Expression In Liver Metastases Research Articles

Abstract C001: Retrospective analysis of TROP2 expression in colorectal cancer (CRC) primary tumors and liver metastases (LM) and its correlation with clinical factors

Abstract Background: Trophoblast cell surface antigen 2 (TROP2) is a transmembrane glycoprotein which is upregulated in CRC, is associated with tumorigenesis, and may serve as a drug target. We aimed to measure TROP2 expression by immunohistochemistry in CRC LM and primary tumors, and to determine how TROP2 expression correlates with prognosis and baseline clinical factors. Methods: TROP2 immunohistochemistry was retrospectively performed by a pathologist (ab214488; Abcam, Cambridge, UK) in paraffin-embedded CRC LM and primary tumor specimens (biopsy or resection, stored at the University of Colorado Surgical Pathology Archive) from adult patients between January 2006 and June 2022. Baseline clinical and survival data was collected. Differences in TROP2 frequency (range: 0-4), intensity (range: 0-3), and frequency x intensity combined scores (range: 0-12) for paired CRC primary and LM specimens were analyzed using a paired t-test. Additional statistical analyses included nonparametric Spearman’s correlation test (correlation of scores between LM and primary tumor), Mann Whitney U test (effect of prior systemic cancer treatment on TROP2 expression), Kaplan-Meier method (survival rates based on TROP2 expression), and log-rank test (differences in survival). Results: TROP2 was expressed in 100% (60/60) of CRC primary and 93% (52/56) of LM specimens, including 50 CRC primary-LM pairs and additional unpaired specimens (10 primary and 6 LM). Among all specimens, primary and LM specimens both had a median intensity score of 2, whereas CRC specimens had a slightly higher frequency score than LM specimens (2 vs. 1, respectively). In paired samples, the TROP2 frequency, intensity, and combined score distribution varied slightly between primary and LM specimens. The median intensity was higher in primary vs LM specimens (3 vs 2, respectively), mean difference was 0.26 (p=0.08), and correlation was weak (Spearman r=0.29, p=0.04). Similarly, the median frequency was higher in primary vs LM specimens (2 vs 1, respectively), mean difference was 0.20 (p=0.21), with moderate correlation (Spearman r=0.55, p<0.0001). The median combined score was higher in primary vs LM specimens (4 vs 3, respectively), mean difference was 0.98 (p=0.04), with moderate correlation (Spearman r=0.51, p=0.0001). Prior systemic cancer therapy did not significantly affect TROP2 intensity, frequency, or combined score in primary or LM specimens (data to be presented). Overall survival was not significantly different between low (<5) or high (≥5) TROP2 combined score (stratified by the median value) for primary (log-rank p=0.67) or LM specimens (log-rank p=0.66). Similarly, there was no difference in overall survival stratified by TROP2 frequency or intensity in CRC primary or LM (data to be presented). Conclusions: TROP2 expression was observed in both primary CRC and LM specimens, and the frequency x intensity combined score was significantly higher in primary CRC specimens compared to paired LM. Higher TROP2 expression in either primary tumors or LM did not correlate with overall survival. Citation Format: Robert W Lentz, Miriam Barnett, Michele Gerber, Jeffrey Kaplan, Adrie van Bokhoven, Kathleen C Torkko. Retrospective analysis of TROP2 expression in colorectal cancer (CRC) primary tumors and liver metastases (LM) and its correlation with clinical factors [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr C001.

Protective effect of miR-18a in resected liver metastases of colorectal cancer and FOLFOX treatment.

Colorectal cancer ranks second in terms of cancer associated deaths worldwide, whereas miRNA play a pivotal role in the etiology of cancer and its metastases. Studying the expression and cellular function of miR-18a in metastatic colorectal cancer and association to progression-free survival. Colorectal liver metastases (N = 123) and primary colorectal cancer (N = 27) where analyzed by RT-PCR and correlated with clinical follow up data. Invasion and migration assays were performed with the liver metastatic cell line LIM2099 after miR-18a knockdown. Cell viability under FOLFOX treatment and knockdown was measured. We found that the expression of miR-18a was increased 4.38-fold in liver metastases and 3.86-fold in colorectal tumor tissue compared to healthy liver tissue and colorectal mucosa, respectively (p ≤ .001). Patients with a high miR-18a expression in liver metastases had a progression-free survival (PFS) of 13.6 months versus 8.9 months in patients with low expression (N = 123; p = .024). In vitro migration of LIM2099 cells was reduced after miR-18a knockdown and cell viability was significantly increased after miR-18a knockdown and treatment with folinic acid or oxaliplatin. Subgroup analysis of PFS revealed significant benefits for patients with high miR-18a expression receiving 5-FU, folinic acid or oxaliplatin. High expression of miR-18a in colorectal liver metastases might have a protective effect after resection of metastases and FOLFOX treatment regarding PFS.

Relationship between SUVmax on 18F-FDG PET and PD-L1 expression in liver metastasis lesions after colon radical operation

PurposeOur study was to investigate the correlation correlation between FDG uptake and PD-L1 expression of liver metastasis in patients with colon cancer, and to determine the value of FDG-PET in predicting PD-L1 expression in liver metastasis of colon cancer.MethodsA total of 72 patients with confirmed liver metastasis of colon cancer were included in this retrospective study. The PD-L1 expression and immune cell infiltrating of tumors were determined through immunohistochemistry staining. The SUVmax of liver metastasis lesions were assessed using 18 F-FDG PET/CT. The correlation between PD-L1 expression and the clinicopathological were evaluated by the Cox proportional hazards model and the Kaplan-Meier survival analysis.ResultsPD-L1 expression was significantly correlated with FDG uptake (SUVmax), tumor size, differentiation, survival and cytotoxic T cells infiltration in liver metastasis of colon cancer (P < 0.05). And liver metastases with high counts of infiltrating cytotoxic T cells showed greater FDG uptake than those with low counts of infiltrating cytotoxic T cells. The SUVmax of liver metastases and the degree of differentiation of metastases were closely related to PD-L1 expression, and were independent risk factors.The combined assessment of SUVmax values and tthe degree of differentiation of metastase can help determine PD-L1 expression in liver metastasis of colon cancer.ConclusionsFDG uptake in liver metastasis of colon cancer was positively correlated with the PD-L1 expression and the number of cytotoxic T cells infiltration. The joint evaluation of two parameters, SUVmax and degree of differentiation, can predict PD-L1 expression in liver metastases.

Deep learning for predicting the human epidermal growth factor receptor 2 status of breast cancer liver metastases based on contrast-enhanced computed tomography: a development and validation study.

This study investigated the value of a deep learning (DL) model based on computed tomography (CT) enhancement for predicting human epidermal growth factor receptor 2 (HER2) expression in patients with liver metastasis from breast cancer. Data were collected for 151 female patients with liver metastasis from breast cancer who underwent abdominal enhanced CT examination in the Department of Radiology at the Affiliated Hospital of Hebei University between January 2017 and March 2022. Liver metastases were confirmed in all patients by pathology. The HER2 status of the liver metastases was assessed and enhanced CT examinations were performed before treatment. Of the 151 patients, 93 were HER2 negative and 58 were HER2 positive. Liver metastases were manually labeled with rectangular frames, layer by layer, and the labeled data were processed. Five basic networks (ResNet34, ResNet50, ResNet101, ResNeXt50, and Swim Transformer) were used for training and optimization, and the model's performance was tested. Receiver operating characteristic (ROC) curves were used to analyze the area under the curve (AUC), as well as the accuracy, sensitivity, and specificity of the networks in predicting HER2 expression in breast cancer liver metastases. Overall, ResNet34 demonstrated the best prediction efficiency. The accuracy of the validation and test set models in predicting HER2 expression in liver metastases was 87.4% and 80.5%, respectively. The AUC, sensitivity, and specificity of the test set model in predicting HER2 expression in liver metastases were 0.778, 77.0%, and 84.0%, respectively. Our DL model based on CT enhancement has good stability and diagnostic efficacy, and is a potential non-invasive method for identifying HER2 expression in liver metastases from breast cancer.

Unveiling the tumor immune microenvironment of organ-specific melanoma metastatic sites

BackgroundThe liver is a known site of resistance to immunotherapy and the presence of liver metastases is associated with shorter progression-free and overall survival (OS) in melanoma, while lung metastases...

Non-invasive detection and complementary diagnosis of liver metastases via chemokine receptor 4 imaging

Noninvasive detection of early-stage liver metastases from different primary cancers is a pressing unmet medical need. The lack of both molecular biomarkers and the sensitive imaging methodology makes the detection challenging. In this study, we observed the elevated expression of chemokine receptor 4 (CXCR4) in uveal melanoma (UM) patient liver tissues, and high CXCR4 expression in liver metastases of UM murine models, regardless of the expression levels in the primary tumors. Based on these findings, we identified CXCR4 as an imaging biomarker and exploited a CXCR4-targeted MRI contrast agent ProCA32.CXCR4 for molecular MRI imaging. ProCA32.CXCR4 has strong CXCR4 binding affinity, high metal selectivity, and r1 and r2 relaxivities, which enables the sensitive detection of liver micrometastases. The MRI imaging capacity for detecting liver metastases was demonstrated in three UM models and one ovarian cancer model. The imaging results were validated by histological and immunohistochemical analysis. ProCA32.CXCR4 has strong potential clinical application for non-invasive diagnosis of liver metastases.

Prognosis-related Factors of Colorectal Cancer Patients with Positive PD-L1 Expression in Liver Metastases After Hepatectomy

Prognosis-related Factors of Colorectal Cancer Patients with Positive PD-L1 Expression in Liver Metastases After Hepatectomy

Expression and prognostic value of epithelial-to-mesenchymal transition and cancer stem cellmarkersin primary lesions and liver metastases of colorectal cancers.

Cancer stem cells (CSCs) and epithelial mesenchymal transition (EMT) markers are considered useful indicators associated with metastasis and prognosis of colorectal cancers (CRCs). However, only a few studies have focused on the expression of these useful markers in metastases. Metastasectomy is widely used in advanced CRCs, and thus the postoperative prognostic factors are worth investigating. The present study investigated the consistency and differences of target proteins between primary and metastatic lesions of colorectal cancer, and discussed the prognostic indicators following resection of colorectal liver metastases. Clinical data of 56 patients with liver metastases from colorectal cancer were collected and the expression levels of target proteins (Ki-67, CD133, CD44, Snail, E-cadherin and β-catenin) were detected in primary tumor and matched liver metastases via immunohistochemistry analysis. Paired comparison between both tissue types was performed. The prognostic values of the target proteins for resectable colorectal cancer liver metastases were assessed. No significant differences were observed between the primary tissues and metastatic tissues. The consistency rates of these protein expression levels ranged from 51.8–78.6%. The maximum diameter of the liver metastases was <5 cm. Low Snail expression in metastases was associated with a longer overall survival (OS) time following resection of colorectal liver metastases. Furthermore, N0 stage and low carcinoembryonic antigen levels were associated with a longer progression-free survival time. Notably, no significant differences were observed in expression levels of the target proteins between the primary tumors and liver metastases. Taken together, the results of the present study suggest that Snail expression in liver metastases may be used as a novel independent prognostic factor for OS following resection of colorectal liver metastases.

PD-L1 expression in liver metastasis: its clinical significance and discordance with primary tumor in colorectal cancer

BackgroundThe outcomes of immune checkpoint inhibitors in cancer patients with liver metastases are poor, which may be related to a different tumor microenvironment in liver metastases from primary tumors. This study was aimed to analyze PD-L1 expression and the immune microenvironment status in liver metastases and compare the differences of PD-L1 expression between primary tumors and liver metastases of colorectal cancer.Methods74 cases of pathologically confirmed colorectal cancer with liver metastasis underwent resection from our hospital were included. Tissue microarrays were used for the interpretation of PD-L1 expression, cluster of differentiation 4 (CD4) and CD8 density by immunohistochemistry. We evaluated the disparity between primary tumor and liver metastasis in PD-L1 expression, CD4 and CD8 density and analyzed the factors associated with obvious PD-L1 disparity.ResultsThe expression of PD-L1 was positively related to the density of CD4 and CD8 in liver metastases. The expression of PD-L1 in liver metastases was higher than in primary tumors in certain subgroups, including patients with concurrent liver metastases (n = 63, p = 0.05), patients receiving concurrent resection of primary and metastatic tumors (n = 56, p = 0.04). The two subgroups generally reflected those without inconsistent external influences, such as treatment and temporal factors, between primary tumors and liver metastases. In these subgroups, the intrinsic differences of microenvironment between primary tumors and liver metastases could be identified. Furthermore, tumor differentiation [moderate vs. poor: OR = 0.23, 95% CI: 0.03–0.99, p = 0.05)] were demonstrated to be associated with obvious discordance of PD-L1 expression between primary tumors and liver metastases.ConclusionsThe expression of PD-L1 in liver metastases was higher than in primary tumors in subgroups, reflecting intrinsic microenvironment differences between primary and metastatic tumors. Obvious discordance of PD-L1 expression between primary tumor and liver metastasis was significantly related to the tumor differentiation.

Targeting BMI-1-mediated epithelial–mesenchymal transition to inhibit colorectal cancer liver metastasis

Liver is the most common metastatic site for colorectal cancer (CRC), there is no satisfied approach to treat CRC liver metastasis (CRCLM). Here, we investigated the role of a polycomb protein BMI-1 in CRCLM. Immunohistochemical analysis showed that BMI-1 expression in liver metastases was upregulated and associated with T4 stage, invasion depth and right-sided primary tumor. Knockdown BMI-1 in high metastatic HCT116 and LOVO cells repressed the migratory/invasive phenotype and reversed epithelial–mesenchymal transition (EMT), while BMI-1 overexpression in low metastatic Ls174T and DLD1 cells enhanced invasiveness and EMT. The effects of BMI-1 in CRC cells were related to upregulating snail via AKT/GSK-3β pathway. Furthermore, knockdown BMI-1 in HCT116 and LOVO cells reduced CRCLM using experimental liver metastasis mice model. Meanwhile, BMI-1 overexpression in Ls174T and DLD1 significantly increased CRCLM. Moreover, sodium butyrate, a histone deacetylase and BMI-1 inhibitor, reduced HCT116 and LOVO liver metastasis in immunodeficient mice. Our results suggest that BMI-1 is a major regulator of CRCLM and provide a potent molecular target for CRCLM treatment.

Metabolic programming of distinct cancer stem cells promotes metastasis of pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) metastasizes to distant organs, which is the primary cause of mortality; however, specific features mediating organ-specific metastasis remain unexplored. Emerging evidence demonstrates that cancer stem cells (CSCs) and cellular metabolism play a pivotal role in metastasis. Here we investigated the role of distinct subtypes of pancreatic CSCs and their metabolomic signatures in organ-specific metastatic colonization. We found that PDAC consists of ALDH+/CD133+ and drug-resistant (MDR1+) subtypes of CSCs with specific metabolic and stemness signatures. Human PDAC tissues with gemcitabine treatment, autochthonous mouse tumors from KrasG12D; Pdx1-Cre (KC) and KrasG12D; Trp53R172H; Pdx-1 Cre (KPC) mice, and KPC- Liver/Lung metastatic cells were used to evaluate the CSC, EMT (epithelial-to-mesenchymal transition), and metabolic profiles. A strong association was observed between distinct CSC subtypes and organ-specific colonization. The liver metastasis showed drug-resistant CSC- and EMT-like phenotype with aerobic glycolysis and fatty acid β-oxidation-mediated oxidative (glyco-oxidative) metabolism. On the contrary, lung metastasis displayed ALDH+/CD133+ and MET-like phenotype with oxidative metabolism. These results were obtained by evaluating FACS-based side population (SP), autofluorescence (AF+) and Alde-red assays for CSCs, and Seahorse-based oxygen consumption rate (OCR), extracellular acidification rate (ECAR), and fatty acid β-oxidation (FAO)-mediated OCR assays for metabolic features along with specific gene signatures. Further, we developed in vitro human liver and lung PDAC metastasis models by using a combination of liver or lung decellularized scaffolds, a co-culture, and a sphere culture methods. PDAC cells grown in the liver-mimicking model showed the enrichment of MDR1+ and CPT1A+ populations, whereas the PDAC cells grown in the lung-mimicking environment showed the enrichment of ALDH+/CD133+ populations. In addition, we observed significantly elevated expression of ALDH1 in lung metastasis and MDR1/LDH-A expression in liver metastasis compared to human primary PDAC tumors. Our studies elucidate that distinct CSCs adapt unique metabolic signatures for organotropic metastasis, which will pave the way for the development of targeted therapy for PDAC metastasis.

Prognostic value of tumor infiltrating lymphocytes combined with PD-L1 expression for patients with solitary colorectal cancer liver metastasis.

BackgroundThe aim of this study was to assess the prognostic value of CD8+ tumor infiltrating lymphocytes (TIL) combined with programmed cell death-ligand 1 (PD-L1) expression for patients with solitary colorectal cancer liver metastasis (SCLM) undergoing R0 resectionMethodsPatients undergoing curative hepatectomy for SCLM were reviewed. Immunohistochemical multiplex technique was used for quantifying CD8+ TIL, and immunohistochemical staining was used for assessing PD-L1 expression. The tumor immune microenvironment (TIME) was classified as strong for high CD8+ TIL and low PD-L1, weak for low CD8+ TIL and high PD-L1, and mild for the rest. Recurrence-free survival (RFS) and overall survival (OS) was compared between these groups.ResultsAmong the 94 patients included, a high CD8+ TIL and high PD-L1 expression was observed in 51 (54.3%) and 47 (50.0%) patients, respectively. Strong, mild, and weak TIME was observed in 24 (25.5%), 42 (44.7%), and 28 (29.8%) patients, respectively. Patients with a high CD8+ TIL had a significant longer RFS than patients with a low CD8+ TIL (3-year RFS rate, 71.6% vs. 55.3%, P=0.018). The 3-year RFS rate in the strong TIME group was significantly higher than that in the mild and weak TIME groups (89.5% vs. 71.7% and 28.8%, P<0.001), as was the 3-year rate of OS (93.8% vs. 81.8% and 61.6%, P<0.001). CD8+ TIL combined with PD-L1 expression showed better predicting accuracy for RFS than CD8+ TIL alone.ConclusionsThe density of CD8+ TIL combined with PD-L1 expression in liver metastasis was a predictor of RFS for patients with SCLM undergoing R0 resection, and therefore can be used for guiding the postoperative treatment of these patients.

HIF-1\u03b1 expression in liver metastasis but not primary colorectal cancer is associated with prognosis of patients with colorectal liver metastasis

BackgroundThe role of hypoxia-inducible factor-1α (HIF-1α) in primary colorectal cancer (CRC) and colorectal liver metastasis (CRLM) has remained unclear. The aim of this study was to investigate HIF-1α expression and its association with prognosis in patients with CRLM with a focus on hepatic stellate cells (HSCs).MethodsColon cancer cells were cultured in HSC-conditioned medium (CM), and HIF-1α expression and cell migration were analyzed. Seventy-five patients with CRLM who underwent an initial curative hepatectomy were enrolled. We examined HIF-1α expressions and patient prognosis between primary CRCs and the matched liver metastatic specimens.ResultsActivated HSCs induced HIF-1α mRNA and protein expression in colon cancer cells (p < 0.01) and promoted cell migration (p < 0.01). The positive rates of HIF-1α expression in primary CRCs and liver metastases were 68.0 and 72.0%, respectively. There were no differences in overall (OS) and disease-free survival (DFS) of HIF-1α expression in primary CRC. However, HIF-1α expression in liver metastasis correlated to poor prognosis in both OS and DFS. Furthermore, patients with HIF-1α positive expression in liver metastasis had poor prognosis.ConclusionHIF-1α expression in liver metastasis determines poor prognosis of CRLM patients. HSCs might play a key role in aggressive phenotypes of tumor cells.

High TKTL1 expression as a sign of poor prognosis in colorectal cancer with synchronous rather than metachronous liver metastases

ABSTRACT Colorectal cancer (CRC) is the third most common cancer in the world. More than half of all affected patients develop liver metastases during the course of the disease, and over half experience recurrence despite radical primary surgery. Transketolase-like protein 1 (TKTL1) is a key enzyme in the glucose metabolism of cancer cells, and its expression in tumor tissue was previously shown to indicate a poor prognosis in colorectal cancer. In this study, we investigated the prognostic significance of TKTL1 in 111 patients with surgically resected colorectal liver metastases, with a minimum follow-up time of 10.3 years. TKTL1 expression was examined in tissue samples of both primary tumors and liver metastases, and compared to clinicopathological parameters, disease-free survival, and overall survival. We show that a high expression of TKTL1 in primary tumor tissue associated with poor disease-free survival in patients with synchronous liver metastases (P = .026, Kaplan-Meier log-rank test), but with better disease-free survival in patients with metachronous metastases, although not statistically significantly (P = .073). We found similar tendencies for TKTL1 expression in liver metastases. Thus, TKTL1 could serve as a candidate marker to identify patients who benefit from liver resection or who need more aggressive perioperative chemotherapy.

Paxillin knockdown suppresses metastasis and epithelial‑mesenchymal transition in colorectal cancer via the ERK signalling pathway.

Paxillin (PXN) is a cytoplasmic protein that plays an important role in regulating focal adhesion, cytoskeletal rearrangements and cell motility. The present study aimed to investigate the role of PXN in the metastasis of human colorectal cancer (CRC) and its possible mechanisms. Immunohistochemical staining of tissues from 102 surgical CRC patients revealed that high PXN expression was positively correlated with tumour-node-metastasis (TNM) stage, lymph node metastasis, distant metastasis, and recurrence at distant sites after radical surgery. In 24 cases of stage IV CRC, PXN expression in liver metastasis was higher than that in the matched primary tumour. The knockdown of PXN inhibited the proliferation, migration and invasion potential of SW480 cells in vitro and in vivo. Transmission electron microscopy revealed the effect of PXN on ultrastructural characteristics, observed mainly in microvilli and desmosomes. The downregulation of PXN decreased the activation of extracellular regulated protein kinase (ERK) and suppressed the epithelial-mesenchymal transition (EMT) process. Following the downregulation of PXN, the addition of an ERK activator or inhibitor restored or further suppressed EMT, respectively, accompanied by corresponding changes in cell migration and invasion. Collectively, the present results confirmed the important role of PXN in CRC metastasis and revealed that PXN regulated EMT progression via the ERK signalling pathway. PXN may represent a future therapeutic strategy to prevent the EMT-associated progression and invasion of CRC.

Overexpression of miR-17 is correlated with liver metastasis in colorectal cancer.

Background:Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in men and women. The presence of systemic disease, with metastatic spread to distant sites such as the liver, considerably reduces the survival rate in CRC. Cancer stem cells contribute to the metastatic potential of CRC. However, the mechanism underlying metastasis in CRC remains unclear. Thus, this study aimed to examine the expression of microRNAs (miRNAs) in CRC stem cells in cases of liver metastases and assess their correlation with clinicopathological features.Methods:miRNAs showing high expression in liver metastases and primary lesions were selected through data mining of gene expression omnibus datasets, and miRNAs characteristic of stem cells were selected through COREMINE medical text mining. Subsequently, paired formalin-fixed paraffin-embedded tissue samples of primary CRC and liver metastasis from 30 patients were examined for the expression of miRNAs common to these lists (hsa-miR-20a, hsa-miR-26b, hsa-miR-146a, hsa-miR-17, hsa-miR-451, hsa-miR-23a, and hsa-miR-29a) using quantitative real-time polymerase chain reaction. Further, miRNA expression was compared between liver metastases and the primary tumor in each patient and the factors associated with differential expression were analyzed.Results:hsa-miR-17 was significantly upregulated in liver metastases (P < .05), but no significant difference in the expression of hsa-miR-26b, hsa-miR-146a, hsa-miR-451, hsa-miR-23a, and hsa-miR-29a was observed between primary tumors and liver metastases. The higher expression of hsa-miR-17 in liver metastases was associated with the administration of neoadjuvant chemotherapy and tumor differentiation (P < .05) but was not associated with age, sex, tumor location, or lymphatic metastasis.Conclusions:High expression of miR-17 may contribute to liver metastasis in CRC. Therefore, an in-depth understanding of its downstream pathways could help in elucidating the mechanisms underlying liver metastases in CRC. However, additional studies are warranted to validate these findings.

The potential role of heat shock protein 27 as a prognostic marker in pancreatic ductal adenocarcinoma

Background: At present no realiable prognostic biomarkers for pancreatic cancer exist. A role of heat shock protein 27 as a potential biomarker is controversial discussed. Heat shock protein 27 occurs as non-phosphorylated (HSP27) and phosphorylated forms (pHSP27) and studies considering all of these forms are lacking. Our study evaluates the significance of non-phosphorylated and phosphorylated forms regarding Serin-site 15, 78 and 82 as a prognostic marker in the biggest patients group by now. Material/Methods: Immunohistochemistry and immunofluorescence was used to determine the expression of HSP27 and pHSP27. We investigated 94 patients with pancreatic ductal adenocarcinoma who went trough surgery at Hospital Barmbek. Furthermore, we evaluated the expression in liver metastasis (n=20) of metastatic patients. Staining was performed using paraffin-embedded tissue with following antibodies: anti-HSP27, anti-pHSP-Ser15, anti-pHSP-Ser78 and anti-pHSP-Ser82. We established a three-graded-system from 0 (no staining) to +2 (maximum staining) for evaluation. All procedures were approved by the local Ethics Committee in Hamburg, Germany. Results: Total expression of HSP27, pHSP27-Ser15, pHSP27-Ser78 or pHSP27-Ser82 was not significantly associated with overall survival (p=0,76 / 0,594 / 0,685 / 0,71). However, we investigated a significant lower HSP27 and pHSP27-Ser82 expression in metastatic patients than in non-metastatic patients (p=0,001 / 0,018). In addition, we found specific patterns regarding the phosphorylated forms of HSP27 with a significant higher expression of pHSP-Ser82 compared to pHSP-Ser15 and –Ser78 (p=5,4×10-11). Same specific patterns exist in liver metastasis (p=0,001). Conclusion: In view of our study, non-phosphorylated HSP27 and all phosphorylated forms of HSP27 cannot serve as a prognostic marker. Interestingly, our Results show a significant lower HSP27 and pHSP-Ser82 expression in patients with presence of metastasis indicating that HSP27 plays a major role in metastatic invasion. The role should be investigated in further studies to elucidate the significance of HSP27 in formation of metastasis.

Prognostic and therapeutic implications of NHERF1 expression and regulation in colorectal cancer.

Na+ /H+ exchanger regulatory factor 1 (NHERF1) has been implicated in the tumorigenesis of several cancer types and is a potential therapeutic target. The current study evaluated the relationship between NHERF1 expression and clinical outcome in colorectal cancer (CRC). NHERF1 expression was evaluated by immunohistochemistry in 167 patients with CRCprimary tumors, 37 patients with no disease, and 27 patients with metastatic CRC (mCRC); and in the orthotopically implanted tumors in mice. NHERF1 expression was manipulated in CRC cells using inducible short hairpin RNAs to determine its biological functions. High expression of NHERF1 correlated with CRC progression and metastasis, as well as significantly worse overall survival, recurrence-free survival, and disease-specific survival. Orthotopic implantation studies demonstrated increased NHERF1 expression in liver metastases. Treatment of CRC xenografts with insulin-like growth factor 1 receptor (IGF1R) inhibitors downregulated NHERF1 expression, indicating NHERF1 is downstream of IGF1R signaling. Knockdown of NHERF1 increased apoptosis and reduced X-linked inhibitor of apoptosis protein (XIAP) and survivin expression, indicating NHERF1 is critical for CRC cell survival. NHERF1 expression levels correlated with worse prognosis in patients with CRC and plays a critical role in CRC cell survival. Together, our findings establish NHERF1 as a novel potential marker for increased risk of CRC-specific mortality and identify NHERF1 as an attractive therapeutic target for mCRC treatment.

Prostate-specific Membrane Antigen Heterogeneity and DNA Repair Defects in Prostate Cancer

BackgroundProstate-specific membrane antigen (PSMA; folate hydrolase) prostate cancer (PC) expression has theranostic utility. ObjectiveTo elucidate PC PSMA expression and associate this with defective DNA damage repair (DDR). Design, setting, and participantsMembranous PSMA (mPSMA) expression was scored immunohistochemically from metastatic castration-resistant PC (mCRPC) and matching, same-patient, diagnostic biopsies, and correlated with next-generation sequencing (NGS) and clinical outcome data. Outcome measurements and statistical analysisExpression of mPSMA was quantitated by modified H-score. Patient DNA was tested by NGS. Gene expression and activity scores were determined from mCRPC transcriptomes. Statistical correlations utilised Wilcoxon signed rank tests, survival was estimated by Kaplan-Meier test, and sample heterogeneity was quantified by Shannon's diversity index. Results and limitationsExpression of mPSMA at diagnosis was associated with higher Gleason grade (p=0.04) and worse overall survival (p=0.006). Overall, mPSMA expression levels increased at mCRPC (median H-score [interquartile range]: castration-sensitive prostate cancer [CSPC] 17.5 [0.0–60.0] vs mCRPC 55.0 [2.8–117.5]). Surprisingly, 42% (n=16) of CSPC and 27% (n=16) of mCRPC tissues sampled had no detectable mPSMA (H-score <10). Marked intratumour heterogeneity of mPSMA expression, with foci containing no detectable PSMA, was observed in all mPSMA expressing CSPC (100%) and 37 (84%) mCRPC biopsies. Heterogeneous intrapatient mPSMA expression between metastases was also observed, with the lowest expression in liver metastases. Tumours with DDR had higher mPSMA expression (p=0.016; 87.5 [25.0–247.5] vs 20 [0.3–98.8]; difference in medians 60 [5.0–95.0]); validation cohort studies confirmed higher mPSMA expression in patients with deleterious aberrations in BRCA2 (p<0.001; median H-score: 300 [165–300]; difference in medians 195.0 [100.0–270.0]) and ATM (p=0.005; 212.5 [136.3–300]; difference in medians 140.0 [55.0–200]) than in molecularly unselected mCRPC biopsies (55.0 [2.75–117.5]). Validation studies using mCRPC transcriptomes corroborated these findings, also indicating that SOX2 high tumours have low PSMA expression. ConclusionsMembranous PSMA expression is upregulated in some but not all PCs, with mPSMA expression demonstrating marked inter- and intrapatient heterogeneity. DDR aberrations are associated with higher mPSMA expression and merit further evaluation as predictive biomarkers of response for PSMA-targeted therapies in larger, prospective cohorts. Patient summaryThrough analysis of prostate cancer samples, we report that the presence of prostate-specific membrane antigen (PSMA) is extremely variable both within one patient and between different patients. This may limit the usefulness of PSMA scans and PSMA-targeted therapies. We show for the first time that prostate cancers with defective DNA repair produce more PSMA and so may respond better to PSMA-targeting treatments.

Hepatocyte transporter expression in liver metastasis: not correlated with the signal intensity pattern on the hepatobiliary phase of gadoxetic acid enhanced magnetic resonance images

Hepatocyte transporter expression in liver metastasis: not correlated with the signal intensity pattern on the hepatobiliary phase of gadoxetic acid enhanced magnetic resonance images