CRISPR/Cas13-Based Platforms for a Potential Next-Generation Diagnosis of Colorectal Cancer through Exosomes Micro-RNA Detection: A Review

Abstract

Colorectal cancer (CRC) is the third most prevalent cancer with the second highest mortality rate worldwide. CRC is a heterogenous disease with multiple risk factors associated, including obesity, smoking, and use of alcohol. Of total CRC cases, 60% are diagnosed in late stages, where survival can drop to about 10%. CRC screening programs are based primarily on colonoscopy, yet this approach is invasive and has low patient adherence. Therefore, there is a strong incentive for developing molecular-based methods that are minimally invasive and have higher patient adherence. Recent reports have highlighted the importance of extracellular vesicles (EVs), specifically exosomes, as intercellular communication vehicles with a broad cargo, including micro-RNAs (miRNAs). These have been syndicated as robust candidates for diagnosis, primarily for their known activities in cancer cells, including immunoevasion, tumor progression, and angiogenesis, whereas miRNAs are dysregulated by cancer cells and delivered by cancer-derived exosomes (CEx). Quantitative polymerase chain reaction (qPCR) has shown good results detecting specific cancer-derived exosome micro-RNAs (CEx-miRNAs) associated with CRC, but qPCR also has several challenges, including portability and sensitivity/specificity issues regarding experiment design and sample quality. CRISPR/Cas-based platforms have been presented as cost-effective, ultrasensitive, specific, and robust clinical detection tools in the presence of potential inhibitors and capable of delivering quantitative and qualitative real-time data for enhanced decision-making to healthcare teams. Thereby, CRISPR/Cas13-based technologies have become a potential strategy for early CRC diagnosis detecting CEx-miRNAs. Moreover, CRISPR/Cas13-based platforms’ ease of use, scalability, and portability also showcase them as a potential point-of-care (POC) technology for CRC early diagnosis. This study presents two potential CRISPR/Cas13-based methodologies with a proposed panel consisting of four CEx-miRNAs, including miR-126, miR-1290, miR-23a, and miR-940, to streamline novel applications which may deliver a potential early diagnosis and prognosis of CRC.

Colorectal Cancer Diagnosis: The Obstacles We Face in Determining a Non-Invasive Test and Current Advances in Biomarker Detection

“…Recent studies have shown that the overall sensitivities can be around 76%, with a similar specificity level, which means there is potential for future use in screening and early detection. There have been numerous candidate miRNAs including miR-21 [113] and miR-23a [114]. Another miRNA candidate, miR-378 has been found to affect signaling pathways that control processes such as cell proliferation and apoptosis, specifically in stage II CRC [115].…”

Use of Omics Technologies for the Detection of Colorectal Cancer Biomarkers

“…On the other hand, some of the most relevant miRNA molecules assessed, such as miR-145, miR-143, miR-135, and miR-17-92, showed a significant diagnostic value for advanced neoplasia [86]. Furthermore, a recent study has used SHERLOCK-based miRNA detection to enhance and facilitate exosome-miRNA detection in blood, showing miR-126, miR-1290, miR-23a, and miR-940 as the best predictive biomarkers for early CRC stages [87]. Other miRNAs have been described as potential biomarkers, such as miR-92a, miR-29a, miR-125b, miR-19a-3p, miR-223-3p, miR-92a-3p and miR-422a, although miR-21 is by far the most studied for CRC [78].…”

Development of clustered regularly interspaced short palindromic repeats/CRISPR-associated technology for potential clinical applications

“…This method could be suitable for personalized cancer modeling and tumor-driven analysis in the future to provide guidance for precision medicine. In recent years, as microRNAs have been reported to be associated with tumorigenesis, diagnosis, and prognosis, the CRISPR/Cas technology promises great potential for an early diagnosis of miRNA-related disease[ 38 , 39 ]. Qiu et al [ 40 ] developed the RCA-CRISPR-split-HRP (RCH) detection system based on dCas9, which facilitated CRISPR/Cas9 technology in miRNA detection for the first time.…”