// Lui Ng 1, * , Timothy Ming-Hun Wan 1, * , Johnny Hon-Wai Man 1 , Ariel Ka-Man Chow 1 , Deepak Iyer 1 , Guanghua Chen 1 , Thomas Chung-Cheung Yau 2 , Oswens Siu-Hung Lo 1 , Dominic Chi-Chung Foo 1 , Jensen Tung-Chung Poon 1 , Wai-Keung Leung 3 , Roberta Wen-Chi Pang 1, 2 , Wai-Lun Law 1 1 Department of Surgery, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China 2 Centre for Cancer Research, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China 3 Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China * These authors have contributed equally to this work Correspondence to: Wai-Lun Law, email: lawwl@hku.hk Roberta Wen-Chi Pang, email: luing@hku.hk Keywords: miR-139-3p, miR-622, CRC, miRNA, biomarker Abbreviations: CRC, colorectal cancer Received: May 16, 2016 Accepted: January 16, 2017 Published: March 14, 2017 ABSTRACT Aberrant levels of circulating microRNAs are potential biomarkers for the early detection of colorectal cancer. The aim of this study was to study miR-139-3p and miR-622 in serum as a non-invasive biomarker for colorectal cancer diagnosis. We applied quantitative polymerase chain reaction to determine the levels of miR-139-3p and miR-622 in 42 pairs of tumor and adjacent non-tumor tissues, and in serum samples of 117 patients and 90 control subjects. Our results showed that miR-139-3p was silenced whereas miR-622 was overexpressed in colorectal cancer. Similarly, serum miR-139-3p level was significantly lower in colorectal cancer patients than in control subjects whereas miR-622 was more frequently detectable in patients. ROC analysis showed that AUC of miR-139-3p was 0.9935, with a sensitivity of 96.6% and specificity of 97.8%. Serum miR-139-3p level showed high sensitivity and specificity for both early and late stage CRCs and proximal and distal CRCs. Detectable serum miR-622 showed a sensitivity of 87.5% and specificity of 63.5% for discriminating CRC patients, but the sensitivity dropped for late stage patients (72.7%). We also included analyses of the blood CEA level for comparing the diagnostic performance of these blood-based biomarkers. The median level in CRC patients (3.6 ng/ml) was significantly higher than that in control (1.8 ng/ml). The AUC value of CEA in diagnosing CRC patients was 0.7515. CEA showed a positive correlation with tumor stage and age of patients and its level was higher in male. Collectively, serum miR-139-3p has strong potential as a promising non-invasive biomarker in colorectal cancer detection.
Bit by bit, over the last few decades, functional genomic tools have been piecing together the molecular puzzle driving tumorigenesis in human patients. Nevertheless, our understanding of the role of several genes and regulatory elements that drive critical cancer-associated physiological processes from disease development to progression to spread is very limited, which significantly affects our ability of applying these insights in the context of improved disease management. The recent advent of clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-based technology and its application in cancer genomics has, however, allowed the generation of a wealth of knowledge that has helped decipher several critical questions associated with translational cancer research. Precisely, the high-throughput capability coupled with a high level of technological plasticity associated with the CRISPR-Cas9 screens have expanded our horizons from a mere struggle to appreciate cancer as a genetic disease to observing the integrated genomic/epigenomic network of numerous malignancies and correlating it with our present knowledge of drugging strategies to develop innovative approaches for next-generation precision cancer medicine. Specifically, within blood cancers, current CRISPR screens have specifically focused on improving our understanding of drug resistance mechanisms, disease biology, the development of novel therapeutic approaches, and identifying the molecular mechanisms of current therapies, with an underlying aim of improving disease outcomes. Here, we review the development of the CRISPR-Cas9 genome-editing strategy, explicitly focusing on the recent advances in the CRISPR-Cas9-based screening approaches, its current capabilities, limitations, and future applications in the context of hematological malignancies.
Change in gene expression is inevitable in cancer development. With more studies demonstrating the contributions of cancer stem cells (CSCs) in colorectal cancer (CRC) development, this study is aimed at investigating whether rectal swab specimen serves as a tool for detection of dysregulation of CSC or stem cell (SC) markers and at evaluating its potential as a new promising screening method for high-risk patients. Expression levels of 15 pluripotency-associated genes were assessed by quantitative PCR in 53 rectal swab specimens referred for endoscopic screening. Dysregulated genes and joint panels based on such genes were examined for their diagnostic potentials for both polyp and CRC. Out of 15 genes, Oct4, CD26, c-MYC, and CXCR4 showed significantly differential expression among normal, polyp, and CRC patients. A panel of Oct4 and CD26 showed an AUC value of 0.80 (p = 0.003) in identifying CRC patients from polyp/normal subjects, with sensitivity and specificity of 84.6% and 69.2%. A panel of c-MYC and CXCR4 achieved CRC/polyp identification with an AUC value of 0.79 (p = 0.002), with a sensitivity of 82.8% and specificity of 80.0%. The sensitivity for polyp and CRC was 80.0% and 85.7%, respectively. Further analysis showed that higher c-MYC and CXCR4 level was detected in normal subjects who developed polyps after 5-6 years, in comparison with subjects with no lesion developed, and the AUC of the c-MYC and CXCR4 panel increased to 0.88 (p < 0.001), with sensitivity and specificity of 84.4% and 92.3%, respectively, when these patients were included in the polyp group. This study suggests that the Oct4 and CD26 panel is a promising biomarker for distinguishing CRC from normal and polyp patients, whereas the c-MYC and CXCR4 panel may identify polyp and CRC from normal individuals.
Piwi-interacting RNAs (piRNAs) represent a novel class of small non-coding RNAs (ncRNAs) that have been shown to have a deregulated expression in several cancers, although their clinical significance in colorectal cancer (CRC) remains unclear. With an aim of delineating the piRNA distribution in CRC, we conducted a systematic discovery and validation of piRNAs within two clinical cohorts. In the discovery phase, we profiled tumor and adjacent normal tissues from 18 CRC patients by deep sequencing and identified a global piRNA downregulation in CRC. Moreover, we identified piR-24000 as an unexplored piRNA that was significantly overexpressed in CRC. Using qPCR, we validated the overexpression of piR-24000 in 87 CRC patients. Additionally, we identified a significant association between a high expression of piR-24000 and an aggressive CRC phenotype including poor differentiation, presence of distant metastases, and a higher stage. Lastly, ROC analysis demonstrated a strong diagnostic power of piR-24000 in discriminating CRC patients from normal subjects. Taken together, this study provides one of the earliest large-scale reports of the global distribution of piRNAs in CRC. In addition, piR-24000 was identified as a likely oncogene in CRC that can serve as a biomarker or a therapeutic target.
3058 Background: Colorectal cancer (CRC) is the most common and treatable gastrointestinal (GI) cancer, yet it remains the second leading cause of cancer associated deaths globally. Cadherin-17 (CDH17) is a biomarker for GI cancers characterized by its overexpression in stomach, liver, and colorectal cancers but controlled expression in normal tissues from healthy adults. Although an improvement in the detection strategy of CDH17 is essential to apply it for large scale CRC screening. We have developed a novel diagnostic immunoassay using a panel of highly specific antibodies (PCT/US2019/032752) that unveiled CDH17 as a potential diagnostic maker with high detection sensitivity and specificity in CRC. Methods: The study recruited 150 patients (Retrospective specimens - 50 colorectal adenomas and 100 colorectal cancer) and 50 asymptomatic 18-80 years old blood donors (prospective specimens). CDH17 expression levels were measured within the individual blood-plasma samples using an enzyme-linked immunoassay. Results: The novel CDH17 diagnostic assay showed a wide operating dynamic range extending from 90ng to a lower limit of detection at 123pg. Using the assay, we observed a progressive and a significant increase in the plasma CDH17 concentration from the asymptomatic individuals to the colorectal adenoma group, while the CRC patients showed the highest expression of the circulating protein (median: 2.5ng/mL vs 4.5ng/mL vs 8.0ng/mL, P<0.0001). Within the CRC group, a higher CDH17 expression was associated with advanced tumor invasion, occurrence of nodal or distant metastases, as well as a clinical presentation of an advanced stage disease. The diagnostic power of the assay in terms of ROC AUC values was higher than 0.9 with an overall sensitivity of 82% that extended to 100% for metastatic CRC, while the specificity was at 94% for all stages combined (P<0.0001). Plasma CDH17 outperformed the clinically applied CEA and CA19-9 markers in differentiating the CRC patients from the asymptomatic population. Conclusions: The CDH17 immunoassay is potentially a diagnostics tool aid for the detection of CDH17 within blood samples prompting the early diagnosis of CRC. [Table: see text]
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