A Light-Inducible Split-dCas9 System for Inhibiting the Progression of Bladder Cancer Cells by Activating P53 and E-cadherin

Overview

Journal Front Mol Biosci

Specialty Biology

Date 2021 Jan 20

PMID 33469550

Citations 4

Authors

Xinbo Huang

Qun Zhou

Mingxia Wang

Congcong Cao

Qian Ma

Jing Ye

Yaoting Gui

Affiliations

Soon will be listed here.

Abstract

Optogenetic systems have been increasingly investigated in the field of biomedicine. Previous studies had found the inhibitory effect of the light-inducible genetic circuits on cancer cell growth. In our study, we applied an AND logic gates to the light-inducible genetic circuits to inhibit the cancer cells more specifically. The circuit would only be activated in the presence of both the human telomerase reverse transcriptase (hTERT) and the human uroplakin II (hUPII) promoter. The activated logic gate led to the expression of the p53 or E-cadherin protein, which could inhibit the biological function of tumor cells. In addition, we split the dCas9 protein to reduce the size of the synthetic circuit compared to the full-length dCas9. This light-inducible system provides a potential therapeutic strategy for future bladder cancer.

Citing Articles

H3 histone methylation landscape in male urogenital cancers: from molecular mechanisms to epigenetic biomarkers and therapeutic targets.

Burlibasa L, Nicu A, Chifiriuc M, Medar C, Petrescu A, Jinga V Front Cell Dev Biol. 2023; 11:1181764.

PMID: 37228649 PMC: 10203431. DOI: 10.3389/fcell.2023.1181764.

Optimization of CRISPR-Cas system for clinical cancer therapy.

Meng X, Wu T, Lou Q, Niu K, Jiang L, Xiao Q Bioeng Transl Med. 2023; 8(2):e10474.

PMID: 36925702 PMC: 10013785. DOI: 10.1002/btm2.10474.

Recent Advances in Genome-Engineering Strategies.

Boti M, Athanasopoulou K, Adamopoulos P, Sideris D, Scorilas A Genes (Basel). 2023; 14(1).

PMID: 36672870 PMC: 9859587. DOI: 10.3390/genes14010129.

Modulating gene expression in breast cancer via DNA secondary structure and the CRISPR toolbox.

Kretzmann J, Irving K, Smith N, Evans C NAR Cancer. 2022; 3(4):zcab048.

PMID: 34988459 PMC: 8693572. DOI: 10.1093/narcan/zcab048.

References

Ma D, Peng S, Xie Z . Integration and exchange of split dCas9 domains for transcriptional controls in mammalian cells. Nat Commun. 2016; 7:13056. PMC: 5063958. DOI: 10.1038/ncomms13056. View

Muller P, Milton M . The determination and interpretation of the therapeutic index in drug development. Nat Rev Drug Discov. 2012; 11(10):751-61. DOI: 10.1038/nrd3801. View

Strumane K, Berx G, VAN Roy F . Cadherins in cancer. Handb Exp Pharmacol. 2010; (165):69-103. DOI: 10.1007/978-3-540-68170-0_4. View

Beerli R, Dreier B, Barbas 3rd C . Positive and negative regulation of endogenous genes by designed transcription factors. Proc Natl Acad Sci U S A. 2000; 97(4):1495-500. PMC: 26462. DOI: 10.1073/pnas.040552697. View

Jusiak B, Cleto S, Perez-Pinera P, Lu T . Engineering Synthetic Gene Circuits in Living Cells with CRISPR Technology. Trends Biotechnol. 2016; 34(7):535-547. DOI: 10.1016/j.tibtech.2015.12.014. View

Wright A, Sternberg S, Taylor D, Staahl B, Bardales J, Kornfeld J . Rational design of a split-Cas9 enzyme complex. Proc Natl Acad Sci U S A. 2015; 112(10):2984-9. PMC: 4364227. DOI: 10.1073/pnas.1501698112. View

Liu Y, Zeng Y, Liu L, Zhuang C, Fu X, Huang W . Synthesizing AND gate genetic circuits based on CRISPR-Cas9 for identification of bladder cancer cells. Nat Commun. 2014; 5:5393. DOI: 10.1038/ncomms6393. View

Mendonsa A, Na T, Gumbiner B . E-cadherin in contact inhibition and cancer. Oncogene. 2018; 37(35):4769-4780. PMC: 6119098. DOI: 10.1038/s41388-018-0304-2. View

Gossen M, Freundlieb S, Bender G, Muller G, Hillen W, Bujard H . Transcriptional activation by tetracyclines in mammalian cells. Science. 1995; 268(5218):1766-9. DOI: 10.1126/science.7792603. View

10.

Santiago-Ortiz J, Schaffer D . Adeno-associated virus (AAV) vectors in cancer gene therapy. J Control Release. 2016; 240:287-301. PMC: 4940329. DOI: 10.1016/j.jconrel.2016.01.001. View

11.

Manshouri R, Coyaud E, Kundu S, Peng D, Stratton S, Alton K . ZEB1/NuRD complex suppresses TBC1D2b to stimulate E-cadherin internalization and promote metastasis in lung cancer. Nat Commun. 2019; 10(1):5125. PMC: 6851102. DOI: 10.1038/s41467-019-12832-z. View

12.

Senis E, Fatouros C, Grosse S, Wiedtke E, Niopek D, Mueller A . CRISPR/Cas9-mediated genome engineering: an adeno-associated viral (AAV) vector toolbox. Biotechnol J. 2014; 9(11):1402-12. DOI: 10.1002/biot.201400046. View

13.

Miller J, Gaiddon C, Storr T . A balancing act: using small molecules for therapeutic intervention of the p53 pathway in cancer. Chem Soc Rev. 2020; 49(19):6995-7014. DOI: 10.1039/d0cs00163e. View

14.

Folcher M, Oesterle S, Zwicky K, Thekkottil T, Heymoz J, Hohmann M . Mind-controlled transgene expression by a wireless-powered optogenetic designer cell implant. Nat Commun. 2014; 5:5392. PMC: 4241983. DOI: 10.1038/ncomms6392. View

15.

Wong S, Fang C, Chuah L, Leong C, Ching Ngai S . E-cadherin: Its dysregulation in carcinogenesis and clinical implications. Crit Rev Oncol Hematol. 2017; 121:11-22. DOI: 10.1016/j.critrevonc.2017.11.010. View

16.

Ran F, Cong L, Yan W, Scott D, Gootenberg J, Kriz A . In vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015; 520(7546):186-91. PMC: 4393360. DOI: 10.1038/nature14299. View

17.

Li L, Hu S, Chen X . Non-viral delivery systems for CRISPR/Cas9-based genome editing: Challenges and opportunities. Biomaterials. 2018; 171:207-218. PMC: 5944364. DOI: 10.1016/j.biomaterials.2018.04.031. View

18.

Sedlmayer F, Aubel D, Fussenegger M . Synthetic gene circuits for the detection, elimination and prevention of disease. Nat Biomed Eng. 2019; 2(6):399-415. DOI: 10.1038/s41551-018-0215-0. View

19.

Lin F, Dong L, Wang W, Liu Y, Huang W, Cai Z . An Efficient Light-Inducible P53 Expression System for Inhibiting Proliferation of Bladder Cancer Cell. Int J Biol Sci. 2016; 12(10):1273-1278. PMC: 5069448. DOI: 10.7150/ijbs.16162. View

20.

Onder T, Gupta P, Mani S, Yang J, Lander E, Weinberg R . Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res. 2008; 68(10):3645-54. DOI: 10.1158/0008-5472.CAN-07-2938. View