M.D-PHD candidates Cynthia Tang and Brian Thomas share their experience applying for the NIH F30 Fellowship. | Photos by Roger Meissen
Brian Thomas got the official letter in the mail Monday after months of waiting.
“It’s a long time coming,” he said, “lots of patience and collaboration.”
Thomas is one of two student scientists at Bond Life Sciences Center to receive F30 fellowships — officially the Ruth L. Kirschstein National Research Service Award (NRSA) — from the National Institutes of Health (NIH) this year, a first for MU.
The agency awards F30 fellowships to MD-Ph.D. students pursuing related areas as they work towards their doctoral degrees. Those awards add up, with up to six years of funding to cover costs of research and clinical training.
When Thomas originally applied, his proposal was rejected, but the second time was the charm. He studies cancer immunology in the lab of Donald Burke at Bond LSC.
“You learn from failure, reflect on it and grow,” he said. “You learn to think critically about your research proposal and organization.”
Cynthia Tang — also working toward her MD-Ph.D. dual degree — got official word about her award ahead of Thomas. It allows her to continue her research in the lab of Henry Wan at Bond LSC. She researches the evolution and spread of Sars-Cov2 — the cause of the Covid-19 pandemic. Her proposal was also rejected the first time, yet she persevered. She encourages other students to apply for the fellowship.
“You should really go for it. It seems like a lot, and it can be really intimidating when you look at the checklist and all the components,” Tang said. “But it is possible, and it is doable.”
Both students are thankful for what the fellowship has done for them. It pushed them to navigate the highly competitive grant application process as well as clearly outline their goals and ambitions.
“We have an incredible grants team and our faculty is amazing,” Tang said. “There are so many resources available at Mizzou.”
Thomas agrees.
“We have fantastic directors doing a wonderful job growing the environment of our programs and tending to our student’s needs,” he said.
This NIH fellowship is an opportunity for students to pursue their passions in science and research while alleviating financial burdens, giving researchers like Tang and Thomas the tools to succeed.
The Tom and Anne Smith MD-Ph.D. Program at the MU School of Medicine is a seven- to nine-year course of study that combines the traditional four years of medical school with the three to five years typically required to earn a doctorate in a scientific discipline. It prepares students for a career in academic medicine.
About Bond LSC At Bond Life Sciences Center, the best answers come from working together. Our building and culture leverage expertise of faculty investigators to develop discoveries that matter. Our researchers represent diverse academic backgrounds with projects focused on infectious diseases, agriculture, informatics, the environment and other areas. By moving beyond the boundaries of departments, our research increases its impact and lays the groundwork for a better world while teaching the next generation of scientists.
With a forceful swing of his badminton racket, Vikranth Chandrasekaran propelled the shuttles across the court. A game with coworkers and friends is the perfect way to wrap up a day in the lab for the postdoctoral fellow. He’s offered to teach his colleagues the strategies of badminton at the University of Missouri Rec Center.
“When I initially embarked on my journey in badminton as a beginner, I received invaluable assistance and guidance from numerous South Korean individuals who graciously taught me the proper techniques,” he said. “Now, I feel compelled to reciprocate this kindness by offering my help to those who aspire to learn badminton.”
As a postdoc in the Bing Stacey lab at Bond LSC, Chandrakaran has the opportunity to help others through soybeans, and his path here started in South Korea with Gary Stacey.
During the joint venture conference between the University of Missouri, USA and Gyeongsang National University, South Korea, Chandrasekaran found Stacey’s research talk fascinating and made him want to visit Bond Life Science center.
“I want to find the answers behind many unknown questions and be the one to make new discoveries,” Chandrasekaran said.
Chandrasekaran is from a land of gold mines: Karnataka, India. Coming from Kolar Gold Fields, he can appreciate searching for the nutrients within our food and his past planted the seed for his future.
“My motivation for what I do comes from my strong determination and passion towards plant science for the betterment of humankind that I have,” Chandrasekaran said. “My childhood aspiration was to achieve the highest degree in the scientific field, a doctoral degree, and with unwavering support from my parents I was able to realize that dream.”
Chandrasekaran deals with soybeans and the macronutrients they provide. Soybeans are an important source of protein for a vegan diet especially, with a single seed consisting of 40% protein and 20% oil. The crop is a leading source for vegetable oil and protein production, providing 60% of global oilseed production and more than 25% of the in food and animal feed worldwide.
Chandrasekaran investigates the phenotypes of the soybean plants and takes care of his plants by doing regular pruning and weeding. | Photo by Sarah Kiefer, Bond LSC
“Engaging with academic theories for research proved to be profoundly challenging because they often neglected the intricate trial-and-error nature inherent in research endeavors,” Chandrasekaran said. “But research entails a myriad of trial-and-error processes, involving a multitude of fine-tuning techniques that extend beyond the scope of conventional academic teachings. These trial-and-error methods lead me to learn more and help me to solve a problem in my research in a more effective way.”
In the Stacey lab he finds the genes responsible for high protein and oil content in order to then edit DNA to increase the amount of protein and oil contained in the plant. Less than 30% of the genes in soybeans and 70% of those in rice have been identified, so Chandrasekaran aims to find more.
“I was first fascinated with research because of the DNA double helix structure and how we analyze things like that,” he said. “I was always focused on the theoretical part of academics, but then all of the sudden I was really curious about the research side as well.”
When he screens the soybean plants for genes, he pays attention to the visible phenotypes, that the plant makes in response to a procedure using fast neutron radiation (ϒ rays or gamma rays), which is often used in cancer patients to treat tumors. For each phenotype there are many genes competing to express themselves, so Chandrasekaran deciphers which traits are likely distinct, or abnormal phenotypes, when compared to normal soybean plant.
Chandrasekaran’s soybean plants are kept at Bradford Research Farm, the largest concentration of plots dedicated to research in Missouri, with 591 acres of land. He regularly goes to weed and take care of the plants, which are uneven in height and thickness because they are mutant plants. | Photo by Sarah Kiefer, Bond LSC
He finds a calmness in this type of work much like the tranquil feeling he experiences when he travels to places such as the Rocky Mountains.
“The drive was about 13 hours and that scared me at first,” he said. “But when I was driving, I was calm and got to experience for the first-time incredibly beautiful scenery that I had never seen before which made the drive worth it.”
He also frequently makes the 405-mile trip to Chicago to visit his childhood friend and their family each month. A long drive gives Chandrasekaran time to think and makes him more motivated about his work when he returns.
“Taking short breaks from continuous lab and field work helps me to feel happier, have more energy and focus more on my research,” Chandrasekaran said.
He uses travelling and a game of badminton with coworkers and friends to wrap up a day in the lab or take time to think on the road he is paving to making scientific discoveries. He hopes to continue his work in the field and secure a position as a crop scientist or to start a research lab of his own one day.
Clayton Kranawetter, a postdoctoral fellow in the Lloyd Sumner lab at Bond LSC, recently received a USDA National Institutes of Food and Agriculture Postdoctoral Research fellowship in which he uses this mass spectrometry machine to study the significance of plant border cells. | photo by Sarah Kiefer, Bond LSC
By Sarah Kiefer | Bond LSC
Some of the most fascinating things in science happen at the border where one organism interacts with its environment.
That’s the case with root border cells, and Clayton Kranawetter is one individual exploring this frontier.
Kranawetter recently received a $223,000, two-year USDA National Institutes of Food and Agriculture Postdoctoral Research fellowship for his project on this group of cells. This fellowship is a part of a larger $12 million investment in multiple institutions by the USDA to expand research in the area of agricultural microbiomes. The University of Missouri is among 17 research institutions to receive funding.
“I’m still shocked by receiving the award. I was already having a good day when I received my award notice. I found a good parking spot in the garage and a quarter on the sidewalk, but then I got the email” said the postdoctoral fellow in the Lloyd Sumner lab at Bond Life Sciences Center. “I thought it was a mistake at first and then I realized, ‘oh, this is real.’”
This fellowship will sustain his work in an influential way as he dives into the secreted metabolites and mechanisms of isolated border cells and their impact on rhizosphere dynamics.
Metabolites are small molecules made or used when a cell breaks down food, reacts to certain signals or completes other essential processes. Some metabolites, such as glucose, are part of an organism’s central or primary metabolism and its basic life functions. Other metabolites, such as secondary or specialized metabolites, are less essential to sustaining life but no less integral as they help with plant defense responses. Identifying these metabolites can help researchers predict how plants will react to various biotic and abiotic conditions, such as high light, stress from salt, symbiotic interactions, and defense against pathogens.
“I love science because everything is a puzzle,” Kranawetter said. “Sometimes the puzzle is a struggle to put together and can look quite different from what you expected, but after everything is complete, it is always rewarding and empowering to see how the pieces fit together.”
Kranawetter’s work focuses on root border cells, which are vital to plant root health. These cells arise from the root epidermis, where the cell wall breaks down and the cells are physically released from the root but still surround the root tips. Border cells are held in place through a water-soluble, complex secreted matrix consisting of a polysaccharide mucilage, comprised of DNA, proteins, and metabolites. As their encasing material is water soluble, upon contact with water they wash away but are rapidly replaced within 24 hours after their removal. After border cells separate from the root tip, Kranawetter conducts large scale genetic reprogramming to divert most of their resources to secretion and specialized metabolism. In doing so, they serve a protective role against infection and environmental stress, but the scope of their functions is far from fully explored.
“Border cells are still very niche, and we don’t fully understand how they’re contributing yet, but we’re starting to see that they are more major players in the rhizosphere than we thought initially,” Kranawetter said.
Root border cells appear across most plant species. They are known influencers of dynamics within the rhizosphere — the layer of soil that is in direct contact with root secretions — but there are still a large number of unknowns about them.
Kranawetter’s previous work dealt with metabolites, or small molecules, and their differential accumulation in root tissues. He collected root border cells and separated individual root tissue types to create a metabolite atlas based on the Arabidopsis eFP browser – a computer program that shows the relative intensity of metabolites by tissue type in a heatmap context. This project builds on his work to further identify the molecules border cells secrete and how they mediate plant-microbe interactions.
Although this project does not directly correlate with his current research in the Sumner lab focused on differential metabolite accumulation in cultivated elderberry plants, it will allow Kranawetter to stay in plant tissue research, build on the foundational mass spectrometry knowledge he accumulated during his Ph.D. and incorporate microbiological techniques to border cells.
“Working on elderberry plants was a good chance for me to start a different project in a distinct plant system, which has been nice,” he said. “And this fellowship will be a great opportunity to apply my current research skills and knowledge base while also developing new abilities.”
For the NIFA fellowship, Kranawetter generated a project narrative, abstract, budget, a logic model, references, and a data management plan, among many other materials. He also utilized a three-member administrative board to support and look over the project in order to assist and give feedback each step of the way.
“This will be a project for me to get out of my comfort zone and gain new expertise compared to what I have done in the lab previously,” Kranawetter said. “It’s nice to have a different perspective when, instead of having a bunch of giant elderberry bushes in the greenhouse, I’m working with three-day-old seedlings.”
The fellowship allots Kranawetter the resources and time he needs to monitor border cell metabolite secretions and their methods of secretion. Kranawetter also observes the bioactivities within an organism living in symbiosis with its pathogens, the microorganisms that cause disease.
“The field of mass spectrometry allows me to apply novel technologies and software systems, while plant sciences allow me to explore the vast diversity of compounds that plants naturally produce,” Kranawetter said.
Kranawetter applies what he knows about border cells in experiments on Medicago truncatula — a close relative to alfalfa — and observes how border cells use their secretions to affect the rhizosphere.
“The question we’re aiming to answer is how are these secretions going into the environment and what role are they playing,” Kranawetter said. “I wanted to get back to my roots with this because as a mostly technical lab, I haven’t done a lot of molecular biology in some instances.”
Kranawetter first removes the seed coat from M. truncatula and sterilizes them in order to protect them from bacterial or fungi contaminants. Next, he places the seeds on a petri dish containing water agar overlaid with sterile filter paper so that the roots remain on top, as opposed to normal methods of plating directly on top of the agar. This ensures that the roots do not penetrate the agar, where border cells would be lost. Although some cells are still lost due to the minimal contact with the filter paper, most remain for later harvest. The final step is to dip the roots in water and harvest border cells — ones that freely come away from their root system — for closer investigation.
As part of his fellowship, Kranawetter is probing for transcripts and proteins associated with metabolite secretion. His results will help determine how border cells affect their environment through their secretions. Kranawetter’s interest in this field stems from his love of plants and technology, both of which he uses in his daily lab work.
“One of the major benefits of being in the Sumner lab is that we’re getting to use very sophisticated instrumentation and applying it to something that we probably would not have been able to examine otherwise,” Kranawetter said.
Kranawetter’s fellowship lasts 2 years, so he hopes this exploration expands our understanding of border cell interactions with the rhizosphere.
“I love this type of research because I get to do a lot of different things! I grow plants, work at the bench, and also use cutting edge instrumentation,” Kranawetter said. “Being able to work in such a diverse manner makes me feel empowered as a scientist.”
The U.S. Department of Agriculture’s National Institute of Food and Agriculture (NIFA) announced an investment of $12 Million to Advance Research in Agricultural Microbiomes. Microbiome research is critical for improving agricultural productivity, sustainability of agricultural ecosystems, safety of the food supply, carbon sequestration in agricultural systems, and meeting the challenge of feeding a rapidly growing world population. Research supported by the Agricultural Microbiomes in Plant Systems and Natural Resources program area priority within the Agriculture and Food Research Initiative (AFRI) will help fill major knowledge gaps in characterizing agricultural microbiomes and microbiome functions across agricultural production systems, and natural resources through crosscutting projects. AFRIis the nation’s flagship competitive grants program for food and agricultural sciences.
Instead of taking on more clear and straightforward science, she dove into vessel regeneration and never looked back as she works on the burning question, ‘can muscles regenerate in the absence of blood vessels and vice versa?’
“Knowing how vessels grow back can one day improve treatment options and help someone who has suffered a traumatic muscle injury and I really like contributing to that, but at the same time I want to know and do more right now” said the D Cornelison lab member.
Diller studies the interaction between regenerating muscle and blood vessels after a traumatic muscle injury and the role of ephrin-B2 (an essential protein for blood vessel formation during development) in blood vessel regeneration and when it is no longer necessary for development. While she studies these questions, she sees the connections in friends, colleagues and hobbies.
“One of my best friends works in the NICU [Neonatal Intensive Care Unit] as a social worker, and it can be hard for us to find common ground in terms of our careers,” Diller said, “but I started working on development and she was like ‘oh we see that in the babies too’ and then all of a sudden we had this crazy crossover between social work and mouse development.”
Recognizing these similarities lets Diller find new research skills and learn from others outside the field.
“It’s crazy how I can talk to an ecologist friend of mine who’s studying something completely different from me, and then they have an idea or say ‘have you tried this,’” she said. “Sometimes that leads to inspiration to help deal with a setback.”
Balance outside work helps with setbacks and obstacles within her research project. Diller tends to spend her free time on activities that prioritize her health, her relationships with her husband and friends, and her dog, Harry. She takes her dog to agility classes, but finds Harry is more in tune with the program than she is.
“I’m constantly being corrected by the trainers because you’re not supposed to step in front of your dog, and Harry just stands there and looks at me like ‘How do you not get this mom?’ But it’s a fun activity for us to do together, so I continue doing it,” Diller said. HIIT and resistance training classes bring more balance and a chance to interact with those outside the research field.
While Diller’s hobbies come in waves, her love for research persists as she expands her work in the realm of muscle regeneration.
Currently, the Cornelison lab is using conditional knockouts—a technique used to eliminate or delete specific genes from the mouse’s DNA—and pharmacological methods to inhibit angiogenesis – the process through which new blood vessels form—to study the impact impaired blood vessel regeneration has on muscle regeneration. The team looks at hints within the vessels and vessel network, such as new growth (i.e., tip cells), vessel dilation, vessel branching, as well as other structural deformities such as anastomoses (i.e., a connection between blood vessels). But Diller is always striving for more with her work.
Resilience is key when often each new discovery raises more questions about how vessels regenerate.
“As a grad student it’s really important to have coping mechanisms for the level of stress that you go through,” Diller said. “When something fails, being able to take a step back and stop thinking about it helps a lot. And once you’ve calmed down, the best way to deal with it is by talking to the people around you.”
Diller recently graduated from the University of Missouri and has accepted an NIH fellowship at the University of Florida to study the effect of hyperbaric oxygen treatment on vessel regeneration in the diaphragm in a spinal cord injury model. Diller is excited to work specifically on diaphragm recovery options and take this patient-focused opportunity.
“Science is for everyone, which sounds cliché, but I think that there is an aspect of science that almost everyone can relate to. It’s not always going to be the same thing for each person, and I like how science connects people through their differences.”
Michaela Beedy, Brian Thomas, and Margaret Beecher work on aptamers in the lab of Donald Burke. | Photo by Beni Adelstein, Bond LSC
Shrinking the Target: Developing Cancer Therapies
As cancer cells multiple and spread, doctors face finding treatments that destroy tumors while doing the least amount of damage.
This search for precision in cancer therapies is for good reason. It takes only a few minutes in a chemotherapy clinic to see the detriment of cancer drugs on the rest of the body.
“The issue with chemotherapeutic drugs is they have a lot of off-target effects,” said Brian Thomas, a MD-Ph.D. candidate working in the Donald Burke lab. “Our goal is to make them more targeted towards cancer cells using aptamers.”
Careful targeting of treatment is one important goal of cancer researchers, and aptamers are one way scientists at Bond Life Sciences Center work toward that goal.
Aptamers are short, single stranded DNA or RNA molecules that can bind to a target when it folds into a 3D structure. They can either serve as a vehicle for a cancer drug — delivering it to cancer cells while avoiding healthy cells — or bind itself to a cell’s receptors to interfere with its natural response.
In the Burke lab, the target of choice is epidermal growth factor receptors (EFGR), a protein that helps cells grow and spread. Mutations — mistakes in the DNA that makes this protein — can increase the number of EFGRs and cause cells to grow out of control, ultimately leading to cancer. The goal is to test how to best ensure aptamers successfully bind to cancer cells so they can be destroyed.
Margaret Beecher and Michaela Beedy worked in the Burke Lab over the summer.
Michaela Beedy, an undergraduate research assistant in the Burke lab works alongside Thomas. She is focusing on pairing aptamers with the chemotherapy drug doxorubicin to better target mutated cancer cells.
“Doxorubicin is really good at killing cells that are fast proliferating, which includes cancer cells and epithelial cells,” Thomas said. “It is also really good at killing cardiomyocytes, which is not a good thing.”
Margaret Beecher points out the tube containing the aptamer treated with doxorubicin.
The goal is to get doxorubicin to kill the cancer cells but to leave healthy cells like those heart muscle cells alone. Ideally, the cancer can be treated without posing too much risk to other vital organ functions.
Additionally, patients who undergo chemotherapy often develop resistance to a drug as newly mutated cancer cells become immune to the old therapy. That is why she is focusing on addressing specific aspects of what causes mutations.
Margaret Beecher, another Burke undergraduate, is testing a different hunch that could work in conjunction with Beedy’s work. Beecher investigates if dimeric aptamers might provide more of a targeted effect.
“Monomeric aptamers only have one spot that can bind to EFGRs while dimeric have two,” Beecher said. “Because there are two spots, this increases its avidity.”
Avidity is the combined strength of binding, and it is critical in getting a drug to interact with its appropriate target. This could be applied to Beedy’s doxorubicin approach.
“If we end up killing more EFGR-mutant positive cancer by using aptamer-doxorubicin treatment we could bring in the dimeric aspect as well to increase the effect of the therapy,” Beedy said.
The treatment in question is aptamer-doxorubicin conjugates, essentially a combination of the three.
Even if Beedy’s approach isn’t as effective at killing more mutant cells, Beecher’s research still comes in handy.
“If we saw less of a targeted effect because the aptamer wasn’t binding as well, maybe adding that dimeric aspect to it can help bring it back up to be on par with other treatments.,” Beecher explained.
New cancer therapies couldn’t come at a better time. Right now, the U.S. is in the midst of one of the worst cancer drug shortages in history due to manufacturing issues, leaving Thomas optimistic about the work in his lab.
“Right now, we’re doing in vitro…doing things in a dish. The ultimate goal is to get these treatments into animal models and, subsequently into humans to then finally create new therapies for patients in need.”
For Elaina Sculley, the word filter means much more than narrowing down your search results on a website.
The second-year animal sciences graduate student spends her days using computer programming tools as part of her bioinformatics studies and her work in Wes Warren’s lab at Bond LSC. Her focus is on the chicken immune response because they serve as invaluable models for studying immunology due to their widespread use in both commercial breeding and scientific research.
Her main objective when studying host immune response is to gain a better understanding of candidate genes and pathways involved in the immune response which is crucial to determine the underlying mechanisms of immunity and the potential in genomic selection schema.
“I think it’s something that is fascinating because it is yet to be fully understood, which sparks my interest,” Sculley said. “There are so many different components and factors that play a role in how chickens respond to a bacterial pathogen.”
Sculley earned a bachelor’s in animal sciences and a certificate in animal nutrition from Colorado State University. Her background in both wildlife animals and agriculture helps her understand the detrimental effects diseases can have on a variety of species.
In the U.S. there are billions of chicks hatched each year and they are among the world’s most important food source, which means their resilience to pathogen infection is an immense economic concern worldwide. Therefore, managing bacterial infections in the poultry industry continues to be important for protecting a global food supply. In recent years, more emphasis has been placed on enhancing host resistance, which has various external factors influencing each bird’s level of immunity. Her goal is to create strategies that enhance the hosts immune response and minimize the infectious disease impact they have on the poultry population worldwide.
She splits her time in the Warren lab working on bioinformatic analysis for 80% of the time and spending the other 20% doing wet lab procedures. Sculley isolates nuclei and high-molecular-weight DNA from tissue samples during her wet lab time and sends the samples off to the University of Missouri Genomics Technology Core for single nuclei RNA sequencing.
Once Sculley receives the snRNAseq data from the Genomics Technology Core, she determines which genes are up or down regulated in response to the bacterial pathogen.
She takes the snRNAseq results and uses a Seurat pipeline – a more specific form of bioinformatics analysis – to create cell type clusters based on similarities between cell types. For example, if a gene is up regulated or down regulated in response to a bacterial pathogen in the chicken spleen, Sculley can draw connections about why that might be based on the cell type identity of the cluster that the gene was found. The insights gained from these findings could contribute to the development of gene candidates that when selected upon offer effective new strategies to enhance immunity against bacterial infection in poultry populations.
Similar to a game of Tetris, each cluster of cells must be sorted into groups and certain elements must be removed in order to piece together the puzzle and win the game – or in this case, break down the chicken’s genetic code.
“This is mostly trial and error, where you manipulate the data in a way that is informative,” Sculley said. “It’s really complex. Sometimes I have to start over and rerun it through the same pipeline to make sure it all aligns in order to get the analysis to work. It takes a lot of patience and willingness to start over.”
As Sculley types away at her desk, she finds that minor edits she makes to the code make all the difference and contribute to an increased understanding of the chicken’s response to a bacterial challenge.
“It can be very draining and stressful when you don’t get informative output and you’ve been working on something for hours,” Sculley said. “You just have to be like, okay, I didn’t get it this time, but I’m going to go back later when I have a fresh mind because pushing myself to a point of burnout isn’t going to fix anything.”
Another way Sculley prioritizes her work-life balance is through a routine at home, free of screen time after a full day on the computer.
“I like this type of work and I work hard, but I enjoy going home, unplugging, and doing my hobbies.” Sculley said. “I love to do yoga and hang out with my French bulldog, who is very playful. She’s my bestie.”
When Sculley was in her first year of graduate school, she had classes and homework on top of laboratory work, filling her days with computer programming. But now, she finds that working with data for publication and having more flexible deadlines helps her have a work life balance.
She plans to apply for a Ph.D. program by the end of the year and continue to study avian single-nuclei studies to expand her knowledge of immune cell types in chickens, as well as their transcriptional responses to bacterial pathogens. The identified gene expression signatures and enriched immune-related pathways in her studies provide a foundation for future research in poultry immunogenetics and disease resistance. Her goal is to develop effective strategies to enhance immune responses and mitigate the impact of infectious diseases in poultry populations worldwide.
“I like science because we’re constantly changing and we’re always having to adapt and learn new things,” Sculley said. “I think some of the best people and leaders are lifelong learners.”
Bond Life Sciences principal investigator and associate research professor of veterinary pathology, Monique Lorson (left) and postdoctoral fellow Gangadhar Vadla (right) worked together to identify the ABT1 modifier in the diseases, spinal muscular atrophy with respiratory distress (SMARD1) and Charcot Marie Tooth 2S (CMT). | Photos by Sarah Kiefer, Bond LSC
By Sarah Kiefer | Bond LSC
It takes a keen detective to sleuth out why and how particular genetic mutations present the severe symptoms seen in neurological diseases.
The labs of Chris and Monique Lorson are one step closer to understanding one piece in the puzzle for spinal muscular atrophy with respiratory distress (SMARD1) and Charcot Marie Tooth 2S (CMT). They recently identified that the protein ABT1 appears to regulate the activity of IGHMBP2 by means of its interaction with it.
Their NIH-supported studies were recently published in JCI Insight with lab member Gangadhar Vadla as first author.
IGHMBP2 is a protein that causes SMARD and CMT2S. In these two rare neurodegenerative diseases, people inherit a mutated IGHMBP2 gene from each parent. The primary clinical presentation for SMARD1 is respiratory distress and without intervention patients typically die within the first two years of life. CMT2S, while less severe than SMARD1, presents with motor function and coordination defects that substantially impact patients’ lives. SMARD1 and CMT2S patients both demonstrate degeneration of the motor neurons within the brain and spinal cord.
The Lorson’s have built a career out of investigative work, looking into spinal muscular atrophy (SMA), spinal muscular atrophy with respiratory distress (SMARD1), and Charcot Marie Tooth types 1A, 2S and 2E.
When it comes to SMARD1 and CMT2S, understanding how cellular processes are altered when IGHMBP2 is mutated, and how particular mutations present in a clinical setting, is key towards developing therapeutic options.
The Lorson labs approach these studies using genetics and biochemistry. They generated a series of mice with Ighmbp2 mutations that correlate with patient mutations, showing traits of SMARD1 or CMT2S, depending on what particular Ighmbp2 mutation is present.
“ABT1 is the first protein found to modify the SMARD1 phenotype in mice through its association with IGHMBP2”, said Vadla, a postdoctoral fellow in the Lorson Lab.
They identified ABT1 as a modifier using an ABT1 gene therapy approach.
“Animals receiving the ABT1 gene therapy showed modest improvements in lifespan and motor function tests, consistent with a modifier, while untreated animals did not, “ Monique Lorson said.
The Lorson lab addressed three primary questions: do IGHMBP2 and ABT1 associate and how stable is this interaction, second, does this interaction change any of the activities associated with IGHMBP2 and, if so, what activities, and, third, can we use this interaction to understand what the role of the IGHMBP2-ABT1 complex is within cells.
“These questions, while seemingly simple, are really important in that is unknown how mutations in IGHMBP2 lead to SMARD1 or CMT2S, “ said Monique Lorson.
Their results demonstrate a strong association between IGHMBP2 with ABT1, and that binding of IGHMBP2 with ABT1 significantly increases biochemical activity of the IGHMBP2 protein. The studies also provided evidence towards the potential role of this complex in 47S pre-rRNA processing. 47S pre-rRNA processing is necessary to generate ribosomes used in the process of translation. Translation is the process cells use to make proteins.
“Because translation is required to make proteins, if you don’t have proper 47S pre-rRNA processing, some aspects of translation will be significantly impacted, putting the cell under a lot of stress,” said Monique Lorson. “The big goal now is demonstrating that 47S pre-rRNA processing and translational defects are present in Ighmbp2 mutant mice and that these defects result in disease.”
Now, the Lorson lab is testing their findings by creating IGHMBP2 that mirror those seen in real patients to study whether the interaction between IGHMBP2 and ABT1 is altered in any of these mutants.
Why is this finding important?
ABT1 is the first protein found that modifies the severity of disease in Ighmbp2 mutant mice. This is one step towards understanding how IGHMBP2 protein functions within cells and how mutations in IGHMBP2 lead to disease. The progress is only possible with involvement from many Lorson lab members and Bond LSC collaborator Kamal Singh.
“Everyone came together and played an important role, “ Monique Lorson said. “The Lorson lab works well as a team because we all try to make each other better scientists.”
This project was special to the Lorson family as it was the first time Chris, Monique and their son, Zachary Lorson, published their findings together.
She finds this work maintains her passion for science because she hopes their contributions can someday make a positive impact on SMARD1 and CMT2S families.
“When you feel like you can make a difference and not only impact people’s quantity of health, but also quality of health,” Monique Lorson said. “That makes coming into work really easy.”
The ocean is a current throughout Lahcen Amor’s childhood memories.
Growing up one block away from the Atlantic Ocean in Rabat-Salé- Morocco, Amor and his friends ventured into the water in search of a good time and some extra spending money.
They would dive down to catch fish, seaweed and mussels, which they each dug out from the ocean floor. When they called it a day, they sold the mussels and seaweed to freezer trucks parked close by. The next stop was the movies or other fun activities to use their hard-earned cash. The experience was not just about the money burning a hole in Amor’s pocket, but about navigating social interaction.
“My childhood helped create my strong and resilient personality and taught me how to act and deal with my peers,” said Amor, a research lab service supervisor at Bond LSC.
While living in Morocco, Amor would often assist his brother with his custom wood turning business. His brother helped instill in him a sense of commitment and action that served Amor later in life.
“When I was growing up, he taught me how to use the equipment, but also how to be responsible,” Amor said.
Lahcen has worked for MU campus facilities for 15 years in various positions. In recent years, his primary responsibility has been focused on heating and cooling systems, maintaining airflow and re-certifying room pressurization systems.
“I feel like I’ve been working here for years because this building was part of my maintenance, and I have been familiar with it for awhile, which makes me comfortable with it,” Amor said.
His present position combines his childhood talents with his knack for ambition he picked up from his brother.
“I have to integrate and enjoy what I am doing. I make it my goal to focus on one task at a time and make sure I learn from it,” Amor said.
He makes the rounds at Bond LSC to check equipment and the overall facility and fields calls from occupants in Bond LSC. Whether it be a researcher with a broken freezer or a staff member with a machine that isn’t working properly, he’s ready to tackle whatever task may come his way.
“I like to learn something new every day and make sure I always grow,” Amor said. “There are a lot of resources on campus that you can use to do so and if you have a goal, you’re going to reach it if you focus on it.”
Lahcen Amor works to repair a centrifuge – a device used to apply a sustained force to a specimen – at Bond LSC as a research lab service supervisor. | Photo by Sarah Kiefer, Bond LSC
Amor also updates the Bond LSC equipment inventory list and currently is developing a preventative care plan to maintain equipment. He gains more knowledge about laboratory equipment every day.
The ebb and flow of Amor’s daily work at Bond LSC gives him the tools he needs to work towards his goals. Although he has experienced several career changes, maintenance combines several of his nautical and tactical abilities all under one roof.
“I didn’t decide to do maintenance, but life just took me in that direction,” Amor said. “You set up objectives in your life that you want to achieve and then you take the opportunity when it’s presented to you to go after them.”
Lorenzo Ceccon wanted a career full of methodical problems for him to try and solve.
“I like science because it is a very logical thing. A + B = C,” he said. “It’s very systematic, so I guess I just like finding the answers to the questions I wonder about.”
The senior biology major put his pursuit of logic to work when he joined the Dawn Cornelison lab as an undergraduate researcher at Bond LSC, but he found himself in that same reasoning mindset growing up.
Originally from Arezzo, Italy, Ceccon attended a high school focused entirely on science and how it is applied. Instead of a traditional U.S. high school where a degree is completed in four years, Italy’s five-year high school experience gave Ceccon the time he needed to figure out what he wanted to pursue. He had the same science professor throughout all five years, which laid the foundational framework for a career in research.
At Bond LSC, a day in the lab exercises that foundation for Ceccon. He uses siRNA – small interfering RNA – to control gene expression with a system that temporarily blocks the copying of mRNA, so that he can target the mechanism of a particular type of cancer, known as Rhabdomyosarcoma (RMS).
Lorenzo Ceccon is using a pipette to take up a balanced salt solution in order to wash a plate of cells away from their medium. This is an early step in Ceccon’s process of recognizing the observable characteristics in the cells. | Photo by Sarah Kiefer, Bond LSC
Ceccon replates cells, begins to count them and makes sure to add siRNA so that once the solution is set a machine can allow the siRNA to get into the cell.After some time, a second experiment is performed to assess if the cells that traded with siRNA still respond to the treatment. This procedure is to understand if the kicked down protein, or the one that is temporarily deactivated, is involved in the process that allows this system to work. He uses an immunofluorescence technique, which utilizes a fluorescence microscope in order to visualize for any antibodies in the cell samples that target specific markers, or a DNA sequence with a particular location in the genome. He uses this tool to make sense of the changes the cells undergo with each treatment. The antibodies used in this process to target the markers of interest are modified so that when a light with a specific wavelength hits them they shine brightly.
Ceccon looks at a group of cells under the microscope to see how confluent, or together, they are in the solution in order to perform tests later. | Photo by Sarah Kiefer, Bond LSC
In the Cornelison lab, the Eph-ephrin system, a signaling system involved in many biological functions, is used to study many different aspects of life. Ceccon uses this system to promote a change of the observable characteristics of the RMS cells and stop the uncontrolled cell reproduction which makes the cancer deadly.
“I like being able to apply the knowledge I’ve learned in my classes to working in the lab because it makes me feel like I didn’t waste time studying all of those hours,” Ceccon said. “Working in the lab is something I’ve never done before, and the thought of achieving something is motivating enough.”
As a whole, the Cornelison lab works to constantly increase their understanding of this cancer’s physiology.
“I love this type of work! I like the problem-solving aspect, and how we always find something new to do,” he said. “I enjoy learning about the processes that allow life to exist.”
Back in Italy, Ceccon played piano or guitar, but he has not been able to ship those items to the U.S., yet. For now, Ceccon enjoys playing sports or going on hikes with his friends when he gets away from the lab. He also likes to delve into philosophy books or read science fiction novels from time to time. These activities help distract him from lab work or studying for his next test.
“Philosophy was one of my favorite subjects in high school,” Ceccon said. “It helped me learn how to reason and produce logical statements, and I like to read about topics that make me think.”
What motivates Ceccon is not just the answers he gains in the lab but also the variables he accounts for along the way. He hopes to be part of a paper on the RMS findings he works toward in the Cornelison lab then he wants to attend medical school to learn even more about this disease and others.
“There’s always a new challenge to face,” he said. “I just have to sit down and think of what’s going wrong when that happens, talk to others in the lab about it, and decide what to do next.”
A strong, stable stem is like Rachel Weber’s career in plant biology.
Weber started in a research lab only a few months after she stepped onto the University of Missouri campus for the first time.
“I was really excited and surprised because I didn’t expect as a freshman to have this opportunity,” Weber said.
Weber began by studying lignin with Jaime Barros-Rios, an assistant professor in the Division of Plant Science & Technology at MU.
Now, she is continuing her lab experience as one of fourteen Cherng Summer Scholarship recipients for a nine-week, full-time program through the MU Honors College. The scholarship supports undergraduate research projects, with a $7,000 award and $1,000 expense account, so Weber can continue her work at Bond LSC. She started under the Barros-Rios lab at Bond LSC in October 2022, where she will continue as a Cherng Scholar.
The Freshman Research in Plants (FRIPS) program gave Weber this chance. The program encourages student involvement in plant biology as soon as they reach campus to start a potential career in the field right away.
“You just have to go for it because it’s really just about how passionate you are and how hard you try,” Weber said. “If you look for the opportunity, you can find it.”
Plants were once fragile and flimsy, unable to hold themselves up, but 450 million years ago lignin and the enzymes that accompany it evolved to stiffen plant cell walls to allow plants to flourish.
Lignin – a large chain polymer – is comprised of three main sub-units that are the product of a complex pathway of compounds. These compounds are being converted into components of living organisms. These sub-units occur in different ratios depending on the plant and impact the chemical properties of the plant’s cell wall. Lignin has proven to be a limiting factor in the production of certain biofuels due to the chemical makeup of the polymer, which is why it is so important to understand how it is constructed.
Along with having an impact on the biofuels industry, “lignin helps plants respond to different stressors, so understanding it has a lot of implications in grasping how the plant survives and how it responds to different environmental factors,” Weber said.
Weber’s research project digs into the way two enzymes interact and how they work together to synthesize lignin. Laccases and peroxidases are enzymes that oxidize the sub-units of lignin to come together and form the larger lignin polymer. Both enzymes use reactive oxygen species (ROS) as a substrate, but their individual role in the polymerization process of lignin is unknown.
The mystery comes down to how 17 laccases and more than 60 peroxidases in Arabidopsis plants work to develop lignin. Weber aims to crack the code on what makes the functions of these enzymes so special. To Weber, this means investigating plant stems and roots.
“Not every gene has been characterized for these enzymes and the fact that there are multiple enzymes with potentially the same function makes it hard to determine their specific roles,” Weber said. “This also means that if you knockout one enzyme, another one may take its place and that may make no difference to the plant, we just don’t know that yet.”
Weber plans to utilize two mutant Arabidopsis plants with one not displaying a certain laccase and the other not containing a peroxidase. This will allow her to observe the enzymes’ individual roles in the plant’s lignin and compare their impact on the plant’s noticeable characteristics. This process brings Weber back to her agriculture roots. Weber spent time at her grandparents’ farm growing up, and she learned to admire the biological complexity of nature. She now sees more of that interwoven fabric with the effects of climate change.
“My grandparents are farmers, so I see how the current drought we are under is adding stress to their lives and impacting their livelihood,” she said. “The fact that many researchers are working on drought resistant crops is really encouraging and exciting to me and I hope to be able to have a similar impact on others.”
Weber spent anywhere from 10 to 15 hours a week in the Barros-Rios lab during the school year, but her part-time research project will now turn into a 40-hour work week. To establish what role each enzyme plays in lignin, Weber will test the waters with protocols to analyze these mutant plants. This includes measuring the composition of subunits of lignin, looking at levels of reactive oxygen species, and utilizing microscopy techniques to visualize the lignin polymer in plants.
Weber chose plant biology to actively alleviate climate change’s effects one step at a time and to fulfill a small role in a large and complex issue within the biofuel and biomaterials industries. This field grants her the resources and support to see how one polymer, or a group of enzymes, can help other researchers grasp the nature of lignin more fully.
“The most important part about science to me is how many possibilities there are to help others. I love being in a place where there are always opportunities to learn new things,” Weber said. “I hope to one day be able to apply what I work on to positively affect the environment.”
