A Commentary on Senolytic Gene Therapies to Target p16 Overexpression

This short commentary discusses the utility of Oisin Biotechnologies' initial strategy for destroying senescent cells, which is to use p16 expression as the determining sign of senescence. Oisin's implementation involves delivering dormant DNA machinery indiscriminately to all cells, and then triggering it only in cells with high levels of p16. This particular implementation is one of many possibilities in the gene therapy space, and thus various other groups are working on their own p16-based approaches as senolytic development as a treatment for aging grows in funding and popularity.

It isn't just senescence and aging in which this is a topic of interest, of course. There is a strong overlap with cancer, and the search for ways to selectively destroy the most aggressive cancerous cells. In many of these forms of aberrant cell the mechanisms of senescence are broken in some way. These cells have at least some of the chemical signatures of senescence, but fail to shut down and cease replication. Thus targeting expression of senescence-associated genes may work fairly well against cancer as well as the contribution of cellular senescence to aging - something that Oisin Biotechnologies is also working on.

p16Ink4a (p16) is an important tumor suppressor which is upregulated in senescent cells and in aged tissues. p16 acts as an inhibitor of the interaction between Cyclin-Dependent Kinases (CDK) 4/6 and CyclinD1 leading to the activation of retinoblastoma protein (RB). Consequently, active RB interferes with the translocation of E2F1 into the nucleus and arrests cells in the G1-S phase of the cell cycle. In cancer cells with mutations in RB or CDK4/6, p16 is normally overexpressed but unable to induce cell cycle arrest. p16-overexpressing cancer cells are found in different types of carcinomas and are considered highly aggressive and invasive.

Several drugs in recent years have been shown to have senolytic properties (i.e. being toxic for senescent cells) and to remove p16+ cells from a variety of tissues. Among these compounds, ABT-737 and its orally-available analogue ABT-263 target the anti-apoptotic proteins BCL-2, BCL-W and BCL-XL, considered essential pro-survival players in senescent cells. The effects of these compounds in mice almost completely overlap with a suicide gene strategy activated by the p16 promoter, thus suggesting specific targeting p16+ cells.

However, when we tested ABT-737 and ABT-263 against p16-overxpressing murine sarcomas we failed to observe any toxicity, despite p16+-cancer cells upregulating both BCL-2 and BCL-XL. These data could be interpreted in 3 ways: 1) ABT compounds are specifically active against non-proliferating p16+-cells; 2) the efficacy of ABTs requires upregulation of BCL-W, which we have recently shown being a common feature of senescent cells; 3) ABTs act independently of p16 expression levels. The latter hypothesis would represent a critical issue, as p16 is used as a major readout for the efficacy of senotherapies.

Since we have recently developed an inducible suicide gene regulated by the full p16 promoter, we have then studied whether the use of this strategy could be effective against p16+ tumors. Indeed, most p16-overexpressing cancer cells were efficiently eliminated by the activation of the suicide gene in both culture and in vivo conditions. These data suggest that p16 upregulation is maintained by active transcription, possibly mediated by emergency signaling pathways attempting to restrain cellular proliferation.

Our study supports the idea that the overexpression of oncosuppressors could be exploited for interventions against cancer. While we studied a specific context in which p16 is present in its wild-type form, this strategy could potentially work in situations of overexpression (by transcriptional regulation) of mutated forms, which is a common feature of cancer cells. On this line, similar strategies against additional oncosuppressors such as p14 and p53 could be effective.

In parallel, it will be of interest to understand whether a p16-based suicide gene therapy could be used in other contexts. Studies in transgenic mouse models have shown that elimination of p16+ cells using suicide genes can significantly delay the onset and progression of a number of age-related pathologies, eventually leading to lifespan extension. Whether a similar strategy could be used for human interventions is still matter of debate.

Link: https://doi.org/10.18632/aging.101422

Comments

Is this Oisin's paper?

Posted by: Ariel at May 28th, 2018 12:12 PM

@Ariel: No, but it is a small community and the author is well aware of Oisin's existence.

Posted by: Reason at May 28th, 2018 1:08 PM

I'm having a little trouble understanding how the Oisín technology precisely distinguishes between healthy and diseased cells. For example, I have read that p53 is always being produced in healthy cells, albeit at low levels. Is it as simple as cells requiring more than a corresponding low level of the suicide protein to trigger apoptosis?

Posted by: Wayne Johnson at May 29th, 2018 11:11 AM

@Wayne - basically, yes. Through adjustment of dose we can spare normal cells while targeting over-expressing ones. We've shown this in tox studies and other in vivo experiments, at least to our satisfaction.

Posted by: Gary at May 29th, 2018 2:17 PM

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