Please provide a brief description of your STEP Signature Project. Write two or three sentences describing the main activities your STEP Signature Project entailed.

Collagen type 4 (Col IV) is a tetrameric protein (polymer consisting of 4 monomers) that forms networks within the basement membrane, a thin layer between epithelial cells and underlying tissue. The goal of my STEP project was to identify the binding regions of Discoidin Domain Receptor 1 (DDR1) to Collagen type 4 (col IV) using Atomic Force Microscopy (AFM). The main activities included sample preparation, AFM imaging, and image analysis.

What about your understanding of yourself, your assumptions, or your view of the world changed/transformed while completing your STEP Signature Project? Write one or two paragraphs to describe the change or transformation that took place.

I was able to challenge two assumptions I had about myself. First, I do not have the skills necessary as an undergraduate to be successful with an independent research project. Second, research papers are too complicated for me to be able to dissect and understand. I also realized at least two things about myself during the STEP project. First, that I could critically think through potential solutions. Second, I could get myself out of my comfort zone and collaborate with other principal investigators across the country. Finally, I was able to get an appreciation for how long innovation takes to get from the bench to the bedside for patients.

What events, interactions, relationships, or activities during your STEP Signature Project led to the change/transformation that you discussed in #2, and how did those affect you? Write three or four paragraphs describing the key aspects of your experiences completing your STEP Signature Project that led to this change/transformation.

I started research in Dr. Gunjan Agarwal’s lab in the fall of my sophomore year. Before getting my own independent project, I got published for my work in AFM of collagen in abdominal aortic aneurysms as well as in the intervertebral disk. For both, I relied on the graduate students in the lab for assistance, especially in sample preparation and image analysis. Also, both projects were ongoing, and I felt that I did not understand it at the same depth I could have if I worked on them from the start. These were what led me to wanting an independent research project. However, I doubted that I had the skills to be successful in an independent project. During my time with the STEP project, I have been able to do all the activities on my own. It also is a project I get to see from start to finish for, which gives me the level of detail I would have liked to have from the other projects.

As part of the background work before the project, I was assigned to read a few collagen IV papers. I had to present these in our lab meetings, which added an extra level of stress. At first, looking at papers that were 12-13 pages long was daunting, but I quickly found that the organizations of the papers made them easy to dissect, especially their results. I still struggle with decoding the methods used, but I still think I have a degree of understanding when reading the papers that I did not expect to have.

The first part of the project was finding the right concentration of col IV for AFM imaging. This part went rather smoothly (Figure 1), but I had difficulty when observing the DDR binding to col IV. Initially, I would just let the concentration of col IV and DDR1 sit in a tube for two hours, hoping that the DDR would bind. Then, I would place this solution on a surface that locks the col IV into place (Mica). However, I noticed that the DDR was not binding; Instead, it looks like the col IV was clumping together (Figure 2). This was not ideal, because I needed to see a clean Col IV molecule bind to DDR to map out the binding. We then tried to increase DDR concentration and place the solution in a rocker for the incubation time. Additionally, the PI and I discussed potentially plating the pure Col IV on mica to make it unmovable and then put the DDR on top of that surface for two hrs. I highlight this example to show the immense amount of troubleshooting that research entails.

Another part of the project I will be working on now is looking at a mutated form of Col IV in collaboration with Dr. Douglas Gould. I would laugh if someone told my sophomore self that I would be able to network with researchers across the country. It has definitely been a unique experience in setting up this collaboration.

Finally, through my Pharmaceutical Sciences major, I first was able to see the length of time it takes for an innovative therapy to get from the bench to bedside. The basic science research I am doing is at the start of this long process, and I will not lose sight of the fact that someone 20 years from now could benefit from my work. This also comes with integrity in research because I should not cut corners. Finally, in the era of COVID-19, I think the story of the vaccines are incredible in how they were able to develop, test and distribute a vaccine within a year. However, I think we should realize that this was an exemption, and that innovation takes time to come to fruition.

Why is this change/transformation significant or valuable for your life? Write one or two paragraphs discussing why this change or development matters and/or relates to your academic, personal, and/or professional goals and future plans.

As I said above, my STEM project has put research into perspective within the larger drug discovery and development process. I look to continue research in medical school and as a physician, and I will not lose sight of the fact that my work could impact a patient’s care. I also now have the self confidence that I can be successful with an independent research project, which would be an expectation as a medical student.

The networking skills I gained is also important for my career. For starters, a new trend for medical students to boost their residency applications is a research year between M2 and M3. Now, this could be something I go down to California and work more directly with Dr. Gould. Also, I have been told by many career development faculties that who I know is as important as what I know to be successful in my future endeavors.

Finally, critical thinking and troubleshooting comes back as a physician as I know a “one size fits all” approach will not work. So, working alongside my patients like I did with the PI to find the best treatment plan will help me provide the most effective care for my patients.

Figure 1: 1.0ug/ml col IV in PBS

Here, this image of Col IV shows the individual monomers coming together to form some tetramers. The collagen IV strands are well defined and I would expect to see DDR binding to these individual strands.

Figure 2: 1.0 ug/ml col IV in PBS

Here, this image is more representative of the collagen IV I have seen more recently. The strands are not neatly defined and it looks like they are clumped together (example in the blue circle). I would not expect to see any DDR binding to this in the counter samples. We suspect that the effect of letting these samples sit in the fridge (to give time for the counter samples with DDR to bind) is a factor in this clumping with itself.

If you have any questions please contact me at my email address (gadde.9@buckeyemail.osu.edu)

Nikhit Gadde