Joint press release by University of Missouri and Hokkaido University
What causes rats without a Y chromosome to become male?
A look at the brains of an endangered spiny rat off the coast of Japan by University of Missouri (MU) Bond Life Sciences Center scientist Cheryl Rosenfeld could illuminate the subtle genetic influences that stimulate a mammal’s cells to develop as male versus female in the absence of a Y chromosome.
The root of the answer is in the chromosomes of this particular mammal. Males of the Amami spiny rat (Tokudaia osimensis) are not like most therian mammals — a name used to group animals that give live birth including placental mammals and marsupials. Unlike in most mammals, these males have no Y chromosome, which has been shed over eons of evolution. And they only have one X chromosome.
“I’d been interested in these rats for many years now, and it’s unclear how sexual differentiation of the gonads and brain occur in this species since both males and females have a single X chromosome,” said Cheryl Rosenfeld, lead author on the study and an MU researcher.
Since most mammals inherit chromosomes from both parents — an X from our mother and either an X or Y chromosome from our father — the only way to develop as male is to inherit a Y chromosome. That male chromosome contains a sex-determining region Y (SRY) gene that stimulates male sexual differentiation in many mammalian species, including humans. SRY triggers the fetus to become male by producing a protein that binds DNA, which leads to development of testes and subsequent production of testosterone. This steroid hormone then stimulates development of the rest of the male reproductive tract. This surge in testosterone from the testes also programs masculinization of the brain.
Naturally, this got Rosenfeld curious about how the absence of a Y chromosome and SRY might affect gene expression differences in male and female Amami spiny rats. In collaboration with Asato Kuroiwa from Hokkaido University Takamichi Jogahara of the Frontier Science Research Center in Japan, and Scott Givan, associate director of the MU Informatics Research Core Facility, her laboratory received brain samples from both males and females of this species.
They took those brains, isolated the RNA from them and sequenced the samples of the male and females to compare transcripts between the two sexes. Transcripts are the step between a gene and a protein, essentially a piece of RNA that encodes the information to make a particular protein. Subtle variations in the resulting RNA can change the final protein, giving rise to alternative forms that can exhibit increased or decreased potency. Investigators found major differences between males and females.
“Several different transcripts or isoforms encode the same gene, and while there might be, for example, 10 transcripts for a particular gene, some transcripts are more potent than others and have more effect in individual cells,” Rosenfeld said. “When we compared males to females, there were several hundred more transcripts upregulated in males than females. Our thought is that since both have the same sex chromosomes, the resulting differences could be originating from the fact that the males might have more of one of the more potent transcripts.”
Expression differences in such transcripts might also arise due to epigenetic changes, which are alterations that affect the turning on/off of certain DNA regions but without affecting the DNA itself. Potentially, in females, these more effective transcripts are not expressed because of such epigenetic changes.
When the team – including MU student Madison Ortega who is in the MU Maximizing Access to Research Careers/Initiative for Maximizing Student Diversity (MARC/IMSD) program — looked closer, they realized many of the transcripts expressed in males encode for various zinc finger protein genes. In males, these genes can be turned on by SRY and are thought to significantly influence sex development.
“What we think might be happening is the males might be turning on all these other zinc finger transcripts that may compensate for the absence of SRY, so they influence undifferentiated gonad to become a testes and help program the brain to be male. Without these zinc finger protein transcripts, female sexual differentiation of the gonad and brain might result,” Rosenfeld said. “It’s possibly it’s more of a potential gradient within these rodents, where you have to turn on all these other zinc finger protein transcripts in males to stimulate male sexual differentiation and compensate for the absence of SRY.”
With the potential extinction of Amami spiny rats on the horizon, furthering this research has a degree of urgency.
“By elucidating more of the mechanisms in this endangered species, I think it might help us save the species or facilitate them being bred in captivity,” Rosenfeld said.
She hopes this research will continue with her team fully sequencing the genome of the Amami spiny rats to look more closely at what’s going on and how that might lead to a better understanding of the nuances of how animals without a Y chromosome undergoes male sexual differentiation.
“If you move outside the mammals, there are all sorts of sex chromosomes and exceptions like the duckbill platypus who has five sets of X and Y, and they can’t find differences between males and females,” Rosenfeld said. “People are focused on sex chromosomes and the SRY gene that they might be forgetting other contributing factors. By understanding these anomalous species, it opens up the idea that the mechanisms regulating gonadal and brain sexual differentiation are quite complex and not fully understood.”
Science and invention are both about discovering the possibilities in something.
Those possibilities can create something new that improves the lives of people and advances our understanding of the world.
It’s no surprise that Gary Stacey, a Bond Life Sciences Center primary investigator, is being recognized this year as one of 148 academic fellows by the National Academy of Inventors.
“I am very proud to welcome another class of outstanding NAI Fellows, whose collective achievements have helped shape the future and who each day work to improve our world,” said Paul R. Sanberg, President of the NAI. “Each of these new NAI Fellows embodies the Academy’s mission through their dedication, creativity and inventive spirit. I look forward to working collaboratively with the new NAI Fellows in growing a global culture of innovation.”
Stacey has spent years focused on the basic science behind biological phenomena including the relationship between bacteria and the roots of nitrogen-fixing plants. His work has explored the specifics of how plants can benefit from interacting with particular bacterium like Bradyrhizobium japonicum.
“This bacterium infects the roots of soybean and established a beneficial, nitrogen fixing symbiosis,” said Stacey, a Curators’ Distinguished Professor of Plant Science in MU’s College of Agriculture, Food and Natural Resources. “The two patents that describe these discoveries formed the basis for the Optimize product, which is now sold by Novozymes.”
When seeds of soybeans or other legumes are treated with Optimize, it encourages what’s natural by giving the future plant the opportunity to build a beneficial relationship with the bacterium. The bacterium infects the roots and creates nodules that “fix” atmospheric nitrogen, providing food for the plant that replaces some need for fertilizer. This improves plant growth and yield in an environmentally-friendly way and leaves us in a better position to feed the world in a sustainable fashion.
While his research has a significant potential economic plus side, Stacey points out that understanding the basic mechanisms underpinning how these plants and bacteria interact.
“Our goals are not to develop intellectual property or products, however, we remain cognizant of any possible applications of our research,” he said. “Beyond the exhilaration of making a basic discovery, it is also gratifying when you see the result of your labors being put to practical use as things move from discovery, through translation to ultimate application. I have been lucky in my career to be able to traverse this full spectrum of research.”
A natural curiosity connects the diverse scientists behind discoveries. That trait started young for Stacey.
“You could see signs of this when I was a small boy constantly turning over rocks just to see what was underneath. In a way, with my research, I continue to ‘turn over rocks’ for the sole purpose of just knowing what is hidden below,” Stacey said. “Although I must on many occasions justify my research in the context of impact or application, the truth is I do science for the sole purpose of satisfying my curiosity, and studies have confirmed that curiosity-driven research is the most effective at making major discoveries and exhibits a much higher return on investment.”
With 13 patents to his name, Stacey’s curiosity has allowed his science to thrive at MU.
“A research lab at a major university is really equivalent to a small business — with 20 employees my lab would qualify in the upper 50 percent of all small businesses in Missouri — and I encounter many of the same issues that any small business would have,” Stacey said. “I relish the challenge of being a scientist in which you are tested in so many ways…as an innovator, organizer, manager, communicator and entrepreneur.”
Election to NAI Fellow status is the highest professional distinction accorded solely to academic inventors who have demonstrated a prolific spirit of innovation in creating or facilitating outstanding inventions that have made a tangible impact on quality of life, economic development and the welfare of society.There are now over 1,000 NAI Fellows, representing more than 250 research universities and government and non-profit research institutes. The 2018 Fellows are named inventors on nearly 4,000 issued U.S. patents.
About the National Academy of Inventors The National Academy of Inventors is a member organization comprising U.S. and international universities, and governmental and non-profit research institutes, with over 4,000 individual inventor members and Fellows spanning more than 250 institutions worldwide. It was founded in 2010 at the University of South Florida to recognize and encourage inventors with patents issued from the U.S. Patent and Trademark Office, enhance the visibility of academic technology and innovation, encourage the disclosure of intellectual property, educate and mentor innovative students, and translate the inventions of its members to benefit society. The NAI publishes the multidisciplinary journal, Technology & Innovation. www.academyofinventors.org
The 2018 NAI Fellows will be highlighted with a full page announcement in the Jan. 25, 2019 issue of The Chronicle of Higher Education and in upcoming issues of Technology & Innovation.
The complete list of NAI Fellows is available on the NAI website.
Like with any family, a new addition brings possibility and excitement.
For Bond LSC, three new faculty promise to enrich research at the University of Missouri by working together across disciplines. Ron Mittler, Wes Warren and Bing Yang all joined Bond LSC recently to continue their research careers.
Bond LSC Interim Director Walter Gassmann said these strategic hires are years in the making and represent how departments and centers come together with a unified goal for MU.
“These are some of the first hires we’ve completed specifically looking at how new faculty could bridge individual research strengths that exist on campus,” he said. “These scientists can bring those strengths together to go in a new direction, and Bond LSC is the perfect place for these faculty to achieve their goals.”
Each researcher comes with a reputation that precedes him. It only takes one look at the framed covers of Nature Magazine lined up in a row in Wes Warren’s office to see evidence of that. The comparative geneticist was recruited from Washington University and has published work in Nature that sequenced the genomes of animals from the orangutan and the gibbon to the elephant shark and the platypus.
“My passion is to crack the black box of the genome and try to find weird and unique traits of various animals,” Warren said. “It’s not enough now to sequence a genome and compare it to others, you have to dig deeper, do wet lab work and try to validate your findings.”
His shared appointment as a Bond LSC investigator and professor of genomics in the Department of Animal Sciences, College of Agriculture, Food and Natural Resources, in conjunction with the School of Medicine, College of Veterinary Medicine and the MU Informatics Institute, gives him the opportunity to more easily move his research forward.
“My thought is that I can act as a liaison between researchers in these areas,” he said. “I want to keep in mind trait evolution and use that divergence in traits to practice evolutionary comparative medicine to think about disease in humans and companion animals.”
Yang brings a different expertise from Iowa State University where he spent years studying bacterial diseases of rice. His joint appointment as a professor of plant sciences comes from a partnership with the Donald Danforth Plant Science Center in St. Louis. His research began by looking at plant disease from the bacterial side, but has evolved to also study its host’s interaction.
“Over half the world’s population eats rice as a staple food and by understanding its basic biology we can engineer better rice varieties with disease resistance and yield improvements,” Yang said. “This joint appointment gives me a bigger scientific community and access to more tools to continue my former research while thinking about some high risk, high reward projects that no one has done before.”
Mittler brings a different expertise in plant science to Bond LSC. Most recently at the University of North Texas, his work focused on cell-to-cell communication and how plants respond to multiple stressors — like heat and drought — at the same time. To do this, Mittler studies proteins with unknown functions. One he identified in plants deals with reactive oxygen species, which is a type of oxygen that becomes activated and can cause damage within cells in its toxic forms.
“We don’t really know what 20 percent of the proteins in our body do and in plants it’s more than that,” he said. “One of them was very interesting to me because it responded to reactive oxygen species in Arabidopsis plants, and we found out that a close relative of this protein is found in humans and accumulates to high levels in cancer cells.”
This has led Mittler to expand his work into mammalian cells and will encourage partnerships outside of plant science. With the cancer connection in mind, Mittler has a joint appointment in the Department of Surgery in the School of Medicine
“This is one of the few places in the U.S. that has a medical school, a veterinary school, college of agriculture and a nuclear reactor, so you have a lot of resources available to you, and here I have access to crop fields I didn’t have before and people who do a lot of crop physiology,” Mittler said. “The biggest thing I’ve found here so far is that there is a big drive for collaboration and the walls and barriers between different departments are low. Not a lot of places have this kind of attitude.”
The emphasis on connections across disciplines is key to the larger research goals within the University of Missouri System.
“It’s the research community, the facilities and resources that convince excellent researchers like these to join our research enterprise, and Bond LSC is the perfect place to have faculty from five colleges bump into each other on a daily basis,” said Mark McIntosh, vice chancellor of research, graduate studies and economic development at MU and vice president of research and economic development for the UM System. “Collaborative research is more competitive when it comes to grant applications to federal agencies, and is more likely to lead to innovations and economic development. Our success with interdisciplinary collaborations — like those in the Bond LSC — is the motivation for our focus on building the Translational Precision Medicine Complex (TPMC). I look forward to seeing how our new Bond LSC investigators can build and nurture these partnerships.”
For scientists, studying a disease presents a puzzle looking for an answer, but there are real people behind the research that may one day cure the illnesses that turned their lives upside down. Chris Lorson and Monir Shababi work on one of these puzzles in Bond LSC.
Find out more about their work and the faces behind SMARD, a rare, often fatal, genetic motor neuron disease in the following story courtesy of the College of Veterinary Medicine.
Monir Shababi, an assistant research professor in veterinary pathobiology, and Christian Lorson, Bond LSC principal investigator, College of Veterinary Medicine professor and associate dean for research and graduate studies, have invested countless hours during the past five years to solving a cruel medical mystery. A family who has endured the agonizing ordeal of having two children born with the same disease has invested funding for the research being conducted at MU’s Bond Life Sciences Center.
The disease is called spinal muscle atrophy with respiratory distress, or SMARD. SMARD is a progressive motor neuron disease that has no treatment or cure. At least, not yet.
Shababi, PhD, and Lorson, PhD, and the Sims family — mother Jill, father Eric, grandparents Grant and Patricia — have teamed up in an effort to change that.
The disease is so rare that it is largely unknown, even to most medical professionals. When you are the parent of a child with SMARD, you are in a daily, nonstop, life-and-death struggle.
It is exhausting. It is frustrating. It is a battle that requires an endless reserve of endurance and willpower. And, it requires cutting-edge, scientific discoveries that are just coming to light at MU’s Bond Life Sciences Center.
Catherine Sims lives on a ventilator and needs around-the-clock care. Yet, now age 5, her life is a victory.
“Our first daughter was born healthy, so we had no idea that we carried such a terrible disease,” Jill Sims says. “Then, our second child, Bobby, — who is named after my dad — was born very small, which was unusual given our family history, and he was very quiet as an infant. Those were the only things I noticed. He was three weeks old and we were driving back from Thanksgiving at my parents’ house. I fed him and put him in his car seat. I checked on him 30 minutes later and he had died. He had aspirated. The disease causes the diaphragm not to work, so he couldn’t breathe and eat at the same time.”
Bobby Sims, born Oct. 31, 2012, died on Nov. 30, 2012. His death was attributed to unknown respiratory failure, and he was considered a victim of Sudden Infant Death Syndrome (SIDS). Catherine Sims was born in August 2013; her diagnosis came four months later.
“I went on to have Catherine next, and then we knew something was up,” Sims says. “Catherine was very similar to Bobby, very small and very quiet. That, of course, led us to figure out something was going on.
“In the period of time when Catherine was having problems and was hospitalized but undiagnosed, Catherine had a test done that put her group of symptoms into a specific category of neuromuscular diseases,” Sims says. “A good friend of mine Googled that category and the search produced a WordPress blog that Lisa Porter Werner had contributed to.”
The blog contained personal stories of families who had children with a disease named SMARD. The goal of the blog was to put SMARD on the radar, for families who didn’t have a diagnosis and needed to find answers as well as find support. Porter Werner had posted her own family’s story.
“My friend forwarded me Lisa’s particular story regarding her two children with SMARD, and the story almost identically matched my own,” Sims recalls.
Porter had read extensively and combed the internet for information and cases similar to those of her children. Porter eventually found a modicum of information about something called SMARD, which had been diagnosed in approximately 60 children.
“Lisa Porter’s blog contained the personal stories of families who had children with SMARD,” Sims recalls. “My friend forwarded me Lisa’s particular story regarding her two children with SMARD, and the story almost identically matched my own.
“The Werner’s first daughter died at six weeks of age. It was called a SIDS case; she just died in her sleep,” Sims says. “They had Silas, their son who is living with SMARD, shortly thereafter and she put him in a sleep study when he was three weeks old. She said, ‘No, my daughter didn’t just die. There was a reason.’ It turned out that Silas was having major breathing problems during sleep.
“I was convinced after reading about Lisa’s family that my two children had SMARD as well, and I asked Catherine’s doctors to test her for it,” Sims says. “Catherine’s test came back positive four weeks later. A year or so later, I connected with Lisa through a Facebook group for families with children with SMARD. We began talking more, once my in-laws funded SMARD research at the Jackson Lab, and continued to talk once we found out about Dr. Shababi’s paper that came out in 2016.”
In order to know what SMARD is, it is important to know what it is not. Despite the obvious similarities in name, spinal muscular atrophy (SMA) and spinal muscular atrophy with respiratory distress have sharp differences.
Both conditions affect the lower motor neuron cells of the spinal cord that control voluntary muscle activities like walking, talking, breathing and swallowing. Both are sometimes characterized as “like ALS in babies.”
SMA, which can range from type 1-4, is caused by mutations in or the absence of the SMN1 gene. SMA typically causes weakness in the core first and the baby or child may present as hypotonic, or having low muscle tone — sometimes called floppy baby syndrome. Babies or children with SMA may eventually develop respiratory compromise over time.
SMA is the leading genetic killer of infants; one in 40 people are carriers of SMA.
SMARD, in contrast, is extremely rare. The exact number of cases is unknown, but it has clearly occurred in more than the approximately 100 children worldwide who now carry that tragic diagnosis. SMARD is branded an “orphan” disease, a term commonly applied to any debilitating medical condition that affects fewer than 200,000 Americans. There is little information and few resources available regarding SMARD.
SMARD is a genetic disease, caused by mutations or loss of the IGHMBP2 gene, Immunoglobulin MU-binding protein 2. The condition is inherited in a recessive pattern, meaning both parents must be carriers of the gene mutation and each parent must pass along a copy of the mutation in order for the child to be affected. In essence, every time two carriers have a baby, there is a one in four chance their child will be affected.
Onset of the disease usually occurs suddenly, in what seems to be an otherwise healthy baby, typically between 6 weeks and 6 months of age. Once the diaphragm is paralyzed, the infant must depend on their accessory muscles to breathe. These muscles also weaken as the disease progresses, until the child needs mechanical ventilation.
Many children die in the first year of life, often in their sleep or from a respiratory illness. Past the age of 1 year, almost all children living with SMARD require a tracheostomy, a ventilator and a wheelchair.
Simply put, SMA usually presents as a hypotonic or “floppy” baby who gradually develops respiratory distress. SMARD presents as a baby in respiratory distress who gradually becomes hypotonic.
SMA and SMARD share a similarity in that both are monogenic disorders, conditions caused by mutations or loss of a single gene. Shababi and Lorson have an established history of working with SMA. Now, their focus is SMARD.
“In 2009 and 2010, a lab at the Ohio State University used a viral vector to introduce the SMN gene in SMA mice,” Shababi, the CVM researcher, says. “The viral vector does not contain the necessary genes required for the virus to cause infectious disease. You can replace viral genes with the specific gene you want and keep only the part of the virus that is required to enter the body, find its receptor and produce the desired protein from the gene it carries.
“They (researchers at Ohio State) put a human SMN gene into a viral vector — adeno-associated virus 9 (AAV9) — that has the potential to pass the blood brain barrier in humans. This virus has the capability to enter into the brain, the spinal cord, muscles and peripheral organs,” Shababi continues. “The AAV9 virus carrying the SMN gene was injected into SMA mice. They were able to rescue the affected mice. That was a huge step toward treating SMA. That vector is currently in Phase 2 clinical trials with AveXis/Novartis.
“With SMARD, there is also a single gene involved in the disease — the IGHMBP2 gene,” Shababi continues. “So, we took a human IGHMBP2 gene, in the form of cDNA, and placed it into the same AAV9 vector and injected it into the brain of SMARD pups that were 2 days of age. Our virus did the job and the SMARD mice were cured.”
“Dr. Shababi posted a paper, I believe in March 2016, that reported the results of her work on SMARD,” Sims says. “Lisa found the paper and contacted Dr. Shababi and had a wonderful reception. They had several very long conversations about what Monir was doing, what she had already been doing, and they immediately had a strong connection.
“Dr. Shababi was very personable over the phone, and was very passionate and very approachable about her work,” Sims relates. “Sometimes, it’s hard to get ahold of people, but Monir answers her own phone, and she was very clear with Lisa about what had already been done, which was pretty cool for us because we didn’t know — we didn’t realize how much work Dr. Shababi and Dr. Lorson had already done on SMARD. We were impressed by how much of a handle they already had on the disease. They were ahead of the game. That was great news for us on the family side; at the time, we were aware of only one other lab in the country — the Jackson Lab in Maine — doing work in this area. We couldn’t believe that, wow, there’s a second lab and they are already in gear, they already have a lot of good things going.
“Then, Lisa got me in the loop with Monir, and I talked to her a few times,” Sims continues. “They were having a funding issue, which is not surprising because of how rare the disease is. When we first learned about the work being done at the Jackson Lab, my in-laws agreed to fund SMARD research at Jackson. After learning what Dr. Shababi and Dr. Lorson were doing, I talked to my in-laws again and asked if they would be interested in funding Monir’s research. My father-in-law and I had a few conversations with Monir and Chris Lorson, and then my in-laws decided to do another fund, this time at Missouri, that started this past December.”
“If you look back a number of years, there has been a gene therapy on the translational side that has had exceptionally powerful results in SMA,” says Lorson. “AveXis now has a Phase 2 clinical trial going for their gene therapy product, which has the potential to be very impactful. It has demonstrated efficacy in SMA, but also provides an important proof of principle for gene therapy as a whole. So, it was really exciting to know that there’s only one gene responsible for each of these horribly devastating diseases, SMA and SMARD. It allows you to consider following a similar path. Knowing that, Monir started developing a project that was gene therapy, gene replacement for SMARD.
“Whenever I talk about this, I give about 110 percent of the credit to Monir,” Lorson explains. “Monir has really been the driver of this entire project. Originally, I said, ‘Monir, I’d really like you to develop this gene therapy for SMARD, I think it’s a really exciting area of research. I’ll check back in about six months.’ When I did, we had the mice, we had the vector and she was doing the experiments. That’s exactly the kind of gumption that you hope to find. She did all of that. My role was to say, ‘Good job, Monir!’
“She was the first author on an important paper in Molecular Therapy published in 2016,” Lorson continues. “Based upon that, and the level of excitement, people found her. Through Facebook and Facebook friends, they started to communicate back and forth. Monir is driving it. Monir is doing it.
“AAV9 is in clinic for a number of other diseases, but every time you put a new gene in, you have to go through the Food and Drug Administration,” Lorson says. “That’s why the process isn’t as simple as it might appear to be. Every single time you change that vector — that gene delivery vehicle — you have to get it approved.”
“My in-laws have been very generous, but you need a lot of capital to do this research,” Jill Sims says. “SMARD is so rare that progress will probably come only from academic research. You really need a lot of support and you need a lot of funding from various sources. Right now, our life continues the same. It’s great that everybody is doing this great research, but you need so much more for a cure. That’s what everybody wants; we want our kids to be normal.
“A day in the life of someone with SMARD is very difficult,” Sims says. “There’s a lot that has to be done to have a normal life, and there are a lot of obstacles to that, so you’re constantly trying to overcome those.
“This disease is devastating,” Sims continues. “It can take away every basic human function: the ability to sit, crawl, stand, walk, talk, swallow, feed oneself, clean oneself, use writing utensils and so on. The disease also makes the person more likely to have respiratory problems since they can’t breathe or even cough on their own. It is hard as a parent. Every day we live with the potential fatality of this disease. If their trach tubes come out, they cannot breathe. These trachs sit in their windpipes, held in by ties, like a tight necklace. It is not secure.
“You may go months without anything happening then, all of a sudden, it’s coming out. When that happens, she may only have 60 or so seconds to live,” Sims says. ”You have to have someone always watching them, either a specially trained nurse or a parent, who is a trained caregiver.
“That’s the hard part that we always live with,” says Sims. “Yes, she looks good, and she goes to school, and she’s in activities, to some degree. We adapt everything so she can do as much as possible. But, she is living with a fatal disease that is non-treatable. We basically just manage her symptoms. We know very well that we could lose a second child. That’s what is hardest on us. Even though there are these great advances, she is alive because of amazing machines. Every day presents the chance that she could die.
“When we take Catherine places, there are always at least 10 machines that go with her,” Sims says. “Everything just takes longer. We have a special van with a lift, because she’s in a wheelchair. You are in the thick of trying to make what is not normal to be normal.
“You can’t just pick up your child and go, you can’t feed them a different way, or put a different outfit on them,” Sims continues. “Those are the silly things I took for granted having had a healthy child before. I just did her hair, brushed her teeth, and put her in whatever, and fed her whatever I wanted. Catherine cannot do that. It’s the small things that you take for granted, and there are so many ‘small’ things. We are fortunate to have excellent in-home nursing care, but this also means that my husband and I have had to sacrifice a lot of our privacy. And, I’ve had to give up a lot of my mothering, because I have someone else that always needs to know what I’m doing. That’s hard.
“So, we want a cure,” Sims states. “We are all in. We are always fighting the disease. Our goal would be to have a cure as fast as possible, because the older the kids get, the less chance you have of curing them. This is a neurologic disease; it is hard to get those nerves back. We realize that our kids may be too old. Catherine will be 5; Lisa’s Silas is 8 or 9. They’re kind of old. The ideal time would be right at birth or shortly thereafter. So, that’s what we want. We want the big places — the big funding sources — to realize how important this is, even though it affects only a small number of people.”
“Our gene therapy vector is a very powerful tool,” Lorson says. “It is early days, in terms of trying to push it to the clinic, but we’re trying to do all the important pre-clinical questions.
“There are a number of questions you have to ask,” Lorson continues. “When do you deliver that kind of vector? Does it work only if you do it right at birth, before disease develops? Can you correct the disease, in other words, once the research animals have the disease, can you bring them back to more of a normal state? Or, once that happens, is it just too late for something like gene therapy? We want to deliver what they want to see, in terms of working hard and getting results out. That is what we are trying to do.
“I want to say, ‘Thank you,’ in the biggest way possible to the Sims family,” Lorson says. “Their generosity is really amazing. We consider this an exceptional honor. We want to be the best stewards they could possibly find, of their trust and of their funds. People go out and raise these funds — in some cases, through car washes and bake sales — so you have to put a particularly high value on those dollars. My fondest hope is that we do that every day.”
If you would like to help in the battle against diseases that could someday be relieved through gene therapy, please visit this page.
After a decade of work, Cheryl Rosenfeld is no stranger to bisphenol A (BPA), and her most recent study challenges the dangers posed by developmental exposure the chemical.
Her results continue to raise concerns about how BPA can potentially turn on or off genes in animals and subsequent effects on that early exposure can have on the development and brains of rats. Their research was published in the journal Epigenetics in July.
Rosenfeld and the University of Missouri joined experts from University of Cincinnati and FDA researchers as part of the Consortium Linking Academic and Regulatory Insights on BPA Toxicity, or CLARITY-BPA Consortium project. This collaboration is one of several across the United States meant to judge the chemical’s effect using standardized protocols established by the FDA to determine whether BPA exposure, especially during perinatal life, leads harmful effects.
“This is the first study published since a February 2018 BPA statement that challenges the FDA assertion that there is no concern for BPA,” Rosenfeld said. “We’ve shown using the FDA models and studies done right there at their facility that, indeed, early life exposure to BPA can result in gene expression and epigenetic changes that persist into adulthood.”
The study looked at gene expression changes in two brain regions — the hippocampus and the hypothalamus. The hippocampus is associated with long-term learning and memory and the hypothalamus plays a large role in hormone production that influence both the endocrine and nervous systems and affects diverse behaviors, including socialization, sexual behaviors, and appetite control.
Partners at the FDA/National Center for Toxicological Research fed Sprague-Dawley groups of rats — a standardized animal model in this research — diets of BPA, the synthetic estrogen present in birth control pills, ethinyl estradiol, or a chemical-free diet during a developmental period.
The brains from these animals were sent to Rosenfeld’s laboratory, who took biopsies from specific regions of the brain. They used these samples to evaluate whether a group of 10 genes, shown to be affected by BPA exposure in other studies, was affected by this exposure. They also examined the DNA methylation patterns for the promoters of three of these genes to determine whether prior BPA exposure led to persistent epigenetic changes. Epigenetic modifications do not affect the DNA sequence itself but gene and/or eventual protein expression.
Investigators determined that for several of the genes examined BPA exposure altered the expression pattern relative to animals not exposed to either chemical. Sex differences in gene expression in these two brain regions exists in normal animals, and such differences might thus contribute to masculinization or feminization of the brain manifesting as differences in various behavioral patterns, such as male or female sexual behavior. However, previous exposure to BPA abolished many of these gene expression differences between males and females, suggesting that it could disrupt male- and female-typical behaviors. For a gene, brain derived neural factor (BDNF), involved in learning and memory, BPA exposure led to increased methylation of its promoter, which could affect the expression of this key gene. Hippocampal expression of several genes was associated with prior performance in a test designed to measure learning and memory.
“It has become increasingly apparent that BPA can act as a weak estrogen, but what we’re seeing in these results is that it can elicit other effects in addition to those mirroring estrogen and likely independent of estrogen receptor pathways,” Rosenfeld said.
Initiatives like this and other CLARITY-BPA studies aim to answer questions that may later inform government regulators on how to limit or balance the health effects of manufactured chemicals that end up in the environment and may affect human and animal growth in previously unknown ways. With more than 15 billion pounds of BPA were estimated to be produced in 2013, its ubiquitous use in making plastics, lining cans and other manufacturing is of concern. Rosenfeld hopes a closer look at its epigenetic effects may lead to better regulation of the chemical.
“When people are thinking about the effects of BPA, they need to be thinking about it on a molecular scale,” she said. “These results might be subtle, but they can lead to dramatic consequences with long-standing, irreversible changes. Once BPA exposure resculpts an animal’s brain through DNA methylation and other epigenetic changes, it may be permanent.”
Research at the undergraduate level offers more than meets the eye. With students from every year of their undergraduate careers working in Bond LSC, it’s a great opportunity to acquire skills and experience.
Linda Blockus, head of the Undergraduate Research office in 150 Bond LSC, advises students to get started early and be proactive.
“I encourage students who are interested in research to talk to people and network,” Blockus said. “Talk to your professors, advisors and other students to find out what is available. Then, pursue those opportunities.”
It isn’t all as intimidating as it might appear. Students have a number of resources available to find out more about research on campus.
“There’s no one way to get involved,” Blockus said. “Students can go directly through our website, undergradresearch.missouri.edu, come to our office or go to their professors.”
That’s exactly what students involved in the Freshman Research in Plant Sciences (FRIPS) program have done. Sarah Unruh, a Ph.D. student who serves as a Graduate Student Coordinator for the program, boasts of the program’s ability to guide research-minded students along their path at Bond LSC.
“They do 10 hours of research in lab,” Unruh said. “We try to give them skills that are helpful moving forward, so things like finding papers and keeping up with a lab notebook.”
Each of the students selected for the program works in a lab they find the most interesting, but the program assists with those relationships to help students adjust to the process.
“Students lead the way in which lab they go to,” Unruh said. “They interview with different faculty, but we facilitate the match-making.”
Those interactions and networking opportunities open doors down the line.
“I think what they get the most out of FRIPS is that they’re actually doing science, so they get an idea of what it looks like,” Unruh said. “They’re making connections on a different level than just the classroom with teaching assistants and professors.”
Jenna Bohler — one of the students involved in FRIPS this year — has benefited from its connection-facilitating.
“Paula McSteen, Norman Best and Jenn Brown have taught me so much this year in particular,” Bohler said. “They’ve been great resources whenever I’ve had questions.”
Bohler is about to finish her FRIPS experience and can attest to the program’s influence on her first year at Mizzou.
“I knew coming into college I wanted to be involved in research, and FRIPS allowed me to get involved really early so I have four years instead of two or three,” Bohler said.
And it’s not only helpful in the lab.
“What I’ve learned from FRIPS has helped with my classes,” Bohler said. “I learn things before I’m taught them in class, which makes them easier to understand.”
Some FRIPS students have even extended their research opportunities beyond their freshman year.
“Students have used their time wisely in the lab and then gone on to do summer research programs,” Unruh said.
For those who aren’t freshman but find themselves interested in research, there are a number of programs available.
The Society of Undergraduate Researchers in Life Sciences (SURLS) is a group of undergraduate researchers who meet twice a month to explore the options they have within their field. It helps participants to network, meet people with similar interests and better understand a number of components of research.
Alec Wilken, a junior bioscience major who works in the Holliday lab in the medical school, served as the vice president and will be the president for his senior year. He’s been part of SURLS since he was a freshman and has seen first-hand how it’s shaped his path in the field of research.
“SURLS helped me find what I was interested in,” Wilken said. “We have professors come in, and we visit labs, which helps undergraduates grasp how interesting research on campus really is.”
SURLS provides students with the opportunity to grow throughout their undergraduate careers.
“I stayed in SURLS after joining my lab because it became a vehicle that helped me be better in my lab,” Wilken said.
The organization’s impact has allowed Wilken to uncover the path he wants his career to take, as he now plans to earn a Ph.D.
“I found a home in research, and SURLS helped me do that,” Wilken said.
For those with plans to pursue a Ph.D. in their future, MU’s Maximizing Access to Research Careers/Initiative for Maximizing Student Diversity (MARC/IMSD) program is the perfect fit.
The grant is funded by the National Institutes of Health (NIH), but at Mizzou there’s the addition of Express to the program’s title. It stands for Exposure to Research for Science Students, which emphasizes the scientific aspect of the program.
Brian Booton, is the undergraduate director for MARC/IMSD-Express at Mizzou, acknowledges the prestige that goes along with being an MARC/IMSD scholar.
“It’s a highly selective grant,” Booton said. “There are only 49 programs in the country.”
With stiff competition for the program at universities across the nation, it’s important to focus on the students’ experiences.
“The ways in which IMSD-Express helps students is more than just research,” Booton said. “We try to expose students to the different pathways where further education can take them.”
Part of that is through the way the weekly meetings breakdown.
“I break programing down into three areas: personal, academic and professional development,” Booton said.
Doing so helps guide students in the right direction because it is set up to further their education by developing skills for success.
But it’s not all lectures and typical meetings. MARC/IMSD-Express offers a peer mentorship program for underclassmen apprentices to be paired with upperclassmen fellows.
“Even if you have a professor you really admire, there’s some distance there,” Booton said. “Someone that’s only two years older than you is more relatable; it’s spending time with your future self.”
The various research opportunities at Mizzou make it possible for students to supplement their classroom learning in a way unlike any other.
“It’s part of your education,” Blockus said. “Taking advantage of research is a great way to set yourself up for the future.”
For more information and to apply for these opportunities, visit:
Endocrine disruptors alter baby mice calls generations later
By Roger Meissen | Bond LSC
The sounds can seem like a mix between a bird tweet and a high-pitched scream to us, but these vocalizations that baby California mice make are essential to how they communicate with their parents and siblings.
Exposure of grandparent mice to bisphenol A (BPA) and related endocrine disrupting chemicals (EDCs) may alter that communication in their grandoffspring, potentially affecting the communication between pups and their parents and the resulting parental care provided to them.
According to a new study, MU Bond Life Science Center’s Cheryl Rosenfeld and an interdisciplinary team of researchers from the US and Germany looked at how this communication alters from normal patterns across multiple generations of California mice.
“We specifically wanted to see if grandparents were exposed, would that affect the communication of the grandoffspring?” Rosenfeld said. “What we saw was that in some cases, some aspects of their vocalizations became even more pronounced. It might be a response to multigenerational exposure to EDCs or they might be calling more because they aren’t receiving sufficient parental care in an effort to say, ‘hey, you’re neglecting me; please pay attention and provide warmth and nutritional support to me.’”
Studies from Rosenfeld previously found that BPA caused lax parenting and neglect in first-generation mice when their parents were developmentally exposed to the chemical. This chemical acts as an endocrine disruptor and mimics the effect of hormones like estrogen in animals, altering their development. BPA is prevalent in the environment because it’s heavily used in manufacturing and leaches out of our plastics, linings of food cans and dozens of other sources.
The study showed female babies tended to make shorter calls out to parents early on after being born, but as they aged they called out more, and male babies made longer calls in early postnatal periods and spoke more as they aged. These patterns were different from controls not exposed to the chemicals.
“Exposure of the their grandparents to EDC’s is altering these grandoffspring behaviors and that could have important ramifications to human babies and how EDCs might affect their initial form of communication, crying,” Rosenfeld said. “This follow up work is clearly important because children with autism have communication deficits, as evidenced even in their early crying patterns, and altered social skills. We’re always trying to find animal models like this that might explain whether exposure to environmental chemicals is increasing the incidence of autism or autistic-like signs in animal models.”
California mice are an especially useful model for studying behavior changes, because these mice are monogamous and both mom and dad are essential in rearing their pups, similar to most human societies. This allows scientists to potentially extrapolate their behavior changes to humans.
In this study, both female and male grandparents were fed one of three diets — A BPA diet that contained an environmentally relevant concentration of this chemical, an ethinyl estradiol diet or diet free of any EDCs. Ethinyl estradiol is another disruptor found in birth control that mimics the effect of estrogen in the body. All offspring were fed the chemical-free food after being weaned off the parents. They had babies, and these grandchildren were the generation scientists looked at to study their communication.
The grandchildren were recorded with special microphones that could pick up the calls of the babies in isolation booths. These sounds range from communications humans can’t even hear as they are high in the ultrasonic range- greater than 20,000 hertz- to communications that begin in the range of human hearing and then project into ultrasonic range. When researchers lowered the frequency of these high-pitched calls to a range we can hear they sound like a mix between owl screeches and bird tweets (how the vocalizations appear and sound are included below for the reader to decide for themselves) . They then compared them to normal mice, looking at the length of each call and the pattern of the calls, what they refer to as “syllables.” Each syllable is akin to an individual sentence or phrase in humans.
These calls from BPA exposed mice were compared to the ethinyl estradiol and the mice not exposed to any chemicals.
“We’re seeing clear traits emerge in this F2 generation with the vocalizations and I think it lends credence to the idea that these things could tamper with vocalization patterns, which are incredibly important in how pups communicate with each other and their parents, whether it’s because they are trying to get more attention from exposed parents or what we call multigenerational effects in that the exposure of their grandparents directly affected their later grandoffspring traits.”
The study, “Multigenerational effects of Bisphenol-A or Ethinyl Estradiol Exposure on F2 California Mice (Peromyscus californicus) pup vocalizations,” was funded by the National Institute of Environmental Health Sciences Grant (5R21ES023150) and was published in the journal PLOS One June 18, 2018.
Answering the unsolved questions is a lifetime commitment for fifth year Ph.D. candidate Rowan Karvas in the Roberts Lab at Bond LSC and Laura Schulz’s lab at the medical school in the Obstetrics and Gynecology department.
Originally from St. Louis, Karvas came to Mizzou and found her keen for science through her undergrad research working on adult muscle cells, but it wasn’t until she became a technician in a radiation oncology research lab at Washington University when she realized she wanted to continue research throughout her life.
“I remember a moment when a grad student, Danny Stark, and I were isolating quail embryos,” Karvas said. “We opened them up and I saw their beating tube hearts and all the details. Just looking under the microscope at them was so fascinating to me, I realized that I wanted to keep doing biological research.”
Karvas believes asking questions is what we often forget to do as a society, and in a scientific world with unlimited questions, she knows the one she’d most like to answer.
“I would like to solve pre-eclampsia,” Karvas said. “It’s a disease that’s most likely been with us since we have been human, and it is a disease that deserves to be solved.”
Karvas works on two projects researching human placental development. The pregnancy disease pre-eclampsia often goes unnoticed until later in pregnancy, but causes dangerously high blood pressure, kidney damage, and a placenta that is underdeveloped and has not invaded the maternal endometrium efficiently.
Karvas was eager to work on this problem and so she started grad school two months early.
“This disease if often very deadly for women and in the most severe forms causes death,” Karvas said. “It is the leading cause of maternal fatality in the developing world and is becoming a problematic issue in the states. There are no rock solid genetic, physiologic factors, or environmental stimulus that accurately predicts who will get this disease.”
And that’s why Karvas spends her time researching how and when pre-eclampsia develops during pregnancy.
In Karvas’s first research project, she is looking at modifications to the genetic code that could be responsible for pre-eclampsia and her second project is finding the answer to the question, “at what stage of pregnancy does our cell model represent?”
When Karvas isn’t in the lab you can find her aiding to bridge the gap between clinical scientists and basic science researchers as president of Interdisciplinary Reproduction and Health Group Trainees. The new group involves graduate students and post-doctoral researchers from animal sciences to biochemistry, all coming together to further explore reproductive science.
“It makes me happy and it is very satisfying to start the group and be a part of the groundwork for its continuation,” Karvas said. “Fostering these relationships with people you may have not met otherwise is important.”
When Karvas isn’t running her group or in the lab, you can find her playing the clarinet. Before discovering her love for science she wanted to be a professional performer.
“It is an opportunity to use the other part of my brain,” Karvas said. “I get to shut off the science part and bring on the musical part.”
For researchers, the shape of molecules gives insight into how cells, viruses and other macromolecular interactions take place.
Getting a clear view of that structure is the hard part, and the new Molecular Interactions Core (MIC) at the Bond Life Sciences Center will now give researchers from many different disciplines one place where state-of-the-art equipment are available for them to use to further science.
Dr. Kamal Singh is excited that goal is being realized.
One of the 10 MU’s core facilities that serve scientists’ needs, the MIC specifically provides training, advising and shared equipment for researchers to take a closer look at molecules.
That’s where Dr. Singh comes in.
He serves as the Assistant Director of the MIC and oversees the day-to-day operations of the facility. Dr. Singh makes sure the machines are operational, communicates with researchers interested in using the facility, trains those who do not yet know how to use the equipment and gives guidance as well as collaborative feedback on things like computer-assisted drug design — his specialty.
The humming of machines is the first thing noticed when walking into the MIC. These instruments allow you to look at 3-dimensional models of the molecules like HIV enzymes or view protein crystals under a microscope before diffracting light through them. It’s a lab where miniscule pieces of life become big and important.
Dr. Mark McIntosh, the vice chancellor of research for all UM system campuses, had the idea to create the MIC.
“It was Dr. McIntosh’s vision to bring everything together; which includes structural biology, molecular interactions, particle size, zeta potential, mass of the nanoparticles, etc. He also wanted to bring peptide synthesis here to have everything at one central location,” Dr. Singh said.
Understanding structure at the molecular level helps scientists figure out how reactions happen, how molecules fit together and serve as signals and how pathogens can invade cells, among other possibilities.
“I’m hoping that we can really facilitate structural and molecular research on campus — structural determination and molecular interactions — and really push boundaries of the current state of the field,” said Dr. Tom Quinn, the Director of the MIC.
Dr. Quinn hopes the state-of-the-art equipment will allow the MIC to be a resource for both research faculty and students to be on the cutting edge in their fields.
Dr. Ritcha Mehra-Chaudhary and Dr. Fabio Gallazzi work within the MIC and provide their expertise. Dr. Mehra-Chaudhary works with X-ray crystallography, dynamic light scattering and custom protein expression, while Dr. Gallazzi is an expert in custom peptide synthesis. Their work can be important for understanding drug design to combat viruses and cancers.
The MIC started with one machine, an X-Ray Diffractometer, in room 442 of the BLSC. It took six months to collect the different machines from different departments in the campus, but in December the MIC became fully operational. The MIC team celebrated with an open house on Jan. 24, 2018.
“It’s kind of crowded, but it’s good,” Dr. Singh said. “We have invited everyone, mainly researchers, but also undergrads. The entire university is welcome to come see what we have. The idea is the advancement of science in our school.”
The MIC won’t only be beneficial for campus researchers, but also researchers from all over and undergrad students who are eager to learn the details of molecular interactions and learn how to use core facilities.
There are many exciting and new technologies in the MIC that will interest outside researchers, according to Dr. Quinn. One of these is the nanodisc technology that Dr. Mehra-Chaudhary works with. This technology allows researchers to study membrane proteins outside of something bigger, like a cell, while also keeping them in a functional and native structural state. The nanodisc project is part of collaboration between the MIC and the Electron Microscopy Core to allow researchers to get high resolution structures of membrane proteins.
While affordable, outside and campus researchers must also pay a price to use the facilities to cover consumables, instrumentation maintenance and staff.
“We definitely want to at least break even. I don’t know how long it will take to get there. However, the major goal is to support the scientists on campus and facilitate their research,” Dr. Singh said.
Bringing this support to campus also means supporting future scientists. Dr. Singh has three undergraduate students working with him who are learning how to use the advanced technology, and he helps to train many more from all different departments.
The goal is to one day expand the MIC to a point where all molecular interactions facilities can be at one place.
“There are certain techniques we don’t have, and I hope that in the future we will get them. We hope to provide all modern techniques to the university community in coming years. Not only linked to that room, we want to expand it,” Dr. Singh said.
Dr. Quinn agrees, and he hopes that as researchers come and use the core. In the process the core can understand future needs and where the research is moving to see what new technology under their umbrella could be added to keep supporting the scientists.
The MIC is a big step for the MU research community, and staff is hopeful that it will continue to grow and produce life-altering research.
It might not sound like a traditional undergraduate experience, but Elizabeth Prenger and Andrew Ludwig found success studying a tiny parasitic worm.
It’s called the soybean cyst nematode (SCN) and it sucks more than a billion dollars a year from American soybean farmers. While farmers have used resistant soybeans and crop rotation to fight against the pest, the nematodes continue to gain ground against increasingly less effective methods to control them.
Working in the lab of Melissa Mitchum, a Professor of Plant Sciences at MU’s Bond Life Science Center, they helped understand how soybeans naturally resist this worm and how SCN evades these protections.
That work recently paid off as they saw their names published in the journal Plant Physiology in November 2017. The study explored the genetic mechanisms behind resistance in order to develop better prevention.
“If scientists can understand how resistance genes work and interact then that information can be applied in breeding and developing soybeans,” said former Mitchum lab member Elizabeth Prenger.
While the findings were published in 2017, for Prenger and Andrew Ludwig the research began several years ago.
Prenger came to college knowing she wanted to improve crops and help farmers like her family, she just wasn’t sure exactly how. She joined Mitchum’s lab as a freshman to begin to find out.
As a freshman and sophomore, Prenger worked to purify, sequence and analyze DNA of various soybeans to help further characterize the SHMT gene, a gene that plays a role in a plant’s ability to resist the pest. She also worked in the greenhouse to identify soybeans with mutations in this gene by infecting them with SCN.
Her fellowship supported by the MU Monsanto Undergraduate Research Program sparked her interest in plant genetics but she also realized she wanted more interaction with plants beyond the lab.
Without this early immersion into the lab, Prenger said it would have taken her longer to find her interests.
Now, as a graduate student, she studies soybean genetics at the University of Georgia.
Ludwig’s position in the lab helped him find his direction in science as well.
He applied for a position while still in high school through the MU Honors College Discovery Fellows Program. The fellowship funds and places undergraduates in labs across campus. His interest in the genetic modification of crops led him to the Mitchum lab.
For three years, Ludwig helped infect different mutants with the nematode and then compare the effect on resistance. This screening helped narrow down the genetic possibilities controlling soybean resistance to a single gene.
“We were hoping the soybeans would have a mutation in one of the resistance genes and then that mutation would cause the gene to cease function so you would see a lot of nematodes on a plant that shouldn’t have any,” he explained.
This experience taught Ludwig how to think like a scientist by developing problem-solving skills.
“I think working in the lab was an immensely valuable experience because I learned so much about what it is to be a scientist and it opened my eyes to a lot more of what the field of plant science really is,” he said.
It also taught him that a traditional lab work environment was not for him. As Ludwig begins to apply for graduate school he is planning to major in horticulture.
His goals changed from wanting to create GMO crops for other countries to now hoping to solve food insecurity closer to home by working with sustainable agriculture and food deserts.
Since joining Mitchum’s lab as undergraduates, both Prenger and Ludwig learned what it means to be scientists and shaped where they are today. The publication of the research that started the path to where they are today was a satisfying conclusion.
“It’s really rewarding to see that all the work exists outside of my lab notebook now,” Ludwig said.
Reflecting on their experience, both students urged other undergraduates to get in a lab as soon as they can to begin discovering themselves and science.
“Go for it. It’s a really helpful experience, it will make you better at what you do even if what you end up doing is different from what you thought you’d do,” Ludwig recommended.