October 17, 2016
I may be a little late to the game, but over the last few weeks I have started consuming episodes of Game of Thrones voraciously. It is such a fun show to watch! And this isn’t the only HBO show I enjoy. Veep, Silicon Valley, and Last Week Tonight with John Oliver all have my attention as well.
You might say that I need HBO, because without it I can’t get my fill of these shows. (Well, there are other routes, but HBO or HBO GO are the easiest). My over watching of these shows has made me dependent on HBO.
Something similar can happen in cancers. Sometimes a key player in keeping a cell cancerous is an overexpressed gene. And just like my binging of Game of Thrones makes me dependent on HBO, so this overexpressed gene (the TV show) makes the cancer cell dependent on another gene (HBO).
A real life example might make this clearer. Most folks have heard of BRCA1 and BRCA2 especially since the Angelina Jolie story. When either of these genes is damaged, you can end up with cancer.
Both of these genes are involved in DNA repair and damaging them means the cell builds up mutations. Making lots of DNA mistakes is a good thing for cancers but only up to a point. Too much damage and the cancer cell dies.
What this means is that these cancer cells are now more dependent on other DNA repair genes. Which means these other DNA repair genes are now targets to go after to selectively kill the cancer cells.
For cancer cells lacking BRCA1 or BRCA2 function, research has shown that these cells are now dependent on a second gene, PARP1. If PARP1 expression is turned down, normal cells survive but BRCA1/BRCA2-dependent cancers die. So, we can kill cancer cells, or end their TV show watching, by going after PARP1, their HBO.
Finding these sorts of genes is not easy unless, of course, you turn to our favorite lab workhorse, the yeast Saccharomyces cerevisiae. Given all of the genetic techniques and tools available with this yeast, it is possible to quickly do a synthetic dosage lethality assay – to look for genes that are lethal only in combination with deleting your gene of interest.
This is just what Reid and coworkers did in a new study just out in GENETICS for CKS1B, a gene that is amplified and overexpressed in many cases of breast, lung, and liver cancers. And they found a more “druggable” target to go after, the kinase PLK1 (the human homolog of yeast CDC5). PLK1 even comes with its own kinase inhibitor, Volasertib.
Reid and coworkers transformed a low copy plasmid containing the CKS1 gene, the yeast homolog of CKS1B, under the control of the galactose promoter into two different yeast strain libraries. The first screen used 9600 yeast deletion strains, each with a single gene deleted in either a MATa or MATα strain. The second screen used strains with temperature sensitive mutants of essential genes. They now looked to see which yeast strains did poorly or couldn’t survive when they were overexpressing CSK1 in the presence of galactose.
In the end they came up with 44 different genes that, when deleted or weakened, had a severe effect on the growth of yeast that overexpressed CKS1. Given that CKS1 plays an important role in cell cycle progression, they focused on the 15 genes that affect mitotic progression. Eventually, through a set of experiments that I don’t have time to go into here, they settled in on CDC5, a polo-like kinase involved in both mitotic entry and exit.
The next step was to see if what they learned about in yeast has any bearing on cancer. It did.
First Reid and coworkers looked at a variety of cancer cells in The Cancer Genome Atlas (TCGA) and found that it was very rare for both PLK1 and CKS1B to be overexpressed in the same cancer at the same time. Next they looked at a data set of short hairpin RNA (shRNA) knockdowns of ~16,000 human genes and found that knocking down PLK1 had negative effects on cancers overexpressing CKS1B. These are consistent with the two genes having a synthetic lethal relationship.
They then took eight breast cancer lines where the shRNA against PLK1 had a negative effect on growth and tested the effects of targeting PLK1 on apoptosis. Did decreasing expression of PLK1 in cells that overexpress CKS1B cause an increase in apoptosis in their hands?
The short answer is yes. They repeated the experiments with the shRNA and also tested the PLK1-specific kinase inhibitor Volasertib and found that both treatments increased apoptosis in CKS1B overexpressing cancer cells. It looks like they may have uncovered a way to go after a subset of cancers using yeast!
Which shouldn’t surprise us. Yeast and other model organisms have been teaching us about cancer at least since the days when Hartwell, Hunt and Nurse first identified cyclins and CDKs (for which they got the 2001 Nobel Prize in Physiology or Medicine), and will continue to school us for years to come.
Hopefully researchers will continue to turn to yeast to continue to better understand and find new treatments for cancer. Yeast has so much more to teach us! #APOYG!
by Barry Starr, Ph.D., Director of Outreach Activities, Stanford Genetics
Categories: Research Spotlight, Yeast and Human Disease
Tags: CKS1, cyclin-dependent kinase, polo-like kinase, synthetic dosage lethal