Marker For Predicting Metastasis Research Articles

MUC16 as a serum-based prognostic indicator of prometastatic gastric cancer

Metastatic gastric cancer (GC) presents significant clinical challenges due to its poor prognosis and limited treatment options. To address this, we conducted a targeted protein biomarker discovery study to identify markers predictive of metastasis in advanced GC (AGC). Serum samples from 176 AGC patients (T stage 3 or higher) were analyzed using the Olink Proteomics Target panels. Patients were retrospectively categorized into nonmetastatic, metastatic, and recurrence groups, and differential protein expression was assessed. Machine learning and gene set enrichment analysis (GSEA) methods were applied to discover biomarkers and predict prognosis. Four proteins (MUC16, CAIX, 5’-NT, and CD8A) were significantly elevated in metastatic GC patients compared to the control group. Additionally, GSEA indicated that the response to interleukin-4 and hypoxia-related pathways were enriched in metastatic patients. Random forest classification and decision-tree modeling showed that MUC16 could be a predictive marker for metastasis in GC patients. Additionally, ELISA validation confirmed elevated MUC16 levels in metastatic patients. Notably, high MUC16 levels were independently associated with metastatic progression in T3 or higher GC. These findings suggest the potential of MUC16 as a clinically relevant biomarker for identifying GC patients at high risk of metastasis.

Early detection and prediction of cancer metastasis – Unravelling metastasis initiating cell as a dynamic marker using self- functionalized nanosensors

The ability of cancer to metastasize to distant organs is an urgent problem to be solved clinically and continue to pose a greater challenge to researchers. Current treatments for cancer are ineffective for metastatic cancer due to inability of conventional imaging techniques to detect at early stages. Additionally, the ability to predict the ability of cancer to metastasize in advance will prove to be a valuable tool to improve patient prognosis. However, current techniques of prediction rely on molecular expression profiles which are highly tumor- specific, patient-specific and does not account for tumor heterogeneity. These factors critically impede the ability to use molecular expression profiles for early diagnosis and prediction of cancer metastasis. Hence, there is a need to concentrate on using tumor cells' phenotypic heterogeneity, which can serve as a diagnostic and predictive marker for cancer metastasis. Here, we have identified a rare subpopulation of cells within the tumor, which can signal metastasis initiation, known as metastasis-initiating cells (MICs) which are cancer non-specific, independent of epigenetic makeup, thereby becoming an excellent, unprejudiced marker for cancer metastasis irrespective of cancer type, the tissue of origin, and cancer stage. However, it is challenging to use MICs as a cellular marker for metastasis from a clinical perspective due to their rare, undetectable nature and the lack of sensitive methods to detect this elusive population of cells. To provide the ultra-sensitivity necessary for the early detection and prediction of cancer metastasis using MICs here we have created a self- functionalised nanosensor which is highly SERS active. The self- functionalized nanosensor enabled the detection of MIC using properties of intracellular biological processes and characters of the tumor microenvironment. The nanosensor enabled a detection sensitivity of 98%. It was able to identify MIC in a heterogeneous population of TICs with an unparalleled specificity of 99.62%. Further, the accuracy of MIC as a predictive marker for metastasis in a heterogeneous population of tumor spheroids was found to be 84.6%. This work contributed to the utilization of cancer cell heterogeneity to identify MIC, which can serve as a universal marker for diagnosis, prediction, and prognosis of tumor metastasis. To the best of our knowledge, this study is the first to design a probe that can provide both the diagnostic signature and predictive signature of cancer metastasis as early as the single cellular stage. This approach holds immense potential in the early diagnosis of metastatic tumors in a clinical setting and considerably improve patient prognosis.

Histone H3 modifying genes may serve as a predictive marker for metastasis in synovial sarcoma.

11058 Background: Synovial sarcomas (SS) are rare, mesenchymal tumors that exhibit heterogeneous clinical behavior. SS is characterized molecularly by a fusion of the SS18 and one of three SSX genes. While fusion type has been shown to play a role in prognosis, little is known regarding the role of secondary genomic variants in SS. We hypothesized that frequently mutated secondary mutations corresponded to altered molecular pathways associated with accelerated disease progression. Here we present further analysis of the largest study of secondary SS genomic alterations and their potential clinical implications particularly in regard to Histone H3 modifying genes. Methods: Comprehensive Genomic Profiling (CGP) from 203 SS18-SSX fusion positive SS subjects were obtained from Foundation Medicine (FMI). Correlation between gene expression and survival endpoints were assessed in two independent datasets (GSE40018, n = 34; GSE40021, n = 58). A histone H3 score (HH3Score) was calculated through summation of expression values of MLL2, MLL3, SETD2. Altered expression was assessed for genes mutated in > 5% of SS in the FMI dataset. Datasets were also combined with appropriate adjustments for multivariate analysis. Cox proportional hazard, and Chi-squared test were used as appropriate. Results: In CGP profiling of SS, only 4 genes were seen in > 5% of subjects ( MLL3 9%, SETD2 7%, MYO18A 6%, MLL2 6%). 3 of these variants are known to regulate histone H3 activity and are collectively altered in 22% (44 of 203) of the FMI SS subjects. Notably, these H3 targeted gene alterations were found to be mutually exclusive of one another (p < 0.001). In 2 independent SS datasets, subjects with high histone H3 modifying gene RNA expression levels (HH3Score) demonstrated worsened outcomes with earlier tumor metastasis (GSE40018: HR = 0.41, 95%CI 0.15-1.07, p = 0.07; GSE40021: HR = 0.22, 95%CI 0.09-0.52, p < 0.001). Multivariate analysis demonstrated that HH3Score remained significantly correlated to metastasis-free survival (HR = 0.41, 95%CI 0.22-0.77, p < 0.01) when subjects’ age (p < 0.001) and fusion type (p = 0.60) were taken into consideration. Conclusions: Altered expression of histone H3 modifying genes may serve as a key driver for SS progression. Further prospective research is necessary to confirm its prognostic importance.

Abstract 177: Increased stiffness of the tumor microenvironment in colon cancer leads to an increase in activin and metastatic potential

Abstract Colorectal cancer (CRC) is the second deadliest cancer in the US and understanding the switch to metastatic behavior and developing therapeutic strategies to target this process are clinical challenges. Stromal cells and cancer associated fibroblasts (CAF) play a critical biophysical role in cancer progression. CAFs are contractile and mechanosensitive which remodel the tumor microenvironment (TME) by cross linking, deforming and stiffening the matrix and release growth factors, which offers a unique niche for the cancer cells to become metastatic. We hypothesize that CAF-generated mechanical forces lead to metastatic spread directly or indirectly via release of growth factors including a member of the TGFb superfamily, namely activin. We will explore the mechanistic link between fibroblast force, activin release, and the induction of EMT and compare it with the activin serum levels of a CRC patient cohort. We measured activin serum levels by ELISA assay from a retrospective cohort of twenty-eight colorectal cases and twenty-eight polyp free controls. Conditioned media from co-culture of human CRC epithelial cells (FET) and stromal cells (CCD18) was examined by ELISA assay for secreted activin and was also used to culture CRC FET cells to measure induction of metastatic behavior through increase in proteins involved in epithelial-to-mesenchymal transition (EMT). CCD18 were seeded on substrates with increasing stiffness (2kPa, 10kPa, 40kPa) and treated for 72 hours with or without TGFb to generate conditioned media. FET cells were incubated with this media and subjected to trans-well migration and metabolic activity assays.Activin levels were up regulated in serum from stage IV CRC patients. High activin levels may be a predictive marker for metastasis in this cohort, with an Area Under the Curve of 0.8163. CCD18 cells secreted a 10-fold higher level of activin than FET cells alone and co-culture of CCD18 and FET cells resulted in an additional 2-fold increase in activin. Treatment of conditional media from TGFb activated stromal cells with the activin ligand trap, follistatin, leads to a decrease in FET cell migration and reversal of EMT indicating that the activin increase is functionally relevant and critical to the pro-oncogenic stromal response by TGFb. We plated CCD18 cells 2, 10 and 40 kPa polyacrylamide (PA) substrates functionalized by fibronectin with and without TGFb. We found that, with increasing gel substrate stiffness, (a) FET became more migratory, (b) CCD18 contractility increased with increasing stiffness, (c) activin in the condition media increased with increasing force. Inhibiting activin with follistatin decreased the TGFb induced activin secretion and migration in FETs. We suggest that increased TME stiffness leads to stromal cells amplification of TGFb pro-oncogenic function through thus far unrecognized induction and utilization of activin signaling. Citation Format: Jessica Bauer, Jonas J. Staudacher, Georgina Mancinelli, Nancy Krett, Emon Bashar, Paul Grippo, M Taher A. Saif, Barbara Jung. Increased stiffness of the tumor microenvironment in colon cancer leads to an increase in activin and metastatic potential [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 177.

Clinical significance of tracing thyroglobulin in predicting metastasis of post-operative patients with differentiated thyroid carcinoma before its first pre-ablation with 131I

Objective To investigate the value of tracing thyroglobulin(Tg)in predicting metastasis of post-operative patients with differentiated thyroid carcinoma(DTC)before its first pre-ablation with 131I. Methods 106 cases with DTC, undergoing total thyroidectomy and lymphadenectomy, were assigned to 2 groups as M0 group(without metastasis)and M1 group(with metastasis). Clinical data including pre-ablation stimulated thyroglobulin(sTg)and pre-operative Tg were determined. sTg, pre-operative Tg, Tg variation(ΔTg), and Tg variation rate (ΔTg/pre-operative Tg)between 2 groups were compared. The ROC curve and the diagnostic critical point(DCP)were analyzed. Results sTg, Pre-operative Tg, Tg variation, and Tg variation rate were significantly higher than those of M0(all P<0.01). The corresponding areas under the ROC curve(AUC)to differentiate the two groups were 0.913, 0.702, 0.773, and 0.943, respectively. The best diagnostic value points(DCP)were 40.60 ng/ml and -72.5%. The sensitivity and specificity were 70.21%, 100.00%, and 89.36%, 88.13%, respectively. Conclusion The pre-ablation sTg seems to be a useful diagnostic marker for predicting metastasis before the first 131I ablation. The sTg value can be effectively corrected by the Tg variation rate, and the sensitivity and accuracy of sTg for metastasis in DTC patients can be improved, finally providing evidence for pre-ablative assessment as well as strategies of 131I therapy. (Chin J Endocrinol Metab, 2018, 34: 102-105) Key words: Differentiated thyroid carcinoma; Thyroglobulin; Radioiodine therapy; Iodine radioisotopes

LEF1, TFE3, and AR are putative diagnostic markers of solid pseudopapillary neoplasms.

The diagnosis of solid pseudopapillary neoplasms (SPNs) is challenging because some SPNs share many similar morphological and immunohistochemical features with other pancreatic neoplasms. In this study, we investigated potential diagnostic markers of SPN.Based on the SPN-specific upregulated genes from a previous DNA microarray and proteome study, we selected six immunohistochemical markers [beta-catenin, androgen receptor (AR), lymphoid enhancer-binding factor 1 (LEF1), transcription factor for immunoglobulin heavy-chain enhancer 3 (TFE3), fused in sarcoma (FUS), and WNT inhibitory factor 1 (WIF-1)]. We also evaluated the Ki-67 proliferative index to investigate its associations with prognosis. To validate these markers, we studied 91 SPNs as well as 51 pancreatic ductal carcinomas (PDC) and 48 neuroendocrine tumors (NET) as controls.We found frequent and diffuse nuclear expressions of β-catenin (98.9%), AR (81.3%), LEF1 (93.4%), TFE3 (74.7%), FUS (84.6%), and cytoplasmic expression of WIF-1 (96.7%) in SPNs. In contrast, PDCs and NETs showed no expression. (P < 0.001). When beta-catenin, LEF1, and TFE3 staining were combined, the sensitivity and specificity were 100% and 91.9%, respectively. Four (4.4%) SPNs showed distant metastasis and these tumors were associated with a relatively high Ki-67 proliferative index (≥ 5%; P = 0.013).We identified LEF1, TFE3, and AR as putative diagnostic markers of SPN, auxiliary to β-catenin. Incorporated into an immunohistochemical panel, these markers could be beneficial to distinguish SPN from PDC and NET. In addition, we suggest that the Ki-67 proliferative index can be a predictive marker of metastasis in SPNs.

Overexpression of cannabinoid receptor 1 in esophageal squamous cell carcinoma is correlated with metastasis to lymph nodes and distant organs, and poor prognosis.

In patients with esophageal squamous cell carcinoma (ESCC), the status of metastasis to lymph nodes is strongly associated with prognosis. Consequently, development of a biomarker to detect the presence of metastasis would be clinically valuable. In this study, we found that overexpression of cannabinoid receptor 1 (CB1R) was applicable as a marker for prediction of metastasis in ESCC. CB1R overexpression was detected immunohistochemically in 54 of 88 cases (61.4%). The intensity of CB1R expression was uniform in both intraepithelial and invasive regions in each case, and was significantly correlated with the status of metastasis to lymph nodes (P = 0.046) and distant organs (P = 0.047). Furthermore, multivariate analysis revealed that CB1R overexpression was independently associated with poor prognosis (P = 0.019). Biological analysis of CB1R overexpression using ESCC cell lines revealed that CB1R activation appeared to promote cell proliferation and invasion. On the basis of these findings, we propose that evaluation of CB1R expression status in biopsy specimens of ESCC using immunohistochemistry might be clinically useful for prediction of metastasis to lymph nodes and distant organs.

Orai3 is a predictive marker of metastasis and survival in resectable lung adenocarcinoma.

Orai3 channel has emerged as important player in malignant transformation. Indeed, its expression is increased in cancer and favors cell proliferation and survival by permitting calcium influx. In this study, Orai3 was overexpressed in lung adenocarcinoma as compared to their matched non-tumour samples and was associated with tumoural aggressiveness. Moreover, its expression was associated with estrogen receptor alpha (ERα) expression and visceral pleural invasion in multivariate analysis. Furthermore, both the overall survival (OS) median and the metastasis free survival (MFS) median of tumors with high Orai3 expression were lower than in low Orai3 expression regardless of cancer stage (35.01 months vs. 51.11 months for OS and 46.01 months vs. 62.04 months for MFS). In conclusion, Orai3 protein level constitutes an independent prognostic marker in lung adenocarcinoma, and a novel prognostic marker that could help selecting the patients with worst prognosis to be treated with adjuvant chemotherapy in resectable stage.

P-015 Detection of gene сopy number variation as predictive markers for metastases in patients with gastric cancer

P-015 Detection of gene сopy number variation as predictive markers for metastases in patients with gastric cancer

Copy number variation of genetic loci: A predictive marker of metastasis in stomach cancer patients.

e23152Background: Copy number variations of genes are critical genetic events that promote development of malignant tumors. The purpose of this work was to study relative copy number of the genetic...

Overexpression of PTP4A3 is associated with metastasis and unfavorable prognosis in bladder cancer.

Bladder cancer (BC) is the most common malignancy in urinary system. The prognosis of metastatic BC is poor, but there remains no reliable marker to early detect metastasis. Dysregulated prenylated protein tyrosine phosphatases (PTPs) are commonly associated with cancer metastasis. From a published BC transcriptome, we identified that PTP IVA3 (PTP4A3) was the most significantly upregulated gene implicated in tumor progression among genes related to prenylated PTPs. We therefore analyzed PTP4A3 expression in our well-characterized cohort of BC. By immunohistochemistry, PTP4A3 expression was determined using H-score. PTP4A3 expression of 295 BCs was compared with clinicopathological parameters, and the effect of PTP4A3 on cancer-specific survival (CSS) and metastasis-free survival (MFS) was also examined. Two independent sets of BCs were used to assess PTP4A3 protein and transcript expression in normal urothelium and different stage tumors. PTP4A3 overexpression was significantly associated with higher pT stage (P<0.001), nodal metastasis (P<0.001), vascular invasion (P<0.001), and perineural invasion (P=0.021). In multivariate analysis, PTP4A3 overexpression was an independent predictor for CSS (P<0.001) and MFS (P=0.007). Notably, the difference in CSS and MFS between high and low PTP4A3-expressing tumors was also significant in muscle-invasive BCs. PTP4A3 protein expression showed significant and stepwise increments from normal urothelium to noninvasive BC, invasive BC, and metastatic foci (P<0.001). PTP4A3 transcript was also obviously upregulated in high-stage BC (P<0.001). PTP4A3 may play a role in BC oncogenesis and is a predictive marker of metastasis. PTP4A3 overexpression represents an independent prognosticator for BC, suggesting its potential theranostic value.

Thrombospondin-1 is highly expressed in desmoplastic components of invasive ductal carcinoma of the breast and associated with lymph node metastasis

Desmoplastic (scirrhous) invasion and lymph node metastasis are critical for the treatment and prognosis of invasive ductal carcinoma of the breast. Despite being an anti-angiogenic therapeutic candidate, Thrombospondin-1 (TSP-1) promotes invasion and metastasis of some carcinomas. To clarify the effect of TSP-1 on invasion and metastasis, we obtained 101 invasive ductal carcinomas of the breast with axillary lymph node resection. All tumors were histologically divided into two categories, carcinomas with, and those with non- /minimal desmoplastic component. Immunohistochemistry for TSP-1 was performed on all primary tumors and axillary lymph nodes with tumor metastasis. Fifty-four (53.5%) of 101 tumors were recognized as positive for TSP-1 in the cytoplasm of tumor cells. Histological study showed that significantly more cancers with desmoplastic components (46/69, 66.7%) manifested TSP-1 expression than did cancers with no- or minimal (less than 20%) desmoplasia (8/32, 25.0%; p<0.001). Axillary lymph node metastasis was significantly higher in TSP-1-positive- (28/54, 51.9%) than TSP-1-negative cancers (11/47, 23.4%; p<0.005). The present study indicates that tumor cells in the desmoplastic component strongly expressed TSP-1 in invasive ductal carcinoma of the breast and TSP-1 participates in invasion of these tumors. Our findings also suggest that TSP-1 promotes lymph node metastasis and TSP-1 potentially could be a predictive marker for metastasis.

MMP-10/Stromelysin-2 Promotes Invasion of Head and Neck Cancer

BackgroundPeriostin, IFN-induced transmembrane protein 1 (IFITM1) and Wingless-type MMTV integration site family, member 5B (Wnt-5b) were previously identified as the invasion promoted genes of head and neck squamous cell carcinoma (HNSCC) by comparing the gene expression profiles between parent and a highly invasive clone. We have previously reported that Periostin and IFITM1 promoted the invasion of HNSCC cells. Here we demonstrated that Wnt-5b overexpression promoted the invasion of HNSCC cells. Moreover, stromelysin-2 (matrix metalloproteinase-10; MMP-10) was identified as a common up-regulated gene among Periostin, IFITM1 and Wnt-5b overexpressing HNSCC cells by using microarray data sets. In this study, we investigated the roles of MMP-10 in the invasion of HNSCC.Methods and FindingsWe examined the expression of MMP-10 in HNSCC cases by immunohistochemistry. High expression of MMP-10 was frequently observed and was significantly correlated with the invasiveness and metastasis in HNSCC cases. Next, we examined the roles of MMP-10 in the invasion of HNSCC cells in vitro. Ectopic overexpression of MMP-10 promoted the invasion of HNSCC cells, and knockdown of MMP-10 suppressed the invasion of HNSCC cells. Moreover, MMP-10 knockdown suppressed Periostin and Wnt-5b-promoted invasion. Interestingly, MMP-10 overexpression induced the decreased p38 activity and MMP-10 knockdown induced the increased p38 activity. In addition, treatment with a p38 inhibitor SB203580 in HNSCC cells inhibited the invasion.ConclusionsThese results suggest that MMP-10 plays an important role in the invasion and metastasis of HNSCC, and that invasion driven by MMP-10 is partially associated with p38 MAPK inhibition. We suggest that MMP-10 can be used as a marker for prediction of metastasis in HNSCC.

ATP11A is a novel predictive marker for metachronous metastasis of colorectal cancer

The adenosine triphosphate-binding cassette transporter-homologous gene ATP11A belongs to an extended family of adenosine triphosphate-binding cassette transporters. We analysed the ATP11A gene in 7 colorectal cancer cell lines and 95 paired cases of colorectal cancer and non-cancerous regions to demonstrate the importance of ATP11A expression in colorectal cancer prognosis. ATP11A was expressed in the 7 colorectal cancer cell lines. ATP11A mRNA expression was higher in colorectal cancer tissue than in corresponding normal tissue (P<0.001). Patients with high ATP11A expression showed a poorer disease-free survival rate compared to those with low expression (P<0.001), thus indicating that increase in ATP11A expression was an independent predictor of metachronous metastasis of colorectal cancer. The present study suggests that ATP11A is a useful predictive marker of metastasis in colorectal cancer patients.

Periostin is frequently overexpressed and enhances invasion and angiogenesis in oral cancer

Oral squamous-cell carcinoma (OSCC) is one of the most common types of human cancer. Typically OSCC cells show persistent invasion that frequently leads to local recurrence and distant lymphatic metastasis. We previously identified Periostin as the gene demonstrating the highest fold change expression in the invasive clone by comparing the transcriptional profile of parent OSCC cell line and a highly invasive clone. Here, we demonstrated that Periostin overexpression enhanced invasiveness in oral cancer cell lines. To know the role of Periostin in invasion, angiogenesis and metastasis in OSCC cases, we first examined the expression of Periostin mRNA in 31 OSCC cases by RT–PCR and Periostin protein in 74 OSCC cases by immunohistochemistry. Then, we compared the Periostin expression with invasion pattern, metastasis and blood vessel density. Periostin mRNA and protein overexpression were frequently found in OSCC cases and Periostin expression was well correlated with the invasion pattern and metastasis. Moreover, blood vessel density of Periostin-positive cases was higher than those of Periostin-negative cases. Interestingly, recombinant Periostin enhanced capillary formation in vitro in a concentration-dependant manner. In summary, these findings suggest that Periostin may promote invasion and angiogenesis in OSCC, and that Periostin can be a strong marker for prediction of metastasis in oral cancer patients.