Increasing detection of reinfections and rediscovering brand new infections within days raises concerns for herd immunity and the durability of vaccine efficacy.
Cynthia Tang working to figure out how COVID-19 reinfections can bring us answers on how the virus is developing at Bond LSC. | photo by Davis Suppes, Bond LSC
By Davis Suppes | Bond LSC
Like many viruses, SARS-CoV-2 continues to develop and evolve with time. As the virus evolves it becomes more infectious and can produce worse symptoms and a higher fatality rate. While more people have been infected, we also learn more about it, including how different variants of the same virus can reinfect someone who already had SARS-CoV-2.
In a recent study led by Cynthia Tang, an MD-PhD student at the University of Missouri (MU) School of Medicine and Institute for Data Science and Informatics, her lab was able to identify a patient who got reinfected with a different strain of SARS-CoV-2 just 19 days after their initial infection. Her Systems Biology Laboratory is headed by Principal Investigator, Dr. Henry Wan, who directs the MU Center for Influenza and Emerging infectious Diseases.
“We found a case of reinfection that occurred during a shorter window of time than what we see in the current US Centers for Disease Control and Prevention (CDC) guidelines,” Tang said, “so we may be missing a subset of those reinfection cases.”
In October 2020, the CDC published investigative criteria for suspected SARS-CoV-2 reinfections. These criteria included: any individuals testing positive over 90 days after their first laboratory-confirmed SARS-CoV-2 infection, or symptomatic individuals testing positive 45–89 days after initial infection with paired respiratory specimens.
For this patient, nineteen days following her initial positive test in March of 2020, she returned for another SARS-CoV-2 test due to return-to-work requirements. Despite her symptoms fading to encompass only productive cough and fatigue she tested positive again. She continued to experience persistent cough, fatigue, and difficulty breathing until 55 days after her initial positive test.
Tang showed that the two samples contained two infectious variants of SARS-CoV-2 viruses. No diverse polymorphisms, or a mixture of different viruses, were identified among the sequences of the viruses from each clinical sample, suggesting true reinfection rather than a coinfection. One good news is that this case also suggests that reinfections may not be worse than the prior infection.
The CDC has encouraged symptom-based strategies for ending isolation rather than viral retesting for asymptomatic individuals or for individuals without new symptoms during 90 days after illness onset due to findings that detectable but noninfectious SARS-CoV-2 RNA can persist in respiratory samples. This means that even if you don’t exhibit symptoms there could still be traces of SARS-CoV-2 in your system, and those traces could be from an original infection or a reinfection.
This study suggested that reinfection could pose a challenge to infection control, especially as the Delta variants are surging around and new variants continue to emerge.
“People showing influenza-like illnesses are recommended to isolate themselves for ten days or wait for their symptoms to go away. We may not be catching cases where individuals contract another COVID-19 infection within a short amount of time,” Tang said.
“The COVID-19 vaccination has been proven to be effective in reducing the chances for people to become infected and reinfected”, Tang added.
Research refines platform to address immune disorders
Dr. Esma Yolcu and Dr. Haval Shirwan, Co-pioneers of ProtEx technology
By Davis Suppes | Bond LSC
The things that protect you can also cause the most harm. That’s especially true when it comes to your immune system, which protects you against infections, but is also responsible for a host of diseases, including autoimmune disorders, such as type 1 diabetes.
More than 1.4 million Americans suffer from the self-harming condition of diabetes without an effective cure, but researchers Haval Shirwan and Esma Yolcu may have the answer.
The pair tackle the problem of diabetes and other autoimmune disorders by targeting the component of the immune system that does harm for modulation. DNA-based gene therapy has traditionally been used as a scheme to modulate the immune system.
“DNA-based gene therapy is a complicated, exhaustive and expensive technology,” said Shirwan, a NextGen Precision Health researcher at the University of Missouri and a professor in the Department of Child Health and Molecular Microbiology and Immunology at the School of Medicine. “We wanted to see if there was a more efficient and practical way to do immunomodulation.”
For conditions like Type 1 diabetes, where the immune system misfires and destroy beta cells producing insulin, they target to replace defective islets with healthy ones from another individual. Insulin is an important hormone that plays a role in regulating your blood sugar level. A major hurdle for islet transplantation is the immune rejection.
Shirwan and Yolcu co-pioneered ProtEx™, a proprietary platform that allows for generation of novel recombinant immune ligands and their positional display as single agents or in combination on biological surfaces, such as islets. This is a practical and safe alternative to gene therapy for localized immunomodulation.
In traditional methods of gene therapy, DNA must first be introduced into the cell, but ProtEx technology bypasses that step. Sticking the protein directly on the cell surface for immunotherapy via a test tube allows for a safer, more effective method than gene therapy. Islets engineered to stick on their surface proteins evade rejection by the immune system.
Shirwan’s translational research program’s goal is to develop safe and practical immunomodulatory approaches with applications to transplantation, autoimmunity, cancer, and infections. On the cancer side, Shirwan and Yolcu team has shown that the immune system of a healthy individual can be trained to surveillance the body for cells that may transform into cancer and eliminate them before cancer takes hold. “This is an exciting finding that has not been reported in the literature and sets the stage for
Shirwan’s focus of learning about the immune system and improving it came to him very early when he was a virologist for University of California, Santa Barbara. He developed an interest in immunology and wanted to learn more about immune defense mechanisms. Before bringing his talents to Mizzou he also spent time working at the California Institute of Technology, Allegheny University of the Health Sciences, University of Louisville, and Cedars-Sinai Medical Center.
He believes preventing these cancers and infections is the route we should be aiming toward, rather than trying to find new ways to treat them after they have already damaged the body to an extent.
“Things like cancer are so effective and harmful because they use the same mechanism as the immune system to evade it,” Dr. Shirwan said, “Just like how a runner can build muscle memory for running, our immune system can learn and build up a memory of these pathogens and infections to create a more effective response.”
“This is just the tip of the iceberg,” Dr. Shirwan said, “I am very excited to continue expanding on what our immune system is capable of.”
This type of research contributes to the University of Missouri System’s NextGen Precision Health focus. The NextGen Initiative unites scientists, government and industry leaders with innovators from across the system’s four research universities in pursuit of life-changing precision health advancements.
Dr. Shirwan and his team are moving to the new NextGen building on Mizzou’s campus in mid-September. He is very thankful for getting the opportunity to work in these facilities with these people, and he is excited for the next step of their research.
Shirwan believes we have only scratched the surface of what our immune system is capable of. Our immune system may not always know what is good and bad for itself but one thing is for sure, Dr. Haval Shirwan is good for it.
After 40 years of hard work, it is finally time for David Pintel to pass the torch.
Dr. David Pintel, retiring after 40 years at Bond LSC, takes in his office during his last week at Bond LSC. | photo by Davis Suppes, Bond LSC
By Davis Suppes| Bond LSC
David Pintel is hanging up his lab coat after 40 years.
“It’s been an honor to be able to do my work at the University of Missouri. I’ve had a great group of colleagues both here and at the medical school,” Pintel said.
The Bond LSC virologist retired July 1 after a storied career. He spent 20 years at the School of Medicine before joining Bond LSC upon its completion more than 15 years ago.
Since then he has aimed to better understand the interaction between viruses and host cells. Pintel specialized in the study of parvoviruses, the smallest of all DNA viruses that infect vertebrates. In addition, the parvovirus adeno-associated virus (AAV) has been developed as a promising gene therapy vehicle.
On his last official day in his office Pintel was busy still packing up old memories while taking everything in.
“I’ve had tremendous students and tremendous colleagues, you know they were the important parts of my career,” Pintel said, “Going through all the old memories, you know, it’s gonna take a little while”
Pintel is a Curators’ Distinguished Professor as well as a Dr. R. Phillip and Diane Acuff Endowed Professor in Medical Research Molecular Microbiology and Immunology. He reflected on fond memories of his work here, but when asked what his favorite memory was his answer was the same, working with all of these people.
“It’s an emotional end, when you do something every day of your life for 40 years,” Pintel said. “I’m so thankful and grateful for the people that have worked with me, people that have been my friends here and allowed me to have a successful career.”
The future for Pintel will come in due time, but for now he is in no hurry and plans to enjoy some time off.
“To be determined,” Pintel said, “I think I will decompress for a while before I make any decisions on the next step.”
Lyndon Coghill is the new Director of Informatics Research Core, and he is already making big moves at Mizzou.
By Davis Suppes | Bond LSC
Lyndon Coghill’s official title may be Director of Informatics for the Informatics Research Core, but his job branches out much wider than just a single label. Even as an undergrad, Coghill wore many different hats.
“I was incredibly excited about the way that the MU Office of Research and Economic Development recruited me,” Coghill said, “With these types of processes you can get an idea as to whether or not an institution is actually committed and excited about building something out.”
With his experience and range of expertise, Coghill was an easy choice for Mizzou to fill the role of Director of the Informatics Research Core located at Bond Life Sciences Center.
Before he achieved his doctorate in biology, he completed his undergraduate degree in zoology with minors in microbiology and geology at Western Illinois University. For his dissertation, the research he conducted was focused heavily on evolutionary genomics. Simply put, he wanted to know how changes in the genome lead to changes in a physical organism that allow them to adapt better to different environments and conditions.
With his doctorate in biology, he would go on to his first postdoc at The Field Museum of Natural History in Chicago in 2013, and then on to Louisiana State University where he began his role as a senior post doctorate in 2015. He continued to diversify his portfolio there working with the department of biology, focusing on computational biology. He was then promoted to research data scientist which had him take on an even more computationally heavy role. With this, he was able to help biologists learn how to talk to computer scientists, and assist them with building collaborative programs together..
As director, Coghill’s mission is to provide bioinformatics and data science support to all researchers across the UM system. He is creating a central hub where faculty whowant toconduct domain- specific biological or life sciences-related research that is computationally heavy can get the help they need to come up with solutions. He does this by helping researchers wrangle incredibly large datasets and by helping them understand what that data is telling them from an information perspective in a meaningful way.
Coghill mentioned how interim Vice Chancellor of Research and Economic Development Thomas Spencer also made a personal effort to make sure Coghill understood his vision going forward on campus “and for me that was enough of a selling point that I wanted to be a part of that,” Coghill said.
In addition to the thorough recruitment process, Mizzou’s facilities and access were other huge factors that Coghill was looking forward to once he got here. With a hospital, vet school and productive biology program all on the same campus instead of in different cities, Mizzou offers a unique opportunity to build the integrated program all in one place.
“We’re trying to reach out to every department on campus to build these relationships because you can’t have true integration of ideas and solutions if you don’t talk to everyone who might be a benefactor or have knowledge about that,” Coghill said.
Coghill believes that to create a phenomenal translational research program, this core must interact with all these programs so that experts of different fields can come together to collaborate.
“Informatics research, especially bioinformatics is a program that really forces you to keep one foot in both worlds of computer science and biology, and there’s a limited number of peoplewho do that kind of work,” Coghill said, “I think that was one of the big pushes for getting my experience here for this position, to bring in somebody who could bring these programs together and integrate across all these different fields.”
Coghill is excited to be working with the variety of researchers and programs across the MU campus and UM System, and learning from them at the same time.
“We may not know their biological system as well as they do and we may not know the high performance computing system as well as a full-time systems administrator, but we know enough of both that we can communicate with both teams and make sure that we can help get the researchers from the starting point to a meaningful result,” Coghill said.
Their goal is to provide Mizzou and sister campuses with research support allowing faculty to build translational research programs using computing power and informatics. This core brings new opportunities for Mizzou students as well.
“We’re going to have programs for students that can rotate through as part of the Informatics and Data Science Institute,” Coghill said.
This means that students who are interested in research fields can get direct experience related to career possibilities outside of Mizzou and academics by working in this program.
“Students can come to us and learn basic coding skills, learn informatics and bioinformatics, and that will help them build a skillset that will make them quite employable,” Coghill said.
Between helping researchers in their labs and analyzing quantities of data they are gathering for the first time, Coghill has a variety of jobs he has to understand and execute.
“So, I am the guy who wears a lot of different hats and allows these researchers from different domains to talk to each other,” Coghill said. “We’re trying to help them get to the point where their work could be as big as they want.”
Mizzou and Coghill know that there is no way to push modern research without computing, especially at the scale research is done today.
It would be extremely rare for someone who has a doctorate and spent their life trying to understand how one particular part of a biological process works to also have a doctorate in computer science, “That’s where we help… we’re providing researchers with the tools to do research at a scale using computing power, and asking questions that for many, might have only been dreamed about at other times in their careers,” Coghill said.
Years at MU lands student turned faculty tenure-track position
By Mariah Cox | Bond LSC
Where can passion, hard work and more than a decade worth of experience get you? They landed Maggie Lange-Osborn her own research lab on the University of Missouri campus.
Lange is starting down that path in Bond Life Sciences Center but will move to a permanent space in either the Medical Science Building or Schweitzer Hall eventually. She’s excited to spread her wings and establish herself independently of her past role in the Bond LSC.
In the meantime, she’s working arduously to build her lab from the ground up. From small materials such as plastics for cell culture, pipettes and pipette tips and chemicals to make buffers to large expensive equipment, Lange will eventually need it all.
“Walking into the lab, you don’t realize all of the things you need to do an experiment. When I walk into my lab space, I literally have nothing,” said Lange, a newly appointed assistant professor in the Department of Molecular Microbiology and Immunology (MMI). “You have to think about all of those things, you know it’s not just the equipment it’s the pipettes, the incubators, the hoods and then also the materials to put in those things.”
Luckily with 10 plus years in the building, Lange knows of all of the shared resources available to her, so she doesn’t have to invest in many expensive machines just yet.
“I cannot wait to do my first experiment with my own equipment,” Lange said.
Lange began her career at MU in 2003 as a Ph.D. student in the Molecular Microbiology and Immunology and Veterinary Pathobiology Graduate Program, whether she knew it at the time or not.
When applying for faculty positions, Lange worried that she would be placed in a box, unable to separate herself from her graduate work if she stayed at MU. With time, she’s found that hasn’t been the case.
“Even though I’ve been here for so long, I’ve been able to surround myself with people who know more than me and who have different areas of expertise than I do,” Lange said. “I can still branch off and learn a lot from other researchers, which is what I found really attractive about Mizzou.”
Her segway into research occurred during her first year as a graduate student in an MMI lab in the School of Medicine. Under Michael Misfeldt at the School of Medicine, Lange studied pattern recognition receptors in the innate immune system, which is the body’s first line of defense against a virus or bacteria.
The specific receptor she worked with recognizes a component of viruses. From there, Lange wanted to understand how a host is able to respond.
“After working on that I felt like I had a really good grasp of the host response side and I wanted to get more of the virus side to understand virus replication and what types of replication mechanisms work to signal the host from that perspective,” Lange said.
That led Lange to join Bond LSC in 2008 as a post-doctoral researcher in Donald Burke’s lab. Known for specializing in HIV research and viral biology, the Burke lab gave Lange the opportunity to understand virus side interactions.
Lange wasn’t quite ready to move on at the end of her post-doc, and the success with her research led Burke to invite her to stay on as an assistant research professor.
“During that time, I was more exposed to leading people and mentoring people in the lab. I was able to get some teaching experience in that role as well in the infection, immunity and advanced virology classes,” Lange said. “The position evolved into really enjoying all of the components that would be required for a tenure track position and it grew from there.”
In her sixth year as an assistant researcher, Lange decided she was ready to run her own lab. However, she knew it would be a challenge to secure a faculty position and even more difficult to stay at MU.
But, her experience in the Burke lab and her proven ability to obtain grant funding worked in her favor.
“It’s really nice to stay in Missouri because mine and my husband’s family are here,” Lange said. “When I was growing up, my dad was in the military and we moved around a lot, so I never got to know my grandparents or cousins. It’s really nice now that my kids get to have those relationships with their extended family.”
Her goal with her new lab is to combine her knowledge of viral interaction in the body and hosts’ response to infection.
Three projects currently in the works for grant submission focus on host-virus interactions and how different host factors and viral proteins interact during replication. One specific project looks at the host factors that are involved in HIV induced death caused by different HIV proteins.
“While HIV has been around for a long time, there are still things we don’t know about it. With the research, I’m getting back to my innate immunity roots and looking at exactly how viruses interact with innate immune receptors and signaling pathways and how that interaction dictates pathogenic outcomes,” Lange said.
Understanding the death pathways for HIV can lead to the development of a strategy to preserve T cells and facilitate the death of the virus. Additionally, it can lead to the development of therapies toward eradication.
Her excitement isn’t just for her new lab, it’s also for her newfound opportunity to provide students with lab experience and open up the possibility of research for those who haven’t had access to it.
“I’m from a rural community and I didn’t even know that a Ph.D. existed when I was in high school,” Lange said. “I’d really like to present those opportunities to people like me who have no idea that they’re even available. There are so many things you don’t realize are possible because of the environment that you’re in whether it be in rural or inner-city communities.”
While unknowingly launching her career at the outset of her Ph.D. program, Lange is grateful her path led her here.
“It’s been really fortunate for me, the way the whole process developed. I love this building and the awesome people I’ve met here, but I’ve been here since 2008, so having a fresh perspective elsewhere could be beneficial. I’ve worked a long time with both Marc Johnson and Donald Burke and being away from the building will allow me to meet new investigators and establish new collaborations,” Lange said. “While we still have very productive collaborations and have promising, active projects, it will help demonstrate that I’m separate from them and have my own interests as well.”
Researchers from MU, the University of Maryland and the Pacific Northwest National Laboratory are building a microscope that doesn’t yet exist.
By Mariah Cox | Bond LSC
Tiny neon dots speckle a black backdrop – and no, this isn’t a Hasbro Lite Brite. Rather, these fluorescent dots indicate something about plants that scientists research and help them see the genes, traits and molecules they study amid thousands of possibilities.
To help in seeing that, a new imaging microscope will allow researchers to better pinpoint molecular interactions in plants they have a hard time highlighting to overcome the obstacle plant scientists face with wavelengths of light they can’t necessarily see.
“When you think about imaging, you think about what you can see with your eyes. But, there are a whole variety of other things you can image that aren’t visible to the human eye,” said Gary Stacey, a Bond Life Sciences Center principal investigator who is working to help develop a new microscope technology to view fluorescent quantum dot markers beyond the range of visible light, into the infrared spectrum.
Stacey, along with collaborators from the University of Maryland and the Pacific Northwest National Laboratory (PNNL), was awarded a combined $2.25 million grant from the Department of Energy (DOE) to develop a novel microscope for ‘multiplexed super-resolution fluorescence imaging in plants.’
The call for the development of this new imaging hardware was borne out of the need for a more precise measurement of enzyme function, tracking of metabolic pathways and monitoring the transport of materials and signaling processes within and among cells in plants. Right now, the emission spectrum of plant pigments limits the usefulness of and the number of fluorescent colors that can be detected in a single experiment.
Stacey and his collaborators were one of six groups to be chosen for a total $13.5 million investment from the DOE for new bioimaging approaches. For bioenergy, using quantum dots in combination with other novel technologies could enhance imaging techniques to allow scientists new ways to re-engineer plants and microbes for bioenergy conservation and production.
Quantum dots are small particles that are only a few nanometers in size, one nanometer equals one billionth of a meter, and are used as fluorescent biological labels in cells. These labels can be tagged to particular molecules, cell parts or genes of interest to a researcher.
“Think about [fluorescence] as a black light. If you have a room that’s completely dark with fluorescent paint on the wall and you turn on a black light, then you will be able to see where the paint is on the wall,” Stacey said. “It’s the same concept for quantum dots. One application is localizing where a virus is or label it and watch it move into a cell to try to understand the mechanism by which it moves.”
However, the mechanics behind quantum dots don’t make it simple. When exciting a single molecule, it will fluoresce and emit light but will do so in a diffuse pattern. This makes it difficult to see the molecule itself.
Additionally, plants absorb 490-700nm of light — essentially covers the entire visible range of light — allowing them to photosynthesize. As a result of absorbing these wavelengths of light, they also auto fluoresce, which is natural emission of light by structures inside plants cells such as chloroplasts.
The problem, then, is that viruses labeled with fluorescent probes in leaves are difficult to see because of the natural fluorescent glow coming from the plant.
For Stacey and his collaborators, the idea is to go beyond the visible light spectrum and use infrared light, which is above the visible light spectrum. Infrared light is most commonly known for its use in heat lamps.
“With infrared light, there would be no autofluorescence and so when you shine infrared light on a leaf, it would appear black,” Stacey said.” If you shine an infrared light on a fluorescent molecule, it would emit light and show up against a black background, making it very easy to see.”
The problem, though, is being able to distinguish one fluorescent molecule from another when they are close together. Because the researchers will be using infrared light, which has a longer wavelength, the imaging resolution decreases.
To get around that obstacle, the researchers will be using super-resolution microscopy to compensate for the resolution loss. The use of this technology will allow them to pinpoint the center of the fluorescence.
“It should be a big breakthrough. We would be able to look at single molecules interacting against a black background without any interference from autofluorescence,” Stacey said.
Stacey’s collaborators each contribute to the project in their unique way.
Zeev Rosenzweig from the University of Maryland, who is an expert in quantum dots, will be making the dots and labeling them with probes that absorb infrared light. Galya Orr from the Environmental Molecular Sciences Laboratory (EMSL), PNNL, in Richmond, Washington, has expertise in fluorescent microscopy and she, and her colleagues, will build the microscope.
An attractive part of submitting a proposal for the grant is the microscope’s prospect of being used as part of the EMSL user facility, which will ultimately allow researchers from all over the world to use the microscope when fully developed.
The researchers are excited to begin work on this project because they’re building a microscope that doesn’t yet exist. The microscope will expand the capabilities of researchers all over the world.
Stacey is appreciative that he gets to work with researchers from multiple disciplines. Namely, because he learns more about science from the expertise of others.
“That’s what makes it exciting because you’re constantly learning. The great thing about science is that you’re learning every day. It’s nice to get into these new areas especially where you don’t feel comfortable and learn new stuff,” Stacey said.
This work is funded by the Department of Energy for innovating new bioimaging approaches for bioenergy. The grant is split among the collaborators at the University of Maryland, University of Missouri and the Environmental Molecular Sciences Laboratory at the Pacific Northwest National Laboratory. Specifically, $1.5 mil. is to be used by researchers at the University of Maryland and the University of Missouri and $750,000 by researchers at EMSL.
Cross-collaborative research team looks to refine delivery of cancer treatments
By Mariah Cox | Bond LSC
“When you want to use a tool to do something in the house, you have to use the right size tool. It does no good to use a large screwdriver to fix the tiny screw on your glasses.”
That’s Donald Burke, Bond Life Sciences Center lead primary investigator, as he begins to explain a project looking to optimize the targeting of cancer cells as part of a large cross-collaborative research team.
And the tools Burke is referring to are aptamers, single-chained synthetic DNA or RNA molecules. Aptamers are tricky molecules. Stemming from the Latin word “aptus,” meaning to fit, and “meros,” meaning part, aptamers must be complementary in size and shape to a certain cell-surface receptor in order to be useful as targeting tools, just like having the right size tool.
Last fall, Burke’s lab was able to identify aptamers as a specialized delivery method that have the potential to carry chemotherapy drugs and imaging agents, or cargo, as little backpacks to diseased sites. An important feature of aptamers is their three-dimensional structures which allow them to bind to target sites with high selectivity. That conceivably means they could deliver a drug to a particular part of the body, like a tumor, and not harm nearby tissues.
Now, the team, comprised of MU experts and researchers who specialize in surgery, radiology, molecular biology and immunology and chemical engineering, is hitting the ground running with a two-year research plan to refine the delivery of cancer treatments. The group was one of 19 innovative research projects across all four UM System campuses to receive a grant from a $20.5 million investment for research and creative works.
“Chemotherapeutics are not very specific and most of them act to block DNA replication or other functions of the cell in both healthy and cancerous cells,” said David Porciani a post-doc who started in the Burke lab in the spring of 2016 after finishing his Ph.D. in molecular biophysics in Italy. “Cells that are dividing are more susceptible to the chemotherapeutic effect, that’s why chemotherapy patients start losing hair.”
In general, therapeutic drugs don’t know to go only to tumors, and they don’t differentiate between cancer cells from healthy cells. The obstacle in only targeting cancer cells is to find specific indicative markers that are unique to tumors. Sometimes researchers have to make do with markers that are overexpressed on tumors cells, but those same markers can also be present in low levels on healthy cells.
The team is working simultaneously towards four major objectives – to identify a comprehensive panel of aptamers that target the majority of tumors, develop molecular tools to enhance the delivery of cargo specifically to cancerous cells, improve imaging for targeted delivery of radiopharmaceuticals, and enhance the efficacy of killing solid tumors through immunotherapy.
Each researcher bringing their unique expertise to the table play an important role in ensuring the project stay on track. Mark Daniels, an associate professor of Molecular Microbiology and Immunology and Surgery from the School of Medicine; Bret Ulery, an assistant professor in the Department of Biomedical, Biological & Chemical Engineering; and Donald Burke, a professor of Molecular Biology and Immunology in the School of Medicine and joint professor of Biochemistry, have been instrumental in the project since its beginning stages.
A collaboration years in the making
“Daniels, Ulery and Burke labs have been collaborating for a number of years, each of us excited about what the other could bring to the collaboration,” said Burke. “[Over the years] we’ve explored several ways of making things move forward, we’ve figured out some productive ways to work together and we’ve identified the key questions that we could pool our respective expertise toward answering,” said Burke.
Daniels’ subgroup of the project has been using flow cytometry, a technique used to detect and measure biochemical and molecular characteristics of tumor cells to see which ones are recognized by a set of different aptamers.
Additionally, Ulery has provided extensive insight into the different ways to package molecules together so that they can move around the body and get to where they need to go.
The grant comes as part of the combined UM system’s and all four campuses mission to supply funding for opportunities that will enhance the ‘well-being for Missouri, the nation and the world through transformative teaching, research, innovation, engagement and inclusion.’
For Burke, the grant couldn’t have come at a better time.
“This team has existed before the announcements were made that there would be these opportunities and so when they announced we said that this was tailor-made for us,” said Burke. “It was just the right time for us. Had they had the same competition three years ago, we weren’t ready for it. Had they had it three years from now hopefully we wouldn’t have needed it anymore and we would’ve already gotten the project to the next level.”
Another key player in the development of this discovery is David Porciani. He has spent three years trying to create ‘smart’ molecules that know exactly where to bind without damaging healthy cells, and he has been working on the project since its beginning.
“There are several ways to target cancer cells. Even on campus, there are different groups that are trying to target cancer cells differently and, in my experience, every strategy has advantages and limitations,” said Porciani. “What I see for this aptamer strategy is that it can provide new molecules that can bind to receptors, and it can also identify new tumor biomarkers.”
Porciani’s portion of the project includes using the advanced microscopy capabilities of the MU Molecular Cytology Core to visualize the kinds of receptors on the surface of cancer cells. This information can be very telling in identifying receptors that are specific only to cancerous cells.
What the future holds
The project now starts down this ambitious road. In the first year, the collaborators are working on discovering their panels of aptamers, developing lung cancer-specific T cells which provide artificial cell receptors for the use of immunotherapy, and beginning testing of the specificity of aptamer-cargo constructs in samples acquired from the American Tissue Cancer Collection. By year two the group hopes to begin testing in biopsy tissues acquired from MU hospital patients acquired under informed consent and in lab mice.
“After we test a specific drug in cell culture samples and see an anti-cancer effect, killing only the cancer cells and leaving the healthy cells, we expect to see the same effect in biopsy tissue samples,” said Porciani. “Having a reduction of the tumor mass and not having side-effects is what we hope to see in biopsy tissues and mice.”
After the end of year two, the team is looking forward to expanding this research to other types of tumors to see if their tumor-targeting method can apply in different cancer types.
“Our team has been trying to piece this together for a long time and it’s been surviving on goodwill up to this point. This is the most substantial funding we’ve had for it to date and we’re really excited that the University of Missouri has chosen to support us on this,” said Burke. “We’re very hopeful that we can use it as the starting point for building a much larger enterprise centered around tumor targeting in general, whether it’s for therapeutics, diagnostics or other purposes.”
Donald Burke is a primary investigator in the Bond Life Science Center and is a professor of Molecular Microbiology & Immunology in the School of Medicine and a joint professor of Biochemistry and Bioengineering. David Porciani is a post-doctoral researcher in the Burke lab. Mark Daniels is an associate professor of Molecular Microbiology and Immunology and Surgery in the School of Medicine. Bret Ulery is an assistant professor in the Department of Biomedical, Biological and Chemical Engineering in the College of Engineering. Other key collaborators on the project include Diego Avella Patino and Jusuf Kaifi in the Department of Surgery and Jeff Smith in the Department of Radiology.
Fourteen days. That’s how long it took Sam McInturf and Ke Gao to put together a root imaging machine named ‘Sun Bear’ at the University of the Western Cape in South Africa this past June.
The pair, a postdoctoral researcher in the lab of Bond LSC’s David Mendoza and a computer science Ph.D. candidate, brought the automated approach to capturing data on root growth abroad as part of a technology transfer collaboration under the University of Missouri African Education Program (UMSAEP) established by the board of curators in 1985.
The machine, which optimizes data collection of root growth and provides more intuitive data about the factors that stunt or spur growth, allows researchers to focus more time on larger and diverse projects. In many research labs, automation is the way of the future.
“It changes the game. Being able to do everything in a high throughput fashion and being able to test lots of things at once allows us to consider the type of experiments we might conduct differently,” said McInturf. “All of a sudden we have access to the physical capacity to ask new types of questions.”
After touching down in Cape Town, the duo was immediately met with their first obstacle — the tools and building materials they shipped over were tied up in customs. Although McInturf and Gao prepared for obstacles with power conversions and tool availability, they weren’t anticipating broken equipment or problems with electrical wiring and motors.
Over 14 days, the pair spent 10-12 hours in the lab constructing the machine in preparation for its unveiling with presidents and administrators of both UWC and MU. The construction of the machine coincided with a visit by the University of Missouri System President Mun Choi.
“We went over there with a hard deadline of presenting the machine to the presidents of both universities and we needed to be ready then,” said McInturf. “The sum total of all of our efforts resulted in 15 minutes with the presidents where we got to meet with them, and we got to show off what we had built and the plans that we have come up with for further collaboration.”
McInturf has been instrumental in the project since its beginning.
In 2014, the project idea was brought to McInturf by his adviser David Mendoza. Since then, the project has morphed through four major prototypes and has been worked on by students and researchers across multiple disciplines including bioengineering, computer science and computer engineering. In all, students and researchers from the Division of Plant Sciences and the Electrical Engineering & Computer Science Department spent five years developing the blueprint for the interface and the algorithm for the machine.
In its beginning stages, the project was part of a bioengineering senior capstone class, and McInturf was the only person working on it in 2017 as he finished his doctoral work.
“We made serious progress starting about two years ago, but we really hit our stride about one year ago in the fall of 2018,” he said.
Gao joined the team about a year ago to help refine an algorithm to process the data. His algorithm recognizes the seed and the root and measures the root growth over time in conjunction with a time stamp.
However, the robot is still undergoing minor tweaks and finishing touches.
“We’re definitely going to be working on the root tracing algorithm for the fall semester. For now, it’s still a prototype. It’s not very optimized and it’s still a little bit slow,” said Gao. “I’ve been working with Sam to make the interface better to show more information like a graph or a table. Right now, we are only measuring the primary root, but there are also a lot of branches on the root. We don’t have an algorithm yet that’s working well for that scenario.”
The robot is currently being used at both UWC and MU to capture detailed information about root growth under a variety of different stresses, conditions and genetic backgrounds. This method of data collection as compared to the traditional methods of manually measuring and recording information is allowing researchers to better understand how different factors affect root growth.
“We are interested in characterizing genes by first breaking them and seeing what happens,” said McInturf. “Once we understand what these genes are doing and how they interact together, then we can go on to engineer crops and conduct targeted breeding strategies to allow plants to survive harsh environments.”
The project is part of a cross-campus collaborative named “The Foundry” funded by CAFNR and housed in the LSC which brings together scientists and engineers from varying disciplines to mechanize biology. Other projects currently being worked on by The Foundry include a hyperspectral camera and a leaf imaging robot.
As for McInturf, this project has opened his eyes to a new realm of possibility in his future career path.
“I didn’t think that I could do this kind of work before this project. Not only did it show me that I have the capacity to do it and that it wasn’t so impossible, but it showed me that I could,” said McInturf. “As I move forward to industry or academia my goal is to be working on the interface between mechanization and plant biology, that is kind of my bread and butter at this point.”
UMSAEP was established in 1985 by the University of Missouri Board of Curators to aid South Africans who were disadvantaged by the then-government’s apartheid policies. In June 1986, a formal memorandum of academic cooperation was signed by then UM President C. Peter Magrath and then UWC Rector Jakes Gerwel. This agreement has the distinction of being the first-ever developed between a nonwhite South African university and an American university.
Situated in the Bond Life Sciences Center is an almost empty research lab on the fourth floor, which to some may look like an end but is really just a new beginning.
Inside you may find a few boxes of gloves, a stack of brightly colored test tube racks and a sole thermocycler sitting on the lab bench. You’ll also find Ashley Meyer, the first researcher hired on by Wes Warren.
“This is a brand-new lab. Dr. Warren came from Washington University in Saint Louis, and he didn’t bring anything with him so all the stuff you see here I just bought in the past two weeks,” Meyer said. “I’m still ordering things and learning all sorts of new protocols and helping to optimize systems.”
Warren hired Meyer to study genes associated with sexual dimorphism. Sexual dimorphism are differences between sexes that present themselves via behavior, physical traits and phenotypes.
“Single-cell RNA sequencing can be really beneficial for understanding diseases. Specifically, there are a lot of diseases that act differently for women and men,” Meyer said. “Women are more prone to get arthritis whereas men are more prone to get colon cancer. If we can understand what genes may be responsible and how they’re different between the sexes, it can be extremely beneficial.”
Meyer wasn’t always interested in genomic research, however. Much like her supervisor, Wes Warren, Meyer went to school in hopes of becoming a veterinarian but eventually found her passion for research.
“I originally wanted to be a vet, that’s a pretty popular answer,” Meyer said. “Once I got into animal science it was more livestock based and I realized that the vet school really wasn’t what I wanted to do and that I really enjoyed science.”
It wasn’t until after she spoke with one of her professors that she realized science and research was the path for her. Meyer completed her graduate degree in animal science focused on reproductive physiology and molecular biology at the University of Missouri.
“I studied reproduction in pigs and I loved it,” Meyer said.
For Meyer, being able to run her own lab will prepare her for her future.
“I like the idea of being able to run a lab. I think that having the opportunity to be the lab supervisor and help start this lab from scratch will benefit me to be able to run a lab pretty effectively in the future,” Meyer said. “The main reason why I took this job is because I really wanted to learn. When you’re a student, you’re narrowed into a specific field. This job is going to give me the chance to learn all sorts of things: bioinformatics, genomic research, cancer research. This job is going to give me the chance to broaden my career.”
As for the future, Meyer doesn’t foresee pursuing a Ph.D., but she has been told that there is nothing wrong with coming back. For right now, she wants a few years of experience under her belt before committing to another four to five years in school.
“I’m a big advocate for science in general and helping the general public understand science and be exposed to it,” Meyer said. “I think it’s easy for people to push it off to the side and say it’s not important when really it’s the basis for everything we have and do, especially in the medical field.”
It takes a lot to move a discovery from lab bench to an application that can provide therapeutic benefits to those suffering from disease.
Bond LSC’s Chris Lorson is making moves to bridge that gap with the start of Shift Pharmaceuticals. With its formation in March 2017, Lorson adds co-founder and Chief Science Officer of the company to his list of titles that include Bond LSC investigator, professor of veterinary pathobiology and associate dean for research and graduate studies.
Shift Pharmaceuticals builds off of years of progress the Lorson Lab has made in understanding spinal muscular atrophy (SMA), which is the leading genetic cause of infant deaths. The disease causes neurons to die, leading to muscle failure, including those that affect walking, arm movement, and respiratory function. While SMA is technically a rare disease, it is remarkably common, affecting nearly 1/10,000 births.
“It’s a devastating disease for patients and families; while the primary defect in is nerves, this leads to problems in muscles, bones, and other vital systems,” Lorson said. “Historically, the majority of kids who develop SMA do not survive beyond 3-5 years.”
Lorson knew his research had potential for drug development, and MU’s Office of Technology Management & Industry Relations (OTMIR) pursued patent protection for the technology. This process helps safeguard the innovations resulting from the research and allows MU to better attract commercial interest to develop and market medical treatments originating from the technology. But it took a partnership with co-founder Steve O’Connor to get the ball rolling. O’Connor is MU’s Entrepreneur in Residence and has significant experience in starting drug development businesses and now serves as CEO of Shift.
“I never knew how to start a company,” Lorson said. “I would argue most academics don’t – this isn’t part of our traditional training. I sat around not knowing what to do, realizing I had this thing that could do a lot, but the practical steps of setting up a biotech start-up were beyond me.”
“Steve thought this technology sounded really cool, so in late March he submitted paperwork and by the end of March we were a company. Shift’s first grant was submitted 3 days later.”
The goal of Shift Pharmaceuticals is to move their lead compound into the clinic for SMA.
“When you look at the disease, it’s not just one cell type and not just one clinical type of patient,” Lorson said. “It really is a disease that is complex and the idea is to bring more options to the fight.”
With the start of any new business, money is always a necessity. Funding from the advocacy group CURE SMA provided the initial funding for the discovery of this compound, while several other foundations including FightSMA, the Gwendolyn Strong Foundation, and Muscular Dystrophy Association have further contributed to the pre-clinical development. MU’s OTMIR negotiated an option agreement with Shift, giving them the exclusive rights to the technology, which helped them obtain a recent $2.73 million grant from the Department of Defense Congressionally Directed Medical Research Program (CDMRP). This will move the company toward the first phase of investigating a new drug.
The root of SMA
Lorson has spent most of his scientific career chasing the underlying causes of SMA.
That search focused in on a few key genes in those suffering from the disease. Two genes — named survival motor neuron-1 and -2 (SMN1 and SMN2, respectively) — are central to SMA development. In patients, SMN1 is mutated and doesn’t process enough of a key protein (SMN) that helps neurons function. While SMN2 acts as a backup gene for this function, a miniscule change in the SMN2 gene causes it to make less SMN protein than required by the body.
In 2016, the Lorson Lab at Bond LSC produced a compound that increases the lifespan of SMA mice . They targeted the back-up gene, SMN2, to produce more functional protein and discovered an increase of protein causing a significant lifespan extension in treated mice. This discovery showed promise for creating a cure for those with SMA.
Shift Pharmaceuticals will be working on developing a drug that builds off Lorson’s work and targets all forms of SMA.
Shift’s first two employees, Mizzou alumnus Paul Morcos and UMKC alumnae Diane Beatty, will help move toward that goal. With Morcos in research and development and Beatty negotiating regulatory affairs, the company hopes to move the drug toward FDA approval.
Thanks to the recent Department of Defense grant, the next step looks to testing in larger animals and at things Lorson did not consider before.
“The experiments we will be doing are not academic in nature, rather, they are focused on the singular goal of preparing our lead compound for an FDA submission. From a traditional academic lab perspective, these might sound rather boring,” Lorson said. “All of these things are not things you do in an academic setting, but that is exactly the point. This is drug development, not the quest for another paper.”
Within the Business Incubator, MU will provide space for the company as well for their research. This sort of partnership is just another part that may one day help translate vital basic research into future treatment.
“Almost everybody has the possibility of doing something that is translational, it is just envisioning it in a different way,” Lorson said.