by Cai Anni
What is the Research Science Institute?
he Research Science Institute (RSI) is a 6-week summer research programme jointly organized by the Centre for Excellence in Education (CEE) and Massachusetts Institute of Technology (MIT). Following a stringent selection process, approximately 50 domestic students and 30 international students are invited every year to complete a research project in their field of interest under the supervision of an eminent scientist. The entire cycle consists of formulating a specific research topic, conducting relevant experiments, consolidating findings, and finally presenting oneís results in the form of a written report as well as an oral presentation. This rigorous approach is intended to kick-start a career in scientific research, and indeed many Rickoids go on to pursue a science or engineering degree in their tertiary education, often in renowned institutions such as Harvard, Princeton, andómost popular of allóMIT itself. Throughout its 28-year history, RSI has produced numerous scientific luminaries, among them Dr. Terence Tao, the 2006 Fields Medallist and one of the youngest mathematicians to be conferred this honour.
My Research Experience, and the Lessons I've Learnt
I had the privilege of working with Dr. Charles S. Hoffman of Boston College, who specializes in yeast genetics. His work centres on glucose sensing, signal transduction and transcriptional regulation in the fission yeast Schizosaccharomyces pombe as a tool to study cyclic nucleotide phosphodiesterases.
Phosphodiesterases (PDE) is an important enzyme involved in cell transduction pathways involving the second messenger cAMP. Spontaneous degradation of cAMP by PDEs allows a cAMP-mediated cellular transduction pathway to be quickly terminated once the initial chemical messenger is no longer present, allowing the cell to respond to a fresh or repeated signal. This feedback loop forms a basis for the tight regulation of metabolic pathways in response to a fluctuating environment. There are 11 different families of PDEs in humans, each with its characteristic substrate specificity and regulatory properties. The fact that different cell types express unique distributions of PDE isoforms which influence distinct cell signalling pathways provides the opportunity to stimulate or inhibit specific pathways linked to disease states without inadvertently inducing a global effect on the body. Isoform-specific PDE inhibitors have already been identified and engineered into commercially viable drugs, the most notable example being sildenafil (Viagra). Though popularly known as a treatment for erectile dysfunction, it is also effective against the rare, but potentially fatal, pulmonary arterial hypertension (PAH).
Sildenafil is a PDE5 inhibitor. My research, however, centred on the PDE4 family, which is implicated in a number of autoimmune diseases including asthma and arthritis. The isoform I investigated, PDE4B, controls the production of TNF-a in both monocytes and macrophages in response to bacterial lipopolysaccharide (LPS) stimulation and is therefore crucial in mounting inflammatory responses. Much effort has been devoted to seeking a PDE4-specific inhibitor, with some degree of success. My mentorís lab previously identified 4 organic compounds which demonstrated selective PDE4B inhibition through a high throughput screen, but the absence of any discernible homology made their success something of a mystery. The task for me, then, was to pinpoint critical amino acids in PDE4Bís primary sequence that interact with the previously identified inhibitors, a crucial step in elucidating the mechanism of selective inhibition.
Our approach seemed straightforward. We first transformed a PDE-deficient strain of S. pombe with randomly mutated PDE4B genes generated via PCR. Subsequent screening of candidates would identify inhibitor-resistant mutant PDEs. After these genes of interest were isolated and sequenced, we would compare them with the wild-type PDE4B gene to pinpoint amino acid substitutions that presumably was responsible for inhibitor resistance, and by extension, interaction with the chemical inhibitor.
Unfortunately, the initial experiment results differed appreciably from what we envisioned. For one, iodine-staining mating assay we performed were supposed to produce distinct staining for inhibitor resistant colonies, but the first batch of transformants all exhibited varying degrees of staining so we could not unambiguously identify target colonies. To avoid losing promising candidates for sequencing, we resorted to inspecting the ~600 colonies picked after a preliminary screening under the microscope one by one, then transferring single cells to a new culture to start a pure colony. It was arduous work, but eventually paid off as the subsequent assays yielded much-improved results.
Similar incidents would recur with frustrating regularity throughout the course of my research. Whenever something goes awry, all I could think of was how it will derail my project. In stark contrast, my mentor was completely unperturbed, immediately searching for ways to circumvent the problem. Astonishingly inventive, he wasnít one to be circumscribed by existing protocols and standard procedure. The steps had not necessarily been optimised, he said, and even if the original inventor had made them optimal for his experiment, in other contexts they must be adjusted to fulfil a different set of aims. So that was my most important lesson: in research there are no golden rules and no clear answers. The researcher has to navigate the murky waters of uncertainty to reach a target that may not even be fixed in place. His mind must be flexible and constantly engaged; his instincts well-honed, to recognize patterns that others would have missed. But even more paramount is resilience, the quality that drives successful scientists to persevere after multiple attempts have ended in abject failure.
For most students our age, a research project is deemed to be complete as soon as publishable results are obtained. Success is defined by reaching an unambiguous conclusion, preferably supported by a plenitude of consistent data. Erratic, anomalous data make for a thoroughly lacklustre project; having no result is even worseóit means that you have wasted all your time at the lab. At the start, every RSI mentor made it unequivocally clear that results are not guaranteed, and indeed many of my peers were stuck in the quandary of having ďnothing to writeĒ at the end of RSI. Therein lies our fundamental misconception that any research venture which did not yield ground-breaking results was not worthy enough to be published. But scientific advancement is an incremental process, often relying on trial and error. Unsuccessful attempts are not as sensational as the occasional breakthrough, yet just as indispensable to progress. Knowing what doesnít work leads us closer to uncovering what actually works. Newton famously said, ďIf I have seen further than others, it is by standing upon the shoulders of giants.Ē Might I also add that all seminal discoveries also rest in part on the shoulders of all researchers past and present who did not attain scientific acclaim, but whose workóeven if it is propounding a theory later proven incorrectóhave contributed to this great enterprise that is science.
Vivid Memories, Lasting Friendships
RSI was truly transformative. I experienced the full gamut of emotions, from abject misery, such as when I came down with flu close to the final deadline, to giddy exhilaration, when I was conferred honours for both the Paper and Presentation.
Of course, research is no sinecure. The perpetual rush to meet looming deadlines and exacting demands, the frequent late nights (we worked till 5am every day in the last week), the strain of working full-day at the labÖall of it pushed me to the limit. But having survived it and risen far above my expectations, I have emerged stronger than before.
Iíve also made great friendsóthe kind Iíll make sure to stay in touch with despite being continents apart. The camaraderie among Rickoids is simply ineffable. We went through trials and tribulations together, played crazy games and sat under the stars watching the fireworks display on Americaís Independence Day. My friends are also an immensely talented bunch. One was the ISEF 2010 Grand Winner, another composed a piano piece overnight, a third is a sophomore at Harvard at age 16. Almost all were top achievers at their respective schools. Everyone had something to teach, and they do it freely, with no hint of condescension. In fact, if RSI was only about meeting these people, it would have been worthwhile already.
When we were asked to describe RSI in an essay, one Rickoid decided to be laconicóhe simply wrote, ďAn epic summer programmeĒ, and that was it. In the same spirit Iíll conclude my experience with a few other words beginning with E: exhilarating, extraordinary, and most of all, enlightening.
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