September 2023

By Beni Adelstein | Bond LSC

Research skills aren’t built in a day, but Cynthia Tang’s diligence brought those skills to bear as she recently received a National Institutes of Health fellowship to further her budding career in science.

“Receiving the F30 fellowship means that the NIH sees value in my research proposal, in my training environment at the University of Missouri, and in my potential to become an independent physician-scientist,” said Tang, who works in Henry Wan’s lab at Bond Life Sciences Center. ­

This F30 predoctoral fellowship supports the research of students pursuing M.D.-Ph.D.’s. These awards help lighten the financial burden of a degree path that takes the better part of a decade for Tang and others pursuing their passions in research.

Tang came to the University of Missouri in 2018, started her journey as an M.D. student, then took a break to pursue her Ph.D. In the dual M.D.-Ph.D., the first two years are spent in pre-clinical studies followed by a full-time Ph.D. for three to five years and then students finish up with clinical training.

So, what project captured NIH’s attention?

Tang focused on how Covid-19 spreads and who gets it in rural areas. The aim is to identify people who have a higher chance of getting sick or being hospitalized so preventive measures can be offered earlier.

Lots of data must be gathered to accomplish this. Scientists sequence the genome of coronavirus variants­ to get a detailed profile and compare how that genetic makeup evolves over time. They do this by testing nasopharyngeal swabs—the Q-tips that are commonly used for covid tests—of people suspected of coronavirus infection. They use electronic health records to see who is getting sick, and add demographic information on age and location to the clues researchers analyze.

Tang focuses on rural populations in the U.S., especially in Missouri, because there is a gap in data from those communities. Once those at highest risk are identified, scientists can prioritize early prevention of COVID-19.

“Our biggest motivation for studying rural populations is so that we can better understand the way the virus changes and how to better serve those communities,” she said.

The process of planning for the grant itself enabled Tang to finetune the quality of her work.

“I feel like it helped speed up my Ph.D.,” she said. “We had to break down the research study into all the little pieces of what needs to be done and what to do when things go wrong. Everything must be so well thought out to put the grant together.”

After Tang’s initial fellowship application was rejected, she challenged herself to put together a completely new study design in under two months for her second proposal.

“It was the best thing that ever happened” said Tang, smiling. “I am extremely grateful to Dr. Wan and my thesis committee.”

This fellowship has provided many opportunities to Tang. She traveled overseas to present her findings at international conferences, which connected Bond LSC research with an international science community. Last year she was in Belfast, Ireland, and this month she will be in Valencia, Spain.

Where is Tang now?

She’s working remotely as she wraps up her Ph.D. and will graduate from Mizzou this upcoming December. Once she secures that Ph.D., she will head out to the University of North Carolina at Chapel Hill in January 2023 to finish her clinical training. Then she will enter the homestretch of her dual degree journey, finishing her M.D. in May of 2026.

Cynthia Tang currently serves as president-elect of the American Physician Scientists Association and will continue in this role next spring.

Tang was recently first author on “SARS-CoV-2 and influenza co-infection: A cross-sectional study in central Missouri during the 2021-2022 influenza season.” in the Journal of Virology in 2022.

By Sarah Kiefer

The spikes that protrude from SARS-CoV-2 present a topography of peaks that drive one MU researcher to ask more questions.

To Kamal Singh, a principal investigator at Bond Life Sciences Center, assistant professor in the MU College of Veterinary Medicine, and the director of the Molecular Interactions Core those spikes are a changing map with every new variant of coronavirus, and they lead his lab to study its constant evolution of mutations and proteins.

The Singh lab recently found an unexpectedly high number of APOBEC-mediated mutations among a patient cohort from South India, who were vaccinated and have taken remdesivir  to treat COVID-19. Using data from patients from South India, Saathvik Kannan, a senior at Hickman High School and a computer programmer and researcher for the Singh lab, tracked changes to the virus’ proteins.

“Viruses are known to evolve under pressure, so what’s happening in this case is that antibodies recognize the virus in a cell, and the virus sees that, ‘okay, there is someone to stop me,’ so it makes mutations,” Sing said.

The virus’ genome often alters, by mutating, inserting, or deleting the building blocks within RNA and DNA known as nucleotides, depending upon the pressure that viruses experience. Only some of these sugar-containing nucleotides are mutated, but these small changes can form a new virus variant.

“A few months ago, we didn’t know the reason behind all of these different mutations in patients because we did not look for APOBEC-mediated changes in viral genome,” Singh said.

Kannan found that between October 2022 to January 2023, the mutations among the patient cohort varied between 114 to 83. With that information, he identified 50% of patients who have diabetes and that when they take insulin the APOBEC protein is also induced. This new discovery leads to more questions about those with underlying conditions like diabetes or hypertension, and how they are affected differently by the drug.

“Ideally if a virus were to evolve you would see a linear progression, but you don’t see that here,” Kannan said. “This shows us that APOBEC proteins are likely one of the causes of this evolution in the virus and the makers of these variants.”

When the number of mutations vary significantly, like in the cohort, it re-affirms that the COVID-19 variants evolve independently.

Viruses don’t stop to wait for researchers to find the root of the problem. They constantly change, and in cases where the patients are vaccinated and take remdesivir, the virus evolves by taking advantage of the APOBEC protein.

Singh started down this path of research early on in the pandemic. Through PCR tests — which converts tiny amounts of RNA into DNA then copies it to a measurable level — he learned more about each mutation by comparing new samples to the original viral strain. Until this point, there had only been around 40 identified mutations in the spike protein in the different XBB sub-variant, but Singh’s lab has identified two more. The details have been recently published in The Lancet Journal. The two mutations found in this process, A27S and T747I, were unique to this cohort.

“For the first time we’re able to get more clues on what is actually happening because we were able to compare it to patient samples in the XBB subvariant and we had a wealth of data to work with,” Kannan said.

The Omicron variant yields new mutations constantly — currently at 43 and counting — and presents plenty of opportunity to look for differences. Singh directs his team to comb through variant sequences to identify differences in their genetic code. Soon they begin to identify trends from one month to another.

“The main difference here was that we have high quality patient data this time, from a patient cohort in Southern India, as opposed to getting it from a database,” Singh said. “The patient cohort tells us that the mutations are more than what was reported before.”

From January to June of 2020, Kannan found mutations that were present in one hundred percent of the sequences. When the Alpha Beta variant arrived in November 2020, they immediately knew what the mutated changes were. Their findings were published in the paper, ‘Clinical characteristics and novel mutations of omicron subvariant XBB in Tamil Nadu, India – a cohort study,’ in April 2023 in the Lancet Regional Health – Southeast Asia Journal.

After new variants — such as the XBB sub-variant of Omicron — materialized, Singh’s lab identified existing drugs and new drug compounds that match structural weaknesses presented by mutations.

One way Singh’s lab keeps abreast of new variants is by utilizing the tools they have available to their advantage with in-house programs either in Python or in R programming languages. Their lab can construct non-infectious, virus-like particles that contain all the same characteristics as the virus minus its genome, or its genetic material makeup.

A collaboration between University of Missouri and University of Nebraska Medical Center helped identify the new drug compound, MU-UNMC-2 in addition to the MU-UNMC-1, previously found in late 2021, as potential coronavirus treatments.

Remdesivir, ribavirin, favipiravir, and molnupiravir were among existing drugs that can be used to treat COVID-19 under certain conditions. Remdesivir — previously ineffective as an Ebola virus treatment — initially proved somewhat useful for COVID-19. Molnupavir only showed moderate promise, and the two drugs were approved for emergency use by the FDA for the treatment of COVID-19. Remdesivir now has full FDA approval.

“To address the problem, you have to find the problem. Where is the virus? What can we address? And then we go after it,” Singh said. “My lab was the first one in the world who suggested when the pandemic came that these are a few drugs that can be used and remdesivir was one of them. I’m very proud that I could contribute to human health in that way.”

With knowledge of this new protein in hand, Singh’s lab now is moving forward to focus most of their attention on the APOBEC protein mediated mutations and how underlying health conditions can affect an individual’s reaction to certain drug compounds.

“The virus has been outsmarting us, outfoxing us, until now. As soon as we get something, it changes itself,” Singh said. “That keeps us motivated as we chase it.”

The Singh lab’s research on the XBB sub-variant mutations was published in the paper ‘Omicron SARS-CoV-2 variant: Unique features and their impact on pre-existing anitbodies’ in the “Computational and Structural Biotechnology Journal” in June 2021 and their updated findings can be found in the paper ‘Clinical characteristics and novel mutations of omicron subvariant XBB in Tamil Nadu, India – a cohort study,’ published in “The Lancet Regional Health – Southeast Asia Journal” in April 2023.

Funding provided by: Bond LSC, Swedish research Council, American Lung Association, National Institute of Dental and Craniofacial Research in collaboration with Prof. Gary Weisman of the Bond Life Sciences center.

By Sarah Kiefer

The hunt for emerging coronavirus variants keeps Torin Hunter busy testing samples from sewer systems across Missouri.

As a part of The Sewershed Surveillance Project, Hunter has spent the last year and a half taking each test tube and carefully filtering the samples to contribute data on how SARS-CoV-2 can be present in our communities.

But Hunter started his journey in virology like many — a student trying to sift through all the different subjects and figuring out what fit him the best. He tried out clinical jobs and considered going into the health professions, but he began to miss the lab and research more than he thought.

“I missed the freedom and the creativity you get from research while still being able to do science and complete it all by yourself. It’s more fun and satisfying to me,” said Hunter, a senior research technician in the Marc Johnson lab at Bond LSC.

Virology, the study of viruses, stuck for Hunter.

“I like the blend of molecular virology and epidemiology here because it is fascinating to see the public health implications of what we are studying,” he said. “Even though our work has focused on coronavirus in the past few years, I would still want to be doing this type of work even if it were involving a different virus.”

As a senior research technician, Hunter enjoys projects where he can think through an experiment and work backward if problems arise. Using his analytical skills, he can determine if the experiment was meant to go in a certain way or if the issues can be boiled down to human errors made along the way.

“Starting an experiment and then getting real results from it that make sense is the most exciting part of my job,” Hunter said. “Knowing that I did it by myself and that I accomplished something I worked hard on is a great feeling.”

Originally from Orange County, California, Hunter moved to Missouri when he was 16 and knew he wanted to attend the University of Missouri not long after that. In high school, he began to take an interest in the inner workings of cells, which is where biochemistry comes into his life.

He went on to earn his bachelor’s degree in health science from Mizzou and his master’s degree in microbiology and cell science from the University of Florida. When Hunter took his first undergraduate class in biochemistry, he became hooked on the subject and its value in a research setting.

“Biochemistry is when everything clicked in the life sciences for me, and it made the most sense to me, but the subject is so broad that I still didn’t know exactly where to go from there.”

Hunter works daily with samples of human waste systems across the state. He extracts viral nucleic acids (such as DNA or RNA) out of the samples using polyethylene glycol or magnetic beads and sends them off to a collaborating lab. The lab then performs digital polymerase chain reaction (dPCR) to quantify, or measure, the viral load of SARS-CoV-2 in the wastewater samples. The process can be compared to a nasal swab coronavirus test, but instead of simply measuring the virus’ presence, the amount of virus in the sample is measured as well.

The amount of each marker is graphed on a chart, and these charts are analyzed and monitored in collaboration with public health officials to compare trends across the state. Hunter finds it interesting to learn about how different viruses cause disease.

“If we can better understand why and how viruses cause harm to people, we can develop more therapeutic or preventative options like vaccines and help a lot of people in the long run,” Hunter said.

Hunter performs these steps often and finds that this type of work takes a certain amount of discipline. He meticulously combs over his notes and studies the details of his experiments and what might have gone wrong in the procedure.

“You go through every possible way that something could go wrong, but if those are still your results, you might just have to go further back in your process or go in a different direction entirely,” Hunter said. “That’s what makes this work so interesting, is that you’re always problem-solving.”

In his time away from the lab, Hunter enjoys playing video games, weightlifting, and cooking to turn his brain off after a day of experimentation.

“It’s important and healthy to have a work-life balance,” Hunter said. “I will often think of new dinner recipes to try and make as my own way of experimenting at home.”

After his research at Bond LSC, he plans to apply for Ph.D. programs this fall to continue his studies in virology. Specifically, he would like to learn more about immune responses to viral infections.

“Everybody has to work, and if you’re going to work hard at something, you might as well do the thing that you enjoy the most,” Hunter said.