Biomarker For Cancer Progression Research Articles

The pro-angiogenesis effect of miR33a-5p/Ets-1/DKK1 signaling in ox-LDL induced HUVECs

Objective: Angiogenesis is involved in multiple biological processes, including atherosclerosis (AS) and cancer. Dickkopf1 (DKK1) plays many roles in both tumors and AS and has emerged as a potential biomarker of cancer progression and prognosis. Targeting DKK1 is a good choice for oncological treatments. Many anticancer therapies are associated with specific cardiovascular toxicity. However, the effects of DKK1 neutralizing therapy on AS are unclear. We focused on how DKK1 affected angiogenesis in AS and ox-LDL-induced human umbilical vein endothelial cells (HUVECs).Methods: ApoE-/- mice were fed a high-fat diet and then injected with DKK1i or DKK1 lentivirus to study the effects of DKK1. In vitro, promoter assays, protein analysis, database mining, dual-luciferase reporter assay (DLR), electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP), and coimmunoprecipitation (co-IP) were used to study the mechanism of DKK1 biogenesis. Cell migration and angiogenesis assays were performed to investigate the function and regulatory mechanisms of DKK1.Results: DKK1 participated in angiogenesis both in the plaques of ApoE-/- mice by knockdown or overexpression of DKK1 and ox-LDL-induced HUVECs. DKK1 induced angiogenesis (increasing migration and capillary formation, inducing expression of VEGFR-2/VEGF-A/MMP) via the CKAP4/PI3K pathway, independent of Wnt/β-catenin. ox-LDL increased the expression and nuclear transfer of Ets-1 and c-jun, and induced the transcriptional activity of DKK1 in HUVECs. Ets-1, along with c-jun and CBP, could bind to the promoter of DKK1 and enhance DKK1 transcription. MiR33a-5p was downregulated in ox-LDL induced HUVECs and aortic artery of high-fat diet ApoE-/- mice. Ets-1 was a direct target of miR33a-5p. MiR33a-5p/Ets-1/ DKK1 axis contributed to angiogenesis.Conclusions: MiR33a-5p/Ets-1/DKK1 signaling participated in ox-LDL-induced angiogenesis of HUVECs via the CKAP4/PI3K pathway. These new findings provide a rationale and notable method for tumor therapy and cardiovascular protection.

LncRNA involvement in cancer stem cell function and epithelial-mesenchymal transitions.

Epithelial to mesenchymal transition (EMT) is a cellular process in which cells composing epithelial tissue lose requirements for physical contact with neighboring cells and acquire mesenchymal characteristics consisting of increased migratory and invasive behaviors. EMT is a fundamental process that is required for initial and later events during embryogenesis. Cancer stem cells (CSCs) possess multipotency sufficient for their differentiation into bulk tumor cells and also have the capacity to undergo EMT. When CSCs initiate EMT programs the resulting cancerous mesenchymal cells become invasive and this migratory behavior also poises them for metastatic activity. Long noncoding RNAs (lncRNAs) are functional RNA molecules that do not encode proteins, yet regulate the expression of protein-coding genes through recruitment or sequestration of gene-regulatory proteins and microRNAs. lncRNA exhibit tissue-specific patterns of gene expression during development and specific sets of lncRNAs are also involved in various cancer types. This review considers the interplay between lncRNAs and the biogenesis of CSCs. We also review function of lncRNAs in EMT in CSCs. In addition, we discuss the utility of lncRNAs as biomarkers of cancer progression, and their potential use as therapeutic targets for treatment of cancer.

Abstract 1984: Investigating circulating tumor DNA as a biomarker of cancer progression and recurrence in sarcoma

Abstract Objective: Circulating tumour DNA (ctDNA) is an active area of research in many types of cancer due to its value as a non-invasive diagnostic tool. Despite making up a small fraction of total cell-free DNA (cfDNA), detection of ctDNA has been made possible given advances in next generation sequencing and Digital Droplet PCR (ddPCR). CtDNA testing has some key advantages compared to tissue biopsy; ease of repetition provides a means of real-time monitoring of treatment response and signs of recurrence in cancer. This has specific relevance in sarcoma, where early detection of recurrence may provide an opportunity for faster intervention and can potentially lead to better prognostic outcomes. The purpose of this study is to determine if ctDNA is detectable in plasma samples collected from sarcoma patients and to validate our pre-analytical procedures and to identify opportunities to optimize our protocols for future use. Methods: 20 ml of blood samples were collected from 270 soft tissue sarcoma and osteosarcoma patients with or without pre-operative adjuvant treatment and processed to separate the plasma. Cf DNA was isolated from 2ml of plasma and the quantity and quality assessment was performed by qRT-PCR and capillary electrophoresis, respectively. Whole exome sequencing (WES) was performed on 6 matched tumor-blood DNA samples to identify tumour specific single nucleotide alterations. Sequence variants identified from WES analysis were used to design primers and probes to detect the tumor specific alterations in the corresponding ctDNA in those 6 cases using ddPCR. Results: Of the 60 patient samples analysed to date, the majority were positive for cfDNA. Quality assessment of 41cfDNA by capillary electrophoresis showed peaks approximately 170 bp in size, characteristic of cfDNA. WES of 6 patients' matched tumor-blood samples identified many tumor specific variants. Six of those sequence variants on SMAD4, COL19A1, DDX3X, ADGRG4, HECW1 and FOXR2 genes (Table 1) were used to design primers and probes to detect the tumour specific alterations in the corresponding ctDNA using dd PCR. We observed the presence tumour specific mutations in all corresponding ctDNAs tested in this study. Table 1:Identification of tumour specific variants by WESCaseGeneSNVSarcoma SubtypeVariant Allele Frequency1SMAD4T1584CMyxofibrosarcoma86%2COL19A1G1252AUndifferentiated pleomorphic sarcoma50%3DDX3XA1038COsteosarcoma54%4ADGRG4T6361CLiposarcoma65%5HECW1C1874AMyxofibrosarcoma25%6FOXR2C130GMyxofibrosarcoma24% Conclusions: Our ongoing study to evaluate the use of ctDNA as a biomarker in sarcoma monitoring shows promise related to the efficiency of our protocols and assays. Further testing is ongoing to assess the clinical value in sarcoma. Citation Format: Nalan Gokgoz, Paige Darville-O'Quinn, Ainaz Malekoltojari, Patrick Prochazka, Peter C. Ferguson, Jay S. Wunder, Irene L. Andrulis. Investigating circulating tumor DNA as a biomarker of cancer progression and recurrence in sarcoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1984.

Serum malondialdehyde (MDA) level as a potential biomarker of cancer progression for patients with bladder cancer.

Bladder cancer is the most common malignancy involving the urinary system. Recent research tends to emphasize the role of oxidative stress products in the carcinogenesis of bladder cancer. The level of oxidative stress can be measured by assessing the MDA levels. This study aimed to evaluate serum MDA levels in patients with bladder cancer, as well as to determine its potential role as a biomarker in the diagnosis of the disease and progression risk considerations. The study was designed as a cross-sectional study and included 90 patients, divided into three groups with 30 patients each: Ta, T1and T2-T4 group, based on histopathological findings after transurethral resection of the tumor. The control group included 30 healthy volunteers. MDA level was determined using the spectrophotometric method. Serum MDA level in patients with bladder cancer [0.86 (0.78-1.05) μmol/L] was significantly higher than the serum MDA level in control group [0.70 (0.69-0.72) μmol/L] (p < 0.001). Serum MDA level in Ta group [0.73 (0.70-1.05) μmol/L], T1 group [0.85 (0.80-1.12) μmol/L] and in T2-T4 group [0.91 (0.84-1.04) μmol/L] was significantly higher than the serum MDA level in control group [0.70 (0.69-0.72) μmol/L] (p < 0.01). MDA level in T1 and T2-T4 group was significantly higher than the MDA level in Ta group (p < 0.01). No significant difference was observed in MDA level between T1 and T2-T4 group (p = NS). A statistically significant positive correlation was found between tumor size and serum MDA level in patients with bladder cancer (rho = 0.254 p < 0.01). The results of the present study suggest that MDA serum level might play a significant role as a biomarker in the diagnosis of bladder cancer, as well as in the monitoring of its progression.

Circulating activated lymphocyte subsets as potential blood biomarkers of cancer progression.

The objective of this study was to predict the value of lymphocyte subsets in cancer progression. Peripheral blood was obtained from 327 untreated patients with cancer and 158 healthy volunteers. Levels of lymphocyte subsets were determined by flow cytometry. There were decreased levels of natural killer (NK) cells, CD8+ T cells, and naïve CD4+/CD4+ T cells in untreated patients with cancer compared to those in healthy controls. Inversely, there were elevated levels of the following T‐cell percentages in cancer patients compared to those in healthy controls: memory CD4+/CD4+, CD8+ T cells, HLA‐DR/CD8+, CD8+ CD38+/CD8+, and CD4+/CD8+. In addition, there are a decreasing trend in terms of CD4+ T‐cell counts and an increase CD8+ HLA‐DR/CD8+ T‐cell and CD8+ CD38+/CD8+ T‐cell percentages in the advanced stage. An increasing trend with advanced tumor stage and the percentages of CD8+ HLA‐DR/CD8+ T cells and CD8+ CD38+/CD8+ T cells was shown in this study. There are a negative correlation for CD4+ T‐cell counts and positive correlation for percentages of CD8+ HLA‐DR/CD8+ T cell and CD8+ CD38+/CD8+ T cells with the lymph node metastasis. In the presence of distant metastatic spread, we observed higher NK‐cell counts, CD8+ HLA‐DR/CD8+ T‐cell percentages, CD8+ CD38+/CD8+ T‐cell percentages, as well as lower CD4+ T‐cell counts than those in the absence of distant metastases spread. Abnormal levels of NK cell, CD8+ T cells, memory CD4+/CD4+, naïve CD4+/ CD4+, CD8+ HLA‐DR/CD8+, CD8+ CD38+/CD8+, and CD4+/CD8+ can be a potential blood biomarkers of cancer development. CD4+ T‐cell counts and percentages of CD8+ HLA‐DR/ CD8+ and CD8+ CD38+/ CD8+ can predict the cancer progression.

Response Assessment With Molecular Characterization of Circulating Tumor Cells and Plasma MicroRNA Profiling in Patients With Locally Advanced Breast Cancer During Neoadjuvant Chemotherapy

BackgroundCells detaching from the primary tumor site are metastasis initiator cells, and the detection of CTC, known as liquid biopsy, is an important test of biomarkers of cancer progression. We investigated the molecular characterization of circulating tumor cells (CTCs), profiled the plasma microRNA (miR) content, and analyzed the relationship with the clinical outcomes by sampling the peripheral blood from patients with locally advanced breast cancer before and after neoadjuvant chemotherapy. Patients and MethodsMarkers of breast cancer, epithelial–mesenchymal transition (EMT), drug resistance, and stem cells were used for CTC isolation and characterization. Plasma miR profiles were obtained from selected patients with CTC positivity determined using next-generation sequencing. ResultsThe proportion of CTC, EMT, and stem cell marker positivity was 16.7%, 8.3%, and 25% before and 18.2%, 15.2%, and 9.1% after treatment, respectively. A significant correlation was found between the pretreatment CTCs and ALDH1 positivity (P = .0245). These CTCs with stemness properties were observed in most hormone receptor–positive, human epidermal growth factor receptor 2–negative cases and were also present with a high incidence in cases of early metastasis. miR-146b-5p and miR-199a-5p, which are involved in metastasis, invasion, and EMT, were accompanied by CTC positivity, and miR-4646-3p was associated with the development of early metastasis. ConclusionsMolecular characterization of CTCs and miR profiling of serial samples from patients with locally advanced breast cancer during neoadjuvant chemotherapy appears to be a very useful in predicting cure and clinical course and might be a key to developing new targeted therapies.

Immunohistochemical expression of p63 in oral premalignant disorders and its correlation with oral squamous cell carcinoma

Introduction: Oral squamous cell carcinoma (OSCC) represents 95% of all head and neck cancers, and its incidence has increased by 50% in the past decade. Accumulation of genetic alterations (mutations, loss of heterozygosity, loss or gain of chromosomes) is the basis for the progression from a normal cell to a cancer cell. OSCCs are frequently preceded by oral potentially malignant disorders (PMDs). It is difficult to predict the malignant transformation of these PMDs. Thus, early detection of genetic changes in PMDs can facilitate detection of those lesions, which may have potential to progress to malignancy. p63 is a protein coded by p63 gene which is a homolog of p53 gene. It has a critical role in cell cycle regulation and is associated with epithelial tumorigenesis. The present study aims to evaluate the role of p63 in carcinogenesis in PMDs (Leukoplakia, Oral submucous fibrosis and Lichen planus) and OSCCs using immunohistochemistry. Aims and Objectives: The aim is to evaluate the expression of the p63 IHC marker in oral premalignant disorders and to compare its expression in OSCC. Materials and Methods: Tissue sections of PMDs (105 cases) and OSCCs (35 cases) were stained for IHC marker p63. The percentage of positive cells and staining patterns were assessed for all the lesions. Results and Conclusions: Progressive accumulation of p63-immunopositive cells from PMDs to OSCC reflects its major role in the development of OSCC according to a multiple-stage model of carcinogenesis and can be a useful indicator of dysplastic change, thus serving as a biomarker for cancer progression.

The Prognostic Value of Small Noncoding microRNA-21 Expression in the Survival of Cancer Patients: A Meta-Analysis.

microRNA-21 (miR-21) is a small noncoding RNA that regulates gene expression in different types of human malignancies. The potential prognostic value of miR-21 in cancer progression is controversial. This meta-analysis includes 76 studies of 10,213 cancer patients to test miR-21 prognostic value in various human cancers. We obtain hazard ratios (HRs) and corresponding 95% confidence intervals (CIs) for overall survival (OS) to assess association strength. In the pooled analysis, high miR-21 expression is associated with poor OS, with a combined HR of 1.59 (95% CI, 1.49-1.70; p < 0.001; random-effects model). Furthermore, subgroup analysis demonstrates that high miR-21 expression is related to shorter OS in patients with digestive system cancers (HR = 1.02; 95% CI, 1.002 to 1.04; p = 0.026), respiratory system cancers (HR = 1.93; 95% CI, 1.48 to 2.51; p < 0.001), and breast cancer (HR = 2.20; 95% CI, 1.78 to 2.73; p = 0.001). These results indicate that miR-21 may be a clinically useful prognostic biomarker for cancer progression.

CD44v3 protein-carrying tumor-derived exosomes in HNSCC patients’ plasma as potential noninvasive biomarkers of disease activity

ABSTRACT The molecular cargo of tumor-cell-derived exosomes (TEX) mimics that of parental tumor cells. Thus, TEX could potentially serve as noninvasive biomarkers of cancer progression. However, separation of TEX from non-TEX in patients’ plasma requires tumor antigen-specific detection reagents. CD44v3 has been of interest as a potential biomarker of disease progression in HNSCC, because its overexpression in tumor cells associates with poor outcome. Here, CD44v3+ TEX immunocaptured from plasma of 44 HNSCC patients and 7 healthy donors (HDs) were evaluated as potential biomarkers of disease activity and stage. Exosomes were isolated from plasma of by size exclusion chromatography. Using anti-CD44v3 or anti-CD3 mAbs on beads, CD44v3+ TEX CD3(-)TEX-enriched exosomes were immunocaptured from supernatants of nonmalignant or HNSCC cell lines and from patients’ plasma. On-bead flow cytometry was used for the detection of FAS-L, PD-L1, TGFF-β. CSPG4 or EGFR on exosomes. The TEX expression profiles were correlated to clinicopathological parameters. Relative florescence intensity (RFI) values for CD44v3 were higher (p < .01) on TEX from HNSCC cell lines or on CD44v3+ CD3(-) plasma-derived exosomes. RFI values of CD44v3 on CD3(-) exosomes were higher (p < .005) in patients than in HDs and correlated (p < .05) with the UICC stage and lymph node metastasis. In HNSCC patients, CD44v3+ exosomes higher levels of immunosuppressive proteins compared to CD44v3(-) exosomes (p < .05-p < .005), and RFI values for these markers correlated with higher disease stages and lymph node metastasis. Isolation of CD44v3+ exosomes by immunocapture allowed for enrichment of TEX which are potentially promising liquid biomarkers of the tumor burden and disease stage in HNSCC.

Plasmatic exosomes from prostate cancer patients show increased carbonic anhydrase IX expression and activity and low pH

Acidity, hypoxia and increased release of exosomes are severe phenotypes of tumours. The regulation of pH in tumours involves the interaction of several proteins, including the carbonic anhydrases which catalyze the formation of bicarbonate and protons from carbon dioxide and water. Among CA isoforms, CA IX is over-expressed in a large number of solid tumours, conferring to cancer cells a survival advantage in hypoxic and acidic microenvironment, but there isn’t evidence that CA IX expression could have a real clinical impact. Therefore, in this study for the first time the expression and activity of CA IX have been investigated in the plasmatic exosomes obtained from patients with prostate carcinoma (PCa). For this purpose, the study was performed through different methodological approaches, such as NTA, western blot analysis, enzyme activity assay, Nanoscale flow cytometry, ELISA, confocal microscopy. The results showed that PCa exosomes significantly overexpressed CA IX levels and related activity as compared to healthy donors. Furthermore, CA IX expression and activity were correlated to the exosome intraluminal pH, demonstrating for the first time that PCa exosomes are acidic. Our data suggest the possible use of the exosomal CA IX expression and activity as a biomarker of cancer progression in PCa.

Epigenetic changes in fibroblasts drive cancer metabolism and differentiation.

Genomic changes that drive cancer initiation and progression contribute to the co-evolution of the adjacent stroma. The nature of the stromal reprogramming involves differential DNA methylation patterns and levels that change in response to the tumor and systemic therapeutic intervention. Epigenetic reprogramming in carcinoma-associated fibroblasts are robust biomarkers for cancer progression and have a transcriptional impact that support cancer epithelial progression in a paracrine manner. For prostate cancer, promoter hypermethylation and silencing of the RasGAP, RASAL3 that resulted in the activation of Ras signaling in carcinoma-associated fibroblasts. Stromal Ras activity initiated a process of macropinocytosis that provided prostate cancer epithelia with abundant glutamine for metabolic conversion to fuel its proliferation and a signal to transdifferentiate into a neuroendocrine phenotype. This epigenetic oncogenic metabolic/signaling axis seemed to be further potentiated by androgen receptor signaling antagonists and contributed to therapeutic resistance. Intervention of stromal signaling may complement conventional therapies targeting the cancer cell.

Extracellular vesicles as a novel source of biomarkers in liquid biopsies for monitoring cancer progression and drug resistance.

Cancer-derived extracellular vesicles (EVs) have been detected in the bloodstream and other biofluids of cancer patients. They carry various tumor-derived molecules such as mutated DNA and RNA fragments, oncoproteins as well as miRNA and protein signatures associated with various phenotypes. The molecular cargo of EVs partially reflects the intracellular status of their cellular origin, however various sorting mechanisms lead to the enrichment or depletion of EVs in specific nucleic acids, proteins or lipids. It is becoming increasingly clear that cancer-derived EVs act in a paracrine and systemic manner to promote cancer progression by transferring aggressive phenotypic traits and drug-resistant phenotypes to other cancer cells, modulating the anti-tumor immune response, as well as contributing to remodeling the tumor microenvironment and formation of pre-metastatic niches. These findings have raised the idea that cancer-derived EVs may serve as analytes in liquid biopsies for real-time monitoring of tumor burden and drug resistance. In this review, we have summarized recent longitudinal clinical studies describing promising EV-associated biomarkers for cancer progression and tracking cancer evolution as well as pre-clinical and clinical evidence on the relevance of EVs for monitoring the emergence or progression of drug resistance. Furthermore, we outlined the state-of-the-art in the development and commercialization of EV-based biomarkers and discussed the scientific and technological challenges that need to be met in order to translate EV research into clinically applicable tools for precision medicine.

Abstract #4317 Peri-operative novel psychological intervention in breast cancer patients aiming to reduce stress responses and improve biomarkers of cancer progression

The way the biological system responds to environmental stress depends on the adaptive response and evolution. Each system responds with a molecular model mediated by oxidative stress known as the phenomenon of hormesis. This response, which is summarized in stimulation at low doses and inhibition at high doses of toxic agents, is the key to survival and the acquisition of fitness against environmental stress. Thus the adaptive response to xenobiotics is a product of the molecular signaling that is present from the first bacteria and is part of the endosymbiotic relationship of mitochondria in eukaryotic cells in mitohormesis. In this way, it is possible to deduce that the microbiota plays a role within the holobiont in the transduction of the hormetic response; also, the reactive oxygen species (ROS) induce the exchange of genetic material through the transposable elements (TEs) between the symbionts and the host. It is in this exchange of information that the production of antioxidant secondary metabolites that regulate the oxidative stress induced under hormesis is possible; the epigenetic regulation of biosynthetic gene clusters (BGCs) for obtaining chemically diverse metabolites also occurs. Thus, the search for homeostasis and coevolution within the biological system for the maintenance of metabolic networks is achieved under the cognition provided by hormesis. It is in this way that this evolutionary model becomes a source of new stressors or hormetins, whose mechanism of action is the maintenance of the structural functioning of the system as well as its replication, and thus becomes a new rational strategy for prospecting for new drugs. Thus, the objective of this chapter is to analyze the phenomena of adaptation to environmental stress and its evolutionary role as an innovative model in the discovery of new medicines.

Abstract 5238: Clinical significance of GET4 expression in colorectal cancer

Abstract Background: Colorectal cancer (CRC) is one of the most common cancers worldwide and the second leading cause of cancer-related deaths in developed countries, with the majority of deaths being attributed to distant metastasis. Therefore, it is necessary to identify a novel biomarker for cancer progression and metastasis, one that could also be a useful therapeutic target in CRC. Amplification of chromosome 7p is frequent in CRC, and it has been considered to harbor driver genes that promote tumorigenesis or tumor progression by the gain of function. We aimed to identify novel candidate driver genes on chromosome 7p and to clarify the clinical significance of their expression in CRC. Material and Methods: 1. We selected the candidate genes that satisfied the following criteria using CRC data from The Cancer Genome Atlas (TCGA). 1) The DNA copy number and mRNA expression are positively correlated with each other, 2) overexpressed in the tumor tissues compared to the normal tissues. 2. The mRNA expression of the candidate genes was measured in 147 surgically-resected CRC tissues and the paired normal tissues in our hospital by quantitative RT-PCR. The differences of mRNA expression between CRC tissues and normal tissues were analyzed by Mann Whitney U-test. 3. Survival analysis between high and low expression group of the candidate genes was performed by the Kaplan-Meier method. Correlation between the mRNA expression of the candidate genes and the clinicopathological factors were analyzed by Fisher’s exact test. 4. Gene Set Enrichment Analysis (GSEA) was performed in CRC data from TCGA to clarify the correlation between the candidate genes and gene sets that are associated with tumorigenesis or tumor progression. Results: The Golgi to ER traffic protein 4 (GET4) was identified as a candidate driver gene. GET4 is known to be one factor of the BCL2-associated athanogene 6 (BAG6) chaperone complex and function as a regulator for the nucleo-cytoplasmic transport of BAG6. The BAG6 complex is implicated in diverse cellular processes including apoptosis, co-chaperone, and DNA damage response. The expression of GET4 was significantly higher in CRC tissues than in normal colon tissues (p=0.03), and it correlated with depth invasion (p=0.02), and lymphatic invasion (p=0.01). The high GET4 expression group had a significantly poorer prognosis than the low expression group (p=0.02). On multivariate analysis, distant metastasis was an independent prognostic factor affecting OS (p&amp;lt;0.05) with hazard ratios (95% CI) of 66.6 (5.46-1649.3) among clinicopathological factors. GSEA showed that GET4 expression was negatively correlated with the pathway associated with the Rb-related protein p130, which belongs to the RB gene family proved to be a tumor suppressor. Conclusions: GET4 could be a promising driver gene of CRC on chromosome 7p possibly through regulating the location of BAG6. GET4 may be a therapeutic target as well as a poor prognostic biomarker in CRC. Note: This abstract was not presented at the meeting. Citation Format: Kensuke Koike, Takaaki Masuda, Kuniaki Sato, Yuushi Motomura, Jyunnichi Takahashi, Dai Shimizu, Shotaro Kuramitsu, Atsushi Fujii, Akihiro Kitagawa, Miwa Noda, Yusuke tsuruda, Hajime Ootu, Yousuke Kuroda, Hidetoshi Eguchi, Takashi Nakagawa, Koushi Mimori. Clinical significance of GET4 expression in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5238.

Rh type C-glycoprotein functions as a novel tumor suppressor gene by inhibiting tumorigenicity and metastasis in head and neck squamous cell carcinoma.

Head and neck squamous cell carcinoma (HNSCC), a major histologic subtype of head and neck cancer, presents great mortality and morbidity worldwide. The aim of this study is to discover new potential biomarkers closely correlated with HNSCC progression. In this study, weighted gene co-expression network analysis was applied to construct a co-expression network, and the brown module was identified as the most correlated with HNSCC progression. Hub gene identification combined with survival analyses determined RHCG as a candidate biomarker for cancer progression and prognosis prediction. Further experimental results proved that RHCG was aberrantly downregulated in HNSCC tissues and cell lines. Moreover, decreased RHCG expression was shown to be associated with advanced stage and dismal prognosis in HNSCC patients. Functional assays revealed that RHCG could inhibit cell viability, clonogenicity, cell migration in vitro and suppress tumor formation in vivo. Further bioinformatics study demonstrated that DNA promoter hypermethylation of RHCG could lead to its downregulation and serve as potential prognostic maker in HNSCC. Our study reveals that RHCG acts as a tumor suppressor gene that plays a crucial role in inhibiting tumorigenicity and metastasis in HNSCC, which will shed light on the potential diagnostic and therapeutic strategies for HNSCC.

Tumor-associated macrophages: role in cancer development and therapeutic implications.

Tumor-associated macrophages (TAMs) are known to play important roles in the initiation and progression of human cancers, as well as in angiogenesis. TAMs are considered as main components of the tumor microenvironment. Targeting TAMs may serve as a therapeutic strategy for the treatment of cancer. In this review, the signaling pathways, origin, function, polarization and clinical application of TAMs are discussed. The role of TAMs in tumor initiation, progression, angiogenesis, invasion and metastasis are also emphasized. In addition, a variety of clinical and pre-clinical approaches to target TAMs are discussed. Clinical therapeutic approaches that show most promise include blocking the extravasation of TAMs along with using TAMs as diagnostic biomarkers for cancer progression. The targeting of TAMs in a variety of clinical settings appears to be a promising strategy for decreasing metastasis formation and for improving patient outcome.

Role of Tissue Factor in Tumor Progression and Cancer-Associated Thrombosis.

It has been long-established that cancer and thrombosis are linked, but the exact underlying pathological mechanism remains to be unraveled. As the initiator of the coagulation cascade, the transmembrane glycoprotein tissue factor (TF) has been intensely investigated for its role in cancer-associated thrombosis and cancer progression. TF expression is regulated by both specific oncogenes and environmental factors, and it is shown to regulate primary growth and metastasis formation in a variety of cancer models. In clinical studies, TF has been shown to be overexpressed in most cancer types and is strongly associated with disease progression. While TF clearly associates with cancer progression, a prominent role for TF in the development of cancer-associated thrombosis is less clear. The current concept is that cancer-associated thrombosis is associated with the secretion of tumor-derived TF-positive extracellular vesicles in certain tumor types. To date, many therapeutic strategies to target TF-both in preclinical and clinical phase-are being pursued, including targeting TF or the TF:FVIIa complex by itself or by exploiting TF as a docking molecule to deliver cytotoxic compounds to the tumor. In this review, the authors summarize the current understanding of the role of TF in both cancer progression and cancer-associated thrombosis, and discuss novel insights on TF as a therapeutic target as well as a biomarker for cancer progression and VTE.

Colorectal cancer-associated ~6kDa hyaluronan serves as a novel biomarker for cancer progression and metastasis.

Low molecular weight hyaluronan (LMW-HA) is believed to accumulate in tumors and to exert protumor effects. This study aimed to identify colorectal cancer (CRC)-associated LMW-HA, precisely determine its MW, and elucidate its role in predicting tumor progression. The MW distribution of HA extracted from CRC and paired noncancerous tissues was evaluated. We found that the level of HA with a MW below 30kDa was markedly elevated in CRC tissues, and we defined HA with a MW of ~6kDa as CRC-associated LMW-HA. In line with this finding, ~6kDa HA was significantly accumulated in cancer tissues relative to total HA, and this LMW-HA played a critical role in tumor metastasis. Moreover, serum ~6kDa HA levels in CRC patients were significantly increased and positively correlated with the levels in matched cancer tissues. Elevated serum ~6kDa HA levels could be used to discriminate patients with or without CRC and was associated with early relapse, advanced tumor-node-metastasis stage, lymphovascular invasion, and lymph node (LN) metastasis. Notably, serum ~6kDa HA levels were significantly reduced after tumor resection. Our study suggests that ~6kDa HA may serve as a new biomarker for estimating tumor progression, predicting LN metastasis, and monitoring tumor recurrence.

Hypoxic BMSC-derived exosomal miRNAs promote metastasis of lung cancer cells via STAT3-induced EMT

BackgroundMetastasis is the main cause of lung cancer mortality. Bone marrow-derived mesenchymal stem cells (BMSCs) are a component of the cancer microenvironment and contribute to cancer progression. Intratumoral hypoxia affects both cancer and stromal cells. Exosomes are recognized as mediators of intercellular communication. Here, we aim to further elucidate the communication between BMSC-derived exosomes and cancer cells in the hypoxic niche.MethodsExosomal miRNA profiling was performed using a microRNA array. Lung cancer cells and an in vivo mouse syngeneic tumor model were used to evaluate the effects of select exosomal microRNAs. Hypoxic BMSC-derived plasma exosomal miRNAs were assessed for their capacity to discriminate between cancer patients and non-cancerous controls and between cancer patients with or without metastasis.ResultsWe demonstrate that exosomes derived from hypoxic BMSCs are taken by neighboring cancer cells and promote cancer cell invasion and EMT. Exosome-mediated transfer of select microRNAs, including miR-193a-3p, miR-210-3p and miR-5100, from BMSCs to epithelial cancer cells activates STAT3 signaling and increases the expression of mesenchymal related molecules. The diagnostic accuracy of individual microRNA showed that plasma exosomal miR-193a-3p can discriminate cancer patients from non-cancerous controls. A panel of these three plasma exosomal microRNAs showed a better diagnostic accuracy to discriminate lung cancer patients with or without metastasis than individual exosomal microRNA.ConclusionsExosome-mediated transfer of miR-193a-3p, miR-210-3p and miR-5100, could promote invasion of lung cancer cells by activating STAT3 signalling-induced EMT. These exosomal miRNAs may be promising noninvasive biomarkers for cancer progression.

AB037. P-05. Evaluation of infrared spectral signature associated with pathology as biomarker for cancer progression in cholangiocarcinoma

Background: Cholangiocarcinoma (CCA) is a cancer of the bile duct epithelium and remains an important public health problem in Southeast Asia, with a particularly in Northeast Thailand due to its high incidence and high fatality rates. The highest prevalence of Opisthorchis viverrini (OV) related CCA found in northeast Thailand. The disease is difficult to early diagnose and is usually fatal because patients usually come to the physician at the advance stage and the effective non-surgical therapy is not currently available. In this study, we aim to focus on periductal fibrosis and bile duct cells initiated into cancer development, which are characterized by Fourier transform infrared (FTIR) microspectroscopy giving specific spectra corresponding to biomarkers as an early diagnosis of CCA.