neck cancer specimens, and reverse transcriptase-polymerase chain reaction and western blotting in four head and neck cancer cell lines (HNCCLs). Cell viability and proliferation assays were performed using 0, 1, 5 and 10 µM of NE and 1 µM of propranolol in four HNCCLs. The expression of β2-AR was positive in the majority of head and neck cancer tissues (55/57, 96.5%); however, it was significantly higher in oral cavity cancer than in pharyngeal cancer (median IRS: 9 vs. 3; P<0.001). All HNCCLs exhibited β2-AR expression, with a higher expression level detected in the oral cavity cancer cell line than in the others. NE stimulated viability (oral cavity, 206%; larynx, 156%; pharynx, 130%; nasal cavity, 137%; 10 µM NE) and proliferation (124, 176, 131 and 127%, respectively) in a dose-dependent manner in all HNCCLs. Conversely, propranolol attenuated such viability (55, 42, 18 and 22%, respectively) and proliferation (22, 40, 61 and 48%, respectively). In conclusion, the viability and proliferation of various head and neck cancers may be directly stimulated by stress and this may be attenuated by β-blockers.

The role of E-Cadherin expression in primary site of breast cancer

PurposeThe tumour’s ability to metastasize is the major cause for fatal outcomes in cancer diseases. In breast cancer, aberrant E-Cadherin expression has been linked to invasiveness and poor prognosis.MethodWe assessed expression of E-Cadherin by immunohistochemistry in primary tumour tissue from 125 female breast cancer patients. Staining intensities were analysed using the immunoreactive score (IRS). We investigated E-Cadherin expression and its associations with clinicopathological parameters (age, tumour size, lymph node status, grade, hormone receptors, Her2 Status) as well as with recurrence and survival.ResultsIncreased, rather than aberrant E-Cadherin expression was found and was associated with poor outcome (p = 0.046). Our data show an association between elevated E-Cadherin in primary tumour tissue and an unfavourable negative prognosis in patients.ConclusionThis association was somehow unexpected as loss of E-Cadherin has long been regarded as a prerequisite for development of invasiveness and metastases. Our findings support the notion that E-Cadherin promotes, rather than suppresses, development of metastasis and invasiveness.

Angiogenesis and proliferation of endothelial cells in hypertrophic and nodular port-wine stain

BackgroundPort-wine stain (PWS) has been classified not as the hyperplasia of cells, but rather, as an expansion of malformed vessels. However, previous studies have reported upregulated expression of proangiogenic factors in PWS. Several studies have indicated that the pathology exhibits proliferation of numerous endothelial cells in hypertrophic/nodular PWS. This study aimed to determine the expression of vascular epithelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9), angiopoietin-2 (ANG-2), and basic fibroblast growth factor (bFGF) in hypertrophic PWS. MethodsImmunohistochemistry was used to analyze skin samples from 33 patients with hypertrophic PWS. Expression levels of VEGF, MMP-9, ANG-2, and bFGF in hypertrophic PWS were determined by multiplying the intensity by the percentage of immunoreactive cells. Immunoreactivity scores were classified as follows: negative (0), low (1), moderate (2, 3, and 4), or high (6). ResultsBased on pathological characteristics, hypertrophic PWS was divided into vascular malformation and pyogenic granuloma (PG) types. VEGF, MMP-9, ANG-2, and bFGF were significantly activated in the blood vessels of PG-type PWS samples compared with their counterparts in blood vessels of vascular malformation-type PWS samples and controls. PG-type hypertrophic PWS, which exhibited proliferation of endothelial cells, showed the strongest activation. ConclusionThe exuberant proliferation of endothelial cells in PG-type hypertrophic PWS may be associated with the regulation of proangiogenic factors during development. These proangiogenic factors that function in the angiogenesis and proliferation of endothelial cells may play an important role in the pathogenesis and progression of PWS. Furthermore, these factors may be dynamic and behave differently in various types of hypertrophic PWS.

Relationship Between O-GlcNAcase Expression and Prognosis of Patients With Osteosarcoma.

Several studies have demonstrated a role of O-GlcNAcylation (O-GlcNAc) in tumorigenesis of various carcinomas by modification of tumor-associated proteins. However, its implication in the pathogenesis of osteosarcoma remains unclear. This study aimed to investigate the levels of O-GlcNAc and the expressions of O-linked N-acetylglucosamine transferase (OGT) and O-GlcNAcase (OGA) in human osteosarcoma tissues, by using immunohistochemistry; and to find correlations between the levels or expressions and several clinicopathologic parameters. There were 109 first diagnosed osteosarcoma patients, including Enneking stage IIB (n=70) and III (n=39). Correlations between the immunoreactive score (IRS) and clinicopathologic parameters, overall survival, and metastasis-free survival were evaluated. A positive correlation was found between the IRS of OGA and the percentage of postchemotherapeutic tumor necrosis (r=0.308; P=0.017). Univariate analysis revealed significantly lower OGA IRS in metastatic patients (P=0.020) and poor chemotherapeutic-responder patients (P=0.001). By multivariate analysis, presence of tumor metastasis (P=0.002) and lower OGA IRS (P=0.004) was significantly associated with shorter overall survival. Subgroup analysis in stage IIB osteosarcoma (n=70) demonstrated that male sex (P=0.019), presence of tumor recurrence (P=0.026), poor chemotherapeutic responder (P=0.022), and lower OGA IRS (P=0.019) were significantly correlated with short metastasis-free survival. But, lower OGA IRS was the only independent predictor for short metastasis-free survival (P=0.006). Our findings suggested that O-GlcNAc pathway, especially OGA, may involve in pathogenesis and aggressiveness of osteosarcoma. Low level of OGA expression may be used as a poor prognostic indicator.

CT-076: Evaluating CD83 Expression on Organ-Specific Normal Tissues to Anticipate Potential Toxicity of Novel Human CD83-Targeted Chimeric Antigen Receptor (CAR) T-Cell

Context and Objective CD83 is expressed on activated CD4+ T-cells, dendritic cells, and acute myeloid leukemia (AML), but its expression level on organ-specific normal tissue is not well investigated. A novel human CD83-targeted CAR T-cell has been developed and shown in preclinical work to treat and prevent graft-versus-host disease (GVHD) and confer graft-versus-leukemia (GVL) activity against AML. The purpose of this study is to evaluate CD83 expression in a panel of normal tissue samples to better characterize CD83 expression. Design and Patients An IRB-approved pilot study utilized specimens collected and maintained by Moffitt after informed consent. Thirty samples of representative normal tissues were utilized, with 3 cores taken from each sample, to construct a tissue microarray (TMA). A second TMA was obtained through the Cooperative Human Tissue Network. CD83 expression was assessed by immunohistochemistry (IHC) after antibody validation. IHC expression was scored by hematopathologists, and immunoreactive scores (IRS) were calculated (product of staining intensity and nominal variable denoting expression percentage). IRS scores of 0–1, 2–3, 4–8, and 9–12 correspond to negative, mild, moderate, and strong expression, respectively. Results TMA constructs included 9 normal tissue types with 6–9 cores each and 51 tissue types with 3 cores each in two separate blocks. Strong expression was seen in plasma cells in lymph nodes and tonsils, pancreas, and parathyroid tissue. Moderate expression was seen in lymph nodes, spleen, tonsil, thymus, lung, mesothelium, kidney, liver, thyroid, bladder, cartilage, cerebral cortex, small bowel, stomach, placenta/amniotic membrane, and endocervix. Mild expression was seen in bone marrow, breast, skin, parotid gland, epididymis/seminal vesicle, esophagus, large bowel, anus, uterus, ovary/fallopian tube, synovium, stomach, gallbladder, aorta, and breast. There was no expression in myocardium, adrenal gland, brain white matter, peripheral nerves, smooth muscle, prostate, or ovarian primary oocyte. Conclusions The present study better characterizes CD83 expression on organ-specific normal tissue. Moderate expression was seen in lymphoid tissue. Further work is needed to determine the impact of CD83-targeted therapy on normal tissue expressing CD83. This data will be used in the development of future clinical trials of novel human CD83-targeted therapies with promising preclinical data.

TRIM47 activates NF-κB signaling via PKC-ε/PKD3 stabilization and contributes to endocrine therapy resistance in breast cancer

Increasing attention has been paid to roles of tripartite motif-containing (TRIM) family proteins in cancer biology, often functioning as E3 ubiquitin ligases. In the present study, we focus on a contribution of TRIM47 to breast cancer biology, particularly to endocrine therapy resistance, which is a major clinical problem in breast cancer treatment. We performed immunohistochemical analysis of TRIM47 protein expression in 116 clinical samples of breast cancer patients with postoperative endocrine therapy using tamoxifen. Our clinicopathological study showed that higher immunoreactivity scores of TRIM47 were significantly associated with higher relapse rate of breast cancer patients (P = 0.012). As functional analyses, we manipulated TRIM47 expression in estrogen receptor-positive breast cancer cells MCF-7 and its 4-hydroxytamoxifen (OHT)-resistant derivative OHTR, which was established in a long-term culture with OHT. TRIM47 promoted both MCF-7 and OHTR cell proliferation. MCF-7 cells acquired tamoxifen resistance by overexpressing exogenous TRIM47. We found that TRIM47 enhances nuclear factor kappa-B (NF-κB) signaling, which further up-regulates TRIM47. We showed that protein kinase C epsilon (PKC-ε) and protein kinase D3 (PKD3), known as NF-κB-activating protein kinases, are directly associated with TRIM47 and stabilized in the presence of TRIM47. As an underlying mechanism, we showed TRIM47-dependent lysine 27-linked polyubiquitination of PKC-ε. These results indicate that TRIM47 facilitates breast cancer proliferation and endocrine therapy resistance by forming a ternary complex with PKC-ε and PKD3. TRIM47 and its associated kinases can be a potential diagnostic and therapeutic target for breast cancer refractory to endocrine therapy.

Early Failure of a Polyvinyl Alcohol Hydrogel Implant With Osteolysis and Foreign Body Reactions in an Ovine Model of Cartilage Repair

Background: Hemiarthroplasty using a polyvinyl alcohol (PVA) hydrogel synthetic implant has been suggested as a good alternative to arthrodesis for the treatment of hallux rigidus. However, failure rates as high as 20% have been recorded. Purpose: To characterize the pathological processes in bone, cartilage, and the synovial membrane after PVA hemiarthroplasty in an ovine model with 6 months of follow-up. Study Design: Controlled laboratory study. Methods: A unilateral osteochondral defect (8-mm diameter × 10-mm depth) was made in the medial femoral condyle in 6 sheep. Animals were randomized to receive a PVA implant (n = 4) or to have an empty defect (n = 2) and were monitored for 6 months. Patellofemoral radiographs were obtained at monthly intervals, and quantitative computed tomography was performed at the end of the study. After death, the joints were macroscopically evaluated and scored. Osteochondral and synovial membrane histological findings were assessed using modified Osteoarthritis Research Society International (OARSI) and aseptic lymphocyte-dominated vasculitis-associated lesion (ALVAL) scoring systems. Immunohistochemistry using Iba1 was performed to evaluate activated macrophage infiltration. Results: Overall, 2 sheep with PVA implants were euthanized at 1 and 5 months because of uncontrollable pain and lameness (failed implants). Quantitative computed tomography showed that sheep with failed implants had 2.1-fold more osteolysis than those with successful implants. The sheep with failed implants had osteoarthritis with extensive glycosaminoglycan loss and cartilage fibrillation of the condyle and opposing tibial surface on histological examination. A foreign body reaction with severe chronic lymphoplasmacytic and granulomatous inflammation with giant cells was detected surrounding the implant. The synovial membrane ALVAL score was 9 of 19 and 14 of 19 in failed implants with synovial hyperplasia and lymphoplasmacytic and macrophage infiltration. In contrast, the synovial membrane in successful implants and empty defects was normal (ALVAL score = 0/19). Immunolabeling for Iba1 in failed implants confirmed extensive and dense macrophage infiltration within the condyle and synovial membrane, with the highest immunoreactive score (9/9). Conclusion: PVA hydrogel implants had a 50% failure rate with uncontrollable pain, severe osteolysis, inflammation, and foreign body reactions. Clinical Relevance: The failure rate and pathological characteristics of the PVA implants suggest that their use should not be continued in human patients without further in vivo safety studies.

Evaluation of 18F-EF5 for detection of hypoxia in localized adenocarcinoma of the prostate

Background A common feature of solid tumours that are resistant to therapy is the presence of regions with low oxygen content (i.e., hypoxia). Oxygen electrode studies suggest that localized prostate adenocarcinoma is commonly hypoxic, although conflicting data have been reported between immunohistochemical detection of hypoxia-induced proteins in biopsy specimens and positron emission tomography (PET) imaging of 18F-labeled hypoxia reporters. Although the 2-nitroimidazole 18F-EF5 is well-established to label hypoxic tumour cells in pre-clinical tumour models and clinical trials of multiple primary tumour sites, it has yet to be tested in prostate cancer. The purpose of this study was to evaluate the feasibility of using 18F-EF5 to detect hypoxia in clinical prostate tumours. Material and methods Patients with localized adenocarcinoma of the prostate were recruited for pre-treatment 18F-EF5 PET scans. Immunohistochemistry was conducted on diagnostic biopsies to assess the expression of glucose transporter 1 (GLUT1), osteopontin (OPN), and carbonic anhydrase IX (CAIX). Immunoreactivity scores of staining intensity and frequency were used to indicate the presence of tumour hypoxia. Results We found low tumour-to-muscle ratios of 18F-EF5 uptake that were not consistent with tumour hypoxia, causing early termination of the study. However, we observed GLUT1 and OPN expression in all prostate tumour biopsies, indicating the presence of hypoxia in all tumours. Conclusion Our data do not support the use of 18F-EF5 PET to detect hypoxia in prostate adenocarcinoma, and suggest the use of immunohistochemistry to quantify expression of the hypoxia-inducible proteins GLUT1 and OPN as indications of prostate tumour hypoxia.

Trace Amine-Associated Receptor 1 (TAAR1) Is a Positive Prognosticator for Epithelial Ovarian Cancer.

Trace amine-associated receptor 1 (TAAR1) is a Gαs- protein coupled receptor that plays an important role in the regulation of the immune system and neurotransmission in the CNS. In ovarian cancer cell lines, stimulation of TAAR1 via 3-iodothyronamine (T1AM) reduces cell viability and induces cell death and DNA damage. Aim of this study was to evaluate the prognostic value of TAAR1 on overall survival of ovarian carcinoma patients and the correlation of TAAR1 expression with clinical parameters. Ovarian cancer tissue of n = 156 patients who were diagnosed with epithelial ovarian cancer (serous, n = 110 (high-grade, n = 80; low-grade, n = 24; unknown, n = 6); clear cell, n = 12; endometrioid, n = 21; mucinous, n = 13), and who underwent surgery at the Department of Obstetrics and Gynecology, University Hospital of the Ludwig-Maximilians University Munich, Germany between 1990 and 2002, were analyzed. The tissue was stained immunohistochemically with anti-TAAR1 and evaluated with the semiquantitative immunoreactive score (IRS). TAAR1 expression was correlated with grading, FIGO and TNM-classification, and analyzed via the Spearman’s rank correlation coefficient. Further statistical analysis was obtained using nonparametric Kruskal-Wallis rank-sum test and Mann-Whitney-U-test. This study shows that high TAAR1 expression is a positive prognosticator for overall survival in ovarian cancer patients and is significantly enhanced in low-grade serous carcinomas compared to high-grade serous carcinomas. The influence of TAAR1 as a positive prognosticator on overall survival indicates a potential prognostic relevance of signal transduction of thyroid hormone derivatives in epithelial ovarian cancer. Further studies are required to evaluate TAAR1 and its role in the development of ovarian cancer.

Correlation between CD44+ cancer stem cell expression and histopathological types of nasopharyngeal carcinoma

Background: Nasopharyngeal carcinoma (NPC) recurrency rate is still high despite patients receiving complete treatment. The response to treatment may vary depending on the type of histopathology and Epstein-Barr virus, however the mechanism remains unclear. Recent studies have found that there is a relationship between response to treatment and the presence of cancer stem cells (CSCs). CD44+ cancer stem cells may cause cancer cells to be resistant to treatment. Therefore, this cross-sectional study aims to determine the correlation between CD44 + cancer stem cell expression and the histopathological types of NPC. Method: Samples were obtained from NPC biopsies of type I, II, III patients (based on WHO histopathology criteria), who had not received prior treatment. CD44+ expression was examined using immunohistochemistry methods by staining CD44+ monoclonal antibodies. The degree of CD44+ cell membrane expression was based on the immunoreactive score scale or the Remmele index scale. Results: Most histopathological types were WHO type III (21 patients, 50%), followed by type II (18 patients, 42.86%), and type I (3 patients, 7.14%). CD44 + expression on type I showed one patient had moderate positive and two patients had a high-positive expression. In type II, 10 were moderate positive and eight were high-positive. In type III, one patient was low-positive, 11 were moderate positive and nine patients were high-positive. Statistical analysis showed that the CD44+ expression difference between the three histopathology types were not statistically significant. Conclusion: There were no correlations between CD44 + expression and histopathological type of NPC.

PSMA Expression in Differentiated Thyroid Cancer: Association With Radioiodine, 18FDG Uptake, and Patient Outcome.

Little is known about prostate-specific membrane antigen (PSMA) expression in patients with cervical involvement of differentiated thyroid cancer (DTC). We investigated PSMA expression in neck persistent/recurrent disease (PRD) using immunohistochemistry and the association with radioiodine (RAI) or 18-fluorodeoxyglucose (18FDG) uptake, and patient outcome. Data from 44 consecutive DTC patients who underwent neck reoperation from 2006 to 2018 in a comprehensive cancer center. Immunostaining was performed with vascular endothelial marker CD31 and PSMA. PSMA expression was quantified using the immunoreactive score (IRS). RAI and 18FDG uptake were assessed before surgery using posttherapeutic RAI scintigraphy and 18FDG positron emission tomography with computed tomography. Mean follow-up after reintervention was 6.5 ± 3.7 years. Thirty patients (68%) showed at least 1 PSMA-positive lesion (IRS ≥ 2) with similar proportions in RAI-positive and RAI-negative patients (75% vs 66%). In RAI-negative patients, however, the proportion of PSMA-positive disease (79% vs 25%, P < 0.01) and the mean IRS (4.0 vs 1.0, P = 0.01) were higher in 18FDG-positive than in 18FDG-negative patients. Furthermore, mean IRS was higher in patients ≥ 55 years, large primary tumors (>40 mm) or aggressive subtypes, and was correlated with structural disease at last follow-up. Strong PSMA expression (IRS ≥ 9) was associated with shorter progression-free survival (PFS). Our findings show that PSMA expression was present in two-thirds of patients with neck PRD, that it was related to poor prognostic factors and that very high expression was associated with poorer PFS. This preliminary study may offer new perspectives for the management of RAI-refractory DTC.

Digital Pathology Scoring of Immunohistochemical Staining Reliably Identifies Prognostic Markers and Anatomical Associations in a Large Cohort of Oral Cancers.

Utilizing digital pathology algorithms for the objective quantification of immunohistochemical staining, this study aimed to identify robust prognostic biomarkers for oral cancer. Tissue microarrays with specimens of a large cohort of oral squamous cell carcinoma (n=222) were immunohistochemically stained to determine the expression of PD-L1, EGFR, and COX-2 and the amount of infiltrating NK cells and CD8-positive T cells. Immunoreactivity scores were assessed using both a classical manual scoring procedure and a digital semi-automatic approach using QuPath. Digital scoring was successful in quantifying the expression levels of different prognostic biomarkers (CD8: p<0.001; NK cells: p=0.002, PD-L1: p=0.026) and high levels of concordance with manual scoring results were observed. A combined score integrating EGFR expression, neck node status and immune cell signatures with a significant impact on overall and progression-free survival was identified (p<0.001). These data may contribute to the ongoing research on the identification of reliable and clinically relevant biomarkers for the individualization of primary and adjuvant treatment in oral cancer.

Correlation between CD44+ cancer stem cell expressionand histopathologicaltypesof nasopharyngeal carcinoma.

Background:Nasopharyngeal carcinoma (NPC) recurrency rate is still high despite patients receiving complete treatment. The response to treatment may vary depending on the type of histopathology and Epstein-Barr virus,howeverthe mechanism remains unclear. Recent studies have found that there is a relationship between response to treatment and the presence of cancer stem cells (CSCs). CD44+ cancer stem cells may cause cancer cells to be resistant to treatment. Therefore, this cross-sectional study aims to determine the correlation between CD44 + cancer stem cell expression and the histopathological types of NPC. Method:Samples were obtained from NPC biopsies of type I, II, III patients (based on WHO histopathology criteria), who had not received prior treatment.CD44+ expression was examined using immunohistochemistry methods by staining CD44+ monoclonal antibodies. The degree of CD44+ cell membrane expression was based on the immunoreactive score scale or theRemmeleindex scale. Results: Most histopathological types wereWHOtype III (21 patients, 50%), followed by type II (18 patients, 42.86%), and type I (3 patients, 7.14%). CD44 + expression on type I showed one patient had moderate positive and two patients had a high-positive expression. In type II, 10 were moderatepositive and eight were high-positive. In type III, one patient was low-positive, 11 were moderate positive and nine patients were high-positive. Statistical analysis showed that the CD44+ expression difference between the three histopathology types were not statistically significant. Conclusion: There were no correlations between CD44 + expression and histopathological type of NPC.