Advancements in Multiomics: MicroRNAs in Tumor Development and Progression using miRveu Technology Developed by Omicsveu

Introduction

MicroRNAs (miRNAs) are endogenous, non-coding RNAs known to regulate gene expression by translational repression or RNA cleavage and play important role in carcinogenesis and metastasis. Due to their well-established selectivity and specificity, miRNAs can represent a useful tool, both in diagnosis and therapy, in forging the path towards the achievement of precision medicine. At OmicsVeu, we have developed Super Sensitive^TM microRNA probes that can be used to identify miRNA expression profile in different grades of cancer, with high sensitivity and specificity. This blog peeks into the world of miRNAs followed by its role in carcinogenesis, development of miRNA probes, their applications using NanoVIP®.

MiRNAs and their role in carcinogenesis

miRNAs regulate transcription by binding to complementary sequences in the target mRNA, usually in the 3’ untranslated region, leading to mRNA degradation or translational repression, thereby influencing protein expression levels. miRNAs have been implicated in a wide range of biological processes such as cell differentiation, proliferation, apoptosis, organogenesis, morphogenesis, immune response, and disease pathogenesis.

Utilizing miRNAs as a potential biomarker to differentiate tumors will allow for specialized treatment regimens and increased clinical care. Numerous studies have demonstrated that aberrantly expressed miRNAs are involved in the pathogenesis of the aggressive triple negative breast cancer (TNBC) phenotype. BRCA mutated breast carcinomas are commonly seen in younger patients and have a more aggressive clinical course. Similarly, miRNA expression profiles were evaluated in a study in formalin-fixed paraffin-embedded (FFPE) cases of different grades of prostate cancer, including paired normal prostate, benign prostatic hyperplasia (BPH) and high-grade prostate cancer. miR-17 was deregulated in prostate cancer IV tissues and miR-125b was strongly upregulated in over 70% of cases of prostate cancer IV and III tissues. In benign prostatic hyperplasia (BPH) tissues, only miR-205 was significantly upregulated. The study findings underscore the importance of visualizing miRNA expression to differentiate cancer and benign cells with spatial differentiation in FFPE tumor tissues.

Super Sensitive^TM miRNA probes

Omicsveu has developed proprietary Super Sensitive^TM miRNA probes that enhance signals from intrinsically low abundant miRNAs. These oligos are synthesized using specially designed bases to give high melting temperatures. The high melting temperature oligonucleotide probes enable stringent washes at elevated temperatures to remove non-specific binding. Our miRNA probes are dual end labelled on 3’ and 5’ with five fluorescent molecules that amplify the signal, giving high resolution stains. These probes enable detection of low expressed miRNAs thus allowing high sensitivity and specificity (Figure 1). Our miRNA probes are currently one of the best products for complex diagnostic assays. Over 220 ready-to-use miRNA probes are now available and more are being developed.

Figure 1. Super Sensitive^TM miRNA probe developed by Omicsveu

NanoVIP® for miRVeu

The high-density fluorophore-labelled miRNA probes combined with our fully automated staining system NanoVIP^® aids in studying the low expressed miRNA populations and assess its expression profiles along with the physiological function.  NanoVIP^® automated staining procedure is complete xylene-free system. It incorporates multiple rounds of staining, stripping, and removal of coverslip to prepare for second multiplex run. miRNA probes combined with the automated processing using NanoVIP^® greatly increases the reliability of the test results.

Apart from the above, our miRVeu detection kits are optimized for miRNA detection with robust amplification of the signal producing clean intense staining. Each kit includes easy-to-follow protocols and ready-to-use reagents. Due to high sensitivity and specificity of the probes, these kits can be used for either manual staining or high-throughput, automated staining. Figure 2 illustrates results of miR-541 in pancreatic tumor tissue using miRveu technology in combination with NanoVIP^®

Figure 2: Expresson of miR-541 on pancreatic tumor tissue detected through miRVeu technology using NanoVIP

miRveu Potential Applications

The miRNA expression profiles are used to assess:

• Classification of cancer subtypes – Due to their well-established selectivity and specificity, miRNAs can represent a useful tool, both in diagnosis and therapy, in forging the path towards the achievement of precision medicine.
• Undifferentiated and poorly differentiated tumors – miRNAs are multifaceted and can be used for differentiation of malignant and benign tumors, early-stage cancer detection marker, identification of cancer of unknown primary, and classifying undifferentiated and poorly differentiated tumors.

Grading and staging of cancer – miRNA expression profiles can be used to unambiguously identify miRNA expression profile in different grades of cancer, with high sensitivity and specificity. It also helps in staging and differentiation of cancer subtypes.