Periodic Reporting for period 1 - RNADIAGON (Excellence in research and development of non-coding RNA DIAGnostics in ONcology)
Okres sprawozdawczy: 2019-02-01 do 2022-07-31
Cancer occurs in more than 2 million individuals every year in Europe alone. It is widely recognized that early diagnosis and monitoring of the disease - during therapy and post-treatment follow-up- is a key step for successful patient management: it helps to offer on-time curative intervention and selecting the most appropriate therapy, improves the quality of life, while contributing to reduce the economic and social burden for both patients and society. There are screening programs available for early detection of some cancers such as colorectal cancer (CRC). Unfortunately, current CRC screening tests suffer with unsatisfactory sensitivity and specificity and low compliance of targeted population. On the other hand, there are cancers with poorly performing biomarkers as in the case of hepatocellular carcinoma (HCC), or no biomarkers at all (e.g. renal cell carcinoma (RCC), which limits not only the screening options but also the diagnosis or monitoring of the disease. Therefore, there is a need for new diagnostic biomarkers and accurate technologies to enable precise detection of asymptomatic tumors in a short time, low costs and, if possible, with minimal invasiveness and risks for the patients. From this perspective, cancer specific small non-coding RNAs (small ncRNAs) circulating in body fluids such as blood serum or plasma present promising diagnostic approach. Project RNADIAGON aims at development of personal skills and knowledge of early-stage and experienced researchers working in the field of small non-coding RNA diagnostics from five European research institutions through their long-term stays at one of the world-leading ncRNAs research centers in United States and traineeships at the education center and manufacturing facilities of industrial partner developing small ncRNAs-based certified diagnostics. This research and innovation staff exchange will increase the scientific excellence and quality of related research in the EU research institutions.
Standard Operating Protocol and the article "Evaluation of miRNA detection methods for the analytical characteristics necessary for clinical utilization" (DOI: 10.2144/btn-2019-0021) prepared by MU and BioVendor have been published meeting the endpoint of WP1 „standardization of small ncRNAs detection in clinical specimens.
Regarding the WP2: Small ncRNAs in Exosomes, two Ph.D. students (UMF, UKE) and a postdoctoral fellow (UMF) visited Calin`s lab to improve their skills in extracellular vesicles biology, methodical aspects, and implications for diagnostics. Furthermore, an experimental workflow has been developed to facilitate combined microRNA, mRNA, and protein detection on a single cell level by combined in situ hybridization (ISH) and immunofluorescent staining.
Regarding the research on small ncRNAs as biomarkers in colorectal cancer performed by MU, a panel of 33 miRNAs had a significantly different expression in the group of responders vs. non-responders to neoadjuvant chemoradiotherapy (CRT; 5-fluoropyrimidines), has been detected, and is currently evaluated on the independent cohort.
Focusing on effective diagnostic biomarkers and biomarkers predictive of sorafenib response in patients affected by liver diseases, miRNA and piRNA signatures specific for patients with differential susceptibility to sorafenib have been detected. Moreover, diagnostic miRNAs were also identified by comparison of circulating miRNAs between cirrhotic vs. early hepatocellular carcinoma patients (HCC), and cirrhotic vs. advanced HCC, respectively.
PANTR1, small ncRNA, has been identified as a useful prognostic biomarker in renal cell carcinoma patients with relevant roles in apoptosis and angiogenesis in RCC tumor cells. Other ncRNAs candidates are evaluated.
Regarding the development of CE-IVD small ncRNAbased diagnostic test in CRC, a feasibility study of two technologies for ncRNA determination (the ELISA-based innovative method for the detection of miRNAs/piRNAs, and amplification-based Two-tailed qRT-PCR method) has been performed.
Regarding the WP2: Small ncRNAs in Exosomes, two Ph.D. students (UMF, UKE) and a postdoctoral fellow (UMF) visited Calin`s lab to improve their skills in extracellular vesicles biology, methodical aspects, and implications for diagnostics. Furthermore, an experimental workflow has been developed to facilitate combined microRNA, mRNA, and protein detection on a single cell level by combined in situ hybridization (ISH) and immunofluorescent staining.
Regarding the research on small ncRNAs as biomarkers in colorectal cancer performed by MU, a panel of 33 miRNAs had a significantly different expression in the group of responders vs. non-responders to neoadjuvant chemoradiotherapy (CRT; 5-fluoropyrimidines), has been detected, and is currently evaluated on the independent cohort.
Focusing on effective diagnostic biomarkers and biomarkers predictive of sorafenib response in patients affected by liver diseases, miRNA and piRNA signatures specific for patients with differential susceptibility to sorafenib have been detected. Moreover, diagnostic miRNAs were also identified by comparison of circulating miRNAs between cirrhotic vs. early hepatocellular carcinoma patients (HCC), and cirrhotic vs. advanced HCC, respectively.
PANTR1, small ncRNA, has been identified as a useful prognostic biomarker in renal cell carcinoma patients with relevant roles in apoptosis and angiogenesis in RCC tumor cells. Other ncRNAs candidates are evaluated.
Regarding the development of CE-IVD small ncRNAbased diagnostic test in CRC, a feasibility study of two technologies for ncRNA determination (the ELISA-based innovative method for the detection of miRNAs/piRNAs, and amplification-based Two-tailed qRT-PCR method) has been performed.
The results obtained so far push the boundaries of knowledge in the use of non-coding RNAs as diagnostic markers, especially in cancer, but many of the findings are also applicable in the field of non-cancer diagnostics. A significant contribution to the general application of small non-coding RNAs in clinical practice was especially the standardization of their preanalytical processing and subsequent quantification, which undoubtedly increased the robustness and repeatability of the results obtained. The possibility of wide application of non-coding RNAs in diagnostics is also significantly increased by the results obtained in their detection using ELISA platforms, which is a commonly available technology even in small diagnostic workplaces. Current problems are also the diagnosis from limited amounts of biological material, which cannot be increased without high risk to the patient, and the well-known heterogeneity of cancer, which often leads to underestimation of the diagnosis. In this area, the introduction of experimental workflows allowing combined analysis of microRNA, mRNA, and protein at the single cell level has significantly contributed to the current state of the art. The results to date on the identification of non-coding RNA diagnostic panels in rectal, renal, and liver cancers are also very promising and will be independently validated by the end of the project period, ready for clinical evaluation and subsequent translation into practice. In addition to the scientific contribution, the results of the project will have a strong clinical impact in the field of patient care and, thanks to activities aimed at disseminating knowledge among the public, should contribute to increased interest in cancer and ultimately to more effective primary and secondary prevention of cancer, leading to improved patient prognosis and ultimately to lower costs of treatment.