Gene Therapy with p14/tBID Induces Selective and Synergistic Apoptosis in Mutant Ras and Mutant p53 Cancer Cells In Vitro and In Vivo

Robert L Fine; Yuehua Mao; Dario Garcia-Carracedo; Gloria H Su; Wanglong Qiu; Uri Hochfeld; Gwen Nichols; Yong-Liang Li; Richard D Dinnen; Anthony Raffo; Paul W Brandt-Rauf

doi:10.3390/biomedicines11020258

Gene Therapy with p14/tBID Induces Selective and Synergistic Apoptosis in Mutant Ras and Mutant p53 Cancer Cells In Vitro and In Vivo

Biomedicines. 2023 Jan 18;11(2):258. doi: 10.3390/biomedicines11020258.

Authors

Affiliations

¹ Experimental Therapeutics Program, Division of Medical Oncology, The Pancreas Center at Columbia, Herbert Irving Comprehensive Cancer Center, NYPH-Columbia University Medical Center, New York, NY 10032, USA.
² Department Pathology and Otolaryngology, Herbert Irving Comprehensive Cancer Center, NYPH-Columbia University Medical Center, New York, NY 10032, USA.
³ Department of Environmental Health Sciences, Mailman School of Public Health, Columbia University, New York, NY 10032, USA.
⁴ School of Public Health, University of Illinois at Chicago, Chicago, IL 60612, USA.
⁵ School of Biomedical Engineering, Science and Health Systems, Drexel University, Philadelphia, PA 19104, USA.

Abstract

Any gene therapy for cancer will be predicated upon its selectivity against cancer cells and non-toxicity to normal cells. Therefore, safeguards are needed to prevent its activation in normal cells. We designed a minimal p14^ARF promoter with upstream Ap1 and E2F enhancer elements and a downstream MDR1 inhibitory element, TATA box, and a transcription initiation site (hereafter p14^ARFmin). The modified p14^ARFmin promoter was linked to bicistronic P14 and truncated BID (tBID) genes, which led to synergistic apoptosis via the intrinsic and extrinsic pathways of apoptosis when expressed. The promoter was designed to be preferentially activated by mutant Ras and completely inhibited by wild-type p53 so that only cells with both mutant Ras and mutant p53 would activate the construct. In comparison to most p53 gene therapies, this construct has selective advantages: (1) p53-based gene therapies with a constitutive CMV promoter cannot differentiate between normal cells and cancer cells, and can be toxic to normal cells; (2) our construct does not induce p21^WAF/CIPI in contrast to other p53-based gene therapies, which can induce cell cycle arrest leading to increased chemotherapy resistance; (3) the modified construct (p14^ARFmin-p14-tBID) demonstrates bidirectional control of its promoter, which is completely repressed by wild-type p53 and activated only in cells with both RAS and P53 mutations; and (4) a novel combination of genes (p14 and tBID) can synergistically induce potent intrinsic and extrinsic apoptosis in cancer cells.

Keywords: Ras; apoptosis; gene therapy; p14; p14ARF promoter; p53; tBID.

Abstract

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