The thing about research is that it’s an exciting and rewarding, but oftentimes complex and infuriating, cycle of asking questions and finding answers. Through some simple, yet expressive “Rage comic” illustrations (adapted from an online source; see further note about the illustrations at the end of this post), I thought I would share some of my research experiences, in an effort to demonstrate its cyclical nature, as well as some other things I learned about research.
Like most projects, mine began with an idea: the idea that if we could compare the genes expressed in primary brain tumours (see Figure 1 below) to brain metastases, we could find genes in the metastasis that would identify the population of cells responsible for initiating the metastasis (think cancer stem cells), and target these cells (or genes) with some sort of therapy that might prevent the metastasis from occurring in the first place.
The simplistic view of the project: compare gene lists, identify the ‘metastasis’ genes, test their functionality, and publish within a year. Sounds pretty straightforward and rewarding, no? But the thing about research is it is rarely a straightforward path.
So with that, I began reading… reading about metastases, cancer stem cells, lab protocols, training documents, animal models… The thing about research is just when you think you’ve read and know it all, there’s something new to look into.
Next step: SCIENCE!
At this point, I began to experience the first of many “FFFFFUUUUUUUs!!!” We used patient samples, rather than cell lines (cells from a singular source [e.g. breast tumour from patient ‘X’] capable of growing indefinitely when in appropriate cell culture conditions; their ‘immortality’ often makes them susceptible to genetic mutations; thus, after a time, they no longer resemble the original culture), so we were dependent upon people getting tumours – which isn’t as frequent (a good thing, really!) as we needed. When a tumour did come in, sample processing left very few cells for experiments. Cue more reading on different culture and processing methods and improving the efficiency of experiments. Here is where I learned the thing about research is that optimal is actually a very subjective term.
The other thing about research is your mistakes are your worst enemy, not some mystical force. Forgot a control? – repeat the experiment. Dumped your sample, instead of the waste, in the bleach? Wait for the next one. Used NaCl (a neutral salt) instead of NaOH (a base) to bring down the acidity of your buffer? Make new buffer. These are only a few of my shining moments of scientific aptitude.
Once the scientific stars aligned, the unexpected results arrived. I then came to several realizations: 1) primary brain tumour experiments shouldn’t be directly applied to brain metastases (they are fundamentally different tumours); 2) brain metastases needed to be separated into origin – breast, lung, colon, etc. – not lumped together; and, 3) optimizing experiments on samples that were different every time, wasn’t really optimizing. The solution: a steady and consistent supply of cells. At this point, many of my lab mates had suitable cell lines; I know – we like patient samples; however, using cell lines proven to model patient samples, and then confirming those findings in patient samples again, is a useful and acceptable practice. I took this as a sign that there must be a cell line for me.
In fact there were two appropriate lines; however, only one grew in our culture conditions, and I still had to show that the cell line was representative of patient brain metastases. But hang on a second, if I could show this, could I have a publication? Could I develop a reliable model system to study brain metastasis from the lung for future work? You’ll notice how this differs from my initial project; the thing about research is there is a necessity to adapt and find the successes buried in the failures and mistakes. And here, finally, I had something. But then what are the markers identifying brain metastasis-initiating cells (i.e. cancer stem cells)? Can these cells re-form the primary lung tumour? How do brain metastases from lung cancer compare to those from other cancers? That’s another thing about research: for every answer there are a dozen more questions.
So I guess the thing about research is that it’s just like every other job – things are rarely simple, some things are beyond our control, mistakes happen, but eventually, something good (albeit, unexpected) comes out. In the end, I didn’t identify any genes; instead, I made a model system that would help the next round of researchers, because ironically, my work ended with a new gene list. That’s the thing about research – it’s a cycle.
Figure 1: The difference between primary brain tumours and brain metastases
A note about the illustrations: This post was inspired, in part, from a science Rage comic, origin unknown, but accessed by the author here; parts of the comic were used to illustrate the story, and were not created from the imagination of the author.
For more funny insights about the day-to-day life in grad school and research, check out PhD Comics online, and follow @WSWCgradschool (#whatshouldwecallgradschool) and @OHMethods (#overlyhonestmethods) on Twitter.