Breaking it Down: The Wet Lab

            So, you might be wondering, what are the nuts and bolts of the work that is being conducted in the Roychowdhury lab? What makes our nuts and bolts so unique? Our interdisciplinary team consists of three components: bioinformatics, clinic/CLIA, and bench research. Today we will focus on bench research. Bench research can sometimes be referred to as “basic science” research, but it is anything but that. It is here where the true innovation and ingenuity of our lab begins.

            It all begins with the cell.

            Cancer infects upwards of 16 million U.S. citizens every year. In broad terms, cancer is a disease that results when cellular changes uncontrolled growth and division of our body’s own cells. Our cells are normally equipped with the equivalent of car brakes. Healthy cells know when they’ve divided enough times and have fulfilled their purpose, at which point they stop multiplying. Cancerous cells, however, are able to bypass or completely eliminate these breaks. They have altered genes that help make the cellular components necessary for limited cell reproduction. As a result, these unchecked cells build up in the body, using oxygen and nutrients that would usually nourish other cells. Cancerous cells can form tumors, impair the immune system and cause other changes that prevent the body from functioning regularly.

            Whether a gene is being used too much, too little, or there’s a mutation within it, something is wrong in a patient’s cells that makes them cancerous. After years of studies, patterns of abnormal genes start to arise. In our bench research, we are able to take human cells and give them the very gene abnormalities we see in a patient’s cancer. After a patient undergoes a tumor biopsy or blood sample, we can see the gene alterations at the source of their cancer. But how does that affect bench research, where we  work with pipettes and plastic dishes?

            The most common experiment performed on patient-simulated cells in our lab is the drug assay. By placing a certain number of the cells in small wells, we are able to test different therapies on the cells. For example, using a 96-well plate, we place 10,000 cells in each well to be tested (it sounds like a lot, but remember these cells are only about 20 micrometers apiece, smaller than the width of a human hair!). Then, across each column of 8 wells, we add an increasing dosage of a therapy drug in testing. Three days later, it’s a thing of beauty to look down at the plate and see the macroscopic changes that occurred. Each column to the right, less and less cells survive as more drug has been added. We can quantify this cell death (unfortunately, science isn’t purely aesthetic) using a staining and wavelength plate reader. All that’s left is to take those numbers and make graphs to show the world which drugs work best on which cancer cells. 

Increasing Drug Concentration on a 96-well Plate

Oftentimes in the cancers we study, patients respond well to initial treatment. However, a small part of the cancer that isn’t killed by the therapy may survive. Think about the hand sanitizers and Clorox wipes that kill 99.9% of bacteria. A small number of bacteria, just 0.1% (or less), survives because of years of evolution that have granted it resistance to our healthy practices. Cancers are no different. As the resistance cells grow on their own, second or third rounds of therapy are less effective.

This is where our drug assays really come into their own. Our assays test all of the available therapies on these stubborn, surviving cells in the hopes of finding a new strategy to a rebounding cancer. This isn’t so easy, though. Drug-resistant cancers are the most difficult to fight, and our field of research hasn’t had much success to use as a foundation. Instead of throwing in the towel, however, we work. Everyone has a voice, and no idea is too crazy when it comes to the possibilities of saving lives.

            There are other experiments, of course; Western Blots, RNA extraction, qPCRs, you name it. All of these, however, are connected back to our goal in bench research and in the Roychowdhury Lab as a whole. We are faced with a difficult task in helping patients with few viable treatment options for their cancers. All of our cell culture dishes, all our pipet tips, all our hours at the lab benches is done knowing we are filling a gap in science, progressing Ohio State as a research institute, and, most importantly, giving patients the hope they deserve.