Shawn Wiederhoeft is a pretty regular guy in his 30s. The Madison native works as a video game developer and maintains an active lifestyle. He’s healthy — in the best shape of his life — and regularly spends time with friends and family in southern Wisconsin.

But it wasn’t always a given that Wiederhoeft would be able to participate in life so fully. In fact, “Uncle Shawn,” as he’s known to family, is healthy today thanks in large part to a new kidney he received in 2020 from his sister, Meagan Hahn, of Wauwatosa.

The transplanted kidney has given Wiederhoeft a new lease on life, and because he and his sister chose to participate in a cutting-edge clinical trial at the University of Wisconsin School of Medicine and Public Health, he’s also able to live without the need for anti-rejection medications.

With no daily medications and only occasional medical checkups, Wiederhoeft says he sometimes almost forgets he’s the recipient of a kidney transplant.

“I have to consciously remind myself that there’s a third kidney in here,” he says. “I feel healthier than I’ve ever been.”

Wiederhoeft’s remarkable journey from serious illness to a medication-free recovery is just one of a growing number of living kidney transplant success stories that an international team of physicians and researchers reported in the July issue of the American Journal of Transplantation.

Led by UW–Madison surgery professor Dr. Dixon Kaufman, who directs the UW Health Transplant Center, the team shared results of a Phase 3 clinical trial that evaluated the effectiveness and safety of a living kidney transplant tolerance protocol that also includes the transplantation of certain stem cells from the kidney donor to the transplant recipient.

These stem cells are injected into the recipient several days after transplantation and take up residence in bone marrow, where they divide and multiply into immune cells that share the genes of the donated organ. The goal is to keep the recipient’s immune system from attacking the new organ, which it would otherwise recognize as a threat.

“This procedure doesn’t replace the immune system, but it complements it with around 5 to 10% of the immune system being from the donor,” says Kaufman.

In doing so, the new method removes the need for costly anti-rejection medications that severely suppress the recipient’s immune system, carrying a whole host of lifelong side effects.

The prospect of participating in a clinical trial that carried such a possibility was surreal for some of the study’s initial participants, including Wiederhoeft and Hahn.

“It felt straight out of a science fiction novel,” says Wiederhoeft.

The feeling of entering uncharted territory was even more pronounced for the trial’s first participants, sisters Barb Okey and Brenda Quale, both of Platteville.

“To say I was nervous — that’s an understatement,” says Okey, who received one of Quale’s kidneys followed by an infusion of her stem cells in 2018. Within months of the procedure, as hoped, tests showed that Quale’s cells had begun multiplying and circulating in Okey’s blood, and Okey was eventually weaned off her anti-rejection medications. Today, the only medication Okey takes is to help control her blood pressure.

“It’s amazing,” Okey says. “To have an opportunity to continue life with a new kidney and without taking medications is pretty fabulous.”

Okey and Wiederhoeft are among the small group of kidney recipients who can forgo anti-rejection medications thanks to UW–Madison’s clinical trial, but there are plans to expand eligibility for the procedure.

The initial phase required donors and recipients to be siblings and a “perfect,” or identical, match, meaning they have compatible blood types and other physiological compatibilities. Kaufman says that the initial trial’s success means that eligibility could soon be extended to living donor transplants of non-identical matches and eventually to recipients of organs from deceased donors.

Kaufman described the trial’s positive results as “immensely gratifying” and a testament to the value of long-term investments in research.

Indeed, before the procedure could be attempted in human patients, researchers at the Wisconsin National Primate Research Center and elsewhere spent years testing its safety and effectiveness in non-human primates. In 2023, the team led by Kaufman reported promising results from its latest primate studies that could pave the way for expanding the procedure to non-identical transplant pairs.

The studies that enabled this clinical trial received substantial support over the years from the National Institutes of Health (NIH). Kaufman credits UW–Madison’s proven track record of executing complex biomedical research for the sustained federal investment.

“The clinical and research environments at UW are outstanding” he says. “I’ve been to other programs, trained at other programs, and been faculty at other programs, and the unique things that make the ability to do complicated clinical trials like this successful at the University of Wisconsin are the strong collaborative culture and fantastic resources. We’ve got a history and a tradition of advancing the field in innovative ways for the benefit of many. We want to keep reaching higher and keep making those advances.”

While the recognition and support from NIH and other funding agencies have underpinned the trial’s success, Kaufman says it’s the patients and donors who are willing to participate in new and potentially risky trials who are ultimately responsible for helping to advance the field.

“Getting [the patients] back on the road to living a happy and fulfilling life is really what this is all about,” he says.

For Hahn, who felt compelled and grateful to participate in the trial as Wiederhoeft’s donor, the results have been immeasurably positive.

“To be able to have Uncle Shawn there for all the family events, and for it to be the most normal version of him — the most healthy version of him — I have to remind myself that this all happened and that this isn’t normal for so many people. But for us, it’s quite a blessing.” ■

Dixon Kaufman, MD, PhD, is the Ray D. Owen endowed professor in the UW–Madison Department of Surgery.

The research that supported this clinical trial occurred over 12 years, first in non-human primates before human trials, with more than $20 million in funding provided by the National Institutes of Health over that period (U01AI102456, MSN150727, T32AI25231, U54). NIH also supports the Wisconsin National Primate Research Center (P51OD011106). The Phase 3 clinical trial described here was sponsored by Medeor Therapeutics (MDR-101) in association with Stanford University.

If the drug proves viable to treat epilepsy in humans, it would be the first to provide lasting relief from seizures even after patients stopped taking it. iStock/cagkansayin

A drug typically prescribed for arthritis halts brain-damaging seizures in mice that have a condition like epilepsy, according to researchers at the University of Wisconsin–Madison.

The drug, called tofacitinib, also restores short-term and working memory lost to epilepsy in the mice and reduces inflammation in the brain caused by the disease. If the drug proves viable for human patients, it would be the first to provide lasting relief from seizures even after they stopped taking it.

“It ticks all the boxes of everything we’ve been looking for,” says Avtar Roopra, a neuroscience professor in the UW–Madison School of Medicine and Public Health and senior author of the study, which was published last week in the journal Science Translational Medicine.

Epilepsy is one of the most common neurological diseases, afflicting more than 50 million people around the world. While there are many known causes, the disease often appears after an injury to the brain, like a physical impact or a stroke.

Some days, months or even years after the injury, the brain loses the ability to calm its own activity. Normally balanced electrical activity through the brain goes haywire.

“The system revs up until all the neurons are firing all the time, synchronously,” says Roopra. “That’s a seizure that can cause massive cell death.”

And the seizures repeat, often at random intervals, forever. Some drugs have been useful in addressing seizure symptoms, protecting patients from some of the rampant inflammation and memory loss, but one-third of epilepsy patients do not respond to any known drugs, according to Olivia Hoffman, lead author of the study and a postdoctoral researcher in Roopra’s lab. The only way to stop the most damaging seizures has been to remove a piece of the brain where disruptive activity starts.

On their way to identifying tofacitinib’s potential in epilepsy, Hoffman and co-authors used relatively new data science methods to sift through the way thousands of genes were expressed in millions of cells in the brains of mice with and without epilepsy. They found a protein called STAT3, key to a cell signaling pathway called JAK, at the center of activity in the seizure-affected mouse brains.

“When we did a similar analysis of data from brain tissue removed from humans with epilepsy, we found that was also driven by STAT3,” Hoffman says.

“[The discovery] ticks all the boxes of everything we’ve been looking for,” says Avtar Roopra (right), seated with research collaborator Olivia Hoffman.

Meanwhile, Hoffman had unearthed a study of tens of thousands of arthritis patients in Taiwan aimed at describing other diseases associated with arthritis. It turns out, epilepsy was much more common among those arthritis patients than people without arthritis — but surprisingly less common than normal for the arthritis patients who had been taking anti-inflammatory drugs for more than five-and-a-half years.

“If you’ve had rheumatoid arthritis for that long, your doctor has probably put you on what’s called a JAK-inhibitor, a drug that’s targeting this signaling pathway we’re thinking is really important in epilepsy,” Hoffman says.

The UW researchers ran a trial with their mice, dosing them with the JAK-inhibitor tofacitinib following the administration of a brain-damaging drug that puts them on the road to repeated seizures. Nothing happened. The mice still developed epilepsy like human patients.

Remember, though, that epilepsy doesn’t often present right after a brain-damaging event. It can take years. In the lab mice, there’s usually a lull of weeks of relatively normal time between the brain damage and what the researchers call “reignition” of seizures. If it’s not really epilepsy until reignition, what if they tried the drug then? They devised a 10-day course of tofacitinib to start when the mouse brains fell out of their lull and back into the chaos of seizures.

“Honestly, I didn’t think it was going to work,” Hoffman says. “But we believe that initial event sort of primes this pathway in the brain for trouble. And when we stepped in at that reignition point, the animals responded.”

The drug worked better than they could have imagined. After treatment, the mice stayed seizure-free for two months, according to the paper. Collaborators at Tufts University and Emory University tried the drug with their own mouse models of slightly different versions of epilepsy and got the same, seizure-free results.

Roopra’s lab has since followed mice that were seizure-free for four and five months. And their working memory returned.

“These animals are having many seizures a day. They cannot navigate mazes. Behaviorally, they are bereft. They can’t behave like normal mice, just like humans who have chronic epilepsy have deficits in learning and memory and problems with everyday tasks,” Roopra says. “We gave them that drug, and the seizures disappear. But their cognition also comes back online, which is astounding. The drug appears to be working on multiple brain systems simultaneously to bring everything under control, as compared to other drugs, which only try to force one component back into control.”

Because tofacitinib is already FDA-approved as safe for human use for arthritis, the path from animal studies to human trials may be shorter than it would be for a brand-new drug. Roopra’s epilepsy research has long been funded by the National Institutes of Health as well as key early support from Madison-based Lily’s Fund for Epilepsy Research and CURE Epilepsy.

The next steps toward human patients largely await NIH review of new studies, which have been paused indefinitely amid changes at the agency.

For now, the researchers are focused on trying to identify which types of brain cells are shifted back to healthy behavior by tofacitinib and on animal studies of even more of the many types of epilepsy. Hoffman and Roopra have also filed for a patent on the use of the drug in epilepsy.

This research was funded in part by grants from the National Institutes of Health (R01NS108756, R21NS093364, R01NS112308, NS112350, R01NS105628, R01NS102937 and R21NS120868).

This image shows a mouse brain cells infected with Venezuelan equine encephalitis virus, which is one of two viruses that cause encephalitis that are neutralized by a compound discovered by UW–Madison researchers. Image courtesy of Dr. Yi Xue

A new antiviral compound designed and synthesized by researchers at the University of Wisconsin–Madison’s School of Pharmacy is highly effective in mice against two types of devastating encephalitis viruses that are harmful to humans.

UW–Madison researchers developed the compound, a quinazolinone known as BDGR-49, in collaboration with cellular virologists at the University of Louisville and researchers at the University of Tennessee Health Science Center who performed animal efficacy studies. The multidisciplinary team found that BDGR-49 protects mice infected with deadly eastern equine encephalitis virus (EEEV) or Venezuelan equine encephalitis virus (VEEV).

The researchers described BDGR-49 and its efficacy against lethal infections of EEEV or VEEV in mouse models in a study published April 12 in Science Translational Medicine.

Jennifer Golden

“Collaboration across disciplines and capabilities was key to this discovery,” says Jennifer E. Golden, a UW–Madison professor of pharmacy and chemistry and synthetic medicinal chemist who led the discovery and optimization effort. Colleen Jonsson, professor at UTHSC, tested the compound in mice. Donghoon Chung, a professor of microbiology and immunology at Louisville’s Center for Predictive Medicine, performed further virology studies.

The team found that BDGR-49 potently inhibited EEEV and VEEV and was well tolerated in mice. The compound provided significant protection to EEEV-infected animals. Meanwhile, it not only fully protected VEEV-infected mice, but could also be used as a therapeutic treatment days after infection.

“We had been working on a different compound structure for years,” says Golden. “Compounds that emerged from that earlier work were pivotal to understanding how to construct a better antiviral compound class that works well against VEEV and EEEV — ultimately providing a roadmap to the design and discovery of BDGR-49.”

An important feature of this antiviral compound is its ability to access the brain where these viruses cause damage, while other critical attributes include its improved stability, potency and efficacy compared to earlier prototypes Golden and her collaborators developed. Based on resistance studies, BDGR-49 efficiently prevents these viruses from copying themselves, implicating that it operates by disrupting the viral machinery needed for replication.

Classified as New World alphaviruses, equine encephalitis viruses are transmitted by the bite of a mosquito and can infect the brain, causing neurological effects, serious illness and death in humans as well as horses. There currently are no FDA-approved vaccines or treatments available specifically for preventing or treating alphavirus infection in humans.

Symptoms of EEEV infection include fever, headache, chills and vomiting. Severe infection can result in seizure, coma and death. About one-third of individuals who develop encephalitis (brain inflammation) from EEEV infection die, and many of those who do recover suffer permanent neurological effects.

Although outbreaks of eastern equine encephalitis are rare, with an average of 11 cases per year in the United States, in 2019 an outbreak across nine states resulted in 38 confirmed cases, 19 deaths and neurological effects in survivors.

Venezuelan equine encephalitis has a much lower mortality rate of 1%, but outbreaks can affect thousands of people, most often occurring in Central and South America. While insect bites are the typical source of these infections, there is also concern the viruses could be used as bioweapons.

The team has been developing and optimizing chemical structures against VEEV and EEEV for more than a decade. Golden, Jonsson and Chung are co-investigators in the Center of Excellence for Encephalitic Alphavirus Therapeutics, based at UTHSC. The center was created to refine the properties and activity of early-stage small molecule compounds discovered in the Golden lab and to develop them into clinical candidates for VEEV and EEEV that could be studied in humans.

“There are no approved prophylactic or therapeutic options in our arsenal for any human alphavirus infection, so our goal is to develop a drug against VEEV and EEEV that is safe and effective in humans,” Golden says. “While there is still much work to be done, the discovery of BDGR-49 is a remarkable achievement in reaching that goal based on the compound’s drug-like characteristics and ability to prevent animals from dying from these infections.”

The team is evaluating BDGR-49 in advanced preclinical studies while expanding the understanding of its antiviral properties. As RNA viruses such as EEEV and VEEV are prone to develop mutations, they can potentially evolve into more lethal or transmissible versions without warning, resulting in widespread infections.

“It is essential that we develop these countermeasures for viruses of pandemic potential so we don’t find ourselves unprepared to respond to an outbreak,” Golden says. “We can do better, and we intend to leverage this discovery as broadly as possible with respect to VEEV, EEEV and other viruses of concern.”

This research was supported by the National Institute of Allergy and Infectious Diseases (U19AI142762 and R01AI118814) and by a grant (S10OD016226) from the Office of the Director of the NIH.

This schematic shows a two-step drug delivery approach to treating cancer tumors developed by UW–Madison researchers. On the left, engineered proteins (shown as red) initiate clotting at the site of a tumor before nanoparticles coated with a blood platelet membrane and loaded with an anti-cancer drug are injected and attracted to the clots within the tumor. Image by Quanyin Hu

University of Wisconsin–Madison researchers have developed a new method for targeting tumors with cancer drugs by exploiting the clotting propensity of blood platelets.

The new approach, first described March 29 in the journal Science Advances, adds to a growing set of innovative drug delivery techniques under development in the lab of Quanyin Hu, a professor in the UW–Madison School of Pharmacy.

One of Hu’s research goals is to improve the effectiveness and safety of cancer immunotherapies. Such treatments have shown promise by bolstering the ability of the immune system to combat cancer cells, but they come with their own set of challenges. One significant obstacle is that the drugs can target normal cells in addition to tumors. This can make the treatments less effective and sometimes lead to serious side effects.

Quanyin Hu

To better target the therapeutics at tumors, Hu and his colleagues considered a mechanism that results in a very specific biological chain reaction — the cellular signals that trigger blood to clot.

“This whole study is purely inspired by nature and the natural features of platelets to participate in clot formation,” says Hu.

First, the researchers used an engineered protein designed to locate and bind to tumor blood vessels and then initiate thrombosis, or clotting, within tumors. They found that in mice that received the engineered proteins via intravenous injections, the proteins led to clot formation almost exclusively within tumors, with only very limited thrombosis occurring elsewhere.

The clot formation is not the therapy itself, but rather creates conditions for efficient, targeted drug delivery to the clot-filled tumor, which Hu describes as a “cellular hive.”

“Once we have this clot formation at the tumor site, we have this so-called cellular hive that can attract these therapeutic drones,” Hu says.

In this case, the drones are blood platelets engineered with immunotherapeutics on their surfaces which the researchers inject once clotting has begun.

In their studies using mice, Hu and his colleagues found that the engineered platelets efficiently delivered a common immunotherapy drug known as an immune checkpoint inhibitor to the tumor sites. The checkpoint inhibitors encourage immune cells known as T cells to eradicate cancer cells.

Mouse models with colorectal tumors that received the treatment saw their tumors shrink and lived longer than mice that received a traditional immunotherapy treatment. Notably, one-third of the mice that received the treatment became completely tumor free.

The researchers then tested the delivery method with blood platelet derivatives — nanoparticles coated with a platelet membrane and loaded with a chemotherapy drug — on mouse models with human breast cancer tumors. Similarly, they found that the chemotherapy drugs were efficiently delivered to tumor sites while avoiding normal cells. The findings open the possibility to more effective and versatile cancer treatments with multiple types of drugs and fewer unintended side effects.

Before the delivery system can go to clinical trials, more testing is required to ensure its safety, especially with respect to clot formation. While their studies in mice have thus far shown that the system creates clots only in tumor sites, a more comprehensive safety evaluation is the next step, Hu says. That process will likely take a couple years.

One of the challenges of treating this disease is that surgeons can’t always remove every bit of tumor or glioma stem cells that might linger in the brain.

Quanyin Hu

“One characteristic of glioblastoma is that the tumor cells are very aggressive, and they will infiltrate the surrounding tissues. So the surgeon can’t clearly feel the boundaries between the tumor and the normal tissue, and you cannot remove as much as possible because all the tissues in the brain are extremely important — you certainly don’t want to remove too much,” explains Quanyin Hu, an assistant professor in the University of Wisconsin–Madison School of Pharmacy’s Pharmaceutical Sciences Division. “So the tumor will come back again, and that sharply decreases the survival rate after treatment.”

But Hu’s Cell-Inspired Personalized Therapeutic (CIPT) Lab has developed a powerful immunity-boosting postoperative treatment that could transform the odds for patients with glioblastoma. Hu and his collaborators published their research on the treatment’s use in mouse models of human glioblastoma this month in the journal Science Translational Medicine.

“It provides hope for preventing glioblastoma relapse,” Hu says. “We prove that it can actually eradicate these glioma stem cells, which can eventually prevent the glioblastoma from coming back. We can significantly improve survival.”

Hu’s lab developed a hydrogel that can be injected into the brain cavity left behind by the excised tumor. The hydrogel delivery method works well because it completely fills the brain cavity, slowly releases the medicine into the surrounding tissue, and promotes the cancer-killing immune response, Hu notes.

“We have a lot of work to do before it can be potentially translated into the clinic, but we feel confident that this is a very promising approach for bringing new hope to patients with glioblastoma so they can recover after surgery.”

Quanyin Hu

The hydrogel is packed with nanoparticles designed to enter and reprogram certain types of immune cells called macrophages. These immune cells normally clean up infectious invaders in the body, but in the tumor environment, they can change into a form that instead suppresses the immune system and promotes cancer growth. And because of the inflammation created by surgery, these rogue macrophages flock to the surgical site, potentially fueling cancer relapse.

“We want to take advantage of these macrophages and turn them from enemy to ally,” Hu says.

To do that, the nanoparticles can engineer the macrophages to target a glycoprotein called CD133, a marker for cancer stem cells. Hu’s team also added an antibody, CD47, that blocks a “don’t-eat-me” signal to promote macrophages to recognize the cancer cells. The preclinical results in mice models show that the hydrogel treatment successfully generated glioma stem cell-specific chimeric antigen receptor (CAR) macrophages — essentially engineering the immune cells on site to target and kill any lingering glioma stem cells.

If effective in humans, the hydrogel treatment could eliminate the need for postsurgical chemotherapy or radiation, reducing toxic side effects while also improving patient outcomes.

Hu’s next step is testing the hydrogel in larger animal models and also monitoring long-term efficacy and toxicity beyond the four- to six-month period he previously studied.

“We have a lot of work to do before it can be potentially translated into the clinic, but we feel confident that this is a very promising approach for bringing new hope to patients with glioblastoma so they can recover after surgery,” Hu says. “We hope we can do our work to be able to advance this technology to the clinic.”

While Hu’s team is initially focused on glioblastoma, the treatment approach could also be applied to other aggressive solid tumors, including breast cancer, he notes. “Our approach is taking advantage of the macrophages in the postsurgical areas and to locally engineer these macrophages,” he says. “In this scenario, we can confidently say that it will apply to the majority of solid tumors with high invasive characteristics.”

Earlier this year, Hu published results on a different cancer-fighting gel, a collaboration with School of Pharmacy Professor Seungpyo Hong and colleagues in the UW School of Medicine and Public Health.

Shaoqin “Sarah” Gong

In new research published in the journal Nature Nanotechnology today, the lab of Shaoqin “Sarah” Gong, a professor with the Wisconsin Institute for Discovery at the University of Wisconsin–Madison, reported a new nanoparticle-based treatment that delivers anti-inflammatory molecules and antibiotics.

The new system saved the lives of mice with an induced version of sepsis meant to serve as a model for human infections, and is a promising proof-of-concept for a potential new therapy, pending additional research.

The new nanoparticles delivered the chemical NAD⁺ or its reduced form NAD(H), a molecule that has an essential role in the biological processes that generate energy, preserve genetic material and help cells adapt to and overcome stress. While NAD(H) is well known for its anti-inflammatory function, clinical application has been hindered because NAD(H) cannot be taken up by cells directly.

“To enable clinical translation, we need to find a way to efficiently deliver NAD(H) to the targeted organs or cells. To achieve this goal, we designed a couple of nanoparticles that can directly transport and release NAD(H) into the cell, while preventing premature drug release and degradation in the bloodstream,” says Gong, who also holds appointments in the Department of Biomedical Engineering and the UW School of Medicine and Public Health’s Department of Ophthalmology and Visual Sciences.

The interdisciplinary work was led by Gong along with Mingzhou Ye and Yi Zhao, two postdoctoral fellows in the Gong lab. John-Demian Sauer, a professor in the Department of Medical Microbiology and Immunology, also collaborated on the project.

Sepsis can be deadly in two phases. First, an infection begins in the body. The immune system responds by creating drastic inflammation that impairs blood flow and forms blood clots, which can cause tissue death and trigger a chain reaction leading to organ failure. Afterward, the body overcorrects itself by suppressing the immune system, which in turn increases infection susceptibility. Controlling complications caused by inflammation is vital in sepsis therapy.

The lipid-coated calcium phosphate or metal-organic framework nanoparticles designed by the Gong lab can be used to co-deliver NAD(H) and antibiotics. Gong’s lab tested the NAD(H)-loaded nanoparticles in multiple mouse models including endotoxemia, multidrug-resistant pathogen-induced polymicrobial bacteremia, as well as a puncture-induced sepsis model with secondary infection by a common illness-causing bacteria called P. aeruginosa.

The nanoparticle treatment performed much better than using NAD(H) alone. For instance, in an endotoxemia mouse model, mice without any treatment or treated with free NAD(H) died within two days. In contrast, mice treated with NAD(H)-loaded nanoparticles all survived. These animal studies demonstrated that the NAD(H) nanoparticles can help maintain a healthy immune system, support blood vessel function and prevent multiorgan injury.

This technology may pave the road for the development of a new clinical therapy for sepsis that could also be applied in other inflammation-related scenarios, such as COVID-19 treatment. An additional benefit of this therapy is the ability to treat infection with lower amounts of antibiotics, which reduces their overuse. Further research in larger animal models will be necessary before clinical trials in people could begin.

“The NAD(H) nanoparticles have the potential to treat many other diseases because NAD(H) is involved with so many biological pathways. There is strong evidence for the use of NAD(H) as an intervention or aid in critical illnesses,” says Gong.

Most MS patients develop a relapsing-remitting disease, in which bouts of numbness, weakness and vision problems come and go. Over time, many of those MS patients develop progressive MS, which just keeps getting worse as it eats away myelin — the insulating coating on axons, the connections between nerve cells — and subsequently, the axons are damaged.

“There are many treatments for people with relapsing-remitting disease which are quite successful in a lot of patients,” says Ian Duncan, a neuroscientist at the University of Wisconsin–Madison’s School of Veterinary Medicine. “And then there are the unlucky people who start out with primary progressive disease, or whose relapsing-remitting MS becomes what’s called secondary progressive MS. There are no effective treatments for them.”

In the normal nerve (a) all axons in the optic nerve are surrounded by thick, dark sheaths of myelin in contrast to the completely demyelinated nerve (b) during active disease (when VEP latency lengthens). After recovery (in c) axons have been remyelinated — and though the sheaths are thin compared to normal, the VEP latency has shortened. Image courtesy of Ian Duncan

At least, there are not yet effective treatments. Progressive types of MS are drawing the attention of pharmaceutical companies working to find drugs that help restore lost myelin, re-growing the insulation that supports proper nerve signaling and protects axons.

Unfortunately, confirming the presence of myelin in the central nervous system has required cutting into nerve tissue to take samples. That sort of destructive method confounds attempts to assess the effectiveness of new treatments in human patients.

Doctors often monitor their MS patients with a noninvasive test called the visual evoked potential, or VEP. Flashing a series of lights into the eye prompts a recognizable electrical signal that travels down the optic nerve from the retina. The signal is recorded in brain activity measured with electrodes on the scalp.

“There’s a time lag between the flashes of light and the brain activity. That’s called the latency,” says Duncan. “When that latency increases, it’s taking longer for the signal from the lights to get from the retina down the optic nerve to the brain. As MS progresses and demyelination of axons in the optic nerve worsens, the latency grows because the axons are not conducting the signal as well as healthy nerves.”

However, what if the latency was decreasing?

“If we could prove that a decrease in latency in the VEP truly reflected remyelination of nerve axons, then you’d have it,” Duncan says. “You’ve got a way to tell if there’s improvement in a patient, an outcome measure that can show whether the drug you are testing is successfully promoting myelin repair.”

Today (Dec. 16, 2019), Duncan and UW–Madison collaborators in ophthalmology, radiology, statistics, surgery and microscopy published in the Proceedings of the National Academy of Sciences a study that shows changes in VEP latency tracking demyelination and myelination in a feline model.

Cats fed irradiated food for several months develop severe myelin loss throughout their nervous systems — especially along their optic nerves. When the cats return to regular diets, nerve function is restored because of extensive myelin repair.

The researchers fed cats irradiated food, measuring VEP latency before the diet had demyelinated the cats’ nerves, during the cat’s neurologic symptoms, and after their recovery.

“The normal latency of the VEP in the cats is between 50 to 60 milliseconds. At the height of disease, it goes up to 90 to 110 milliseconds,” Duncan says. “And then, at recovery, it comes back down to around 60 to 70 milliseconds.”

Tissue samples confirmed the return of myelin along the nerve axons as the latency times decreased.

“Latency doesn’t fully recover because the myelin sheath in remyelination remains thinner than the original myelin,” says Duncan, whose work is supported by the National Multiple Sclerosis Society. “But we know from previous studies that thin myelin is enough to restore function and sufficient to protect nerve fibers in the long run.”

And now it’s clear that visual evoked potential is a true measure of remyelination.

“If you’ve got a drug to promote myelin repair in MS patients, we now have a proven outcome measure,” says Duncan. “VEP can accurately quantify the drug’s remyelinating effect in the optic nerve — likely reflecting remyelination throughout the central nervous system — and can help begin to sort the wheat from the chaff in potential remyelinating therapies for people with progressive MS.”

Materials science and engineering professor Xudong Wang fits a new wound dressing around the wrist of graduate student Yin Long. The device stimulates healing using electricity generated from the body’s natural motions. Photo by Sam Million-Weaver

A new, low-cost wound dressing developed by University of Wisconsin–Madison engineers could dramatically speed up healing in a surprising way.

The method leverages energy generated from a patient’s own body motions to apply gentle electrical pulses at the site of an injury.

In rodent tests, the dressings reduced healing times to a mere three days compared to nearly two weeks for the normal healing process.

Materials science and engineering professor Xudong Wang, left, holds a prototype wound-healing device that generates energy from body movements. Graduate student Yin Long, seated, and visiting radiology professor Hao Wei collaborated on development of the new device. Photo by Sam Million-Weaver

“We were surprised to see such a fast recovery rate,” says Xudong Wang, a professor of materials science and engineering at UW–Madison. “We suspected that the devices would produce some effect, but the magnitude was much more than we expected.”

Wang and collaborators described their wound dressing method today (Nov. 29, 2018) in the journal ACS Nano.

Researchers have known for several decades that electricity can be beneficial for skin healing, but most electrotherapy units in use today require bulky electrical equipment and complicated wiring to deliver powerful jolts of electricity.

“Acute and chronic wounds represent a substantial burden in healthcare worldwide,” says collaborator Angela Gibson, professor of surgery at UW-Madison and a burn surgeon and director of wound healing services at UW Health. “The use of electrical stimulation in wound healing is uncommon.”

In contrast with existing methods, the new dressing is much more straightforward.

“Our device is as convenient as a bandage you put on your skin,” says Wang.

A new device powered by energy harvested from the body’s natural motions accelerates wound healing by delivering gentle electric pulses to an injury site. Photo by Sam Million-Weaver

The new dressings consist of small electrodes for the injury site that are linked to a band holding energy-harvesting units called nanogenerators, which are looped around a wearer’s torso. The natural expansion and contraction of the wearer’s ribcage during breathing powers the nanogenerators, which deliver low-intensity electric pulses.

“The nature of these electrical pulses is similar to the way the body generates an internal electric field,” says Wang.

And, those low-power pulses won’t harm healthy tissue like traditional, high-power electrotherapy devices might.

In fact, the researchers showed that exposing cells to high-energy electrical pulses caused them to produce almost five times more reactive oxygen species — major risk factors for cancer and cellular aging — than did cells that were exposed to the nanogenerators.

Also a boon to healing: They determined that the low-power pulses boosted viability for a type of skin cell called fibroblasts, and exposure to the nanogenerator’s pulses encouraged fibroblasts to line up (a crucial step in wound healing) and produce more biochemical substances that promote tissue growth.

“These findings are very exciting,” says collaborator Weibo Cai, a professor of radiology at UW-Madison. “The detailed mechanisms will still need to be elucidated in future work.”

In that vein, the researchers aim to tease out precisely how the gentle pulses aid in healing. The scientists also plan to test the devices on pig skin, which closely mimics human tissue.

And, they are working to give the nanogenerators additional capabilities—tweaking their structure to allow for energy harvesting from small imperceptible twitches in the skin or the thrumming pulse of a heartbeat.

“The impressive results in this study represent an exciting new spin on electrical stimulation for many different wound types, given the simplicity of the design,” says Gibson, who will collaborate with the team to confirm the reproducibility of these results in human skin models.

If the team is successful, the devices could help solve a major challenge for modern medicine.

“We think our nanogenerator could be the most effective electrical stimulation approach for many therapeutic purposes,” says Wang.

And because the nanogenerators consist of relatively common materials, price won’t be an issue.

“I don’t think the cost will be much more than a regular bandage,” says Wang. “The device in itself is very simple and convenient to fabricate.”

This research was supported by grants from the National Institutes of Health (R01EB021336 and P30CA014520).

###

— Sam Million-Weaver, (608) 263-5988, millionweave@wisc.edu

Mason Konsitzke, 7, plays in his bedroom at home in Stoughton. Photo: Jeff Miller

Mason Konsitzke is 7. He loves food (especially when he can share it with others) and anything military (both of his grandfathers served). He likes to fly kites and play with his 5-year-old sister, Alexandra. But Mason was born with a disease called neurofibromatosis type 1, or NF1, and each day can present new challenges for him and his family.

NF1 is a genetic disease caused by changes, or mutations, to a single gene in the human DNA library. Roughly one out of 3,000 babies born in the United States have the disease. That’s more than three times as many as have cystic fibrosis. Yet few people have ever heard of NF1.

Mutations in the NF1 gene cause defects in the neurofibromin 1 protein, which acts as a tumor suppressor. Children with NF1 can develop painful tumors along their nerve tracts, including their skin and in their eyes. Sometimes this renders them blind. They are often diagnosed with autism spectrum disorder, though not all children with NF1 are also autistic, and they are sometimes diagnosed with attention deficit hyperactivity disorder. They may have soft bones that bend and break. They are at a higher risk for cancer. And there is no cure.

It was not a disease Mason’s parents, Charles and Malia Konsitzke, had ever heard of. As a newborn, he was healthy. But when Mason was 6 months old, the couple began to suspect something was wrong. Mason developed coffee-and-cream-colored spots all over his body. His father later learned these were a hallmark of NF1. Mason received a genetic diagnosis of the disorder just before his first birthday.

“We were like deer in the headlights,” Malia says. “We were in shock, wondering, what does this mean for us? What does it mean for Mason?”

At 18 months, Mason began to lose his ability to speak. He was falling over, screaming constantly and deliberately banging his head. That’s when an MRI revealed a tumor called a plexiform neurofibroma in a mesh of nerves in the left side of his face. It was growing fast.

A father turns to science

Charles “Chuck” Konsitzke is the associate director of UW-Madison’s Biotechnology Center, a sort of one-stop shop for scientists in need of DNA sequencing, genome editing and other services.

Upon Mason’s diagnosis he began to delve into published NF1 research. He wanted to know where it was happening, who was doing it and how he might be able to help. He sought opinions from experts, wondering how the field could be improved. Many identified the same bottleneck: the lack of a good research model.

In biology, research models are animals, cells, plants, microbes and other living things that allow scientists to study biological processes and recreate diseases in order to better understand them. Good models yield information relevant to humans, but the right model can sometimes be difficult to find.

Seen through a microscope, a researcher guides a micro-needle (at right) to inject DNA into a pig embryo at UW–Madison’s Biotechnology Center. Photo: Jeff Miller

NF1 is especially complex, affects many systems of the body and touches many areas of scientific inquiry, from cancer research to neurobiology. Chuck began to search for a better model and in 2013, when Mason was 3, he settled on pigs. Pigs are similar to humans in many ways that other common research animals, such as mice and flies, are not. That includes their size, which means drugs and devices that work on humans can also be tested on pigs. They have a robust immune system, which rodents lack. And they’re intelligent, so scientists can study changes to their cognition.

Chuck then went on the hunt for researchers who studied swine.

Braving the risks

Dhanansayan “Dhanu” Shanmuganayagam, a nutrition and animal sciences professor in the UW–Madison College of Agricultural and Life Sciences, has spent most of his career using swine to study human diseases, particularly heart disease. In fact, he and colleagues in the animal sciences department created the Wisconsin Miniature Swine, a pig that, like people, can develop heart disease under the right conditions.

Dhanansayan “Dhanu” Shanmuganayagam speaks about his NF1 research at a symposium for patients and families at UW-Madison in May. Photo: Jeff Miller

Dhanu’s office was a few blocks from Chuck’s but they’d never met until a few years ago, when they bumped into one another while helping campaign for a new building on campus. They got to know one another and Chuck asked Dhanu whether he had ever heard NF1. He hadn’t. Chuck told him about Mason, about the need for a better model, about the promise that pigs offered to help understand and treat the disease. Would Dhanu join forces to help create that model, Chuck asked.

Dhanu took some time to think about it. He consulted the members of his laboratory. All would be helping to forge this new path. His risks would be their risks. A pig model could fail, leading them all down a blind alley. That kind of outcome can derail a scientist’s entire career.

Dhanu told Chuck he was in.

The risks remain significant, he says, “but I’ve come to terms with it and it’s fine. I’ve been lucky in my career to work on things that have gone to clinic. If it works it’s going to be impactful.”

Meanwhile, Chuck consulted a legal team to ensure he was clear of conflicts of interest, and took steps to ensure his involvement was ethical and not problematic for his staff at the Biotechnology Center.

There aren’t many places in the world where this kind of work – melding basic science with clinical research and a large animal model like swine – is possible. UW–Madison has large biomedical research centers, the capacity for high-powered basic science, and a 1,500-pig research facility called the Swine Research and Teaching Center (SRTC), based in Arlington, a 35-minute drive from campus.

“It’s a brave new frontier, to move into swine,” says David H. Gutmann, a physician and researcher at the Washington University School of Medicine in St. Louis, considered one of the foremost NF1 experts in the world. “I’m glad they’re doing this work at UW–Madison because the combination of specialized resources and expertise are found in very few places worldwide.”

Like scissors for genes

Dhanu and Chuck determined the course they wanted to chart included gene editing, using a powerful new tool known as CRISPR, which stands for Clustered Regularly Interspaced Short Palindromic Repeats.

The genetic technology is reshaping basic biological research. Like a pair of molecular scissors, CRISPR enables scientists to target a stretch of cellular DNA for alteration. They can cut out pieces of DNA or swap out letters in the genome, changing the message it encodes or shutting off genes entirely.

The two set their sights on creating pigs that carry the NF1 mutations they and other researchers are most interested in studying. “But we had to figure out where to start,” says Dhanu. “It’s like learning to fly a space shuttle.”

With Dhanu’s lab manager and lead scientist Jen Meudt at the helm, the team dove in. But the challenges were many.

They had to learn about swine reproduction, about CRISPR and gene editing, how to perform the necessary surgeries on pigs, how to time events so no step of the process failed and ruined all the efforts before it. Again and again, they hit roadblocks.

It took more than a year, but finally, they came up with a plan: The researchers would use artificial insemination to impregnate a female pig carefully primed to produce more eggs than she naturally would. Shortly after fertilization, they would remove the embryos, whisk them to the Biotechnology Center and inject them with a solution containing the gene-editing CRISPR. This would have to be done quickly, while the embryos were still a single cell. When the single cell divided, all the subsequent cells would contain the NF1 mutation. Inject too late and the pig would develop into a mosaic of cells that contain the mutation and those that do not.

Researchers at UW–Madison created a breed of pig called Wisconsin Miniature Swine to better model and understand human diseases. Photo by: Jeff Miller

Researcher Kathy Krentz uses a microscope and micro-needle to inject DNA into an embryo at the UW–Madison Biotechnology Center. The embryos are then rushed to Arlington where the host mother awaits. Photo by: Jeff Miller

A closeup of the micro-needle. Photo by: Jeff Miller

Dhanu Shamuganayagam and Jen Meudt prepare to transplant the genetically edited pig embryos into the host mother. Photo by: Jeff Miller

Implanting the embryos. Photo by: Jeff Miller

Larry Britzman and his 11-year-old daughter, Mackenzie, talk to advocate Lindsay Geier during a campus event for patients and their families in May. Photo by: Jeff Miller

Siblings Alexandra, 5, and Mason Konsitzke, 7, work on colored drawings while playing at the Konsitzke family home in Stoughton, Wis., on May 26, 2017. Photo by: Jeff Miller

Then it would be off to the surrogate mother, a pig chosen to reproductively match the embryo-donating pig. The researchers would perform surgery to implant the CRISPR embryos into her womb. If all went well, months later she would give birth to piglets, at least some of which would carry the desired NF1 mutations.

A few months passed. On November 7, 2016, Chuck and Dhanu were meeting in Madison with a group from the Neurofibromatosis (NF) Network, which supports research and clinical care for NF1. They were sipping coffee when a text came in from Jen: “The mom carrying NF piglets is delivering right now.”

The piglets – eight in all, and four with the NF1 mutation – were a living embodiment of the team’s hard work. They had proved that they could create pigs genetically-engineered to carry the disease. It was an emotional experience for the scientists, involving tears and prayers. They immediately went out to celebrate.

Why are pigs used in research? Learn more

Then they set to work building on that success. One of the four piglets with the mutation was a male. Mason named him Tank. His job is to sire more piglets with the mutation since the changes conferred by CRISPR were designed to be passed on from generation to generation.

The team took the process they’d developed and applied it to other NF1 mutations, including some related to cancer. And they set an even more ambitious goal: precision medicine. A pig personalized for every child with NF1.

With CRISPR, the researchers believe they can take the genetic fingerprint of an individual child’s NF1 mutation and create a pig with that same mutation. They can then test potential medications and treatments and see if they’ll work. Can tumors, like the one that afflicts Mason, be shrunk?

The promise of precision

By the time Mason reached pre-kindergarten, the tumor in his face had grown into his cranial sinus. His parents were told he could lose his sight and his ability to taste.

Surgery wasn’t an option. It was too risky and could leave Mason in even greater pain, permanently. “He’s literally been in pain his whole life,” Malia says.

Then, for reasons doctors couldn’t explain, the tumor stopped progressing. He regained his speech and no longer screamed or struggled to stay upright. His doctors keep a close watch on the tumor with MRI scans. And they continue to work to determine the best medication regimen for the other symptoms that come with his particular variant of NF1. His treatment must be continuously modified. Because of NF1’s unique manifestations, each child is an experiment unto himself.

Pigs develop faster than children do so they offer the possibility of helping predict how NF1 might affect a particular child, enabling parents, doctors, teachers and others to prepare. Earlier intervention for a child who develops autism could lead to better outcomes. Doctors could start working to find drugs to treat tumors before they grow too large.

“Precision medicine is more than matching the right drug to the right gene. With NF1 it’s more complicated and involves searching for the factors that make each individual with NF1 unique,” says Washington University’s David Gutmann. “This is an amazing opportunity to find the risk factors that put an affected child at risk for developing a brain tumor, a bone defect, or another serious complication of NF1.”

Washington University researcher David Gutmann, center, speaks with UW–Madison’s Dhanu Shanmuganayagam, left, and Neha Patel. Photo: Jeff Miller

Dhanu, Chuck and Jen are not doing this work on their own. The team now includes many talented individuals like Biotechnology Center scientists C. Dustin Rubinstein, Kathy Krentz and Michael Sussman, along with Jamie Reichert and his team at the Swine Research and Teaching Center. And there’s now a broader research group, the UW NF1 Translational Research team, that includes Thomas Crenshaw, an animal sciences professor and department chair, and Marc Wolman, a professor of integrative biology.

They have also enlisted the skill and knowledge of Neha Patel, a pediatrician at the University of Wisconsin School of Medicine and Public Health who treats about 150 children with NF1 in Wisconsin and surrounding regions.

Dhanu hopes to make the NF1 pigs accessible to other researchers around the country, charging only what it costs to produce them. And the team plans to use the pigs to help identify metabolic and cellular pathways common to the variety of NF1 mutations, to help target and develop better drugs.

But to accomplish all of this requires funding.

“We’re at a critical moment,” Dhanu says. “We have to turn our successes into funding opportunities.”

Parents and health advocates Lindsay and Ryan Geier participate in a panel discussion during the May event for patients and families. Their daughter Lauren has NF1. Photo: Jeff Miller

The UW NF1 Translational Research team has bootstrapped most of its work so far, relying primarily on funding and donations from the NF Network. Most of that comes from an annual charity golf tournament the Konsitzkes and four other families help organize and run. Called Links for Lauren, the tournament honors Lauren Geier, an 8-year-old girl in Madison with NF1.

Families can play a surprisingly influential role in the fight against rare diseases.“They often provide critical resources and financial support at the earliest stages of a high-risk project, when funding from federal agencies is not possible,” says David Guttman. “Our families, they inspire us because they ask us to do things that are really meaningful and take risks by taking the roads not frequently traveled. Through their involvement they can move the field forward in ways that no one else can.”

Where there’s research, there’s hope

Larry Britzman had no idea there were pigs at UW–Madison that might one day help children like his 12-year-old daughter Mackenzie. He learned that, and much more, in May when he traveled to campus from La Valle, Wisconsin, for a symposium for patients and families.

“I didn’t realize each child is specific,” he says. “I didn’t realize UW has swine research and there aren’t too many facilities in the country researching NF1.”

The NF1 team hopes to host the symposium each year, to invite families to learn more about the science of NF1, to give them a chance to meet researchers and clinicians, and to ask questions and meet other families living with the disease.

“We’ve gone very far in two years because it hasn’t been just about building a model, it’s also been about creating a community around it,” says Dhanu.

The opportunity to work so closely with and on behalf of the people who may ultimately benefit from his work is not something he’d ever experienced. And it’s been profoundly rewarding.

Not long ago, he invited a family whose college-aged daughter has NF1 into his lab. They’d been donors to NF1 causes for years but had never talked to a researcher. “It meant a lot to them and my first thought was: ‘How can we do more of this?’”

He and his lab members now participate in running events like the Madison Half Marathon, often with The NF Team organization, to raise money for NF1 research and to increase awareness. The runners sport neon yellow performance shirts with bold, black lettering. They also participate in the annual charity golf tournament.

“As scientists, we don’t often see the payoff of what we’re working on,” Dhanu says. “It redefines our research priorities and it also aids discovery. The best people to note observations are the people who live with it.”

To him, success can be measured by individuals. “Even if our research just raises awareness and someone gets treated because of what we do, that alone is big,” he says.

Chuck believes the disease is underdiagnosed because very few people are genetically tested for it and most physicians are not familiar with it. So they may diagnose patients with autism or a behavioral disorder and miss the broader picture.

That has frustrated Danielle Wood, a teacher and mother of two who lives in Reedsburg, Wisconsin. Her daughter, Bernadette, is 2 and was diagnosed with NF1 as an infant. Along with springy blonde curls and an arresting smile, Bernadette has a weak abdominal wall, which causes her pain and may require surgery. She wears braces to support her frail ankles.

Danielle, too, has NF1. Her mother had it and so did her grandmother. Though her condition is mild – she simply wears glasses for poor vision caused by a tumor on her optic nerve – deciding whether to have children was hard. Because it is a dominant mutation, Danielle and her husband had at least a 50 percent chance of giving birth to a baby with the disease. Having grown up with NF1, Danielle felt she had a good idea of what to expect. She now sees herself as an advocate for Bernadette.

“While things never move as fast as we want them to, there’s a tremendous amount of exciting progress in this field and where there’s research, there’s hope,” says David Gutmann. UW–Madison is “in a really great position because (it has) young faculty who are excited and a patient community that is challenging them to improve the lives of people with NF1 through research.”

This is what drives Chuck, Dhanu and the rest of the UW NF1 Translational Research team, which is working to establish a NF1 Center for Excellence at UW–Madison. Not only is this possible, David Gutmann says, it is necessary. “There is no established therapy for NF1 and no magic bullet that works for all kids or adults. The challenge for us is to learn more about this disorder so that personalized and effective treatments emerge.”

Moreover, he says, what NF1 teaches researchers will inform their approaches to other conditions. And he’s excited to see what the future holds.

“All of us in the NF field get up every morning and are excited to get to work. What we learn from our colleagues and our families each day brings us one step closer to that better future for children and adults with NF,” he says. “I can imagine getting up every morning and running to work to see what’s happening with those pigs.”

For Mason, the pigs don’t play much of a role in his daily life today. Rather, he relies on regular visits to therapists and other professionals both in and out of school to help him manage his symptoms. He also benefits from the support of his family, from Chuck and Malia to aunts and uncles who have learned all they can about NF1. And the family dog, Donatella, is his packmate, Malia says.

At 7, Mason can still take all that for granted. He can focus on what he loves best, like sharing the tastiest mini pizzas he can make. You should try the pepperoni.

Mason at home with his parents and sister Alexandra, 5. Photo: Jeff Miller

Mason Konsitzke, 7, plays in his bedroom at home in Stoughton. Photo: Jeff Miller

Mason Konsitzke is 7. He loves food (especially when he can share it with others) and anything military (both of his grandfathers served). He likes to fly kites and play with his 5-year-old sister, Alexandra. But Mason was born with a disease called neurofibromatosis type 1, or NF1, and each day can present new challenges for him and his family.

NF1 is a genetic disease caused by changes, or mutations, to a single gene in the human DNA library. Roughly one out of 3,000 babies born in the United States have the disease. That’s more than three times as many as have cystic fibrosis. Yet few people have ever heard of NF1.

Mutations in the NF1 gene cause defects in the neurofibromin 1 protein, which acts as a tumor suppressor. Children with NF1 can develop painful tumors along their nerve tracts, including their skin and in their eyes. Sometimes this renders them blind. They are often diagnosed with autism spectrum disorder, though not all children with NF1 are also autistic, and they are sometimes diagnosed with attention deficit hyperactivity disorder. They may have soft bones that bend and break. They are at a higher risk for cancer. And there is no cure.

It was not a disease Mason’s parents, Charles and Malia Konsitzke, had ever heard of. As a newborn, he was healthy. But when Mason was 6 months old, the couple began to suspect something was wrong. Mason developed coffee-and-cream-colored spots all over his body. His father later learned these were a hallmark of NF1. Mason received a genetic diagnosis of the disorder just before his first birthday.

“We were like deer in the headlights,” Malia says. “We were in shock, wondering, what does this mean for us? What does it mean for Mason?”

At 18 months, Mason began to lose his ability to speak. He was falling over, screaming constantly and deliberately banging his head. That’s when an MRI revealed a tumor called a plexiform neurofibroma in a mesh of nerves in the left side of his face. It was growing fast.

A father turns to science

Charles “Chuck” Konsitzke is the associate director of UW-Madison’s Biotechnology Center, a sort of one-stop shop for scientists in need of DNA sequencing, genome editing and other services.

Upon Mason’s diagnosis he began to delve into published NF1 research. He wanted to know where it was happening, who was doing it and how he might be able to help. He sought opinions from experts, wondering how the field could be improved. Many identified the same bottleneck: the lack of a good research model.

In biology, research models are animals, cells, plants, microbes and other living things that allow scientists to study biological processes and recreate diseases in order to better understand them. Good models yield information relevant to humans, but the right model can sometimes be difficult to find.

Seen through a microscope, a researcher guides a micro-needle (at right) to inject DNA into a pig embryo at UW–Madison’s Biotechnology Center. Photo: Jeff Miller

NF1 is especially complex, affects many systems of the body and touches many areas of scientific inquiry, from cancer research to neurobiology. Chuck began to search for a better model and in 2013, when Mason was 3, he settled on pigs. Pigs are similar to humans in many ways that other common research animals, such as mice and flies, are not. That includes their size, which means drugs and devices that work on humans can also be tested on pigs. They have a robust immune system, which rodents lack. And they’re intelligent, so scientists can study changes to their cognition.

Chuck then went on the hunt for researchers who studied swine.

Braving the risks

Dhanansayan “Dhanu” Shanmuganayagam, a nutrition and animal sciences professor in the UW–Madison College of Agricultural and Life Sciences, has spent most of his career using swine to study human diseases, particularly heart disease. In fact, he and colleagues in the animal sciences department created the Wisconsin Miniature Swine, a pig that, like people, can develop heart disease under the right conditions.

Dhanansayan “Dhanu” Shanmuganayagam speaks about his NF1 research at a symposium for patients and families at UW-Madison in May. Photo: Jeff Miller

Dhanu’s office was a few blocks from Chuck’s but they’d never met until a few years ago, when they bumped into one another while helping campaign for a new building on campus. They got to know one another and Chuck asked Dhanu whether he had ever heard NF1. He hadn’t. Chuck told him about Mason, about the need for a better model, about the promise that pigs offered to help understand and treat the disease. Would Dhanu join forces to help create that model, Chuck asked.

Dhanu took some time to think about it. He consulted the members of his laboratory. All would be helping to forge this new path. His risks would be their risks. A pig model could fail, leading them all down a blind alley. That kind of outcome can derail a scientist’s entire career.

Dhanu told Chuck he was in.

The risks remain significant, he says, “but I’ve come to terms with it and it’s fine. I’ve been lucky in my career to work on things that have gone to clinic. If it works it’s going to be impactful.”

Meanwhile, Chuck consulted a legal team to ensure he was clear of conflicts of interest, and took steps to ensure his involvement was ethical and not problematic for his staff at the Biotechnology Center.

There aren’t many places in the world where this kind of work – melding basic science with clinical research and a large animal model like swine – is possible. UW–Madison has large biomedical research centers, the capacity for high-powered basic science, and a 1,500-pig research facility called the Swine Research and Teaching Center (SRTC), based in Arlington, a 35-minute drive from campus.

“It’s a brave new frontier, to move into swine,” says David H. Gutmann, a physician and researcher at the Washington University School of Medicine in St. Louis, considered one of the foremost NF1 experts in the world. “I’m glad they’re doing this work at UW–Madison because the combination of specialized resources and expertise are found in very few places worldwide.”

Like scissors for genes

Dhanu and Chuck determined the course they wanted to chart included gene editing, using a powerful new tool known as CRISPR, which stands for Clustered Regularly Interspaced Short Palindromic Repeats.

The genetic technology is reshaping basic biological research. Like a pair of molecular scissors, CRISPR enables scientists to target a stretch of cellular DNA for alteration. They can cut out pieces of DNA or swap out letters in the genome, changing the message it encodes or shutting off genes entirely.

The two set their sights on creating pigs that carry the NF1 mutations they and other researchers are most interested in studying. “But we had to figure out where to start,” says Dhanu. “It’s like learning to fly a space shuttle.”

With Dhanu’s lab manager and lead scientist Jen Meudt at the helm, the team dove in. But the challenges were many.

They had to learn about swine reproduction, about CRISPR and gene editing, how to perform the necessary surgeries on pigs, how to time events so no step of the process failed and ruined all the efforts before it. Again and again, they hit roadblocks.

It took more than a year, but finally, they came up with a plan: The researchers would use artificial insemination to impregnate a female pig carefully primed to produce more eggs than she naturally would. Shortly after fertilization, they would remove the embryos, whisk them to the Biotechnology Center and inject them with a solution containing the gene-editing CRISPR. This would have to be done quickly, while the embryos were still a single cell. When the single cell divided, all the subsequent cells would contain the NF1 mutation. Inject too late and the pig would develop into a mosaic of cells that contain the mutation and those that do not.

Researchers at UW–Madison created a breed of pig called Wisconsin Miniature Swine to better model and understand human diseases. Photo by: Jeff Miller

Researcher Kathy Krentz uses a microscope and micro-needle to inject DNA into an embryo at the UW–Madison Biotechnology Center. The embryos are then rushed to Arlington where the host mother awaits. Photo by: Jeff Miller

A closeup of the micro-needle. Photo by: Jeff Miller

Dhanu Shamuganayagam and Jen Meudt prepare to transplant the genetically edited pig embryos into the host mother. Photo by: Jeff Miller

Implanting the embryos. Photo by: Jeff Miller

Larry Britzman and his 11-year-old daughter, Mackenzie, talk to advocate Lindsay Geier during a campus event for patients and their families in May. Photo by: Jeff Miller

Siblings Alexandra, 5, and Mason Konsitzke, 7, work on colored drawings while playing at the Konsitzke family home in Stoughton, Wis., on May 26, 2017. Photo by: Jeff Miller

Then it would be off to the surrogate mother, a pig chosen to reproductively match the embryo-donating pig. The researchers would perform surgery to implant the CRISPR embryos into her womb. If all went well, months later she would give birth to piglets, at least some of which would carry the desired NF1 mutations.

A few months passed. On November 7, 2016, Chuck and Dhanu were meeting in Madison with a group from the Neurofibromatosis (NF) Network, which supports research and clinical care for NF1. They were sipping coffee when a text came in from Jen: “The mom carrying NF piglets is delivering right now.”

The piglets – eight in all, and four with the NF1 mutation – were a living embodiment of the team’s hard work. They had proved that they could create pigs genetically-engineered to carry the disease. It was an emotional experience for the scientists, involving tears and prayers. They immediately went out to celebrate.

Why are pigs used in research? Learn more

Then they set to work building on that success. One of the four piglets with the mutation was a male. Mason named him Tank. His job is to sire more piglets with the mutation since the changes conferred by CRISPR were designed to be passed on from generation to generation.

The team took the process they’d developed and applied it to other NF1 mutations, including some related to cancer. And they set an even more ambitious goal: precision medicine. A pig personalized for every child with NF1.

With CRISPR, the researchers believe they can take the genetic fingerprint of an individual child’s NF1 mutation and create a pig with that same mutation. They can then test potential medications and treatments and see if they’ll work. Can tumors, like the one that afflicts Mason, be shrunk?

The promise of precision

By the time Mason reached pre-kindergarten, the tumor in his face had grown into his cranial sinus. His parents were told he could lose his sight and his ability to taste.

Surgery wasn’t an option. It was too risky and could leave Mason in even greater pain, permanently. “He’s literally been in pain his whole life,” Malia says.

Then, for reasons doctors couldn’t explain, the tumor stopped progressing. He regained his speech and no longer screamed or struggled to stay upright. His doctors keep a close watch on the tumor with MRI scans. And they continue to work to determine the best medication regimen for the other symptoms that come with his particular variant of NF1. His treatment must be continuously modified. Because of NF1’s unique manifestations, each child is an experiment unto himself.

Pigs develop faster than children do so they offer the possibility of helping predict how NF1 might affect a particular child, enabling parents, doctors, teachers and others to prepare. Earlier intervention for a child who develops autism could lead to better outcomes. Doctors could start working to find drugs to treat tumors before they grow too large.

“Precision medicine is more than matching the right drug to the right gene. With NF1 it’s more complicated and involves searching for the factors that make each individual with NF1 unique,” says Washington University’s David Gutmann. “This is an amazing opportunity to find the risk factors that put an affected child at risk for developing a brain tumor, a bone defect, or another serious complication of NF1.”

Washington University researcher David Gutmann, center, speaks with UW–Madison’s Dhanu Shanmuganayagam, left, and Neha Patel. Photo: Jeff Miller

Dhanu, Chuck and Jen are not doing this work on their own. The team now includes many talented individuals like Biotechnology Center scientists C. Dustin Rubinstein, Kathy Krentz and Michael Sussman, along with Jamie Reichert and his team at the Swine Research and Teaching Center. And there’s now a broader research group, the UW NF1 Translational Research team, that includes Thomas Crenshaw, an animal sciences professor and department chair, and Marc Wolman, a professor of integrative biology.

They have also enlisted the skill and knowledge of Neha Patel, a pediatrician at the University of Wisconsin School of Medicine and Public Health who treats about 150 children with NF1 in Wisconsin and surrounding regions.

Dhanu hopes to make the NF1 pigs accessible to other researchers around the country, charging only what it costs to produce them. And the team plans to use the pigs to help identify metabolic and cellular pathways common to the variety of NF1 mutations, to help target and develop better drugs.

But to accomplish all of this requires funding.

“We’re at a critical moment,” Dhanu says. “We have to turn our successes into funding opportunities.”

Parents and health advocates Lindsay and Ryan Geier participate in a panel discussion during the May event for patients and families. Their daughter Lauren has NF1. Photo: Jeff Miller

The UW NF1 Translational Research team has bootstrapped most of its work so far, relying primarily on funding and donations from the NF Network. Most of that comes from an annual charity golf tournament the Konsitzkes and four other families help organize and run. Called Links for Lauren, the tournament honors Lauren Geier, an 8-year-old girl in Madison with NF1.

Families can play a surprisingly influential role in the fight against rare diseases.“They often provide critical resources and financial support at the earliest stages of a high-risk project, when funding from federal agencies is not possible,” says David Guttman. “Our families, they inspire us because they ask us to do things that are really meaningful and take risks by taking the roads not frequently traveled. Through their involvement they can move the field forward in ways that no one else can.”

Where there’s research, there’s hope

Larry Britzman had no idea there were pigs at UW–Madison that might one day help children like his 12-year-old daughter Mackenzie. He learned that, and much more, in May when he traveled to campus from La Valle, Wisconsin, for a symposium for patients and families.

“I didn’t realize each child is specific,” he says. “I didn’t realize UW has swine research and there aren’t too many facilities in the country researching NF1.”

The NF1 team hopes to host the symposium each year, to invite families to learn more about the science of NF1, to give them a chance to meet researchers and clinicians, and to ask questions and meet other families living with the disease.

“We’ve gone very far in two years because it hasn’t been just about building a model, it’s also been about creating a community around it,” says Dhanu.

The opportunity to work so closely with and on behalf of the people who may ultimately benefit from his work is not something he’d ever experienced. And it’s been profoundly rewarding.

Not long ago, he invited a family whose college-aged daughter has NF1 into his lab. They’d been donors to NF1 causes for years but had never talked to a researcher. “It meant a lot to them and my first thought was: ‘How can we do more of this?’”

He and his lab members now participate in running events like the Madison Half Marathon, often with The NF Team organization, to raise money for NF1 research and to increase awareness. The runners sport neon yellow performance shirts with bold, black lettering. They also participate in the annual charity golf tournament.

“As scientists, we don’t often see the payoff of what we’re working on,” Dhanu says. “It redefines our research priorities and it also aids discovery. The best people to note observations are the people who live with it.”

To him, success can be measured by individuals. “Even if our research just raises awareness and someone gets treated because of what we do, that alone is big,” he says.

Chuck believes the disease is underdiagnosed because very few people are genetically tested for it and most physicians are not familiar with it. So they may diagnose patients with autism or a behavioral disorder and miss the broader picture.

That has frustrated Danielle Wood, a teacher and mother of two who lives in Reedsburg, Wisconsin. Her daughter, Bernadette, is 2 and was diagnosed with NF1 as an infant. Along with springy blonde curls and an arresting smile, Bernadette has a weak abdominal wall, which causes her pain and may require surgery. She wears braces to support her frail ankles.

Danielle, too, has NF1. Her mother had it and so did her grandmother. Though her condition is mild – she simply wears glasses for poor vision caused by a tumor on her optic nerve – deciding whether to have children was hard. Because it is a dominant mutation, Danielle and her husband had at least a 50 percent chance of giving birth to a baby with the disease. Having grown up with NF1, Danielle felt she had a good idea of what to expect. She now sees herself as an advocate for Bernadette.

“While things never move as fast as we want them to, there’s a tremendous amount of exciting progress in this field and where there’s research, there’s hope,” says David Gutmann. UW–Madison is “in a really great position because (it has) young faculty who are excited and a patient community that is challenging them to improve the lives of people with NF1 through research.”

This is what drives Chuck, Dhanu and the rest of the UW NF1 Translational Research team, which is working to establish a NF1 Center for Excellence at UW–Madison. Not only is this possible, David Gutmann says, it is necessary. “There is no established therapy for NF1 and no magic bullet that works for all kids or adults. The challenge for us is to learn more about this disorder so that personalized and effective treatments emerge.”

Moreover, he says, what NF1 teaches researchers will inform their approaches to other conditions. And he’s excited to see what the future holds.

“All of us in the NF field get up every morning and are excited to get to work. What we learn from our colleagues and our families each day brings us one step closer to that better future for children and adults with NF,” he says. “I can imagine getting up every morning and running to work to see what’s happening with those pigs.”

For Mason, the pigs don’t play much of a role in his daily life today. Rather, he relies on regular visits to therapists and other professionals both in and out of school to help him manage his symptoms. He also benefits from the support of his family, from Chuck and Malia to aunts and uncles who have learned all they can about NF1. And the family dog, Donatella, is his packmate, Malia says.

At 7, Mason can still take all that for granted. He can focus on what he loves best, like sharing the tastiest mini pizzas he can make. You should try the pepperoni.

Mason at home with his parents and sister Alexandra, 5. Photo: Jeff Miller