CRISPR in Targeted Therapy and Adoptive T Cell Immunotherapy for Hepatocellular Carcinoma

Figures & data

Table 1 The Summary of Studies Using CRISPR Tools to Target HCC

CRISPR Method	Target Gene	HCC Model	Delivery route and method	Effects of CRISPR	Ref.
Cas9 nuclease	Survivin	Orthotopic liver tumor	IV injection of lactose-derived branched cationic biopolymer carrying pCas9-gRNA and sorafenib	CRISPR efficiently inhibited tumor growth. A synergistic effect was observed for the CRISPR + sorafenib combination.	[37]
Cas9 nuclease	Survivin	Orthotopic liver tumor	IV injection of nanocomplexes with cationic shell and heparin nanoparticle core carrying the pCas9-gRNA and sorafenib	Tumor growth substantially decreased. A synergistic effect was observed for the CRISPR + sorafenib combination.	[38]
Cas13a, RNA-targeting	TERT EZH2 RelA	Subcutaneous xenograft	IV injection of AAVs coding for Cas13 and crRNAs. The Cas13 was expressed from a tumor-specific promoter.	The apoptosis rate of tumor cells increased, and cell growth decreased significantly. Cas13a expression was shown to be specific for the tumor tissues. The weight, volume, and markers of tumors decreased significantly.	[39]
Cas9 nuclease	EGFR	Subcutaneous xenograft	IV injection of aptamer-coated hollow mesoporous silica nanoparticles enabled the co-delivery of pCas9-gRNA and sorafenib	CRISPR efficiently inhibited tumor growth. A synergistic effect was observed for the CRISPR + sorafenib combination.	[40]
Cas9 nuclease	HIF‑1α	Subcutaneous xenograft and orthotopic liver tumor	In the xenograft model, intratumoral injection of lentivirus coding Cas9 and gRNA was performed. Orthotopic tumor-bearing mice were generated using lentivirally injected HCC cells.	In the xenograft model, CRISPR caused a substantial decrease in HIF‑1α in tumor cells. In the orthotopic tumor model, a synergistic effect was observed with the combination of CRISPR + hepatic artery ligation, which mimics transarterial chemoembolization (TACE). The combination therapy reduced tumor growth and angiogenesis, induced apoptosis, and significantly prolonged the survival of HCC-bearing mice.	[41]
Cas9 nuclease	VEGFR2	Xenograft tumor	IV injection of pH-responsive chitosan-based nanoparticles enabled the co-delivery of pCas9-gRNA and paclitaxel	CRISPR efficiently inhibited tumor growth. A synergistic effect was observed for the CRISPR + paclitaxel combination, which provided the best antitumor effect among all groups.	[42]
Cas9 nuclease	WNT10B	Subcutaneous xenograft; tumor organoids are also used ex vivo	IV injection of aptamer-coated extracellular vesicles carrying Cas9-gRNA RNP complexes	CRISPR halted tumor development in vivo, and widespread apoptosis was observed in tumor organoids.	[43]
Cas9 nuclease	NFE2L2	Subcutaneous xenograft	IV injection of lipid nanoparticles carrying Cas9-gRNA RNP complexes and hematoporphyrin monomethyl ether (HMME)	Ultrasound-controlled nanoparticles decreased tumor growth substantially. A synergistic effect was observed for the CRISPR + sonodynamic therapy combination.	[44]
Cas9 nuclease	IQGAP1, FOXM1	Subcutaneous xenograft	IV injection of anti-GLP3 antibody-coated extracellular vesicles containing Cas9 protein and dual gRNA expression plasmid	CRISPR efficiently inhibited tumor growth. A synergistic antitumor effect was observed for the CRISPR + sorafenib combination, which was given intraperitoneally.	[45]
Cas9 nuclease	IQGAP1	Subcutaneous xenograft	Intratumoral injection of extracellular vesicles containing Cas9 protein and gRNA expression plasmid	Tumor growth was inhibited in CRISPR-treated xenograft models. A synergistic effect was observed for CRISPR + sorafenib combination in vitro.	[46]
Cas9 nuclease	KAT5	Orthotopic liver tumor	IV injection of exosomes carrying Cas9-gRNA RNP complex	CRISPR-treated mice had smaller tumors and significantly prolonged survival.	[47]
Cas9 nuclease	PLK1	Subcutaneous xenograft	Intratumoral injection of lipid nanoparticles carrying pCas9-gRNA	CRISPR efficiently induced apoptosis and inhibited tumor growth.	[48]
Cas9 nuclease	ADSL	Mice with IV–injected tumor-initiating vectors for c-Myc or CTNNB1-N90 overexpression, or TP53 knockout	Mice were also IV–injected with CRISPR plasmid targeting ADSL.	While multiple liver tumors were observed in the control group, only microscopic tumor nodules developed in the liver of ADSL-targeted mice.	[49]
Cas9 nuclease	NSUN2	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumors were smaller for the NSUN2-deficient group than the control groups.	[50]
Cas9 nuclease	Enhancer of C/EBPβ	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumor growth was significantly lower in both homozygous and heterozygous C/EBPβ enhancer mutants.	[51]
Cas9 nuclease	CXCR4	Subcutaneous xenograft developed from CRISPR-treated cells	—	Knockout of CXCR4 resulted in increased sensitivity of cancer cells to cisplatin in vitro. Tumors were smaller for the CXCR4-deficient group than for the control group.	[52]
Cas9 nuclease	NSD1	Subcutaneous xenograft developed from CRISPR-treated cells	—	In vitro studies showed suppressed cell proliferation, migration, and invasion. Tumor weight and volume were significantly lower in those generated from NSD1 knockout HCC cells.	[53]
Cas9 nuclease	TRRAP, KAT5	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumor weight and volume were significantly lower in those generated from TTRAP or KAT5 knockout HCC cells.	[54]
Cas9 nuclease	PTPMT1	Orthotopic liver tumor developed from CRISPR-treated cells	—	Tumors were smaller and less pathologically aggressive for PTPMT1-knockout groups. Lung metastasis was also reduced.	[55]
Cas9 nuclease	G9a	Orthotopic liver tumor developed from CRISPR-treated cells	—	Tumor development and lung metastasis were substantially lower for orthotopic tumors derived from G9a-knockout HCC cells compared to the control group.	[56]
Cas9 nuclease	SEPT11	Mice with IV–injected CRISPR-treated cells	—	Tumor metastatic nodules were not observed in the lungs of mice injected with SEPT11-knockout HCC cells. The group also had reduced angiogenesis in histology.	[57]
Cas9 nuclease	SQSTM1	Mice with IV–injected CRISPR-treated cells	—	In mice with CRISPR-treated HCC cells, no metastatic lesions were seen in the liver, and tumor cells were much lower in the lung.	[58]
Cas9 nuclease	CDK5	Mice with IV–injected CRISPR-treated cells	—	Cdk5 knockout by CRISPR impaired HCC cell dissemination in vivo.	[59]
Cas9 nuclease	LMNA	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumors developed from LMNA-knockout HCC cells had lower tumor weight and volume.	[60]
Cas9 nuclease	ZNF384	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumors developed from ZFN384-knockout HCC cells were smaller than those of the control group.	[61]
Cas9 nuclease	GLS1	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumors developed from GLS1-knockout HCC cells had smaller volumes and reduced tumor-initiating capacity.	[62]
Cas9 nuclease	ZIC2, BPTF	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumors developed from ZIC2-knockout HCC cells had smaller volumes and reduced tumor-initiating capacity. Knockout of BPTF impaired sphere formation in vitro.	[63]
Cas9 nuclease	miR-23a-3p	Orthotopic liver tumor developed from CRISPR-treated cells	—	Tumors developed from miR-23a-3p-knockout HCC cells were significantly lower when combined with sorafenib therapy.	[64]
Cas9 nuclease	SOX4	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumors developed from SOX4-knockout HCC cells had lower growth rates, improved PET results, and reduced extracellular matrix support and angiogenesis.	[65]
Cas9 nuclease	BOLA2	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumor development was halted when BOLA2-knockout cells were used. Also, intracellular iron levels were lower in these tumors.	[66]
Cas9 nuclease	FAM122A	Subcutaneous xenograft developed from CRISPR-treated cells	—	Tumors developed from FAM122A-knockout HCC cells had reduced cell proliferation and tumor growth.	[67]
Cas9 nuclease	Loci with cancer-specific mutations	Tumor organoids transfected with CRISPR-encoded lentivirus	—	Organoid number was substantially reduced when cell-line specific mutations were targeted via CRISPR, co-treated with DNA repair inhibitors.	[68]
Cas9 nuclease	ASPH	—	—	Knockout of ASPH significantly reduced cell proliferation and colony formation, and induced tumor cell senescence.	[69]
dCas9-KRAB transcriptional repressor	GRN	—	—	Silencing of GRN reduced cell proliferation, invasion, and tumor sphere formation abilities of HCC cells.	[70]
dCas9-VPR transcriptional activator	HHIP, MT1M, PZP, and TTC36	—	—	Epigenetic activation of tumor suppressor genes decreased cell proliferation, viability, and migration ability of HCC cells.	[71]
dCas9 transcriptional repressor	lncRNA SNHG9	—	—	Knockdown of SNHG9 inhibited the proliferation, migration, and invasion of HCC cells, and caused cell cycle arrest.	[72]
Cas9 nuclease	GPC1	—	—	The proliferation of HCC cells was significantly attenuated.	[73]
Cas9 nuclease	PHGDH	—	—	Knockout of PHGDH significantly suppressed HCC cell proliferation in the presence of sorafenib.	[74]
Cas9 nuclease	TXNDC9	—	—	Cell proliferation inhibition, cell cycle arrest, and induction of apoptosis were observed in TXNDC9-knockout HCC cells.	[75]
Cas9 nuclease	MDM2	—	—	Knockout of MDM2 reduced HCC cell growth and invasion.	[76]
Cas9 nuclease	EEF2	—	—	Knockout of EEF2 reduced HCC cell proliferation and altered the cell morphology.	[77]
Cas9 nuclease	HGF	—	—	Knockout of HGF decreased HCC cell proliferation, invasion, and migration while inducing apoptosis.	[78]
Cas9 nuclease	NCOA5	—	—	CRISPR inhibited proliferation, tumor microsphere formation, and migration abilities of HCC cells by suppressing epithelial to mesenchymal transition.	[79]

Table 2 CRISPR-Mediated Adoptive T Cell Immunotherapy Studies in HCC

Production of Adoptive T Cells	Resulting T Cells	Results of the Study	Ref.
T cells with lentiviral expression of GPC3-targeted CAR were electroporated with Cas9 protein and 2 gRNAs targeting PD-1	PD-1-knockout, GPC3-targeted CAR T cells	PD-1 disruption enhanced antitumor efficacy, persistence, infiltration, and pro-inflammatory cytokine production of GPC3-CAR T cells in vivo.	[89]
CIK cells were electroporated with Cas9-gRNA RNP complex targeting PD-1 followed by lentiviral transduction of hTER	PD-1-knockout, hTERT-expressing CIK cells	PD-1 knockout improved IFN-γ secretion capacity and antitumor efficacy of hTERT-transduced CIK cells.	[91]
T cells with lentiviral expression of HBsAg-targeted TCR and a sgRNA targeting TRBC1/2 were electroporated with cytosine base editor (BE3) mRNA	TRBC1/2-knockout, HBsAg-targeted TCR T cells	Knockout of the TCR β chain eliminated endogenous TCR expression and prevented mispairing of endogenous TCR with recombinant TCR chains.	[93]
T cells were electroporated with 2 vectors: one for expression of Cas9 and gRNA targeting AAVS1 locus; and the other carrying AAVS1 homology arms and CD105-targeted CAR cassette	CD105-targeted CAR T cells	CRISPR-based knock-in of anti-CD105 CAR expression cassette into AAVS1 locus enabled potent and stable expression of anti-CD105 nanobody.	[94]

Table 3 Clinical Trials Using CRISPR in Adoptive Cell Therapy for HCC

Immunotherapy Approach	Clinical Trial Identifier	Phase	Current Status	Co-Treatment	Estimated Participants	Estimated Primary Completion	Ref.
Autologous PD-1-knockout T cells to treat advanced HCC	NCT04417764	I	Recruiting	TACE	10	Dec 2024	[95]
Autologous PD1-knockout TILs/CAR-TILs that secretes anti-PD-1/CTLA-4 scFvs and CARs* to treat advanced solid cancers including liver cancer	NCT04842812	I	Recruiting	N/A	40	Dec 2025	[96]
Autologous CISH-knockout TILs to treat metastatic gastrointestinal malignancies	NCT04426669	I/II	Recruiting	Aldesleukin, Cyclophosphamide, Fludarabine	20	Sep 2023	[98]

Notes: *Against various antigens, including HER2, Mesothelin, PSCA, MUC1, Lewis-Y, GPC3, AXL, EGFR, Claudin18.2/6, ROR1, GD1, or B7-H3.

Figure 1 Overview of CRISPR-based gene editing technologies. Created with BioRender.com. (A) CRISPR-mediated approaches for gene disruption (NHEJ) and precise editing (HDR). (B) Double nickase strategy employing two Cas9 nickases for controlled and precise DSBs, minimizing off-target effects. (C) CRISPRa and CRISPRi systems for transient regulation of gene expression using deactivated Cas9 and transcription activator/inhibitor domains. (D) CRISPRoff system for durable gene repression by combining deactivated Cas9 with DNA methyltransferase domains and a transcriptional repressor domain. (E) Cytosine and adenine base editing systems that alter DNA bases without inducing double-strand breaks. (F) Prime editing method enabling precise installation of single base mutations and small indels using a Cas9 nickase and reverse transcriptase. (G) RNA-targeting Cas13 system for the manipulation of RNA molecules in cells.

Figure 2 CRISPR-based enhancements for CAR T and TCR T cell therapies in HCC. Created with Biorender.com. (A) Autologous T cells are transduced with a lentivirus carrying a GPC3-targeting CAR expression cassette, followed by electroporation of Cas9-gRNA RNPs to knockout genes encoding the PD-1 and TCR α and β chains. These modifications prevent T cell exhaustion and enhance specificity. The engineered CAR T cells effectively bind GPC3 on HCC cells via GPC3-targeted CARs, triggering a release of cytotoxic proteins (perforin, granzyme B) and pro-inflammatory cytokines (IFN-γ, TNF-α), which collectively contribute to the targeted destruction of tumor cells. (B) Development of Hepatitis B surface antigen (HBsAg)-targeted TCR T cells involves lentiviral delivery of recombinant TCR α and β chains to T cells. The presence of endogenous TCR chains can lead to the formation of mispaired TCRs with unpredictable antigen specificity, posing a potential safety risk. For enhancing safety and efficacy, the cytosine base editor (CBE) is introduced as mRNA to perform knockout of endogenous TCR genes and PD-1. The specific interaction between the HBsAg-targeted TCR and HBsAg presented by MHC-I on HCC cells activates the TCR T cells, promoting robust antitumor activity.

Qi Y, Liu Y, Yu B, et al. A Lactose-Derived CRISPR/Cas9 Delivery System for Efficient Genome Editing In Vivo to Treat Orthotopic Hepatocellular Carcinoma. Adv. Sci. 2020;7(17):2001424. doi:10.1002/advs.202001424

 Web of Science ®Google Scholar

Nie -J-J, Liu Y, Qi Y, et al. Charge-reversal nanocomolexes-based CRISPR/Cas9 delivery system for loss-of-function oncogene editing in hepatocellular carcinoma. J Control Release. 2021;333:362–373. doi:10.1016/j.jconrel.2021.03.030

 PubMed Web of Science ®Google Scholar

Gao J, Luo T, Lin N, Zhang S, Wang J. A New Tool for CRISPR-Cas13a-Based Cancer Gene Therapy. Molecular Therapy - Oncolytics. 2020;19:79–92. doi:10.1016/j.omto.2020.09.004

 PubMed Web of Science ®Google Scholar

Zhang B-C, Luo B-Y, Zou -J-J, et al. Co-delivery of Sorafenib and CRISPR/Cas9 Based on Targeted Core–Shell Hollow Mesoporous Organosilica Nanoparticles for Synergistic HCC Therapy. ACS Appl. Mater. Interfaces. 2020;12(51):57362–57372. doi:10.1021/acsami.0c17660

 PubMed Web of Science ®Google Scholar

Liu Q, Fan D, Adah D, et al. CRISPR/Cas9‑mediated hypoxia inducible factor‑1α knockout enhances the antitumor effect of transarterial embolization in hepatocellular carcinoma. Oncol Rep. 2018;40(5):2547–2557. doi:10.3892/or.2018.6667

 PubMed Web of Science ®Google Scholar

Zhang B-C, P-Y W, Zou -J-J, et al. Efficient CRISPR/Cas9 gene-chemo synergistic cancer therapy via a stimuli-responsive chitosan-based nanocomplex elicits anti-tumorigenic pathway effect. Chem Eng J. 2020;393:124688. doi:10.1016/j.cej.2020.124688

 Web of Science ®Google Scholar

Zhuang J, Tan J, Wu C, et al. Extracellular vesicles engineered with valency-controlled DNA nanostructures deliver CRISPR/Cas9 system for gene therapy. Nucleic Acids Res. 2020;48(16):8870–8882. doi:10.1093/nar/gkaa683

 PubMed Web of Science ®Google Scholar

Yin H, Sun L, Pu Y, et al. Ultrasound-Controlled CRISPR/Cas9 System Augments Sonodynamic Therapy of Hepatocellular Carcinoma. ACS Cent. Sci. 2021;7(12):2049–2062. doi:10.1021/acscentsci.1c01143

 PubMed Web of Science ®Google Scholar

He C, Jaffar Ali D, Qi Y, et al. Engineered extracellular vesicles mediated CRISPR-induced deficiency of IQGAP1/FOXM1 reverses sorafenib resistance in HCC by suppressing cancer stem cells. J Nanobiotechnol. 2023;21(1):154. doi:10.1186/s12951-023-01902-6

 PubMed Web of Science ®Google Scholar

He C, Jaffar Ali D, Xu H, et al. Epithelial cell -derived microvesicles: a safe delivery platform of CRISPR/Cas9 conferring synergistic anti-tumor effect with sorafenib. Exp. Cell. Res. 2020;392(2):112040. doi:10.1016/j.yexcr.2020.112040

 PubMed Web of Science ®Google Scholar

Wan T, Zhong J, Pan Q, Zhou T, Ping Y, Liu X. Exosome-mediated delivery of Cas9 ribonucleoprotein complexes for tissue-specific gene therapy of liver diseases. Sci Adv. 2022;8(37):eabp9435. doi:10.1126/sciadv.abp9435

 PubMed Web of Science ®Google Scholar

Li C, Yang T, Weng Y, et al. Ionizable lipid-assisted efficient hepatic delivery of gene editing elements for oncotherapy. Bioact. Mater. 2022;9:590–601. doi:10.1016/j.bioactmat.2021.05.051

 PubMed Web of Science ®Google Scholar

Jiang T, Sánchez‐Rivera FJ, Soto‐Feliciano YM, et al. Targeting the De Novo Purine Synthesis Pathway Through Adenylosuccinate Lyase Depletion Impairs Liver Cancer Growth by Perturbing Mitochondrial Function. Hepatology. 2021;74(1):458.

 PubMed Web of Science ®Google Scholar

Sun Z, Xue S, Zhang M, et al. Aberrant NSUN2-mediated m5C modification of H19 lncRNA is associated with poor differentiation of hepatocellular carcinoma. Oncogene. 2020;39(45):6906–6919. doi:10.1038/s41388-020-01475-w

 PubMed Web of Science ®Google Scholar

Xiong L, Wu F, Wu Q, et al. Aberrant enhancer hypomethylation contributes to hepatic carcinogenesis through global transcriptional reprogramming. Nat Commun. 2019;10(1):335. doi:10.1038/s41467-018-08245-z

 PubMed Web of Science ®Google Scholar

Wang X, Zhang W, Ding Y, Guo X, Yuan Y, Li D. CRISPR/Cas9-mediated genome engineering of CXCR4 decreases the malignancy of hepatocellular carcinoma cells in vitro and in vivo. Oncol Rep. 2017;37(6):3565–3571. doi:10.3892/or.2017.5601

 PubMed Web of Science ®Google Scholar

Zhang S, Zhang F, Chen Q, Wan C, Xiong J, Xu J. CRISPR/Cas9-mediated knockout of NSD1 suppresses the hepatocellular carcinoma development via the NSD1/H3/Wnt10b signaling pathway. J Exp Clin Cancer Res. 2019;38(1):467. doi:10.1186/s13046-019-1462-y

 PubMed Web of Science ®Google Scholar

Kwan S-Y, Sheel A, Song C-Q, et al. Depletion of TRRAP Induces p53-Independent Senescence in Liver Cancer by Down-Regulating Mitotic Genes. Hepatology. 2020;71(1):275–290. doi:10.1002/hep.30807

 PubMed Web of Science ®Google Scholar

Bao MH-R, Yang C, Tse AP-W, et al. Genome-wide CRISPR-Cas9 knockout library screening identified PTPMT1 in cardiolipin synthesis is crucial to survival in hypoxia in liver cancer. Cell Rep. 2021;34(4):108676. doi:10.1016/j.celrep.2020.108676

 PubMed Web of Science ®Google Scholar

Wei L, Chiu DK-C, Tsang FH-C, et al. Histone methyltransferase G9a promotes liver cancer development by epigenetic silencing of tumor suppressor gene RARRES3. J Hepatol. 2017;67(4):758–769. doi:10.1016/j.jhep.2017.05.015

 PubMed Web of Science ®Google Scholar

Fu L, Wang X, Yang Y, et al. Septin11 promotes hepatocellular carcinoma cell motility by activating RhoA to regulate cytoskeleton and cell adhesion. Cell Death Dis. 2023;14(4):280. doi:10.1038/s41419-023-05726-y

 PubMed Web of Science ®Google Scholar

Lu J, Ding Y, Zhang W, et al. SQSTM1/p62 Knockout by Using the CRISPR/Cas9 System Inhibits Migration and Invasion of Hepatocellular Carcinoma. Cells. 2023;12(9):1238.

 PubMed Web of Science ®Google Scholar

Ardelt MA, Fröhlich T, Martini E, et al. Inhibition of Cyclin‐Dependent Kinase 5: a Strategy to Improve Sorafenib Response in Hepatocellular Carcinoma Therapy. Hepatology. 2019;69(1):634.

 Web of Science ®Google Scholar

Liu H, Li D, Zhou L, et al. LMNA functions as an oncogene in hepatocellular carcinoma by regulating the proliferation and migration ability. J Cell & Mol Med. 2020;24(20):12008–12019. doi:10.1111/jcmm.15829

 PubMed Web of Science ®Google Scholar

He L, Fan X, Li Y, et al. Overexpression of zinc finger protein 384 (ZNF 384), a poor prognostic predictor, promotes cell growth by upregulating the expression of Cyclin D1 in Hepatocellular carcinoma. Cell Death Dis. 2019;10(6):444. doi:10.1038/s41419-019-1681-3

 PubMedGoogle Scholar

Li B, Cao Y, Meng G, et al. Targeting glutaminase 1 attenuates stemness properties in hepatocellular carcinoma by increasing reactive oxygen species and suppressing Wnt/beta-catenin pathway. EBioMedicine. 2019;39:239–254. doi:10.1016/j.ebiom.2018.11.063

 PubMed Web of Science ®Google Scholar

Zhu P, Wang Y, He L, et al. ZIC2-dependent OCT4 activation drives self-renewal of human liver cancer stem cells. J Clin Invest. 2015;125(10):3795–3808. doi:10.1172/JCI81979

 PubMed Web of Science ®Google Scholar

Lu Y, Chan Y-T, Tan H-Y, et al. Epigenetic regulation of ferroptosis via ETS1/miR-23a-3p/ACSL4 axis mediates sorafenib resistance in human hepatocellular carcinoma. J Exp Clin Cancer Res. 2022;41(1):3. doi:10.1186/s13046-021-02208-x

 PubMed Web of Science ®Google Scholar

Tsai C-N, S-C Y, Lee C-W, et al. SOX4 activates CXCL12 in hepatocellular carcinoma cells to modulate endothelial cell migration and angiogenesis in vivo. Oncogene. 2020;39(24):4695–4710. doi:10.1038/s41388-020-1319-z

 PubMed Web of Science ®Google Scholar

Luo J, Wang D, Zhang S, et al. BolA family member 2 enhances cell proliferation and predicts a poor prognosis in hepatocellular carcinoma with tumor hemorrhage. J Cancer. 2019;10(18):4293–4304. doi:10.7150/jca.31829

 PubMed Web of Science ®Google Scholar

Zhou Y, Shi W-Y, He W, et al. FAM122A supports the growth of hepatocellular carcinoma cells and its deletion enhances Doxorubicin-induced cytotoxicity. Exp. Cell. Res. 2020;387(1):111714. doi:10.1016/j.yexcr.2019.111714

 PubMed Web of Science ®Google Scholar

Jiang J, Chen Y, Zhang L, et al. i-CRISPR: a personalized cancer therapy strategy through cutting cancer-specific mutations. Mol Cancer. 2022;21(1):164. doi:10.1186/s12943-022-01612-x

 PubMed Web of Science ®Google Scholar

Iwagami Y, Huang CK, Olsen MJ, et al. Aspartate β‐hydroxylase modulates cellular senescence through glycogen synthase kinase 3β in hepatocellular carcinoma. Hepatology. 2016;63(4).

 PubMed Web of Science ®Google Scholar

Wang H, Guo R, Du Z, et al. Epigenetic Targeting of Granulin in Hepatoma Cells by Synthetic CRISPR dCas9 Epi-suppressors. Mol Ther Nucleic Acids. 2018;11:23–33. doi:10.1016/j.omtn.2018.01.002

 PubMedGoogle Scholar

Sgro A, Cursons J, Waryah C, et al. Epigenetic reactivation of tumor suppressor genes with CRISPRa technologies as precision therapy for hepatocellular carcinoma. Clin epigenetics. 2023;15(1):73. doi:10.1186/s13148-023-01482-0

 PubMed Web of Science ®Google Scholar

Ye S, Ni Y. lncRNA SNHG9 Promotes Cell Proliferation, Migration, and Invasion in Human Hepatocellular Carcinoma Cells by Increasing GSTP1 Methylation, as Revealed by CRISPR-dCas9. Original Research. Front Mol Biosci. 2021;8. doi:10.3389/fmolb.2021.649976

 Web of Science ®Google Scholar

Cheng F, Hansson VC, Georgolopoulos G, Mani K. Attenuation of cancer proliferation by suppression of glypican-1 and its pleiotropic effects in neoplastic behavior. Glypican-1; TCGA; bladder carcinoma; hepatocellular carcinoma; glioma. Oncotarget. 2023;14(1).

 Google Scholar

Wei L, Lee D, Law C-T, et al. Genome-wide CRISPR/Cas9 library screening identified PHGDH as a critical driver for Sorafenib resistance in HCC. Nat Commun. 2019;10(1):4681. doi:10.1038/s41467-019-12606-7

 PubMed Web of Science ®Google Scholar

Chen D, Zou J, Zhao Z, et al. TXNDC9 promotes hepatocellular carcinoma progression by positive regulation of MYC-mediated transcriptional network. Cell Death Dis. 2018;9(11):1110. doi:10.1038/s41419-018-1150-4

 PubMedGoogle Scholar

Wang W, Hu B, Qin -J-J, et al. A novel inhibitor of MDM2 oncogene blocks metastasis of hepatocellular carcinoma and overcomes chemoresistance. Genes Dis. 2019;6(4):419–430. doi:10.1016/j.gendis.2019.06.001

 PubMed Web of Science ®Google Scholar

Pott LL, Hagemann S, Reis H, et al. Eukaryotic elongation factor 2 is a prognostic marker and its kinase a potential therapeutic target in HCC. Oncotarget. 2017;8(7).

 PubMedGoogle Scholar

Lee HK, Lim HM, Park S-H, Nam MJ. Knockout of Hepatocyte Growth Factor by CRISPR/Cas9 System Induces Apoptosis in Hepatocellular Carcinoma Cells. Journal of Personalized Medicine. 2021;11(10):983.

 PubMed Web of Science ®Google Scholar

He J, Zhang W, Li A, Chen F, Luo R. Knockout of NCOA5 impairs proliferation and migration of hepatocellular carcinoma cells by suppressing epithelial-to-mesenchymal transition. Biochem. Biophys. Res. Commun. 2018;500(2):177–183. doi:10.1016/j.bbrc.2018.04.017

 PubMed Web of Science ®Google Scholar

Guo X, Jiang H, Shi B, et al. Disruption of PD-1 Enhanced the Anti-tumor Activity of Chimeric Antigen Receptor T Cells Against Hepatocellular Carcinoma. Front Pharmacol. 2018;9. doi:10.3389/fphar.2018.01118

 Web of Science ®Google Scholar

Huang K, Sun B, Luo N, Guo H, Hu J, Peng J. Programmed Death Receptor 1 (PD1) Knockout and Human Telomerase Reverse Transcriptase (hTERT) Transduction Can Enhance Persistence and Antitumor Efficacy of Cytokine-Induced Killer Cells Against Hepatocellular Carcinoma. Med Sci Monit. 2018;24:4573–4582. doi:10.12659/msm.910903

 PubMed Web of Science ®Google Scholar

Preece R, Pavesi A, Gkazi SA, et al. CRISPR-Mediated Base Conversion Allows Discriminatory Depletion of Endogenous T Cell Receptors for Enhanced Synthetic Immunity. Mol Ther Methods Clin Dev. 2020;19:149–161. doi:10.1016/j.omtm.2020.09.002

 PubMed Web of Science ®Google Scholar

Mo F, Duan S, Jiang X, et al. Nanobody-based chimeric antigen receptor T cells designed by CRISPR/Cas9 technology for solid tumor immunotherapy. Signal Transduction and Targeted Therapy. 2021;6(1):80. doi:10.1038/s41392-021-00462-1

 PubMed Web of Science ®Google Scholar

TACE Combined With PD-1 Knockout Engineered T Cell in Advanced Hepatocellular Carcinoma. Available from: https://clinicaltrials.gov/study/NCT04417764. Accessed December 25, 2023.

 Google Scholar

Engineered TILs/CAR-TILs to Treat Advanced Solid Tumors. Available from: https://clinicaltrials.gov/study/NCT04842812. Accessed December 25, 2023.

 Google Scholar

A Study of Metastatic Gastrointestinal Cancers Treated With Tumor Infiltrating Lymphocytes in Which the Gene Encoding the Intracellular Immune Checkpoint CISH Is Inhibited Using CRISPR Genetic Engineering. Available from: https://clinicaltrials.gov/study/NCT04426669. Accessed December 25, 2023.

 Google Scholar