Targeting mRNA-coding genes in prostate cancer using CRISPR/Cas9 technology with a special focus on androgen receptor signaling

Tabibian, Mobina; Moghaddam, Fahimeh Salasar; Motevaseli, Elahe; Ghafouri-Fard, Soudeh

doi:10.1186/s12964-024-01833-1

Cell Communication and Signaling

Table 2 CRISPR-mediated modification of PI3K/AKT/mTOR pathway in prostate cancer

From: Targeting mRNA-coding genes in prostate cancer using CRISPR/Cas9 technology with a special focus on androgen receptor signaling

Target gene	Study type	Cell line	Vector	Screening/verification	Experimental data	Ref
DEPTOR	In vitro	DU145, 22RV1	PX459 plasmid	Puromycin selection	Enhanced cell proliferation, survival, migration, and invasion, activated AKT-dependent EMT, and nuclear translocation of β-catenin	[69]
BLM	In vitro	PC-3	Plasmid	Immunohistochemistry, western blotting, T7 endonuclease I assay, PCR	Lower level of phosphorylated AKT (Ser473) and PRAS40 (Thr246)	[143]
ELOVL5	In vitro, in vivo	C4-2	Plasmid	Western blotting, qRT-PCR	Reduced AKT-mTOR signaling activity	[144]
SIRT5	In vitro	PC-3	Plasmid	Western blotting, FACS	Elevated IL-1β expression and increased PI3K/AKT/NF-κB signaling	[145]
SIRT5	In vitro	PC-3	Plasmid	Western blotting, FACS, wound-healing assay, transwell assay, CCK-8 assay	Increased cell invasion capability	[146]
AKT 1/2	In vitro, in vivo	CWR22rv1	Lentiviral, plasmid	Puromycin screening, western blotting, sequencing	Suppressed metastasis	[147]
KDM1A (LSD1)	In vitro	22Rv1	Lentiviral	Puromycin screening, immunoblotting	Inhibited PI3K/AKT signaling	[148]
ATF3	In vitro	LNCaP, DU145, PC3	Plasmid	Western blotting, Surveyor mutation detection assay, qRT-PCR, zymography	Promoted AKT activation, resulting in enhanced cell proliferation	[149]
CKB	In vitro	PC3	PX458 plasmid	Sanger sequencing, western blotting, flow cytometry	Activation of AKT, driving EMT and cell migration	[150]
ARID4B	In vitro, in vivo	PC3, DU145	Plasmid	Western blotting, MTT assay	Identification of ARID4B as a key regulator in the PTEN-PI3K pathway	[151]
NKX3.1	In vivo	C57BL/6 mice	Cas9 protein and sgRNA	Western blotting, IF staining, IHC staining	Inhibition of the PI3K/AKT/mTOR pathway	[73]
PSMA	In vitro	TRAM,22RV1	Lentiviral, plasmid	Puromycin selection, sequencing, TIDE	Decreased PI3K-AKT signaling, with increased MAPK-ERK1/2 signaling	[152]
PPARG	In vitro, in vivo	PC3-M	Plasmid	Puromycin selection	Identification of a functional impact of PPARG on AKT3, associated with a more aggressive disease phenotype	[153]
PTEN	In vitro, in vivo	MSK-PCa1, MSK-PCa2, MSK-PCa3, MSK-PCa8, MSK-PCa11, MSK-PCa12, MSK-PCa15, MSK-PCa16, LNCaP, DU145, CWR22Pc, CWR22Pc-EP	Lentiviral	Sequencing	Effective pan-PI3K inhibition, with upregulation of IGF1R and promoted resistance	[154]
PTEN	In vitro	2924 V	PX459 plasmid	DNA sequencing, western blotting, MTT assay	Enhanced phosphorylation of RAC alpha serine/threonine protein kinase and increased cyclin D1 level	[155]
PTEN, Fos, Jun	In vivo, in vitro	BPH-1, PC-3, DU145, murine prostate epithelium	Lentiviral, plasmid, AAV	Validation of guide RNA efficiency, Histology, IHC, qRT-PCR, sanger sequencing	Loss of Fos contributing to disease progression through accelerated proliferation and invasiveness, partly via Jun activation	[156]
PTEN, α6-integrin, β4-integrin, plectin	In vitro, in vivo	RWPE1, DU145, PC3, 22Rv1, LNCaP, VCaP, JIMT-1	Lentiviral, plasmid	Western blotting, qRT-PCR, proliferation assay, soft agar assay, RNA-Seq	Activation of EGFR/PI3K/Akt and FAK/Src	[157]
WHSC1	In vitro, in vivo	PC3	Plasmid	Western blotting, qRT-PCR, ChIP	Condensed chromatin structure at the RAC1 gene locus, leading to reduced transcription	[158]
Giα2	In vitro	PC3	Plasmid	Sanger sequencing, western blotting, flow cytometry	Impaired lamellipodia establishment at the leading edge of migrating cells and attenuation of TGFβ1 signaling	[159]
KDM5B	In vitro	LNCaP, C4–2B, PC3	Lentiviral, plasmid	QPCR, western blotting, IF staining,	Significant reduction in P110α and PIP3 levels, leading to decreased cell proliferation	[160]
PLK-1	In vitro	PC-3	Plasmid	Western blotting	Suppression of tumor growth	[161]
PLK-1	In vitro, in vivo	LNCap, PC-3	Aptamer-liposome-CRISPR/Cas9 chimera	Apoptosis assay, MTT assay	Noticeable regression of prostate cancer	[70]
ATM	In vitro	DU145, 22Rv1, LNCaP	Lentiviral, plasmid	Immunoblotting, NGS	Altered DNA damage response signaling	[162]
		C4-2	Lentiviral, plasmid	Puromycin selection, western blotting	Increased sensitivity to Enzalutamide	[163]
		22Rv1	Plasmid	Immunostaining, sanger sequencing	Heightened sensitivity to ATR inhibition	[164]
MCL1	In vitro, in vivo	LNCaP	Plasmid	Western blotting	Significant sensitization of cancer cells to apoptosis induced by navitoclax, but not by venetoclax	[165]
GPRC5A	In vitro, in vivo	PC3, DU145	Guide-it™ CRISPR/Cas9 System	Real-Time RT-PCR, MTT and BrdU assays, cell cycle assay, RNA-seq	Decreased cell proliferation	[166]

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