Author Archives: Michael Lee

Only the beginning

Eight weeks is almost nothing in the world of science, where experiments are marked in months and results in years. At most, I can say I caught a fleeting glimpse, a snapshot of a singular moment in the scientific process. But even this ephemeral glimpse was saturated with incredibly valuable lessons that do a little more in clearing the uncertainty shrouding my future. In just these short eight weeks, I was able to experience both the highs and lows of scientific research. I got a taste of the beauty and freedom in asking your own questions and creatively crafting your own solutions, while encountering many unforeseen obstacles along the way. Through this process, I am grateful to my mentors Dr. Ru-Rong Ji and Dr. Chris Donnelly for their close mentorship. I am also immensely grateful for Dr. Grunwald and Anna for their support in this journey of discovering what science means to me.  

This summer was truly an immersive experience I had initially hoped for. In the beginning, I could barely differentiate a microglia from and an astrocyte, but now I can explain the distinct roles they play in the pain system as a whole. I got the opportunity to learn many new techniques, such as behavioral tests and immunohistochemistry, and strategies in general that scientists use to elaborately and seemingly effortlessly answer some of the most mind-boggling questions. Through trials and failures, I was also able to experience the struggles of research: time-consuming results and elusive perfection. However, what I enjoyed the most this summer more than learning any technique is thinking like a scientist, from asking questions, to communicating science to diverse audiences, to assessing the validity of scientific methods. Events such as the chalk talks and poster session have shown me that science is always an ongoing conversation that I’d be excited to be a part of. 

What gave me an even greater sense of what science could mean to me were the faculty talks. It was really helpful to hear the distinct perspectives of different faculty and how many different paths can all lead to the same desired result. From their experiences, I have learned to see scientific research as a humbling relationship, where no one can really outlearn science. These scientists are some of the most knowledgeable people in their fields, yet they have the humility to admit they know nothing when in search of new knowledge gaps to investigate. And so, I feel like I am definitely farther along the “chase” of science I had mentioned the first week but I am nowhere near the end. 

Trust the Process

Few people are recognized by the HHMI as exceptional scientists who deserve a lifetime of nearly unconditional funding– scientists who are entrusted to do good science no matter what, as long as they have the money to do so. It is not surprising that nobel laureate Dr. Robert Lefkowitz has earned this trust, having spearheaded work on G-protein coupled receptors involved in one-third of all medical drugs today. Despite Dr. Lefkowitz’s impressive list of accolades and contributions to science, his career in scientific research came rather late and unexpectedly. In fact, he could have never imagined a research career in his early days. 

Dr. Lefkowitz was always a physician by training, used to dealing with patients daily and seeing quick results–the antithesis of basic science research. So how did he find himself in the role of one of the most well-regarded research scientists? Dr. Lefkowitz’s first immersion in research was not exactly the product of deliberate will. The US military initiated a draft of doctors to aid in the Vietnam War, and Dr. Lefkowitz decided to instead serve as a Clinical and Research Associate at the NIH to avoid participation in the war. Surrounded by a group of talented colleagues, Dr. Lefkowitz quickly caught a passion for basic science research. 

This ultimately gave way to a dilemma in choosing between medicine and basic science research, one I could imagine left him especially torn because medicine and research are two vastly different, yet intimately entwined fields that build off of each other. Research is a thrilling journey to reinvent medicine, but medicine is often a deep-rooted calling to make humane interactions. Of most humane interactions were the raw and empowering moments of Dr. Lefkowitz with his father. The medical complications of his father only made it increasingly difficult for Dr. Lefkowitz to justify a full commitment to basic science research. But after his time at NIH, Dr. Lefkowitz realized that what he missed was the day to day activity of experimental results. Dr. Lefkowitz would eventually commit to fully pursue research over clinical hours. What I found most interesting was the nature of this trajectory– unconventional and sudden, yet very much fulfilling. It exemplifies the malleability of our interests sometimes and the limitations of planning.

One underlying principle that Dr. Lefkowitz seemed to emphasize throughout his talk was “serendipity”, attributing many of his life events to this elusive force. Dr. Lefkowtiz’s emphasis on the prevalence of serendipity and the importance of being prepared to take chance opportunities that come is, I think, a valuable piece of advice that comes at a critical moment in our lives. It helps find some solace in a daunting future, in the unplanned, but at the same time encourages us to fearlessly tread into the unknown. 

Abstract

The activation of type I interferons has long been reported to regulate pain, but the precise mechanism has remained unclear. We tested the hypothesis that type I interferons are critical pain regulators and function via a neuronal mechanism. Sensory, motor, and behavioral testing in addition to immunohistochemistry were performed on type I interferon receptor knockout (Ifnar1 KO) and wild type (WT) mice. Heightened sensitivity in KO mice suggests an importance of type I interferons in regulating pain under basal conditions. The lack of any difference in motor performance and quantity of nerve fibers in tissue of WT and Ifnar1 KO, along with RNA scope, support a neuronal-specific mechanism of interferons in regulating pain. These findings suggest that type I interferons are critical in regulating both pain and neuroinflammation by a neuronal mechanism, even under normal, healthy conditions. These findings redefine our understanding of the role of interferons in pain and could translate to improved treatment of a heterogeneous array of chronic pain.

A Morning Blend of Thoughts

On most mornings, I’m lulled into a new day at lab by the hum of a Krueger machine spouting out warm, ripe coffee. As decisions run through my head and I reach for the hazelnut creamer cups, a flurry of brief philosophical conversations with my mentor flutters into my head: Does free will really exist? Are decisions really ours to make? Maybe in what I thought was a free will decision in choosing to put creamer in my coffee was merely the manifestation of an innate evolutionary urge for creamy food, a predetermined course of action I am just biologically hard-wired to. Like this, I feel like I am learning more than just techniques but different modes of thought everyday in the lab.

However, before any intellectual discourse with my mentor, I must tend to start any experiments that run on a rather rigid schedule. To set up the experiments for the day, I gear up with a full outfit of personal protective equipment and head into the animal room to retrieve mice. I then transfer them to a private suite, where I randomly place mice of all cages into separate enclosures on top of a perforated stand. For an hour or so, they are left to roam freely in their enclosures as they habituate to new scents and sights of the room. When it is my turn to test the sensitivity of these mice, I frantically evade their droppings and track their most subtle movements as they scurry around in their enclosure. Towards the end of the day, I will usually analyze and display the data I’ve collected on an Excel spreadsheet and run multiple statistical tests. Science is about robust data and multiple lines of evidence. And so, on select days I would complete immunohistochemistry to visually verify the behavior of certain cells of interest. To do so, the first step is to section different tissue using a cryostat, almost like a deli slicer. In the next step, I would stain for different types of cells by marking different cells with colorful tags using antibodies. When it comes time for these tissue to be imaged, I would get to operate a powerful imaging scope to take multi-layered photos of the samples. 

It is in between these experiments when my mentor would test my understanding of the fundamental science and then proceed to thoroughly explain the purpose and thought process behind each experiment. Currently, I am working on an experiment that involves siRNA, which knocks down IFN receptors in mice temporarily to eliminate any developmental issues as confounding factors that could accompany knockout models of mice. To help aid my understanding, my mentor doodles precisely illustrative diagrams to accompany each explanation. It is also during this time when I get to ask any questions on my mind or discuss any philosophical inquiries that somehow relate to our project. Day after day, lab feels like an intellectual playground, and I am grateful for the freedom my mentor allows for me to wonder. From the elaborate and clarifying explanations my mentor consistently provides, I am more than ever opened up to the intricate beauty and cleverness of scientific research. 

Fight Fire With Fire

I really enjoyed hearing everyone’s story of their ongoing scientific journey this past week and was able to gain a better glimpse into the multifaceted world of biology. But in light of the one of the most dire epidemics in America–the opioid crisis–I thought that John’s project was particularly inspiring and relevant. The current state of dealing with this issue in our country is at best tenuous: 11.4 million people have misused prescription opioids, and more than 130 have died every day from opioid-related drug overdoses (NCHS). In response to this, instead of developing a drug to tackle a certain disease, John is developing a drug to tackle a phenomenon that has often received less and much needed attention: addiction. I thought John’s question of whether or not certain drugs can be developed to curb addiction to pain-relieving drugs was both innovative and compelling, with far-reaching potential in helping a wide array of people.

In investigating his hypothesis, John’s goes about in a well-founded scientific manner. Rats are placed into a skinner box, in which they are trained to press a lever that then triggers an intravenous injection with a powerful synthetic opioid called Remi-fentanyl. To test the efficacy of anti-addiction drugs, the behavior of addicted rats previously injected with opioid are compared with control rats injected with saline. If the addicted rats exhibit less of a need for Remi-fentanyl after being administered with the drug that John is trying to develop, then it could have potential to fight off addiction and the opioid crisis. This experimental process I thought was a simple, yet effective strategy in isolating the effects of anti-addiction drugs. With an interesting and robust experimental set-up, I have become really curious as to how drugs are chosen to be tested and what types of drugs have already been shown to have some desired effect. 

As I reflect more on John’s exciting and intriguing project, a plethora of questions flood my mind in regards to the implications of this work. On the most basic level, I am wondering about the mechanistic pathways of these drugs. Even if these drugs suppresses a mental urge for opioids, can it also eliminate a physical dependence on these drugs that develops in addicted patients? On a similar note, these drugs have a very noble goal in mitigating addiction in order to help people, but how can they be administered if certain patients resist in receiving these drugs, especially if this anti-addiction drug isn’t able to provide nearly as many relieving effects as opioids? Furthermore, opioid addiction is caused by increased tolerance, but could these anti-addiction drugs have problems with tolerance and resistance themselves? I’m sure many of these questions will be answered and addressed if this project progresses into the drug development process, and I am definitely excited to be there for that and to hear more about it. 

The Audacity to Imagine and Create

In the world of non-neuronal cell contribution to pain, there is one name that does not escape the mind: Dr. Ru-Rong Ji. Currently chief of pain research within Duke Anesthesiology, Dr. Ji’s humble roots extend back to China, where he went to Nanjing University for undergraduate studies in the biology department and studied human physiology. Afterwards, he found himself at the Shanghai Institute of Physiology for graduate studies and earned a Ph.D. in neuroscience. Further in his academic journey, he studied neurobiology at the Karolinska Institute, the Royal Institute of Medicine in Sweden. It was after this when he first came to the US, initially at Johns Hopkins Medical School. He then spent his next 14 years at Harvard and the Massachusetts General Hospital, where he went from an instructor to assistant professor to associate professor and received his first NIH independent grant before coming to Duke. Upon arrival to Duke for the first time, what first stuck out to him as special and necessary in science was the environment–one that was collaborative and supportive.

Throughout this extensive scientific background, his goals have mostly remained the same: to understand the mechanisms by which pain operates and create translational potential to help patients dealing with chronic pain. However, over time, his interests have shifted from the role of neural circuits to non-neuronal cells in pain, and now he is one of the leading experts in the non-neuronal cell contribution to pain.

Biology, however, was not always his calling. In high school, Dr. Ji first gravitated towards math or physics since biology at that time was not so popular. Before his year, biology was not even taught in high school. However, he had an amazing teacher that was able to open him up to the wonders of the world of biology. From then, his interest snowballed and led to an entire career in neurobiology. Dr. Ji has since very much enjoyed science because it is a creative and innovative craft, with every project leading to a new journey. Through the seminars and talks he has given all over the world, he has very much enjoyed communicating with other scientists to spark new ideas in each other and to eliminate any kind of boundaries science might have. One of the most rewarding aspects of his career he thinks is mentoring students and fostering in them a love for pain research and science itself.

In recollecting what first got him into pain research, Dr. Ji remembers being very fascinated with how acupuncture was able to produce pain relieving effects. He was interested in how it happened and what neural pathways were activated by acupuncture. When he first started as a research scientist, the focus of pain research was on how neurons themselves contributed to pain. Over the years, Dr. Ji and others have discovered that non-neuronal cells such as microglia or astrocytes release mediating molecules that regulate pain, elucidating the interaction between the pain system and the immune system.
When asked what he hopes will happen in the field of pain research, he expressed the need for more effective and personalized therapeutics for pain relief. Currently, every drug and medicine has its own limitations: there is no one magical treatment. However, he’s optimistic that his research can increase our knowledge base of the mechanisms of pain, ultimately to increase our options of treatment and address a wider array of diseases.

Though widely recognized today, Dr. Ji has personally experienced the difficulty in taking unconventional routes, often away from what the public deems to be the most exciting or popular topics of science. But he believes that even if what you are doing is not the biggest, most showy story, you must not let go of your personal belief of what will make an impact in the future. Communication with others is important but you should always maintain your independent thinking. For young scientists today, his biggest advice is to be creative, stay open-minded, and remain persistent and patient. He believes that successful science doesn’t require raw intelligence. With enough persistence and focus on specific goals, you can eventually be recognized as a leader in whatever field you pursue. The world might be very different in the next 10 to 15 years with the rapid progress and innovation in medicine, and he is very optimistic that the young generation will pioneer this progress.

With all that, I am immensely grateful for the excellent mentorship I have received in the Ji Lab from mentors such as Dr. Ru-Rong Ji and Dr. Chris Donnelly and the opportunity to be a part of this exciting, ongoing journey.

A Stinging Pain

Cancer pain delivers a message. A vile, biting message to patients that life itself is seeping through their fingers. A disheartening, damning message to medical specialists that even their powers are limited. It thrusts itself as a physical manifestation of the unseen, of your interiors being eaten away. It is common to mistaken pain as merely pain neurons being stuck in the ‘on’ position, but it is much more complex than that: pain that develops from an acute injury is actually the result of hyperactivation of the peripheral pain neurons responsible for inflammation. Previously thought of as merely bystanders, these pain neurons have been revealed to actively participate in the progression of the carcinogens. With cancer pain being one of the most intense and difficult brands of pain to treat, further research is warranted more than ever.

My project this summer in the Ji lab branches off of my mentor’s ongoing project on the role of STING, a protein that activates interferons, in suppressing pain and reducing neuroinflammation. STING holds a central role in the innate immunity–it produces a protective response by inducing mediators such as cytokines or chemokines. These molecules can then either produce or suppress neuroinflammation. One of the most important mediators that STING activates is type I interferons, which are proteins that boost the immune system in fending off viruses or cancer. As a result, activating STING has potential to not only decrease tumor growth but also dampen neuroinflammation, the underlying etiology of pain.

My mentor and I have been running experiments on mice to not only affirm the role of STING in producing interferons that decrease sustained pain but to also eliminate the possibility that other mechanisms are contributing to this effect. Mice that experience chronic pain have a heightened ability to feel pain from that stimuli that wouldn’t induce pain in normal conditions. To determine the exact threshold where these mice are first able to sense pain, I have been conducting a test called the Von Frey assay. A factor that can lead to false signs of pain in mice is anxiety, so I have also been conducting an open field test to track the movement of mice and then analyze signs of anxiety, ultimately to show that these mice are not any more anxious than normal. So far, the hypothesis that activating STING decreases pain has been supported, but further experiments are necessary to identify the exact mechanism by which STING is able to do so.

Abstract

On the first day of lab, I was greeted by the familiar weight of the heavy steel-rimmed doors, the luminous glass windows lining the lobby, and a never-changing view from the sixth floor of the picturesque Chapel, but something felt different. Walking down the hall towards the Ji Lab, my fingers felt a buzz and my heart felt tingly because I knew something was going to be different. Despite the rush of familiarity, there was a new, expansive world of science out there that I needed to see. Scientific discovery was never something meant to be reached or completed, only approached asymptotically, but I was excited for the chase this summer more than ever.

With research as a singular goal, I expect this summer to be a time of deep immersion. During the school year, I was involved a bit already in Dr. Ji’s neurobiology lab. However, I was limited to shorter, more flexible tasks. With research as a focus now, I am excited to try more time-intensive and complicated experiments, such as animal behavior experiments, this summer to contribute to the lab in novel ways. This past week for the first time, I was able to perform Von Frey assay, which measures the ability of mice to detect pain using thin hair filaments as stimulants. More than just learn how to perform new lab techniques, I hope to gain an understanding of the science behind the methods and why they are used, and I want to not only learn how to answer scientific questions but also how to ask them. By working closely with my mentors, I can hopefully understand their thought processes and how they fundamentally approach science.

Professor Patek’s faculty discussion on her journey in finding her role in science really invited me to think deeply and critically about my own relationship with science. How can worth be measured in the world of science? Is an inherent interest not enough to justify a particular field of work? Can countless hours of transferring liquid really amount to any real substance? And so, a personal, yet essential goal I have for this summer is to continue to redefine what science means to me and to discover new meaning in the science I do. What initially drew me to pain research was the agonizing pain that cancer patients often experience, a constant reminder of the fear and stigma surrounding cancer–that cancer equates to death. This heightened sense of mortality is frightening, but also deeply humanistic. Understanding the intersection of cancer and pain holds power to address emotional dimensions of care and pave way for humane treatments. As I learn more about microglia or ion channels and the potential and limitations of science this summer, I expect these reasons to shift and change but hopefully, eventually I can find personal meaning in the work I do that excites me as both a scientist and human being.