Diagnosing Liver Cancer Through Amplification of Mutational Extracellular mRNA: A Novel Approach

Authors

  • Dr. Ansar Hussain Chongqing Precision Medical Industry Technology Research Institute, 400000 Chongqing, China
  • Musavir Abbas Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.
  • Mehwish Kanwal Department of Horticulture and Pomology Fruit Quality & Storage of Horticulture crops lab, Anhui Agriculture University, Hefei, China.
  • Hafiz Muhammad Yasir Chongqing Precision Medical Industry Technology Research Institute, 400000 Chongqing, China.
  • Ghulam Mustafa Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.
  • Zain-Ul-Abideen Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China.
  • Ahmad Hayat Department of Zoology, The Islamia University of Bahawalpur, Punjab, Pakistan.
  • Muhammad Qasim Department of Zoology, The Islamia University of Bahawalpur, Punjab, Pakistan.
  • Yousaf Raza Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.
  • Muhammad Bilal Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.
  • Wasim Shah Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.

DOI:

https://doi.org/10.33687/hzqsk539

Keywords:

Liquid biopsies, Hepatocellular carcinoma (HCC), Extracellular vesicles (EVs), SCOPE platform, Biomarkers, Early cancer detection, EV-based mRNA profiling

Abstract

Hepatocellular carcinoma (HCC) remains the predominant cause of cancer-related mortality. Traditional diagnostic methodologies are invasive and exhibit limited sensitivity for early detection. Non-invasive alternatives, particularly liquid biopsies utilizing extracellular vesicles (EVs), have emerged as promising approaches. EVs contain crucial biomarkers, including mRNA, proteins, and nucleic acids. However, the limited abundance of EV mRNA in liquid biopsies has impeded its clinical application. To address this limitation, researchers have developed the Self-amplified and CRISPR-aided Operation to Profile EVs (SCOPE) platform. This innovative system integrates CRISPR-Cas13 for RNA target identification with replication and signal amplification, achieving subattomolar detection sensitivity. SCOPE offers high specificity with single-nucleotide resolution in a single-step assay. Investigators have validated probes targeting key mutations in KRAS, BRAF, EGFR, and IDH1 genes and developed an automated device for multi-sample analysis. SCOPE has demonstrated efficacy in identifying early-stage lung cancer in animal models, monitoring tumor mutational burden in colorectal cancer, and classifying glioblastoma patients. In HCC, EV mRNA exhibits potential for non-invasive detection of recurrence and monitoring disease progression. Current studies indicate that EV-based mRNA profiling holds significant promise for early detection, treatment monitoring, and recurrence prediction in HCC, offering valuable clinical applications. The integration of advanced platforms such as SCOPE with EV analysis could transform liquid biopsies in oncology, providing a rapid, highly sensitive, and non-invasive method for cancer detection and management.

Downloads

Download data is not yet available.

Author Biographies

  • Musavir Abbas, Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.

    Anhui Province Biomedical Sciences and Health Laboratory, First Affiliated Hospital of USTC, Hefei National Laboratory for Physical Sciences at Microscale, the CAS Key Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China; Division of Reproduction and Genetics.

  • Mehwish Kanwal, Department of Horticulture and Pomology Fruit Quality & Storage of Horticulture crops lab, Anhui Agriculture University, Hefei, China.
    Department of Horticulture and Pomology Fruit Quality & Storage of Horticulture crops lab, Anhui Agriculture University, Hefei, China.
  • Hafiz Muhammad Yasir, Chongqing Precision Medical Industry Technology Research Institute, 400000 Chongqing, China.
    Chongqing Precision Medical Industry Technology Research Institute, 400000 Chongqing, China.
  • Ghulam Mustafa, Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.

    Anhui Province Biomedical Sciences and Health Laboratory, First Affiliated Hospital of USTC, Hefei National Laboratory for Physical Sciences at Microscale, the CAS Key Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China; Division of Reproduction and Genetics.

  • Zain-Ul-Abideen, Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China.
    Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China.
  • Ahmad Hayat, Department of Zoology, The Islamia University of Bahawalpur, Punjab, Pakistan.
    Department of Zoology, The Islamia University of Bahawalpur, Punjab, Pakistan.
  • Muhammad Qasim, Department of Zoology, The Islamia University of Bahawalpur, Punjab, Pakistan.

    Department of Zoology, The Islamia University of Bahawalpur, Punjab, Pakistan.

  • Yousaf Raza, Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.

    Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.

  • Muhammad Bilal, Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.

    Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.

  • Wasim Shah, Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China.

    Laboratory of Innate Immunity and Chronic Disease, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230027, China; Division of Reproduction and Genetics.

References

Aalberts, M., van Dissel-Emiliani, F. M., van Adrichem, N. P., van Wijnen, M., Wauben, M. H., Stout, T. A., & Stoorvogel, W. (2012). Identification of distinct populations of prostasomes that differentially express prostate stem cell antigen, annexin A1, and GLIPR2 in humans. Biology of reproduction, 86(3), 82, 81-88.

Abudayyeh, O. O., & Gootenberg, J. S. (2021). CRISPR diagnostics. Science, 372(6545), 914-915.

Arraud, N., Gounou, C., Linares, R., & Brisson, A. R. (2015). A simple flow cytometry method improves the detection of phosphatidylserine‐exposing extracellular vesicles. Journal of Thrombosis and Haemostasis, 13(2), 237-247.

Arraud, N., Linares, R., Tan, S., Gounou, C., Pasquet, J. M., Mornet, S., & Brisson, A. R. (2014). Extracellular vesicles from blood plasma: determination of their morphology, size, phenotype and concentration. Journal of Thrombosis and Haemostasis, 12(5), 614-627.

Block, T., Zezulinski, D., Kaplan, D. E., Lu, J., Zanine, S., Zhan, T., Doria, C., & Sayeed, A. (2022). Circulating messenger RNA variants as a potential biomarker for surveillance of hepatocellular carcinoma. Frontiers in oncology, 12, 963641.

Chandler, W., Yeung, W., & Tait, J. (2011). A new microparticle size calibration standard for use in measuring smaller microparticles using a new flow cytometer. Journal of Thrombosis and Haemostasis, 9(6), 1216-1224.

Cheng, L., Zhang, L., Wang, X., Wang, Y., Yu, J., Li, M., Ma, Z., Ho, P. C.-L., Chen, X., & Wang, L. (2024). Extracellular vesicles in the HCC microenvironment: Implications for therapy and biomarkers. Pharmacological research, 107419.

Daane, M., Vigneri, P., Sieuwerts, A., Dits, N., van Royen, M., Jenster, G., Sleijfer, S., Lolkema, M., Martens, J., & PHM, J. M. (2022). Detection of tumor-derived extracellular vesicles in plasma from patients with solid cancer. Evaluation of Liquid Biopsy Workflows and ESR1 Alterations in Breast Cancer, 21, 85.

de Lima, L. T. F., Broszczak, D., Zhang, X., Bridle, K., Crawford, D., & Punyadeera, C. (2020). The use of minimally invasive biomarkers for the diagnosis and prognosis of hepatocellular carcinoma. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, 1874(2), 188451.

Dixson, A. C., Dawson, T. R., Di Vizio, D., & Weaver, A. M. (2023). Context-specific regulation of extracellular vesicle biogenesis and cargo selection. Nature reviews Molecular cell biology, 24(7), 454-476.

Everaert, C. (2020). Development of extracellular RNA sequencing technologies and applications for cancer diagnosis and monitoring Ghent University].

Ferlay, J., Colombet, M., Soerjomataram, I., Mathers, C., Parkin, D. M., Piñeros, M., Znaor, A., & Bray, F. (2019). Estimating the global cancer incidence and mortality in 2018: GLOBOCAN sources and methods. International journal of cancer, 144(8), 1941-1953.

György, B., Szabó, T. G., Pásztói, M., Pál, Z., Misják, P., Aradi, B., László, V., Pállinger, E., Pap, E., & Kittel, A. (2011). Membrane vesicles, current state-of-the-art: emerging role of extracellular vesicles. Cellular and molecular life sciences, 68, 2667-2688.

Heijnen, H. F., Schiel, A. E., Fijnheer, R., Geuze, H. J., & Sixma, J. J. (1999). Activated Platelets Release Two Types of Membrane Vesicles: Microvesicles by Surface Shedding and Exosomes Derived From Exocytosis of Multivesicular Bodies and-Granules. Blood, The Journal of the American Society of Hematology, 94(11), 3791-3799.

Horstman, L. L., Jy, W., Jimenez, J. J., Bidot, C., & Ahn, Y. S. (2004). New horizons in the analysis of circulating cell-derived microparticles. The Keio journal of medicine, 53(4), 210-230.

Ignatiadis, M., Sledge, G. W., & Jeffrey, S. S. (2021). Liquid biopsy enters the clinic—implementation issues and future challenges. Nature reviews Clinical oncology, 18(5), 297-312.

Jo, A., Green, A., Medina, J. E., Iyer, S., Ohman, A. W., McCarthy, E. T., Reinhardt, F., Gerton, T., Demehin, D., & Mishra, R. (2023). Inaugurating High‐Throughput Profiling of Extracellular Vesicles for Earlier Ovarian Cancer Detection (Adv. Sci. 27/2023). Advanced Science, 10(27), 2370180.

Kaminski, M. M., Abudayyeh, O. O., Gootenberg, J. S., Zhang, F., & Collins, J. J. (2021). CRISPR-based diagnostics. Nature Biomedical Engineering, 5(7), 643-656.

Kebschull, J. M., & Zador, A. M. (2015). Sources of PCR-induced distortions in high-throughput sequencing data sets. Nucleic acids research, 43(21), e143-e143.

Killingsworth, B., Welsh, J. A., & Jones, J. C. (2021). EV translational horizons as viewed across the complex landscape of liquid biopsies. Frontiers in cell and developmental biology, 9, 556837.

Kim, D. Y., & Han, K.-H. (2012). Epidemiology and surveillance of hepatocellular carcinoma. Liver cancer, 1(1), 2-14.

Lacroix, R., Judicone, C., Poncelet, P., Robert, S., Arnaud, L., Sampol, J., & DIGNAT‐GEORGE, F. (2012). Impact of pre‐analytical parameters on the measurement of circulating microparticles: towards standardization of protocol. Journal of Thrombosis and Haemostasis, 10(3), 437-446.

Llovet, J. M., Kelley, R. K., Villanueva, A., Singal, A. G., Pikarsky, E., Roayaie, S., Lencioni, R., Koike, K., Zucman-Rossi, J., & Finn, R. S. (2021). Hepatocellular carcinoma (primer). Nature Reviews: Disease Primers, 7(1), 6.

Makrygianni, E. A., & Chrousos, G. P. (2023). Extracellular vesicles and the stress system. Neuroendocrinology, 113(2), 120-167.

Martins, I., Ribeiro, I. P., Jorge, J., Gonçalves, A. C., Sarmento-Ribeiro, A. B., Melo, J. B., & Carreira, I. M. (2021). Liquid biopsies: applications for cancer diagnosis and monitoring. Genes, 12(3), 349.

Moldogazieva, N. T., Zavadskiy, S. P., & Terentiev, A. A. (2021). Genomic landscape of liquid biopsy for hepatocellular carcinoma personalized medicine. Cancer Genomics & Proteomics, 18(3 Suppl), 369-383.

Noerholm, M., Balaj, L., Limperg, T., Salehi, A., Zhu, L. D., Hochberg, F. H., Breakefield, X. O., Carter, B. S., & Skog, J. (2012). RNA expression patterns in serum microvesicles from patients with glioblastoma multiforme and controls. BMC cancer, 12, 1-11.

Nomura, S., Shouzu, A., Taomoto, K., Togane, Y., Goto, S., Ozaki, Y., Uchiyama, S., & Ikeda, Y. (2009). Assessment of an ELISA kit for platelet-derived microparticles by joint research at many institutes in Japan. Journal of atherosclerosis and thrombosis, 16(6), 878-887.

Pantel, K., & Alix-Panabières, C. (2019). Liquid biopsy and minimal residual disease—latest advances and implications for cure. Nature reviews Clinical oncology, 16(7), 409-424.

Parikh, A. R., Leshchiner, I., Elagina, L., Goyal, L., Levovitz, C., Siravegna, G., Livitz, D., Rhrissorrakrai, K., Martin, E. E., & Van Seventer, E. E. (2019). Liquid versus tissue biopsy for detecting acquired resistance and tumor heterogeneity in gastrointestinal cancers. Nature medicine, 25(9), 1415-1421.

Park, J., Park, J. S., Huang, C.-H., Jo, A., Cook, K., Wang, R., Lin, H.-Y., Van Deun, J., Li, H., & Min, J. (2021). An integrated magneto-electrochemical device for the rapid profiling of tumour extracellular vesicles from blood plasma. Nature Biomedical Engineering, 5(7), 678-689.

Pickar-Oliver, A., & Gersbach, C. A. (2019). The next generation of CRISPR–Cas technologies and applications. Nature reviews Molecular cell biology, 20(8), 490-507.

Potapov, V., & Ong, J. L. (2017). Examining sources of error in PCR by single-molecule sequencing. PLoS One, 12(1), e0169774.

Prince, D., Liu, K., Xu, W., Chen, M., Sun, J.-Y., Lu, X.-J., & Ji, J. (2020). Management of patients with intermediate stage hepatocellular carcinoma. Therapeutic Advances in Medical Oncology, 12, 1758835920970840.

Revenfeld, A. L. S., Bæk, R., Nielsen, M. H., Stensballe, A., Varming, K., & Jørgensen, M. (2014). Diagnostic and prognostic potential of extracellular vesicles in peripheral blood. Clinical therapeutics, 36(6), 830-846.

Shao, H., Im, H., Castro, C. M., Breakefield, X., Weissleder, R., & Lee, H. (2018). New technologies for analysis of extracellular vesicles. Chemical reviews, 118(4), 1917-1950.

Singal, A. G., Pillai, A., & Tiro, J. (2014). Early detection, curative treatment, and survival rates for hepatocellular carcinoma surveillance in patients with cirrhosis: a meta-analysis. PLoS medicine, 11(4), e1001624.

Skog, J., Würdinger, T., Van Rijn, S., Meijer, D. H., Gainche, L., Curry Jr, W. T., Carter, B. S., Krichevsky, A. M., & Breakefield, X. O. (2008). Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers. Nature cell biology, 10(12), 1470-1476.

Song, J., Cho, M. H., Cho, H., Song, Y., Lee, S. W., Nam, H. C., Yoon, T. H., Shin, J. C., Hong, J.-S., & Kim, Y. (2024). Amplifying mutational profiling of extracellular vesicle mRNA with SCOPE. Nature Biotechnology, 1-11.

van de Haar, J., Ma, X., Ooft, S. N., van der Helm, P. W., Hoes, L. R., Mainardi, S., Pinato, D. J., Sun, K., Salvatore, L., & Tortora, G. (2023). Codon-specific KRAS mutations predict survival benefit of trifluridine/tipiracil in metastatic colorectal cancer. Nature medicine, 29(3), 605-614.

Van der Pol, E., Böing, A. N., Harrison, P., Sturk, A., & Nieuwland, R. (2012). Classification, functions, and clinical relevance of extracellular vesicles. Pharmacological reviews, 64(3), 676-705.

Van Der Pol, E., Van Gemert, M., Sturk, A., Nieuwland, R., & Van Leeuwen, T. (2012). Single vs. swarm detection of microparticles and exosomes by flow cytometry. Journal of Thrombosis and Haemostasis, 10(5), 919-930.

Van Der Vlist, E. J., Nolte-'t Hoen, E. N., Stoorvogel, W., Arkesteijn, G. J., & Wauben, M. H. (2012). Fluorescent labeling of nano-sized vesicles released by cells and subsequent quantitative and qualitative analysis by high-resolution flow cytometry. Nature protocols, 7(7), 1311-1326.

Vitale, S. R., Helmijr, J. A., Gerritsen, M., Coban, H., van Dessel, L. F., Beije, N., van der Vlugt-Daane, M., Vigneri, P., Sieuwerts, A. M., & Dits, N. (2021). Detection of tumor-derived extracellular vesicles in plasma from patients with solid cancer. BMC cancer, 21, 1-17.

Wei, Z., Batagov, A. O., Schinelli, S., Wang, J., Wang, Y., El Fatimy, R., Rabinovsky, R., Balaj, L., Chen, C. C., & Hochberg, F. (2017). Coding and noncoding landscape of extracellular RNA released by human glioma stem cells. Nature communications, 8(1), 1145.

Witwer, K. W., Buzás, E. I., Bemis, L. T., Bora, A., Lässer, C., Lötvall, J., Nolte-‘t Hoen, E. N., Piper, M. G., Sivaraman, S., & Skog, J. (2013). Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. Journal of extracellular vesicles, 2(1), 20360.

Wolf, P. (1967). The nature and significance of platelet products in human plasma. British journal of haematology, 13(3), 269-288.

Zarà, M., Guidetti, G. F., Camera, M., Canobbio, I., Amadio, P., Torti, M., Tremoli, E., & Barbieri, S. S. (2019). Biology and role of extracellular vesicles (EVs) in the pathogenesis of thrombosis. International journal of molecular sciences, 20(11), 2840.

Zonneveld, M. I., Brisson, A. R., van Herwijnen, M. J., Tan, S., van de Lest, C. H., Redegeld, F. A., Garssen, J., Wauben, M. H., & Nolte-'t Hoen, E. N. (2014). Recovery of extracellular vesicles from human breast milk is influenced by sample collection and vesicle isolation procedures. Journal of extracellular vesicles, 3(1), 24215.

Ricos Biology Journal Vol. 3, No. 3

Downloads

Published

16-03-2025 — Updated on 15-04-2025

Data Availability Statement

The authors confirm that the data presented in the article and additional data can be provided by the corresponding author upon request.

How to Cite

Diagnosing Liver Cancer Through Amplification of Mutational Extracellular mRNA: A Novel Approach. (2025). Ricos Biology, 3(3), 52-65. https://doi.org/10.33687/hzqsk539

Similar Articles

21-30 of 66

You may also start an advanced similarity search for this article.