CRISPR技术在肿瘤学精准诊疗领域的研究进展
Research Progress of CRISPR Technology for Accurate Diagnosis and Treatment in Oncology
摘要: CRISPR基因编辑技术通过精准定位和编辑特定基因序列,可以识别与肿瘤细胞生长、增殖、分化、侵袭和耐药性相关的基因,简化并高效推进肿瘤的分子机制研究,被广泛地应用于精准肿瘤学研究当中,包括用于检测肿瘤源性的生物标志物、肿瘤异质性分析、工程化改造免疫细胞,以及构建肿瘤疾病模型等。尽管CRISPR/Cas基因编辑技术在肿瘤诊疗领域展现出了巨大潜力,但仍需面对脱靶效应、递送效率低等瓶颈。文中就目前CRISPR技术在肿瘤学精准诊疗领域的最新进展进行综述。
Abstract: CRISPR gene editing technology can identify genes associated with tumor cell growth, proliferation, differentiation, invasion, and drug resistance by precisely targeting and editing specific gene sequences, simplifying and efficiently advancing the study of molecular mechanisms of tumors, and has been widely used in precision oncology research, including for detecting tumor-derived biomarkers, analyzing tumor heterogeneity, engineering modified immune cells, and constructing tumor disease models, etc. Although CRISPR/Cas gene editing technology has shown great potential in the field of tumor diagnosis and treatment, it still needs to face bottlenecks such as off-target effects and low delivery efficiency. The paper provides an overview of the current state-of-the-art of CRISPR technology in the field of accurate diagnosis and treatment in oncology.
文章引用:邓继婷, 韩媛媛, 肖威, 曹东林. CRISPR技术在肿瘤学精准诊疗领域的研究进展[J]. 临床医学进展, 2025, 15(1): 787-796. https://doi.org/10.12677/acm.2025.151106

参考文献

[1] Zhang, Y.F. and Lu, M. (2024) Numerical Simulation of Thermal Therapy for Melanoma in Mice.
https://pmc.ncbi.nlm.nih.gov/articles/PMC11273475/pdf/bioengineering-11-00694.pdf
[2] Liu, L., Kuang, L. and Ji, Y. (2020) Multimodal MRI Brain Tumor Image Segmentation Using Sparse Subspace Clustering Algorithm. Computational and Mathematical Methods in Medicine, 2020, 1-13. [Google Scholar] [CrossRef] [PubMed]
[3] Yoshizawa, N., Yamada, M., et al. (20240 Nephritis-Associated Plasmin Receptor (NAPlr): An Essential Inducer of C3-Dominant Glomerular Injury and a Potential Key Diagnostic Biomarker of Infection-Related Glomerulonephritis (IRGN).
https://pmc.ncbi.nlm.nih.gov/articles/PMC9456382/pdf/ijms-23-09974.pdf
[4] Barkley, D., Moncada, R., Pour, M., Liberman, D.A., Dryg, I., Werba, G., et al. (2022) Cancer Cell States Recur across Tumor Types and Form Specific Interactions with the Tumor Microenvironment. Nature Genetics, 54, 1192-1201. [Google Scholar] [CrossRef] [PubMed]
[5] Tang, Y., Gao, L., Feng, W., Guo, C., Yang, Q., Li, F., et al. (2021) The CRISPR-Cas Toolbox for Analytical and Diagnostic Assay Development. Chemical Society Reviews, 50, 11844-11869. [Google Scholar] [CrossRef] [PubMed]
[6] Bravo, J.P.K., Liu, M., Hibshman, G.N., Dangerfield, T.L., Jung, K., McCool, R.S., et al. (2022) Structural Basis for Mismatch Surveillance by CRISPR-Cas9. Nature, 603, 343-347. [Google Scholar] [CrossRef] [PubMed]
[7] Bhatia, S., Pooja, and Yadav, S.K. (2023) CRISPR-Cas for Genome Editing: Classification, Mechanism, Designing and Applications. International Journal of Biological Macromolecules, 238, Article 124054. [Google Scholar] [CrossRef] [PubMed]
[8] Knott, G.J. and Doudna, J.A. (2018) CRISPR-Cas Guides the Future of Genetic Engineering. Science, 361, 866-869. [Google Scholar] [CrossRef] [PubMed]
[9] Gier, R.A., Budinich, K.A., Evitt, N.H., Cao, Z., Freilich, E.S., Chen, Q., et al. (2020) High-Performance CRISPR-Cas12A Genome Editing for Combinatorial Genetic Screening. Nature Communications, 11, Article No. 3455. [Google Scholar] [CrossRef] [PubMed]
[10] Yang, H. and Patel, D.J. (2024) Structures, Mechanisms and Applications of RNA-Centric CRISPR-Cas13. Nature Chemical Biology, 20, 673-688. [Google Scholar] [CrossRef] [PubMed]
[11] Biagioni, A., Chillà, A., Del Rosso, M., Fibbi, G., Scavone, F., Andreucci, E., et al. (2021) CRISPR/Cas9 UPAR Gene Knockout Results in Tumor Growth Inhibition, EGFR Downregulation and Induction of Stemness Markers in Melanoma and Colon Carcinoma Cell Lines. Frontiers in Oncology, 11, Article 663225. [Google Scholar] [CrossRef] [PubMed]
[12] Singh, A., Irfan, H., Fatima, E., Nazir, Z., Verma, A. and Akilimali, A. (2024) Revolutionary Breakthrough: FDA Approves CASGEVY, the First CRISPR/Cas9 Gene Therapy for Sickle Cell Disease. Annals of Medicine & Surgery, 86, 4555-4559. [Google Scholar] [CrossRef] [PubMed]
[13] Yoshimi, K., Takeshita, K., Kodera, N., Shibumura, S., Yamauchi, Y., Omatsu, M., et al. (2022) Dynamic Mechanisms of CRISPR Interference by Escherichia Coli CRISPR-Cas3. Nature Communications, 13, Article No. 4917. [Google Scholar] [CrossRef] [PubMed]
[14] Stella, G. and Marraffini, L. (2024) Type III CRISPR-Cas: Beyond the Cas10 Effector Complex. Trends in Biochemical Sciences, 49, 28-37. [Google Scholar] [CrossRef] [PubMed]
[15] Makarova, K.S., Wolf, Y.I., Iranzo, J., Shmakov, S.A., Alkhnbashi, O.S., Brouns, S.J.J., et al. (2019) Evolutionary Classification of CRISPR-Cas Systems: A Burst of Class 2 and Derived Variants. Nature Reviews Microbiology, 18, 67-83. [Google Scholar] [CrossRef] [PubMed]
[16] Yan, W.X., Hunnewell, P., Alfonse, L.E., Carte, J.M., Keston-Smith, E., Sothiselvam, S., et al. (2019) Functionally Diverse Type V CRISPR-Cas Systems. Science, 363, 88-91. [Google Scholar] [CrossRef] [PubMed]
[17] Hillary, V.E. and Ceasar, S.A. (2022) A Review on the Mechanism and Applications of CRISPR/Cas9/Cas12/Cas13/Cas14 Proteins Utilized for Genome Engineering. Molecular Biotechnology, 65, 311-325. [Google Scholar] [CrossRef] [PubMed]
[18] Zhou, Y., Tao, L., Qiu, J., Xu, J., Yang, X., Zhang, Y., et al. (2024) Tumor Biomarkers for Diagnosis, Prognosis and Targeted Therapy. Signal Transduction and Targeted Therapy, 9, Article No. 132. [Google Scholar] [CrossRef] [PubMed]
[19] Wang, M., Chen, K., Wu, Q., Peng, R., Zhang, R. and Li, J. (2019) RCasFISH: CRISPR/dCas9-Mediated in Situ Imaging of mRNA Transcripts in Fixed Cells and Tissues. Analytical Chemistry, 92, 2468-2475. [Google Scholar] [CrossRef] [PubMed]
[20] Shaik, M.R., Sagar, P.R., et al. (2023) Liquid Biopsy in Hepatocellular Carcinoma: The Significance of Circulating Tumor Cells in Diagnosis, Prognosis, and Treatment Monitoring. International Journal of Molecular Sciences, 24, Article 10644.
[21] Sivapalan, L., Murray, J.C., Canzoniero, J.V., et al. (2023) Liquid Biopsy Approaches to Capture Tumor Evolution and Clinical Outcomes during Cancer Immunotherapy. The Journal for ImmunoTherapy of Cancer, 11, e005924.
[22] Zhang, Q., Gao, X., Ho, Y., Liu, M., Han, Y., Li, D., et al. (2024) Controllable Assembly of a Quantum Dot-Based Aptasensor Guided by CRISPR/Cas12A for Direct Measurement of Circulating Tumor Cells in Human Blood. Nano Letters, 24, 2360-2368. [Google Scholar] [CrossRef] [PubMed]
[23] Guan, X.T., Zhao, J.R., et al. (2024) CRISPR/Cas12A and Aptamer-Chemiluminescence Based Analysis for the Relative Abundance Determination of Tumor-Related Protein Positive Exosomes for Breast Cancer Diagnosis. Biosensors and Bioelectronics, 259, Article 116380.
[24] Guan, L., Peng, J., Liu, T., Huang, S., Yang, Y., Wang, X., et al. (2023) Ultrasensitive Mirna Detection Based on Magnetic Upconversion Nanoparticle Enhancement and CRISPR/Cas13A-Driven Signal Amplification. Analytical Chemistry, 95, 17708-17715. [Google Scholar] [CrossRef] [PubMed]
[25] Zhang, Y., Wei, X., Teng, X. and Chen, G. (2023) P53 Aberration and TFE3 Gene Amplification May Be Predictors of Adverse Prognosis in Epithelioid Angiomyolipoma of the Kidney. Diagnostic Pathology, 18, Article No. 14. [Google Scholar] [CrossRef] [PubMed]
[26] Wang, T., Ding, K., Wang, X., Wang, Z., Liu, G., Zang, Y., et al. (2024) Dual Amplification Dynamic DNA Network System for CRISPR/Cas12A Based P53 Gene Detection. Analytica Chimica Acta, 1321, Article 343048. [Google Scholar] [CrossRef] [PubMed]
[27] Zhang, H., Gao, H., Mu, W., Que, L., Gu, X., Rong, S., et al. (2024) Electrochemical-Fluorescent Bimodal Biosensor Based on Dual CRISPR-Cas12A Multiple Cascade Amplification for ctDNA Detection. Analytical Chemistry, 34, 14028-14035. [Google Scholar] [CrossRef] [PubMed]
[28] Martinez, S., Wu, S., Geuenich, M., Malik, A., Weber, R., Woo, T., et al. (2024) In Vivo CRISPR Screens Reveal SCAF1 and USP15 as Drivers of Pancreatic Cancer. Nature Communications, 15, Article No. 5266. [Google Scholar] [CrossRef] [PubMed]
[29] Alayoubi, A.M., Khawaji, Z.Y., Mohammed, M.A. and Mercier, F.E. (2023) CRISPR-Cas9 System: A Novel and Promising Era of Genotherapy for Beta-Hemoglobinopathies, Hematological Malignancy, and Hemophilia. Annals of Hematology, 103, 1805-1817. [Google Scholar] [CrossRef] [PubMed]
[30] Boumelha, J., de Castro, A., Bah, N., Cha, H., de Carné Trécesson, S., Rana, S., et al. (2024) CRISPR-Cas9 Screening Identifies KRAS-Induced COX2 as a Driver of Immunotherapy Resistance in Lung Cancer. Cancer Research, 84, 2231-2246. [Google Scholar] [CrossRef] [PubMed]
[31] Barkley, D., Rao, A., Pour, M., França, G.S. and Yanai, I. (2021) Cancer Cell States and Emergent Properties of the Dynamic Tumor System. Genome Research, 31, 1719-1727. [Google Scholar] [CrossRef] [PubMed]
[32] Guo, T., Chen, G., Li, X., Wang, M., Liu, K., Yang, X., et al. (2023) Small Extrachromosomal Circular DNA Harboring Targeted Tumor Suppressor Gene Mutations Supports Intratumor Heterogeneity in Mouse Liver Cancer Induced by Multiplexed CRISPR/Cas9. Genome Medicine, 15, Article No. 80. [Google Scholar] [CrossRef] [PubMed]
[33] Wu, M., Zhang, T., Gao, C., Zhao, T., Wang, L. and Sun, G. (2024) Assessing of Case-Cohort Design: A Case Study for Breast Cancer Patients in Xinjiang, China. Frontiers in Oncology, 14, Article 1306255. [Google Scholar] [CrossRef] [PubMed]
[34] Chamberlain, C.A., Bennett, E.P., Kverneland, A.H., Svane, I.M., Donia, M. and Met, Ö. (2022) Highly Efficient Pd-1-Targeted CRISPR-Cas9 for Tumor-Infiltrating Lymphocyte-Based Adoptive T Cell Therapy. Molecular Therapy Oncolytics, 24, 417-428.
[35] Pandey, S., Lee, M., Lim, J., Park, S., Choung, Y., Kim, J.E., et al. (2023) SMO-CRISPR-Mediated Apoptosis in CD133-Targeted Cancer Stem Cells and Tumor Growth Inhibition. Journal of Controlled Release, 357, 94-108. [Google Scholar] [CrossRef] [PubMed]
[36] Wang, L., Liu, C., Wang, X., Ma, S., Liu, F., Zhang, Y., et al. (2023) Tumor-Specific Activated Nano-Domino-CRISPR to Amplify Intrinsic Oxidative and Activate Endogenous Apoptosis for Spatiotemporally Specific Therapy. Biomaterials, 295, Article 122056. [Google Scholar] [CrossRef] [PubMed]
[37] Wang, S., Gao, C., Zheng, Y., Yi, L., Lu, J., Huang, X., et al. (2022) Current Applications and Future Perspective of CRISPR/Cas9 Gene Editing in Cancer. Molecular Cancer, 21, Article No. 57. [Google Scholar] [CrossRef] [PubMed]
[38] Morton, L.T., Reijmers, R.M., Wouters, A.K., Kweekel, C., Remst, D.F.G., Pothast, C.R., et al. (2020) Simultaneous Deletion of Endogenous Tcrαβ for TCR Gene Therapy Creates an Improved and Safe Cellular Therapeutic. Molecular Therapy, 28, 64-74. [Google Scholar] [CrossRef] [PubMed]
[39] Stadtmauer, E.A., Fraietta, J.A., Davis, M.M., et al. (2020) CRISPR-Engineered T Cells in Patients with Refractory Cancer. Science, 367, eaba7365.
[40] Nie, D., Guo, T., Yue, M., Li, W., Zong, X., Zhu, Y., et al. (2022) Research Progress on Nanoparticles-Based CRISPR/Cas9 System for Targeted Therapy of Tumors. Biomolecules, 12, Article 1239. [Google Scholar] [CrossRef] [PubMed]
[41] He, C., Jaffar Ali, D., Li, Y., Zhu, Y., Sun, B. and Xiao, Z. (2020) Engineering of HN3 Increases the Tumor Targeting Specificity of Exosomes and Upgrade the Anti-Tumor Effect of Sorafenib on Huh-7 Cells. Peer Journal, 8, e9524. [Google Scholar] [CrossRef] [PubMed]
[42] Ye, Y., Zhang, X., Xie, F., Xu, B., Xie, P., Yang, T., et al. (2020) An Engineered Exosome for Delivering sgRNA: Cas9 Ribonucleoprotein Complex and Genome Editing in Recipient Cells. Biomaterials Science, 8, 2966-2976. [Google Scholar] [CrossRef] [PubMed]
[43] Yu, D., Li, Y., Wang, M., Gu, J., Xu, W., Cai, H., et al. (2022) Exosomes as a New Frontier of Cancer Liquid Biopsy. Molecular Cancer, 21, Article No. 56. [Google Scholar] [CrossRef] [PubMed]
[44] Ipsen, M.B., Sørensen, E.M.G., Thomsen, E.A., Weiss, S., Haldrup, J., Dalby, A., et al. (2022) A Genome-Wide Crispr-Cas9 Knockout Screen Identifies Novel PARP Inhibitor Resistance Genes in Prostate Cancer. Oncogene, 41, 4271-4281. [Google Scholar] [CrossRef] [PubMed]
[45] Ludwig, M., Birkeland, A., Smith, J., et al. (2024) A Genome Wide CRISPR Profiling Approach Identifies Mechanisms of Cisplatin Resistance in Head and Neck Squamous Cell Carcinoma.
https://www.researchsquare.com/article/rs-3922565/v1
[46] Deng, L., Yang, L., Zhu, S., Li, M., Wang, Y., Cao, X., et al. (2023) Identifying CDC7 as a Synergistic Target of Chemotherapy in Resistant Small-Cell Lung Cancer via CRISPR/Cas9 Screening. Cell Death Discovery, 9, Article No. 40. [Google Scholar] [CrossRef] [PubMed]
[47] Fazeli, Z., Rajabibazl, M., Faramarzi, S., et al. (2021) Correlation of TCF4, GSK, TERT and TERC Expressions with Proliferation Potential of Early and Late Culture of Human Peripheral Blood Mesenchymal Stem Cells. Cell Journal (Yakhteh), 22, 431-436.
[48] George, S.L., Lorenzi, F., King, D., Hartlieb, S., Campbell, J., Pemberton, H., et al. (2020) Therapeutic Vulnerabilities in the DNA Damage Response for the Treatment of ATRX Mutant Neuroblastoma. E Bio Medicine, 59, Article 102971. [Google Scholar] [CrossRef] [PubMed]
[49] Hansen, S.L., Larsen, H.L., Pikkupeura, L.M., Maciag, G., Guiu, J., Müller, I., et al. (2023) An Organoid-Based Crispr-Cas9 Screen for Regulators of Intestinal Epithelial Maturation and Cell Fate. Science Advances, 9, eadg4055. [Google Scholar] [CrossRef] [PubMed]
[50] Knott, G.J. and Doudna, J.A. (2018) CRISPR Cas Guides the Future of Genetic Engineering. Science, 361, 866-869.
[51] Kawamata, M., Suzuki, H.I., Kimura, R. and Suzuki, A. (2023) Optimization of Cas9 Activity through the Addition of Cytosine Extensions to Single-Guide RNAs. Nature Biomedical Engineering, 7, 672-691. [Google Scholar] [CrossRef] [PubMed]
[52] Zuo, Z., Babu, K., Ganguly, C., Zolekar, A., Newsom, S., Rajan, R., et al. (2022) Rational Engineering of CRISPR-Cas9 Nuclease to Attenuate Position-Dependent Off-Target Effects. The CRISPR Journal, 5, 329-340. [Google Scholar] [CrossRef] [PubMed]
[53] Kitada, T., DiAndreth, B., Teague, B. and Weiss, R. (2018) Programming Gene and Engineered-Cell Therapies with Synthetic Biology. Science, 359, eaad1067. [Google Scholar] [CrossRef] [PubMed]
[54] Zheng, Y., Li, Y., Zhou, K., Li, T., VanDusen, N.J. and Hua, Y. (2024) Precise Genome-Editing in Human Diseases: Mechanisms, Strategies and Applications. Signal Transduction and Targeted Therapy, 9, Article No. 47. [Google Scholar] [CrossRef] [PubMed]