My journey to becoming a Laidlaw Scholar was somewhat backwards. As a curious, science-loving person, all I knew was that I wanted to get involved with research in some capacity. I began searching for labs to join in the Boston area and reflecting on what I was interested in studying, which led me to Dr. Levy’s Headache Research Lab at Beth Israel Deaconess Medical Center. I’ve suffered from migraines for many years, and so I felt a personal connection to Dr. Levy’s work trying to understand the pathophysiology of migraines. It was only after I had identified this particular lab that I discovered the Laidlaw Scholars program, and I immediately felt conflicted. I knew that I wanted to be involved in the program, because I’m deeply passionate about both research and leadership, however I initially struggled seeing how my migraine research could fit into the scope of the Laidlaw program. I somehow thought that studying migraines wasn’t “big” or “grand” enough to participate in a global program like Laidlaw. However, after some reflection and more thought, I realized that my research itself didn’t matter as much as my ability to think about its broader implications, and my own goals as a researcher and leader. I began thinking about migraine less as just a headache disorder, and more as a microcosm of global healthcare inequities. I learned that migraines affect women on average 3 times more than they do men, and that diseases and disorders that predominantly impact women have historically been understudied in a clinical setting. With this broader perspective in the back of my mind, I eagerly embarked on my summer research journey. 

Migraines are a complex neurovascular pain disorder linked to the meninges, the protective tissue surrounding the brain, that affect roughly 1 billion people globally yet the exact mechanisms by which they arise remain unknown to scientists. Symptoms of migraines include severe headache, nausea, photophobia (light sensitivity), phonophobia (sound sensitivity), and allodynia, a condition that causes pain from stimuli that normally don't cause pain, such as a light breeze or touch. Scientists believe that certain neuropeptides are involved in the pathophysiology of migraine, including calcitonin gene-related peptide (CGRP), which is what many of the current migraine drugs on the market are designed to target. While these drugs do exist, they are only effective in roughly 50-60% of migraine patients, indicating that there is still much to discover about the nature of migraines and that better treatment methods are needed. 

This is the ultimate goal of the Levy lab: to understand migraine pathophysiology with the hopes of one day being able to contribute to the development of a successful migraine therapeutic treatment. Mouse models are used in the Levy lab to study migraines, however mice don’t naturally develop them, and so a large challenge in studying them has been knowing whether the animals used in the studies are actually experiencing something analogous to a migraine. It is important that mice are actually experiencing migraine-like symptoms so that they are appropriate proxies for what humans with migraine disorders experience. Thus, the broad overarching question that fueled my research this summer was: How do you trigger a migraine-like response in a mouse?

The project that I worked on was aimed at measuring the behavioral response of the animal in response to stimuli aimed at triggering migraines. Specifically, we measured the light-evoked squinting of the animals as a migraine-pain proxy, since mice squint when they experience pain. There were two different methods that we used to evoke a squinting response from the mice: direct activation of the sensory innervation of the meninges and systemic injection of nitroglycerin. Both methods used a behavioral apparatus that included a head-fixed mouse freely locomoting on a wheel exposed to 300 lux light while a camera recorded its right eye. For sensory innervation of the meninges, the mice used in this particular experiment had been genetically modified to express a specific pain channel in the brain when exposed to infrared light. Additionally, they underwent a surgery to remove a small portion of skull above the sinus and place a small piece of glass over the hole, called a cranial window. The purpose of the cranial window was to allow the infrared light to activate the sensory innervation of the meninges in a more direct way rather than through the bone. 

After the mouse habituated on the wheel for 30 minutes, its baseline behavior was then recorded for an additional 30 minutes. Next, the mouse was exposed to one hour of optogenetic  stimulation of the meningeal sensory innervation under infrared light and the behavioral response was recorded during and 30 minutes post stimulation. The recording of the mouse’s eye was then analyzed through a machine-learning algorithm (DeepLabCut) to determine the level of squinting. 

The thought process behind this experiment was that in response to the infrared stimulation, the light-evoked pain channel would activate in these genetically modified mice, and simulate migraine-like pain. Thus, if the mice squinted during and post infrared stimulation, that would be considered an indication that the mouse was experiencing something close to a migraine. The experiment that I personally worked on used mice with the cranial window at a stimulation frequency of 10hz. As mentioned previously, our idea was that mice with cranial windows would have a greater squinting response to the infrared light than mice without the cranial window, however our data showed that to not exactly be true. The data we collected showed that the mice we tested did not squint at all during nor post stimulation, whereas previously collected data showed that squinting had been detected in mice without cranial windows and at a lower stimulation frequency. We hypothesized that the reason for these unexpected results could be due to the cranial window allowing for the nerves to be overstimulated at the higher frequency, and essentially killing the nerves so that there was no activation of the pain channel. Still, more testing with cranial windows must be done, and at different frequencies, to better understand the results that we got. 

The second method we used to trigger a migraine-like response in mice was injecting nitroglycerin. Nitroglycerin is a powerful vasodilator that is used in migraine studies involving humans. When people diagnosed with a migraine disorder are injected with nitroglycerin, it triggers them to get a migraine. Interestingly, scientists don’t actually understand how nitroglycerin triggers migraines, just that it does. The mice we used in this experiment were not genetically modified because we were not attempting to activate the same light-evoked pain channel. 

The protocol for the nitroglycerin experiment was fairly similar to the optogenetic stimulation experiment. The mice were placed in the same apparatus, and allowed 30 minutes to habituate on the wheel. Next, they were injected with 1 mg/kg of nitroglycerin, and their behavioral response was recorded for 2 hours post injection. Three days later, the same mice underwent the same protocol, but instead were injected with 10 mg/kg of nitroglycerin. We were anticipating that the mice would exhibit a greater squinting response after the 10mg injection versus the 1 mg injection, however our data did not provide robust evidence to support this. After both injections, squinting was detected, but there was not a large enough difference between the two to extrapolate a concrete relationship. 

Immediate next steps for this project are to repeat the nitroglycerin experiment to see if different results can be achieved, and to test mice with the cranial window at a lower frequency of optogenetic stimulation to see what kind of squinting response they have. Additionally, the optogenetic experiment was only done acutely, and so the next logical step would be to complete the experiment chronically. This would entail exposing the mice to the infrared light over a period of five days rather than one. 

Ultimately, neither method of triggering a migraine-like response in mice, activation of the sensory innervation of the meninges nor injection of nitroglycerin, provided sufficient data for establishing a reliable protocol for studying migraines with mice. A large challenge is understanding why some mice squint, and some don’t, and while these results do not tell a complete story, they will further inform future experiments. Establishing a reliable protocol to trigger a migraine within a mouse is vital for knowing how migraines work and ultimately

how to cure them. While this project is very much ongoing and my work this summer was just one piece of a much larger exploration, it certainly revealed to me how complex studying migraine pathophysiology is, and broader, the scientific process in general.

I view my research experience this summer as a window into the world of science, and in particular medical research. I learned a lot about the day to day pace of research, and how things always seem to move slower than you anticipate. I also came to the realization that the majority of results you get in the lab are negative ones, which led me to improve my relationship with failure. No matter how many unusual or disappointing results we got in the lab this summer, my mentors never appeared upset or dejected, and instead simply got to work on figuring out what went wrong. It was incredibly valuable for me to witness this, and truly emphasized to me that there is no such thing as scientific progress without failure. 

Naturally, at times I felt very removed from the “impact” of my research since things were moving slowly, if at all. I never felt that I was getting closer to curing migraines. However, I also realized that the skills I gained over the course of the summer will greatly help me make a tangible impact as I embark on my leadership in action project. Mainly, the skill of connecting with people and sharing complex ideas in a digestible way. I developed this skill in two ways throughout the summer. The first way was through an assignment given to me by my Principal Investigator. He gave me a complex paper on migraines and tasked me with presenting it to our entire lab. Initially, I barely understood a word of the paper, however after a lot of work and studying on my own, I was able to confidently present the ideas of the paper to my lab and even could highlight some of its limitations. The second way was through presenting my research in a poster at the end of the program. At first, it was difficult to even begin trying to explain “optogeneic stimulation” and “light-evoked squinting” to people who knew nothing about migraines, but the more I practiced, the easier it became to explain these complex topics in a way that made sense to people. 

I also experienced significant personal growth as a scientist. When I first started my research at the beginning of the summer, I was incredibly nervous, barely knew what was going on, and was scared to even ask questions. In short, I simply felt like I didn’t belong and that I was just slowing down the work of the people around me. However, throughout the course of the summer, I actively pushed myself to step outside of my comfort zone to ask difficult questions and learn as much as I could. By the end, I was able to have meaningful scientific conversations with the mentors in my lab, understand exactly what I was doing and why, and overall contribute positively to the lab environment, all because I pushed myself to learn and grow. 

As I look ahead to my leadership in action project, I know these skills will be incredibly useful in sharing information with populations that might not be familiar with certain technical or scientific language. As I said before, I have thought a lot about how my research connects to women’s health and healthcare inequity. Around the world, women face barriers to accessing healthcare or even adequate education about their own bodies, and it is my hope that next summer I will be able to make a positive impact and effort towards rectifying these issues.