By Cara Penquite | Bond LSC
Wendy and Bill Picking tackle a stomach-churning area of science.
Fascinated by the bacterium responsible for bacillary dysentery, Bill studies its structure and function, while Wendy aims to use information on that same bacterium for healing.
“I do the vaccine stuff, but he’s a protein chemist. So the proteins are what keep us together,” Wendy said.
As two of Bond LSC’s newest investigators, the couple brings pathogenic microbiology expertise to the center. Their hiring is part of MizzouForward, an investment that aims to elevate and promote the University of Missouri as one of the best research universities in the nation.
The bacterium they study, known as Shigella flexneri, has a structure on the surface that is shaped like a needle and syringe — a structure that makes it easy to inject proteins into intestinal cells. It essentially reprograms the cells, which allows it to continue infection. The Picking’s labs study the same bacterium, but each investigator has their own projects.
“Bill has his structure work, I have my vaccine work. That’s how it’s split, but it’s so intertwined and the labs are intertwined that there would be no way to divorce us,” Wendy said.
Wendy uses proteins from the tip of the bacteria to create vaccines. Her goal is to get her work into clinical trials.
“There’s what’s called the valley of death,” Wendy said, “from the time that you have proof of concept to the time it goes to the clinic. There’s at least 10 million dollars in there,” Wendy said.
Bill’s research is focused on how the bacteria is structured — an interest that developed during his protein chemistry graduate work at the University of Kansas. Bill’s experiments involve mutating the bacteria and analyzing how the “needle and syringe” system — known as a type III secretion system — works.
“It’s purely curiosity and being able to break things and put them back together,” Bill said.
The two labs often collaborate as each lab’s work sometimes fills in the gaps for the other. The two investigators often work on the same projects.
“We have really complementary expertise,” Bill said. “Those are the best collaborations, when she does things that I don’t do, and I do things that she doesn’t do.”
Bill and Wendy most recently came to MU from the University of Kansas. The couple also spent time at the University of Texas and St. Louis University. Both earned degrees at KU and met while they were students.
“Bill and I met on the softball field . . .,” Wendy said.
A few years behind Bill in her studies, Wendy started off working in his lab. Her work quickly outgrew the scope of his research and the two split into their respective specialties and began working as collaborators.
The couple’s only impression of Columbia for years was the drive home to Kansas from St. Louis University.
“It really wasn’t until we came here and started actually getting into the town or the city of Columbia that we got an appreciation for just what all is here. It’s actually really nice,” Bill said.
Wendy finds herself often occupied in the lab, but she quilts when she gets a chance.
Bill hopes to utilize Columbia’s many fishing sites, and he also brings microbiology into his home as he pickles his own food.
“I just think bacteria are pretty cool,” Bill said. “People don’t realize just how much microbiology impacts everybody’s life.”
Wendy and Bill Picking join Bond LSC thanks to MizzouForward. Both Wendy and Bill Picking are professors in the College of Veterinary Medicine.
MizzouForward is a transformative, $1.5 billion long-term investment strategy in the continued research excellence of the University of Missouri. Over 10 years, MizzouForward will use existing and new resources to recruit up to 150 new tenure and tenure-track faculty to address some of society’s greatest challenges. Investments also will enhance staff to support the research mission, build and upgrade research facilities and instruments, augment support for student academic success, and retain faculty and staff through additional salary support.
Pew Charitable Trusts may be best known for its non-partisan think tank subsidiary that focuses on demographic and social science issues, but its mission to improve public policy and inform bleeds over into support for science.
David Mendoza — a Bond Life Sciences Center principal investigator and associate professor of plant sciences in the College of Agriculture, Food and Natural Resources — received a $200,000 grant for biomedical sciences from the Pew Charitable Trusts.
“To have people you need plants, so if we manage to engineer better and more nutritious plants, the chances of improving human health are high,” Mendoza said.
Mendoza was one of six pairs of scientists awarded grants for collaboration. Mendoza will team up with Clarissa Nobile, an associate professor at the University of California Merced, to look at plant-microbe interactions within the context of iron intake. Both Mendoza and Nobile are previous Pew scholars/ fellows, making them eligible for this Pew Innovation Fund grant to work together on cutting-edge science.
Most of Mendoza’s work involves growing plants in the lab, looking at how plants manage essential nutrients including heavy metals like iron. With this grant, he plans to take the next step to grow plants in a less sterile environment and exploring the role of the microbiome on nutrient acquisition.
Bacteria and other microscopic organisms surrounding plants are known as the microbiome. These microorganisms don’t just sit passively around the plant, they interact and affect plant’s health and behavior.
“That communication between plants and microorganisms led to the discovery of growth-promoting bacteria, which could be used as biofertilizers,” Mendoza said. “We [want to] find which bacteria really helps the plant to fight against pathogens, then instead of spraying them [with fertilizers], maybe we can just spike them with the right combination of bacterial communities.”
Mendoza’s expertise lies in plant nutrition, and Nobile brings the microbiology expertise. Nobile will send collections of bacteria known as synthetic communities, or SynComs, for the Mendoza lab to use on plants to study how they affect plant nutrition.
“Pew people are eager collaborators, so Clarissa replied immediately, and said, ‘yes, absolutely,’ and we got into a Zoom call,” Mendoza said. “We wrote the grant together, and we submitted together.”
The grant allows for flexibility during the research process. If Mendoza and Nobile start their research on one path and discover another path they want to pursue, they are allowed to change their plans.
“You get judged by the idea in general,” Mendoza said. “So, we have a lot of flexibility when it comes to execution and changes, if we have to. It’s like a priming type of award to get you to the next stage.”
Getting to the next stage of Mendoza’s project would mean learning more about how SynComs can improve plant nutrition.
“It was not about cancer, it was not about a disease, it’s about plant nutrition for human nutrition,” Mendoza said.
Pew Innovation Fund 2022 projects range from cancer and neurology to schizophrenia and animal regeneration. Find out more about all six 2022 collaborations at pewtrusts.org.
Bond Life Sciences Center’s Ron Mittler was recently named Curators’ Distinguished Professor by the University of Missouri System Board of Curators.
This top honor is bestowed on professors for outstanding scholarship who have established substantial reputations within their field.
“I am honored. Mizzou is such an amazing, supportive, and collaborative research environment and I feel lucky being here,” Mittler said. “I enjoy every moment working at Bond LSC.”
Mittler’s research substantially focuses on the role reactive oxygen species (ROS) play in the regulation of different biological processes. While ROS can be destructive within cells, he discovered it also plays a role in how plants systemically respond to environmental threats. He is a nationally recognized expert in this field of study.
“My work covers many biological systems and organisms with a focus on how they respond to stress and the role of ROS in their responses,” Mittler said. “I also study how organisms respond to a combination of different stresses, a problem that we are already facing in nature and in field environments that will get worse due to global warming and climate change.”
Mittler came to Mizzou in 2018 from the University of North Texas. Now a Bond LSC principal investigator and professor of plant sciences in the College of Agriculture, Food and Natural Resources, he joins eight other current and former Bond LSC principal investigators who received this honor in previous years.
“Dr. Mittler’s path-breaking research on cell signaling by reactive oxygen species and its role in how organisms cope with stress combinations is a nice example of cross-disciplinary research in Bond LSC on contemporary problems that require life sciences solutions, Bond LSC Director Walter Gassmann said. “That his insights have relevance also for cancer research in humans supports Bond LSC’s concept of bringing researchers from different disciplines together. Ron is exceedingly deserving of the Curators’ Distinguished Professorship and is a fitting example of the research excellence the Bond LSC strives to enable and stimulate.”
Individuals are nominated for this honor based on performance, service and their teaching record, and the title can be renewed every five years.
Visit Show Me Mizzou to see all of the Mizzou faculty members who received distinguished professor honors at the Sept. 7 Board of Curators meeting. Previously named Curators’ Distinguished Professors at Bond LSC include Chris Lorson, Dong Xu, Gary Stacey, Chris Pires, David Pintel, John Walker, Gary Weisman and Michael Roberts.
Kamlendra Singh sat down in his fourth floor office as a Bond LSC Investigator for the first time on September first after nearly 14 years at the Bond Life Sciences Center studying HIV, COVID-19 and how the right molecule can interact to fight disease.
Designer compounds are Singh’s specialty, and as a new principal investigator he directs four projects on microscopic treatments for various diseases. He will also continue his work as Molecular Interactions Core Director alongside his new role.
“Everyone [in the center] is such a tight community … That’s why this building is a benefit to be here,” Singh said.
Bond LSC director Walter Gassmann recalls Singh’s arrival at the center as an assistant research professor.
“He developed independent ideas very quickly, but there was no tenure track position for him,” Gassmann said. “That changed in January of this year.”
As a research center, the LSC does not have tenure-track positions and partners with academic units for those. Once Singh obtained a tenure-track position in the Department of Veterinary Pathobiology, faculty members in the LSC voted Singh into the LSC Investigator position after he presented his research plan during a chalk talk.
“He’s done it. He’s done great work, and he deserves it,” Gassmann said.
As Singh pivots into his new role, he tackles major projects designing molecules as solutions to diseases that may pave the way for future treatment options.
A drug is only as good as its ability to avoid resistance.
That’s the case with HIV, which is manageable with medication but evolves and mutates to avoid getting caught. Singh’s job is to be smarter.
Singh’s idea is to find new HIV antivirals that block the virus from infecting host cells. That goal could lead to novel antivirals that work for mutated HIV subtypes in underdeveloped countries.
“Most of the HIV treatments are developed where there is money, that is in developed countries,” Singh said.
HIV treatments are often made for the version of the virus that exists in first-world countries where the research takes place. Singh, on the contrary, concentrates his efforts on overlooked strains of HIV that often exist in underdeveloped countries, trying to understand the structural components of the virus to improve the treatment.
“We have to develop the drugs such that they work everywhere, but to do that you have to know the mechanism first,” Singh said.
Too much of a good thing can become bad, especially in living organisms. That’s where Singh’s second project focuses, tackling copper concentrations.
Too much copper facilitates cancer growth and spread. Singh works with Bond LSC researcher Michael Petris to find ways to hijack molecules that transport copper in and out of the cell. With funding from the National Cancer Institute, they have already found two potential molecules.
“Copper is a trace metal, and if it accumulates too much into the cell … then those cells become cancerous cells,” Singh said.
Singh’s preliminary trials sifted through millions of drug candidates and then tested 10 compounds to find one that could bind to the copper transporters and block copper’s path into the cell. Out of the initial 10, two were successful.
“That’s a lot. In this kind of drug discovery if you can get two out of a million that’s a success,” Singh said.
He plans to continue searching for viable compounds that could be used as cancer treatment, but his compounds may also be multi-purposeful.
Certain bacteria have similar compounds to the copper transporters, and just as viruses mutate to avoid antivirals, bacteria rarely go down without a fight.
Resistance bacteria mutate to survive antibiotic treatments, but Singh’s compounds may bind to compounds in the bacteria and have antibacterial roles.
“So, now we want to develop antibiotics using those drugs but making subtle changes. We want to make it specific for bacteria,” Singh said.
In the face of the COVID-19 pandemic, Singh’s third project tackles another set of viruses.
“The idea was to discover an inhibitor that not only blocks this coronavirus, but it can also be used for other viruses,” Singh said.
Tackling a diverse portfolio of viruses with a single method sounds impossible with vast differences between each pathogen. Singh’s solution — look for a common compound.
The enzyme RNA polymerase might be just that.
It’s role is to copy the virus’ genetic material to create more virus within the host. By targeting RNA polymerase, Singh hopes to prevent the virus from replicating.
“Not only can it be effective against Coronavirus … but other viruses, let’s say Zika virus for example,” Singh said.
Returning to Roots
Singh’s fourth project brings him back to his postdoctoral work with a fresh perspective and decades of more experience. He turns to DNA polymerases now with the goal of cancer treatment.
Similar to RNA polymerase in viruses, DNA polymerase’s job is to replicate the genetic material for new cells in humans.
Every day, cells replicate to replace damaged or dead cells. Think about your skin — skin cells are constantly exposed to the elements and suffer wear and tear. When skin cells die, nearby cells divide to create new cells in their place. To create new cells, the first step is to duplicate the genetic material in a cell to be used in the new cell.
But with cancer cells, DNA polymerase replicates faulty genetic material and creates more cancer cells. If DNA polymerase can be stopped in cancer cells, the faulty genome will not be replicated.
“So that’s a brand new project, and I’m really excited about it,” Singh said.
Gene-editing is the pinnacle of a biologist’s toolbox, but often left unexplained it seems more magic than science.
Growing rice from a small cluster of cells to 4-foot stalks can take six-months or more of planning and careful nurture. But how do scientists change the intricate genetic material in each cell of the plants?
The CRISPR-Cas9 gene editing tool changes a plant’s DNA. As Ph.D. student Ajay Gupta knows firsthand through work altering plants for the Bing Yang lab.
“CRISPR is relatively new. It’s like 10 years old only and still we are working to modify it and improve it in plants,” Gupta said.
The process starts with a few cells from seeds capable of maturing into any type of cell to form a cell cluster, known as a callus.
But how do scientists insert instructions into a cell?
In the Yang lab, they use particle bombardment to insert DNA, fundamental instructions used to build everything within a cell. These inserted DNA strands tell the cells how to build the CRISPR tools to change their own DNA.
To shoot the DNA molecules into the cells, researchers use high pressure helium gas operated gene gun to bombard the cells with DNA coated gold particles.
Once the DNA instructions are in the cell, they build the CRISPR-Cas9 tool using the plant’s genetic machinery.
CRISPR-Cas9 has two parts: the Cas9 protein and an RNA molecule that only binds to the DNA section scientists want to change. This RNA strand guides the Cas9 to the correct location.
But the parts of the guide RNA do two things. First, the spacer part — usually about 20 base pairs only binds to the section of DNA researchers want it to. Second, a part called the scaffold — nearly four times the size of the spacer is required for attaching to the Cas9.
When the spacer of guide RNA binds to the DNA, the Cas9 enzyme acts like a pair of scissors, cutting that section of DNA.
Now we have DNA broken in two, and the plant’s built-in repair system starts its job. However, this repair system is flawed, and it usually messes up while rebuilding the DNA’s bases, or building blocks. Its errors vary every time, so many different mutations are possible.
“The repair machinery which is employed is not perfect,” Gupta said. “When it repairs those double-strand breaks, it causes some errors. The error could be a single base, and it could be 10 bases or it could be a thousand bases. So that is unpredictable.”
Think of it this way: imagine you have a ten-page instruction manual, and you rip out pages two and three. You could replace them with blank, yellow or red pieces of paper and the book would have ten pages again, but the instruction manual can’t be used again. Just like that, it doesn’t matter which error happens, because any mutated change in the gene results in it not working.
“This is how the normal CRISPR-Cas9 works, when you just want to shut that gene down,” Gupta said.
This puts the gene’s instructions out of commission and is commonly called a knockout gene. That helps scientists determine what the particular gene does. They then compare “normal” plants to plants with the knocked out gene to determine what happens differently.
Now the waiting happens. Researchers grow the plants using different hormones to mature the callus cells into roots and stems.
“Once they have shoots and roots then you transfer them to soil and grow the complete plants out of that,” Gupta said.
When the plants become adults, the researchers check the knocked out changes by extracting its DNA in a process known as genotyping.
While traditional CRISPR-Cas9 is used by researchers to learn more about plant genomes, Gupta and others in the Yang lab also work to edit parts of DNA and replace it with specific mutations. This process is known as prime editing.
“The exciting part about this [prime editing] is that theoretically it can do anything,” Gupta said.
Prime editing takes traditional CRISPR-Cas9 editing a step further, and the Yang lab’s research into it opens the door for future collaborations with labs around the world in applied and basic sciences.
What started as an email correspondence between two aptamer enthusiasts rapidly snowballed into a hat trick of authorships for Donald Burke.
“I was contacted by a student in India asking if I would be an external advisor for her Ph.D. committee,” said Burke, a principal investigator at MU’s Bond Life Sciences Center.
Burke’s extensive research with sticky molecules called aptamers — totaling about 60 publications over 30 years — makes him an expert in the field of aptamer technologies. When Ph.D. student Shringika Soni at Amity University in Noida, India, near New Delhi, began characterizing the use of these molecules for drug testing in a literature review, she turned to Burke for advice on her work. The connection soon turned into a mentorship through regular email correspondence.
“I told her that I was interested in the kinds of things that she does, and I would be happy to interact with her about her science,” Burke said.
That project led Burke to collaborate on a project that attempts to create a rapid test to detect various drugs. This test would provide quicker information than waiting for the results of a blood or urine sample to be returned from a laboratory.
Starting out as a literature review, it soon turned into experimental research at Amity University. Burke’s role from halfway around the world was to ask tons of questions.
“There were a lot of back-and-forth emails around the general idea of defining precisely what her message was, what did she want to say about these systems, and as a particular case study might be mentioned, how much detail should be talked about in the review article,” Burke said.
While the research is not ready to be used commercially, Burke suggests potential applications could be for police officers who suspect drug use or for medical personnel trying to respond quickly to an affected individual.
“Maybe you’re suspicious that [someone is] on a particular drug. Having them spit onto a stick is just a whole lot less invasive and quicker,” Burke said.
Detection of the drugs falls to sticky molecules called aptamers. Essentially a chain of nucleotides folded into a particular shape, aptamers are selective in what they stick to, and each aptamer sticks to a specific compound. The researchers engineered their own aptamers that can stick to particular compounds found in certain drugs.
“The two major layers to this technology are designing the sensor components so that it will bind to the [compounds] really well and specifically,” Burke said. “The second component is to somehow turn that binding event into a detectable signal.”
But, aptamers just sticking to a drug is not enough because the researchers also need a signal to know the aptamer detected it.
To sense the chemical interaction, the researchers focused on electrical currents. With no drugs present, the aptamers do not stick to anything, and the electrical current flows. In contrast, when there are drugs present the aptamers stick to the compounds and block the electrical current from flowing. This allows the researchers to measure the drugs in the solution.
Due to Burke’s help on the project, the research team at Amity University offered to list Burke as a co-corresponding author.
“I just didn’t think that was appropriate, they were the ones leading the charge,” Burke said. “My role was to ask them questions from time to time and to push them to make sure they rounded out their arguments.”
This project adds to Burke’s long list of aptamer research fueled by his fascination with the properties of molecules driving living things.
“Just about every aspect of biology is driven by this choreography of who’s dancing with whom, by which molecules dance with which other molecules,” Burke said. “But why is it that some of the players refuse to dance with certain other players and others are just immediately drawn to certain other players?”
Burke’s research tries to answer these questions and determine why some molecules interact with specific compounds and why others do not.
“When you look at molecules the way they interact, it’s just so fun to watch the dance that they actually do,” Burke said.
Cynthia Tang’s academic career is marked by her propensity to multitask. From earning a major and three minors during her undergrad to making a documentary while getting lab and clinical experience, she makes the most of her time.
Recently Tang received the Excellence in Public Health Award from the United States Public Health Service, and a $181.734 National Institutes of Health grant to be used over four years . . . all while getting an M.D. and Ph.D. simultaneously.
The funding from the grant goes towards Tang’s research on SARS-CoV-2, the virus responsible for the COVID-19 pandemic, and its effect in rural areas. By comparing DNA sequences of the virus from patients in rural and urban areas, Tang looks for differences in variants. Her goal is to understand how the virus has been evolving and why patients in rural areas experience more severe symptoms.
“I’ve always found infectious diseases really interesting,” Tang said. “This pandemic came up and it was exciting to be in the position to help contribute to the knowledge base of it.”
In a family of immigrants, Tang saw the challenges in health literacy and cultural differences affecting patient care in the U.S first-hand, and she also caught a glimpse into a global view of healthcare.
“I had an opportunity to travel to Vietnam and actually see some of the health disparities of different health care structures in different countries,” Tang said. “We were visiting my grandfather who was in the hospital, and it was alarming to see the contrast between the quality of health care and availability of health care there compared to some of the hospitals that we have here.”
Tang recalls noticing lack of supplies, physicians and space for patients. She saw similar issues in the U.S., and shined light on public health disparities here by creating a documentary about the challenges immigrants face in the U.S. healthcare system. She also held competitions across Washington University’s campus to teach students more about public health issues while working as a clinical research coordinator.
“I wanted to help improve health equity,” Tang said.
A bachelor’s in chemistry and minors in philosophy, international development and pre-health professions at the College of Idaho may start to explain Tang’s public health focus and her path to bench and clinical research.
“My minor in international development was really useful for me because that gave me a lot more exposure to the different economies of different countries of the world, their health status and how politics, economics and health care all interrelate in different countries,” she said.
Her research at Washington University led Tang to an interest in its clinical applications.
“I really enjoyed the aspect of working with patients, actually getting to talk to patients and hearing their stories,” Tang said. “I felt like with clinical research, you can see … slightly quicker results from bench to bedside compared to bench research, and that was also when I decided I was interested in becoming a physician.”
Part of an eight-year joint M.D.-Ph.D. program at Mizzou, Tang started with two years of pre-clinical medical school before transitioning to Ph.D. research and then will return to finish the clinical years of medical training.
When she came to Mizzou, Tang continued fighting health disparities for immigrants.
“We organized a group of volunteers that were able to translate COVID-19 information to different non-English speaking communities in Boone County,” Tang said.
Always one to tackle several projects at once, Tang plans to pursue a career as a pediatric physician scientist interfacing with patients while continuing lab research.
“I like the idea of doing both. As a physician, you get that one-on-one contact and you get to make a very direct contribution to someone’s life,” Tang said. “And then with research, you don’t have as much contact with patients, but what you do can affect populations.”
If the world can be taxing on a person as pressure mounts, just think about how stress must feel to plants.
Humans can add a layer of clothing when cold or get a glass of water when thirsty, but plants do not share this simple luxury and must endure whatever environment they sprout in.
As climate change, pollutants, and extreme weather patterns escalate, this poses a serious global threat to plants and our food supply.
Ron Mittler, a principal researcher at the Bond Life Sciences at the University of Missouri, recently looked at how this piling up of multiple stressors at once can significantly decrease plant survival.
“The principle is that you can have a combination of several different stressors, each by itself has no effect on the plant but when they come together, they’re causing severe effects,” Mittler said.
Studying plant response to stress isn’t a new thing, but Mittler’s focus on the compounded effects of stressors may give us a better idea of the threshold of stress plants can endure in our changing climate. Multifactorial stressors combine three or more stress factors simultaneously impacting plants. Four categories of stress— biotic, climate, anthropogenic, and soil threats — all become worse as climate change and environmental pollution progress, subsequently decreasing plant quality of life.
Biotic threats relate to enemies like pathogenic bacteria and insects. Similarly, soil threats are determined by poor nutrient soil and salinity. Climate threats include extreme temperatures and drought. Anthropogenic threats are man-made as humans use harmful pesticides and create microplastics.
Arabidopsis thaliana seedlings, a model plant used in experiments, were placed side by side on a plate and received a combination of stress conditions such as heat, salt, excess light, acidity, heavy metal, and oxidative stress. Researchers studied the growth, survival, and molecular responses of the seeds. Seedlings were grown on plates rather than in soil to isolate and study the impact of multifactorial stress.
Seedlings grew on separate plates and experienced different individual or combined stresses. Results showed that each individual stressor applied to the seedling had a minimal effect on the plants but with the increase in the complexity and number of stress factors affecting the plants, survival, root growth, and chlorophyll content declined. Similar results were also found for seedlings grown in soil.
Ecosystems are already seeing these impacts in Florida and Germany. Multifactorial stress of heat and pollution prompted algae blooms to grow exponentially. This toxic overgrowth led to thousands of manatees dying. Entire forests in Germany are experiencing massive storms followed by long periods of drought, insect attacks, and fires.
Mittler said things may not look dire now, but we will eventually reach a point of no return where plants die off in mass quantities or even go extinct.
“The harmful effects of stress on the nation can serve as a dire warning for society. We may not see the effects now but 10, 20, 40 years down the road we will be having severe problems with our food chain,” Mittler said.
Since there are multiple factors at stake, predicting negative impacts on agriculture and ecosystems is tricky as researchers are unsure how this domino effect may unravel. What they do know for certain is that there will be severe consequences and we are already seeing them today.
These sporadic weather extremes are weakening the plants, making them more vulnerable to insect predators and other stressors.
The research yielded alarming results, and Mittler highlights that this is a dire problem that people need to take seriously. Once people understand the severity of this problem, he hopes individuals and policymakers will take action before the consequences become irreversible.
The Mittler lab is working on several fronts to address this problem and is trying to find a solution to it. They are currently studying multifactorial stress combination in different crop species, such as soybean, rice, and tomato. In addition, also identifying key plant regulators that are activated during multifactorial stress combination. These will be used in future breeding efforts to increase the tolerance of different crops to multiple stresses.
This study is now an international collaboration between the Mittler and Zandalinas laboratories, as Dr. Sara Zandalinas took on a faculty position in Spain.
Investigators at Bond LSC take steps to apply basic research
By Cara Penquite | Bond LSC
Scribbling in a lab notebook and planning experiments tucked between shelves of equipment, it’s easy to fixate on day-to-day lab operations. But scientists also face the challenge of finding how research can improve the world around us.
“The direction, the vision of the lab, ultimately comes from the principal investigator that bridges the research into applied directions,” said Jay Thelen, biochemistry professor and Bond LSC principal investigator
Despite the focus on basic research within the Bond LSC, many principal investigators choose to take their research to the next level with commercial partnerships.
Thelen’s lab researches ways to increase oil production in seeds and has three patents licensed to Yield 10 Bioscience, a sustainable crop innovation company who applies Thelen’s research to commercial crops. While seed oils like canola and soybean oil are known for their use in cooking, Thelen explains that increased production of these oils could play a larger role in sustainable fuel sources such as biodiesel and sustainable aviation fuel.
“We have to make more oil to balance out our need to eat it [and] our need to wean ourselves off of fossil fuels,” Thelen said. “To do that, we need to either plant more acres of oil seeds, or we have to raise the oil in existing oil seeds.”
Thelen researches enzymes with that potential application in mind. One is acetyl-CoA carboxylase, the enzyme which initializes the production of fatty acid chains found in plant oils.
“We’ve known this is an important enzyme, and we know that any tinkering you do with it has an impact on the oil production,” Thelen said. “In this case we’ve made new discoveries that permitted us to rationally engineer this enzyme to make it more active.”
Thelen suggests thinking of the enzyme as a “gatekeeper” to oil production which initializes the production of fatty acids and increases oil production. Thelen’s lab identified two different gene families that influence the activity of the enzyme in Arabidopsis and camelina plants. Yield 10 then applies these discoveries in other commercial plants.
While Thelen works closely with his commercial partner — having served on their scientific advisory board for three years and now stays in contact with Yield 10’s CEO to develop research projects — some labs stick with short-term arrangements.
Kamlendra Singh — assistant director of the Molecular Interactions Core at Bond LSC and Veterinary Pathobiology research assistant professor — studies HIV treatments. His lab identified a compound licensed by a commercial partner that targets the shell containing the virus’ genetic information.
Singh’s work in HIV started in 1994 with basic research investigating the enzyme that makes the viral DNA.
“I wasn’t into [studying] the drugs when I started working on HIV, I was mostly trying to understand how HIV enzymes works,” Singh said. “Once you know how the enzyme works, then you can target these enzymes for discovering the drugs.”
After years of studying how the enzyme works, Singh switched to HIV treatment. The first step to develop a treatment is to look for structures in the virus that the drug could potentially target to stop the viral replication. Singh targeted the shell around the virus’ genetic information known as the HIV capsid.
Building on previous research, Singh’s lab developed a compound able to bind the HIV capsid and prevent it from releasing the contained genetic information. Even with the licensing of his compound, Singh plans to continue researching ways to improve it.
“There are two reasons to keep working on it. One, well it’s my brainchild,” Singh said. “The second reason is as the company grows, we grow. We get more recognition and more funding. You can use it to [study] different viruses or use the same funding to improve upon it.”
While Singh plans to remain looking towards the applied side of his HIV research, he does not forget his roots in basic research.
“You have to put in time … [to] understand the system first, which is basic science, before you go to applied science,” Singh said.
Michael Roberts, a Chancellor’s Professor Emeritus of animal sciences and biochemistry who has had several patented projects, focuses on improving basic science projects and applies for patents if warranted.
“I don’t deliberately go into anything for commercial purposes,” Roberts said. “If I see something that I think does have commercial application, I’m happy to do it, but that is usually after you do [basic sciences].”
Whether starting a project with applications in mind or focusing on basic research, knowledge gained through research can be building blocks for the future.
“Science is simple. Even the most applied research project has its genesis in basic biology and basic research,” Thelen said.
One step into the Advanced Light Microscopy Core (ALMC) sounds an automated bell prompting Alexander Jurkevich, the core’s assistant director, to step out of his corner office into the open square room. With a friendly smile, Jurkevich coordinates biologists across MU’s campus to reveal the wonders of the microscopic world.
“Our mission is to provide researchers campus-wide with advanced microscopy instrumentation,” Jurkevich said. “We not only provide access to instrumentation, but we also train, advise users and support them during their early research at the core.”
The core hosts an annual image contest celebrates MU researchers’ microscopic imaging throughout the year. After being canceled for the past two years, this year contestants submitted their best images for consideration.
Christie Herd, a postdoctoral fellow in the Alexander Franz Veterinary Pathobiology lab won the Best of Show Award for her image of the La Crosse Virus in mosquito ovaries.
David Porciani, assistant research professor under the supervision of Bond LSC’s Donald Burke, won Director’s Award for Best Technically Challenging Image with his image showing epidermal growth factor receptors in two types of resolution.
Janlo Robil, a graduate student in the Bond LSC’s Paula McSteen lab won Experts’ Choice Award with his image of hormones in a corn leaf.
A wonderful surprise
Herd was “blown away” when she got her colorful image that won the Best of Show Award. Not only had she tried imaging other viruses with less success, she also worked on different La Crosse samples with no luck.
“That image in particular, I was not expecting to see because the day before I was there for three hours and had to quit,” Herd said. “So, when I saw that image, I was surprised because I did not think I would see that much detail.”
Herd’s image shows the La Crosse virus in developing eggs within mosquito ovaries. Herd uses La Crosse, a type of bunyavirus, as a model to study virus transmission from a female mosquito to her larvae to determine how viruses can remain transmitted within generations of mosquito populations. While La Crosse infects a small amount of humans a year, it transmits quickly, making it the perfect model to learn more about other bunyaviruses like Zika and Chikungunya.
“With bunyaviruses, they’re so multifaceted, and I like being able to research different aspects of them,” Herd said. “I just feel like they’re medically important.”
Herd’s image includes different colors to label different parts of the ovaries as well as the virus so she can see where in the ovaries the virus is traveling. Using a technique that allows her to see different focal planes, Herd can see where the virus is in every dimension, but it can be tricky to get high quality images.
“It does require a few hours of playing around, looking at the microscope,” Herd said with a chuckle. “It requires an investment of time, and sometimes there are days where it’s just not what you wanted and it doesn’t work … and you wasted days.”
Even with the challenges, the ability to see multiple dimensions of a sample at once is valuable for research projects like Herd’s.
“You just have to persevere and try again with a new set of samples,” Herd said.
From side project to passion
Although a deviation from his primary research, David Porciani’s project to image cell surface receptors slowly took over his focus.
“The biggest surprise was that I really had fun,” Porciani said. “This was not my primary project, but it became my primary project for a while.”
Porciani, an assistant research professor under the supervision of Donald Burke, studies molecules on the surface of cancer cells which are receptors for growth factors. These receptors act as a lock, with growth factors as a key. When the growth factors and receptors come together, the cells divide and create more cells. In lung cancer, there are more of these receptors, which can lead to uncontrolled tumor growth.
“This receptor, EGFR, has been widely studied,” Porciani said. “But for me there is definitely an interest because it’s one of the markers in lung cancer.”
Porciani tags those receptors with small molecules called fluorophores that glow under the light of the microscope, so he can see where the receptors are and how they move. However, the fluorophores cannot attach to the receptors alone, so he used aptamers — synthetic keys created by researchers that can bind receptor locks with specificity similar to the natural growth factors. Ultimately, the aptamers clip the fluorophores to the receptors.
“If you can follow the motion of the receptors, these receptors are kind of dancing,” Porciani said.
In his image, each dot is a different receptor made visible by the attached fluorophore.
However, fluorophores can become bleached, rendering them invisible after being exposed to the laser from the microscope for a while. If the synthetic keys, or aptamers, are still bound to those receptors, they cannot be imaged any longer. To address this, Porciani developed an aptamer which attaches to the receptor for a shorter time and then detaches so that even if the fluorophore bleaches, another aptamer can replace it and so receptors can be imaged for longer time.
“We engineered an aptamer with lower affinity that could work with this approach,” Porciani said. “By having lower affinity aptamers we can still determine localization of a high number of receptors and their motion.”
For his winning image, Porciani split the image to show the difference between single molecule resolution on the bottom right and a lower resolution image on the top left.
“With [lower] resolution, you don’t have a single molecule solution. If there are two molecules close together you will see them as just one single dot,” Porciani said. “But with the image, after software analysis with the image on the bottom right, then you have single molecule resolution.”
With this technique — made possible with microscopes at ALMC in Bond LSC — Porciani saw his efforts come together.
“At the beginning, I was just focused on the aptamer engineering from a high affinity aptamer to low affinity aptamer, and making them was the fun part to play with the structure,” Porciani said. “But when we started doing the imaging experiments at the Bond Life Sciences Center I realized that it was not just fun, but it was actually meaningful and this approach could have lots of biomedical applications.”
The artist’s touch
Although passionate about biology and microscopy, Janlo Robil decided to submit his image based on aesthetics.
“I chose this one because I am also a graphic artist, and I appreciate the color and composition,” Robil said.
Robil’s image shows a developing corn leaf with different colors labeling different plant hormone response proteins involved in stimulating growth. His unique image of an entire leaf, which is just under a millimeter in length, required piecing together images of sections of the leaf.
“About 0.75 millimeter, that’s still big in the microscope,” Robil said. “This is kind of difficult to make because it means that you need to tile several images [together] and sometimes it takes up to an hour just to [get] an image.”
The experiment requires planning ahead since the plants containing the fluorescent proteins must be crossed with plants with genetic mutations to determine the roles of the hormones in the leaf development.
To tag the plant proteins with fluorescent proteins requires planning ahead since the plants must be grown with genetic mutations.
“The beautiful image is actually a result of the expression of fluorescent proteins that are tagging the hormone response protein and also the hormone transport protein,” Robil said.
Initially from the Philippines where the agricultural staple is rice, Robil came to Mizzou interested in genetic mechanisms to make rice plants more productive. One way to enhance the rice plants is to make the rice leaves more similar to corn leaves, so Robil found interest in the McSteen lab’s project understanding the role of hormones in corn leaves.
“This project is perfect for me because I am studying the leaf and also integrating genetics,” Robil said. “And I love microscopy so much. I had quite a good amount of training starting in 2021 on confocal microscopy, and that’s why I was able to image this.”