MU researchers are getting a deeper understanding how mosquito-borne viruses infect the mosquito – an important step toward eliminating the transmission of mosquito-borne diseases

They approach with the telltale sign – a high-pitched whine. It’s a warning that you are a mosquito’s next meal. But that mosquito might carry a virus, and now the virus is in you. Now, with the help of state-of-the-art technology, researchers at the University of Missouri can see how a virus moves within a mosquito’s body, which could lead to the prevention of mosquitoes transmitting diseases.

“Previously, the common understanding was that when a mosquito has picked up a virus, it first needs some time to build up inside the midgut, or stomach, before infecting other tissues in the mosquito,” said Alexander Franz, an assistant professor in the Department of Veterinary Pathobiology in the MU College of Veterinary Medicine and the study’s corresponding author. “However, our observations show that this process occurs at a much faster pace; in fact, there is only a narrow window of 32 to 48 hours between the initial infection and the virus leaving the mosquito’s stomach. For this field of research, that revelation is eye opening.”

In this study, Franz and the team of researchers observed a mosquito infected with the chikungunya virus, which originates in Africa and was first found in the Americas in 2013. There is no vaccine to prevent or treat this virus, and while most common symptoms include fever and joint pain, they can be severe and disabling. The researchers used three separate electron microscopes to view the virus traveling through the mosquito, beginning with its midgut, or stomach. The first two microscopes provided different two-dimensional views of a single layer of tissue in the mosquito’s stomach. The third, a focused ion beam electron microscope, allowed researchers to see multiple layers of tissue.

“We’re now visualizing a real virus with a three-dimensional model, at scale,” said DeAna Grant, a researcher with the Electron Microscopy Core Facility at the University of Missouri and a co-author on the study. “We can take a three-dimensional image showing the inside of a mosquito’s stomach and say that this dot is a virus particle; there is no guessing to what that dot is. In addition, with this technology we were able to track, in three-dimension, the virus traveling through the mosquito at 24, 32 and 48 hour intervals, and within 48 hours or less, we could see the virus particles leaving the mosquito’s midgut.”

Researchers hope to one day inhibit the genes involved with the release of the virus from within the mosquito’s stomach to prevent future transmission of mosquito-borne diseases. This study is the first life sciences work published using the focused ion beam electron microscope that the University of Missouri purchased through the MU Office of Research in 2016.

The study, “Ultrastructural analysis of chikungunya virus dissemination from the midgut of the yellow fever mosquito, Aedes aegypti,” was published in Viruses. Funding was provided by grants from the National Institutes of Health-National Institute of Allergy and Infectious Diseases (R01AI091972 and R01AI134661), and the University of Missouri award for “Excellence in Electron Microscopy”. The content is solely the responsibility of the authors and does not necessarily represent the official views of the funding agencies.

The study is part of the doctoral project of Asher Kantor, a graduate student in Franz’s lab. In addition to Kantor, Grant and Franz, the publication was co-written by Velmurugan Balaraman of the MU Department of Veterinary Pathobiology, along with Tommi White of the MU Department of Biochemistry and Electron Microscopy Core Facility at the University of Missouri.

Story Contact(s):
Eric Stann, stanne@missouri.edu, 573-882-3346

Researchers mimic swallowing problems in rodents that will allow more targeted study of the disease’s progression

Immunology has changed biological research with an improved understanding of how to manage, treat and prevent diseases. At the Laboratory for Infectious Disease Research (LIDR), we foster research for the advancement of vaccines and therapeutics to combat diseases caused by some of the world’s most deadly pathogens. A mainstay in LIDR’s portfolio is the immunology facility offering cytometry and imaging of fixed or live BSL-1, BSL-2, and BSL-3 samples.

Jeffrey Whyte, PhD, LIDR Immunology research specialist, and Paul Anderson, PhD, LIDR associate director, assist investigators with identifying and fulfilling their projects’ immunological needs. To determine the host response to infection in researchers’ studies, Whyte and Anderson utilize a range of instrumentation including a MoFlo XDP, multiple epifluorescence microscopes, and a new Lionheart FX ― an automated microscopy system with imaging capabilities for fluorescence, brightfield, color brightfield, and phase contrast, as well as an environmental control chamber that allows long-term live-cell imaging applications. The expertise and equipment provided by LIDR Immunology help investigators analyze the cellular response to infectious agents. These techniques yield novel insights into the molecular mechanisms of infection that lead to pathology and potential ways to prevent or treat infections.

Such data and facilities allow investigators to vie successfully for highly competitive federal funding. The LIDR BSL-3 facility supports university grant applications, like the current Centers of Excellence for Translation Research program submission, for proposed research on Tier 1 and Non-Tier 1 select agents, arboviruses, and other diseases of public concern. Within the College of Veterinary Medicine (CVM), Alexander Franz, PhD, Veterinary Pathobiology assistant professor, received a National Institutes of Allergy and Infectious Diseases (NIAID) award to conduct studies at the LIDR on how arboviruses exit the mosquito midgut to infect hosts.  Likewise, Guoquan Zhang, DVM, PhD, Veterinary Pathobiology associate professor, recently received NIAID funding to conduct research into prophylactics and therapeutic strategies against Coxiella burnetti (Q fever) at the LIDR. Other CVM investigators, including Jerod Skyberg, PhD, and Deborah Anderson, PhD, have recent NIAID grants that specifically utilize LIDR’s immunology services.

“The LIDR Immunology core has some resources that are unique to BSL3” said Skyberg, Veterinary Pathobiology assistant professor, “[they] have been pivotal for our research.” Skyberg utilizes LIDR’s immunology services to sort live cells from infected animals under BSL3 conditions to identify cell types that contain live Brucella during chronic infection. In the course of this research, Skyberg’s team identified a non-canonical cellular reservoir of chronic Brucella infection. He plans to continue use of LIDR Immunology to investigate the reservoir’s role in the development of arthritis in his recently funded study.

Deborah Anderson, Veterinary Pathobiology associate professor, works with LIDR Immunology to understand the interactions of Yersinia pestis and host immune cells. She specifically seeks to identify how host immune cells contribute to the development of pneumonic plague, as well as the bacterial virulence factors required for gaining access to, and growing in, the airway. Using fluorescent markers, flow cytometry, and image analysis offered by LIDR Immunology, Anderson is able to characterize the cellular immune response in lungs after exposure to Y. pestis.

Investigators have access to additional services at the LIDR to support their research. In Deborah Anderson’s newly funded project, she plans to use LIDR Aerobiology to test whether the stimulation of heme breakdown in the lungs stimulates a more effective immune response that can prevent pneumonia.

For additional information about LIDR Immunology, click here.

Story Contact:

Krista Rodgers, rodgerskl@missouri.edu, 573-882-8860

Shift Pharmaceuticals developed by team of MU researchers, Office of Technology Management and Industry Relations

Approximately one out of every 40 individuals in the United States is a carrier of the gene responsible for spinal muscular atrophy, a neurodegenerative disease that causes nerves to die and muscles to weaken over time. In 2014, Chris Lorson, an investigator in the Bond Life Sciences Center and associate dean of research in the University of Missouri College of Veterinary Medicine, and his team developed a molecule that was found to be highly effective in animal models exhibiting SMA. Now, testing of that compound has led to the founding of a new company, Shift Pharmaceuticals, and the possibility of the development of drugs that will improve outcomes for people with SMA.

“More than a decade of research has guided our fine-tuning of a potential therapeutic for SMA and what it does,” Lorson said. “We identified a genetic ‘switch’ that turns bits and pieces of the genetic code on and off. Using this information, we then developed a molecule called an antisense oligonucleotide, or ASO, that essentially is a synthetic string of nucleic acid that binds to a specific sequence in the SMN2 gene.”

In individuals affected by SMA, the survival motor neuron-1 (SMN1) gene is mutated and lacks the ability to produce a key protein that helps neurons function. Muscles that control walking or even lifting an arm often are profoundly affected as well as muscles important for breathing. Fortunately, humans have a nearly identical copy of the gene called SMN2; however, SMN2 normally only makes a small amount of the correct SMN protein. Lorson’s compound targets SMN2 and effectively “turns the volume” up for the replacement gene, allowing it to make more of the correct SMN protein.

“Our current treatment helps the body induce a backup mechanism to combat the disease and extends survival in mice with SMA giving them more mobility,” Lorson said. “This compound helps produce the right form of SMN, the one that was only produced at very low levels before. The mice are able to move around and live fairly normal lives.”

Early results of this research are promising. If additional studies are successful, researchers at Shift Pharmaceuticals hope to begin human clinical trials with the goal of developing new treatments for SMA. To move their compound closer to the clinic, Shift has recently received a Technology and Therapeutic Development Grant from the Department of Defense, through the Congressionally Directed Medical Research Program.

Therapeutics produced by Shift highlight the University of Missouri’s impact on the state’s economic development efforts, workforce development and job growth, quality of life improvements for residents, and commercialization of technologies through the MU Office of Technology Management and Industry Relations. During the past five years, companies commercializing MU technologies have secured hundreds of millions of dollars in investments and grants to advance their commercialization efforts.

Cure SMA, a 501c non-profit, provided initial funding for the discovery of this compound, as well as additional support from FightSMA, the Gwendolyn Strong Foundation, and the Muscular Dystrophy Association.

This research also highlights the power of translational precision medicine and the promise of the proposed Translational Precision Medicine Complex at the University of Missouri. The TPMC will bring together industry partners, multiple schools and colleges on campus, and the federal and state government to enable precision and personalized medicine. Scientific advancements made at MU will be effectively translated into new drugs, devices and treatments that deliver customized patient care based on an individual’s genes, environment and lifestyle, ultimately improving health and well-being of people.

Editor’s Note: For more on the story, please see: “A shift in focus: Lorson moves basic research to drug development”

Story Contact(s):
Jeff Sossamon, sossamonj@missouri.edu, 573-882-3346