Those natural enzymes are called ATPases, and their activity’s reliance on ATP makes these catalysts a desirable model for drug discovery. For example, a naturally derived ATPase that’s smaller and more malleable than a protein could be delivered to cancer cells to direct ATP’s energy toward killing those cells, or targeted to block the release of energy that stimulates harmful cellular processes.

Researchers at The Ohio State University have gotten one step closer to that possibility by identifying the first noncoding RNA derived from human cells that can bind to ATP. The team is looking now at that RNA’s function, with hopes that its affinity to ATP resembles an ATPase function that could form the basis of a therapeutic nanoparticle.

“The concept of having a noncoding RNA function as an ATPase is still really new. No one has been able to find ATPase activity in an RNA yet,” said senior study author Peixuan Guo, professor in the Division of Pharmaceutics and Pharmacology at Ohio State.

“Many scientists think human proteins handle all ATP activity in the body, but I think there’s no way it’s limited to only a protein function,” he said.

The study was published recently in the journal Non-coding RNA Research.

Guo is a pioneer in the discovery of functional noncoding RNAs – tiny RNA molecules lacking instructions for building proteins – that constitute over 98% of the human genome. His lab has spent years refining nanoparticle technology with versatile RNA molecules that cooperate with the body, most recently designing molecular clusters called RNA micelles that deplete metastatic colorectal cancer tumors in mice.

He is optimistic that there is a noncoding RNA to be found in the genome that functions as an ATPase, and foresees making use of such a molecule for a new class of therapeutic nanomaterials. 

This new work builds on previous work in Guo’s lab that led to the identification of a viral RNA that could bind to ATP. 

“I was essentially doing a continuation of that study, looking to see if there’s anything in human RNAs that can bind to ATP,” said first author Margaret Bohmer, a PhD student in Guo’s lab. 

Bohmer ran multiple rounds of screening total RNA extracted from human cells, looking for segments that could connect with ATP. She removed non-binders between rounds and amplified the presence of ATP until she had an enriched pool of RNAs that bound only to ATP. She then sequenced what was left, confirming identification of a single noncoding RNA that made a strong attachment to ATP. 

The structure of this RNA is called a G-quadruplex, which has an advantage of being very compact and highly stable, Bohmer said. 

The team is following up by investigating the mechanism of exactly how this noncoding RNA binds to ATP. 

There’s a decent chance, she said, that this noncoding RNA could be part of an ATPase motor that assists in decomposing ATP – a prediction based on the lab’s previous finding that the viral RNA they found years ago that bound to ATP had a functional role in the motor. 

In another recent Non-coding RNA Research paper, Guo’s team also found evidence that viral molecules called packaging RNAs that bind ATP structurally remained very similar or the same while changing their coding sequences over the course of evolution to perform motor-related biological functions – a sign that they, like enzyme proteins, could trigger similar catalytic reactions in cells. 

“We are curious to see if this G-quadruplex RNA is an RNA that functions in a similar way,” Bohmer said. 

This work was supported by the National Institutes of Health, an Ohio State President’s Research Excellence Catalyst Award and an Ohio State Center for RNA Biology Fellowship.

Kai Jin, a PhD student in Guo’s lab, was also a co-author of the paper.

These particles, called extracellular vesicles, are known to safely carry signaling cargo as a communication method between cells in bodily fluids and within tissue, and to influence health and disease. Understanding how the properties of these vesicles differ in normal versus diseased tissue could make them outstanding biomarkers for early disease detection, researchers say. 

Scientists at The Ohio State University have made inroads into understanding one side of that equation, finding that extracellular vesicles’ range of travel in a cancer tumor environment is limited almost entirely to nearby recipient cells – which is related to how densely packed cells are in a tumor. 

“This is the first assessment of how far a vesicle can move in physiological conditions. We didn’t perturb the system. We just measured on a single-cell basis how far a vesicle can go,” said senior study author Emanuele Cocucci, associate professor of pharmaceutics and pharmacology at Ohio State, who is studying these particles – and how physiological conditions affect them – to weigh their potential as biomarkers for pancreatic cancer. 

“There are no clear biomarkers for pancreatic cancer at the moment. Our goal is to define a novel biomarker,” said Cocucci, also a member of the Leukemia and Hematologic Malignancies Program at The Ohio State University Comprehensive Cancer Center. “But until we know how extracellular vesicle exchange occurs in tissue before and during cancer onset, it’s impossible to understand whether extracellular vesicles could be the next answer for early diagnosis of pancreatic cancer.” 

The study was published recently in the Journal of Extracellular Vesicles. 

This work involved a series of cell culture experiments in two cancer cell types, first showing that increasing the density of cells in a petri dish led to a decrease in the number of vesicles released per cell – suggesting either the cell-to-cell interface inhibits release of vesicles, or that neighboring cells are gobbling the particles up. 

Researchers then used a technique that tagged donor cells – those releasing extracellular vesicles – with one color of dye and all other surrounding cells with a different color, and observed the vesicle activity at a single-cell level using flow cytometry. Results showed that the surrounding cells engulfed many of the vesicles and, in some cases, degraded them. Though any signal exchange was not measured, the results clearly showed that the vesicles didn’t stray very far from their donor cells. 

“I was expecting we would see a huge exchange over a distance, and that no matter what, the vesicles would diffuse away, so the chance for being taken up nearby or far away should have been the same,” Cocucci said. “Instead, that seems not to be the case. When cells are close together, they share more vesicles with each other.” 

To analyze what would happen in living tissue, tumors were developed in mice and then injected with one vesicle donor cell for every 3,000 acceptor cells, all dyed so they could be identified through confocal microscopy imaging. After introducing a chemical to prevent any degradation of the particles, the researchers measured how far extracellular vesicles dispersed from the donor cells. 

The findings showed that 80% of the vesicles stayed within 40 microns – or 40 thousandths of a meter – of the donor cell, and 95% of these carriers traveled no farther than 70 microns away, a distance roughly equal to the thickness of a human hair. 

Computational modeling predicted the same result.

“If we interpret this in a tissue situation, then donor cells will release vesicles, and the cells most affected by vesicle release are the surrounding cells,” Cocucci said. “When you go two to three cells away, these cells will affect the neighboring cells – suggesting that the majority of the effect of vesicles is constrained within 70 microns from the cell of origin.” 

Cocucci’s lab is now developing tools for use in animal models that would allow the team to investigate and define the contribution of each tissue to the circulating pool of extracellular vesicles. 

“If the release of the vesicle changes according to the physiological state of the tissue, this means the contribution of each tissue to the circulating extracellular vesicle pool changes according to the status of the tissue itself,” he said. “If you are able to define the contribution of each tissue in normal or disease conditions to the circulating pool of vesicles, then when we analyze vesicles from a patient, we may be able to suggest, without bias, a diagnosis without further analysis.” 

This research was supported by the National Cancer Institute, the Ohio State University Comprehensive Cancer Center, Ohio Cancer Research via the McCurdy/Kimball Midwest Research Fund, the American-Italian Cancer Foundation and a Pelotonia Postdoctoral Fellowship. 

Additional co-authors were Federico Colombo, Kartik Nimkar, Erienne Grace Norton and Francesca Lovat, all of Ohio State.

Researchers constructed tiny molecular clusters called RNA micelles, loaded them with potent chemo drugs and an RNA molecule that blocks cancer survival, and placed a tumor targeting molecule on their outer wall that attaches to receptors on cancer cell surfaces to enhance delivery. 

Treatment with these RNA micelles almost completely depleted metastatic colorectal cancer tumors in mouse lungs within 26 days. The tumors in mice mimicked colorectal cancer that metastasizes to the lung in humans, which comes with a poor prognosis: Only 16.2% of patients survive five years after diagnosis. 

“Developing RNA therapeutic technology is key to treating colorectal cancer lung metastasis because there is no cure,” said senior study author Peixuan Guo, Sylvan G. Frank Endowed Chair professor in the Division of Pharmaceutics and Pharmacology at The Ohio State University. “We’ve developed a nanoparticle that can treat it efficiently without toxicity – the particle spontaneously targets the cancer and no toxicity is detected.” 

The study was published Jan. 20 in the journal Advanced Functional Materials. 

The research coincides with a Nature Protocols publication from Guo’s lab published Feb. 3, in which he and colleagues provide detailed instructions on how to construct RNA nanoparticles for targeted delivery of therapeutics combined in one package to produce synergistic treatment effects. 

The scientists describe Lego-like architectural structures composed mainly of versatile RNA molecules that have rubber-like properties, all factors that foster cooperation with the body and, as a result, enabling spontaneous tumor targeting and rapid excretion by the kidneys. 

“The main idea of the micelle studies is that we can literally prove part of the idea we talk about in the Nature Protocols paper,” said Kai Jin, first author of the colorectal cancer study and a PhD student in Guo’s lab. “The micelle paper is an example of how a different kind of RNA nanoparticle can be achieved.” 

In the study, the researchers doubled the cancer-killing power of the RNA micelles by combining multiple copies of the chemo drug gemcitabine with a molecule called a small interfering RNA to silence the gene encoding for survivin, resulting in treatment of the colon cancer lung metastasis and termination of cancer cell growth, respectively. 

“It’s a two-pronged approach. The chemotherapy is killing the cells and the small interfering RNA is blocking survival gene expression,” said co-author Daniel Binzel, a research assistant professor of pharmaceutics and pharmacology at Ohio State. “The two together help kill the cells in multiple different pathways.” 

The team also attached a molecule called a ligand to the micelles to enhance their attraction to receptors on cancer cell surfaces.

The team initially found in cell culture experiments that delivering gemcitabine and survivin siRNA packaged together in micelles induced DNA damage and programmed cell death in a human colorectal cancer cell line. 

In the animal experiments, a six-dose micelle therapy over three weeks started five days after mice were injected with metastatic colorectal cancer cells. Compared to the tumor status in groups of control mice receiving micelles containing either chemo or siRNA alone, the mice receiving micelles loaded with the combined therapeutics were nearly cancer-free. Attaching the ligand to the micelles significantly improved the tumor reduction. 

Results showed the micelles attacked the cancer in two ways, accumulating in tumor blood vessels and entering the cells with the ligand’s help. 

“We showed that one nanoparticle can carry a drug and therapeutic RNA at the same time, and also used an RNA ligand – these three things are put together so the particle recognizes the cancer cells, binds to them and delivers the particles into the cancer cells,” said Guo, also an investigator in The Ohio State University Comprehensive Cancer Center. 

Publishing the RNA nanoparticle methodology in Nature Protocols, which publishes only protocols proven to work, comes almost four decades after Guo, as a student in the 1980s, published a paper in Science determining that short segments of noncoding RNA not only existed, but had important functions in cells. 

“The recent boom of RNA therapeutics companies and the FDA approval of RNA therapeutics have indicated that my prediction that RNA was the third milestone of pharmaceutical drug development has been realized,” he said. 

The work in both publications was supported by the National Institutes of Health, the National Cancer Institute and an Ohio State President’s Research Excellence Catalyst Award. 

Binzel is the first author of the Nature Protocols paper, co-authored by Guo, Jin and Tesla Yudhistira, all of Ohio State. Additional co-authors of the Advanced Functional Materials paper include Piotr Rychahou and Mark Evers of the University of Kentucky. 

RNA Nanobiotics, a company based in Cambridge, Massachusetts, holds exclusive global licenses to multiple patents covering RNA nanoparticle platforms for targeted cancer therapy and RNA exosome technologies developed by Guo at Ohio State and the University of Kentucky. 

Kroetz, also a professor of pharmaceutics and pharmacology at Ohio State, was recognized by the academy for her scientific contributions in pharmacogenetics – among them, identifying genetic markers that predict drug toxicity in patients – and for her exceptional mentorship and academic leadership. 

“Mentoring has always been a key component of what I do, because I have always been involved in graduate education,” she said. “I’ve considered job No. 1, when you’re on the research faculty, is to train the next generation of scientists.” 

The NAM was founded in 1970 as the Institute of Medicine and is one of three academies that make up the National Academies of Sciences, Engineering and Medicine in the United States. The academies are private, nonprofit institutions that work outside of government to provide objective advice on matters of science, technology and health. 

“The National Academy of Medicine has been instrumental in shaping U.S. and global policy around the promotion of health for decades. I’m thrilled to congratulate Dr. Kroetz on her election to this prestigious and dedicated community of scholars,” said John M. Horack, vice president for research at Ohio State. 

With her background in both the pharmacy profession and the science around drug development, Kroetz looks forward to engaging in national conversations about the health sciences as a new member of the NAM. 

“I’m very honored and really humbled to be elected,” Kroetz said. “And I do want to take it as an opportunity to be involved where I can, and to promote the profession, making sure pharmacy and all of the health sciences have a voice at the table.” 

Kroetz describes her research program as “bedside to bench,” taking human tissue samples to the lab to investigate the molecular mechanisms behind problems physicians see in the clinic. 

Her initial genetics research focused on a family of proteins called ABC transporters that perceive drugs as unfamiliar molecules and pump them away from tumors and other disease targets, limiting the drug’s effectiveness and exposing sensitive surrounding tissue to toxic compounds. She and colleagues explored genes that regulated the transporters’ functions with the aim of designing agents that could block their activity. 

Among the toxicities associated with ABC transporters is their role in chemotherapy-induced sensory peripheral neuropathy, which causes pain, numbness, tingling or burning in the hands and feet, loss of function in the fingers and increased temperature sensitivity. A primary focus of Kroetz’s lab in recent years has included genome-wide association studies to identify genes that contribute to individuals’ increased risk for this toxic side effect and molecular studies to understand how these genes are involved in the neurotoxicity. 

These investigations advanced from the earliest days of human genetics to the rapid genomic sequencing available today through the Pharmacogenetics Research Network funded by the National Institutes of Health for 15 years, Kroetz said. 

“Multiple centers were working on pharmacogenetics and every group across the country was working on a specific class of drugs. We got together several times a year to share resources and help each other,” she said. “This never would have happened without the NIH umbrella funding across these groups.” 

She is a rare example of a pharmacy scholar in academia, combining training as a practicing pharmacist with a scientific doctoral degree and research-heavy career exploring molecular mechanisms behind drugs’ therapeutic – and potentially toxic – effects and how the human body responds to both. 

“I’m still keeping my lab going, but I also wear a different hat. I see pharmacy both from the profession side and the research side, both of which are really equally important,” she said. “So this is an opportunity to help give back to the profession as well.” 

Kroetz earned a bachelor’s degree in pharmacy at Ohio State and a PhD in pharmaceutics at the University of Washington, and completed a postdoctoral fellowship at the National Cancer Institute Laboratory of Carcinogenesis. 

She was appointed dean of Ohio State’s College of Pharmacy in 2023 after spending the previous 30 years at the University of California, San Francisco, as a faculty member and eventually chair of the School of Pharmacy Department of Bioengineering and Therapeutic Sciences. 

Kroetz has been elected a fellow of the American Association for the Advancement of Science (2018), the American Heart Association (2002) and the American Association of Pharmaceutical Scientists (2008). She has also received mentoring awards from numerous professional societies. 

The academy elected 90 regular members and 10 international members during its annual meeting. The newly elected members bring NAM’s total membership to more than 2,500, which includes more than 200 international members. New members are elected by current members through a process that recognizes individuals who have made major contributions to the advancement of the medical sciences, health care and public health.  

Kroetz is one of 10 current Ohio State faculty members who have been elected to the NAM.

Jianrong Li, professor of virology in the College of Veterinary Medicine Department of Veterinary Biosciences, and Peixuan Guo, holder of the Sylvan G. Frank Endowed Chair in Pharmaceutics and Drug Delivery in the College of Pharmacy, are among the 169 academic inventors to be named National Academy of Inventors (NAI) fellows this year. They are the 15th and 16th Ohio State inventors to be chosen as NAI fellows. 

In 2021, Guo was named Ohio State’s Innovator of the Year, and Li, along with colleague Stefan Niewiesk, was among the finalists for the award.

Jianrong Li 

Li has led the development of a number of vaccine candidates for respiratory viruses and enteric illnesses – those that affect the respiratory and gastrointestinal systems. His lab worked swiftly to develop vaccines against the SARS-CoV-2 virus that causes COVID-19 in response to the pandemic, developing separate candidates using the measles and mumps viruses as platforms for a COVID-19 vaccine. 

“The idea is that because children have been safely vaccinated for measles and mumps for decades, we could incorporate this vaccine into the MMR vaccine to prevent four diseases simultaneously: measles, mumps, rubella and COVID-19,” Li said. 

These platforms also enable delivering a “needle-free” vaccine through the nose, which generates both a system-wide antibody response – the same response as that produced by an injection in the arm – as well as a mucosal immune response in the linings of airways, where viral particles attach to host cells. All currently approved COVID-19 vaccines are delivered intramuscularly but are unable to induce sufficient mucosal immunity, the body’s first line of defense against respiratory viruses. The White House’s effort to develop the next generation of COVID-19 vaccines includes an intranasal vaccine approach. 

In each candidate, the team’s use of specific snippets of the “prefusion” version of the SARS-CoV-2 spike protein – the shape the protein is in before it infects a cell – and strategic insertion of those genes into modified measles and mumps virus genomes produced strong neutralizing antibody, mucosal antibodies and T cell immune responses and protected against lung damage in animals. 

The SARS-CoV-2 vaccine candidates and a third vaccine candidate against respiratory syncytial virus (RSV), the No. 1 cause of human respiratory infections that can also cause serious illness in infants and the elderly, have been licensed to an India-based vaccine and pharmaceutical company. 

Vaccines developed in Li’s lab have also been commercialized for use in protecting piglets from two deadly viruses, porcine epidemic diarrhea virus and porcine deltacoronavirus. Previous viral outbreaks have caused significant economic losses for the pork industry in the United States and worldwide. 

A member of Ohio State’s Infectious Diseases Institute, Li has been awarded eight patents and has eight pending patent applications, and has entered into a total of eight licensing agreements with companies that are commercializing vaccine products developed in his lab. 

Li, who joined Ohio State’s faculty in 2008, attributed his lab’s success to Ohio State’s creative and collaborative environment and support from his college and department. 

“I am humbled and truly honored that my peers have considered me for this award,” he said. “This honor really belongs to the postdoctoral fellows, graduate students and research assistants in my laboratory. In the past 14 years at Ohio State, I have had the privilege to work with them to generate innovative ideas, make new scientific discoveries, and translate basic virology research into the development of novel vaccines against viral pathogens that threaten human and animal health.” 

Peixuan Guo 

The many RNA-related discoveries attributed to Guo, also director of the College of Pharmacy Center for RNA Nanotechnology and Nanomedicine, date back to his days as a student in the 1980s, when he determined that short segments of noncoding RNA not only existed, but had important functions in cells. In findings published in Science, he called them “small RNA” or sRNA. Among those he proposed at the time were the microRNAs that have been found since to have a powerful influence over genes’ protein-building functions. 

Guo’s discoveries involve some of the most intricate cellular processes – many of which have opened the door to the development of therapies, particularly for several types of cancers. He is a member of the Ohio State University Comprehensive Cancer Center Translational Therapeutics Program. 

Guo was a pioneer in the field of RNA nanotechnology, first proving RNA strands could self-assemble into a nanoparticle, and since advancing understanding of RNA particles and properties that make them useful vehicles for targeted delivery of anticancer drugs. In 2014, he proposed that RNA therapeutics would be the third milestone in pharmaceutical drug development. 

In 2020, his lab showed in animal studies that RNA nanoparticles have rubbery and amoeba properties that help explain why they target cancer tumors so efficiently and how they keep their toxicity low – by quickly leaving the body. 

He reported in 2021 on a promising liver cancer therapy: an RNA nanoparticle designed to carry both a chemotherapy drug and a molecule to prevent the pumping of the drug out of liver cells. In animal studies, these nanoparticles targeted tumor cells effectively and inhibited tumor growth. 

“RNA can be therapeutic, but it has taken some time for the field to get to pharmaceutical development,” he said. “The mRNA vaccines have opened up the public’s understanding of RNA. At one time, people didn’t believe it was possible – but now we know it is.” 

Guo also points to his first construction of a viral DNA packaging motor. He discovered the third type of biomotors that use a revolving mechanism without rotation to transport the lengthy dsDNA genome. A biomotor is a biological machine that drives movement to help with the energy-related transport of proteins and other molecules – key processes behind such activities as walking and breathing, because transport is essential to the completion of numerous cellular functions. The use of the revolution mechanism without the use of rotation avoids the coiling or tangling during the transportation of the lengthy genome. 

The finding eventually led to applications of this motor to perform “single pore sensing,” including the detection of individual peptides – short chains of amino acids – to advance cancer diagnostics.  

Guo, too, took on new projects at the start of the pandemic, including development of an at-home diagnostic test for COVID-19 infection. An invention of his dating back more than 30 years made an indispensable contribution to the mRNA vaccines against SARS-CoV-2: a novel way of producing a molecule of vaccinia virus mRNA capping enzyme that prevents mRNA from degradation and ensures proper translation of mRNA protein-manufacturing instructions. 

Guo has been a member of Ohio State’s faculty since 2015. He has filed 70 patent applications, with 13 granted and others pending. Most of these technologies have been licensed for commercialization. 

“Of all the recognitions I have received, this is the highest honor in the field of innovation,” Guo said. “I am happy to be recognized as an NAI fellow.” 

The NAI Fellows Program highlights academic inventors who have demonstrated a spirit of innovation in creating or facilitating outstanding inventions that have made a tangible impact on the quality of life, economic development and the welfare of society. 

“This year’s class of NAI fellows represents a truly outstanding caliber of innovators,” said Paul R. Sanberg, president of the NAI. “The breadth and scope of their inventions is truly staggering. I am excited to see their creativity continue to define a new era of science and technology in the global innovation ecosystem.” 

The 2022 class of fellows will be inducted at the annual meeting of the National Academy of Inventors in June 2023 in Washington, D.C.

The review, due to be completed in January 2023, will also consider how to alter the agency’s approach to support appropriate use of opioid painkillers in an expanded public health context, said Patricia Zettler, associate professor in Ohio State’s Moritz College of Law, who is the subject matter lead for the team review.

“The focus of our review is forward-looking,” Zettler said. “How could the FDA use the authority it has to better serve its public health mission, and what additional legal authorities might be useful to improve opioid regulation?” 

An average of 44 people in the United States died each day from prescription opioid painkiller overdoses in 2020, accounting for almost 18% of all opioid overdose deaths that year, according to the Centers for Disease Control and Prevention. The 16,000-plus overdose deaths in 2020 represented a single-year 16% increase in prescription opioid-involved deaths. 

A report on national strategies to address the opioid epidemic issued by the National Academies of Science, Engineering and Medicine (NASEM) in 2017 recommended that the FDA seek this external review. The NASEM report, which was requested by the FDA, generally recommended that the FDA use a comprehensive approach for incorporating public health considerations into the regulatory framework for prescription opioids, including accounting for benefits and risks to patients, households and society as well as incorporating such considerations at the clinical development stage. 

“One of the really challenging things is that these are not products without benefits,” Zettler said. “Prescription opioids have important benefits for a lot of patients under many different circumstances. So there is a need to figure out how to best maximize those benefits while also minimizing the really serious risks of the products.” 

Ohio State is home to a collection of scholars with the right mix of expertise in law, public health, pharmacy and medicine to address the many policy considerations at issue, said review team member Micah Berman, associate professor in the College of Public Health and Moritz College of Law. Both Zettler and Berman have worked at the FDA in the past, and Zettler was a consultant to the NASEM committee that issued the 2017 report. 

“We have the interdisciplinary skills to look at the whole process all the way from the clinical trial to design to some of the post-marketing issues, but also to look at questions of legal authority and how the FDA might want to reconsider the standards it uses for approvals or any new authority that it might need,” Berman said. “It’s a pretty broad mandate to look at all of those things – the process the agency used and the decisions that it came to, and the broader lessons that can be gleaned from the FDA’s past experience regulating opioids – and to approach those questions from the research side, the clinical side and the legal side all at the same time.” 

Ohio State scholars joining Zettler and Berman for the review are Kathy Lancaster, associate professor of epidemiology in the College of Public Health; Macarius Donneyong, assistant professor of outcomes and translational sciences in the College of Pharmacy and College of Public Health; and Marty Fried, clinical assistant professor of internal medicine in the College of Medicine. 

In a recent blog post, FDA Commissioner Robert Califf described the upcoming review. 

“While the FDA’s previous strategies have largely focused on opioid use and overdoses, the evolving nature of the overdose crisis calls for both a new approach and honest reflection about what the FDA can do differently moving forward. … We have initiated this review with the intended goal of understanding what revisions are needed to support appropriate use of opioid analgesics. Our ‘lessons learned’ will actively inform our future approach,” he wrote. 

The opioid crisis was determined in 2017 to be a public health emergency, and that designation is still in effect, Califf wrote, noting that illicit opioids – particularly heroin, fentanyl and compounds with similar chemical structures – are huge contributors to the problem of misuse and overdose deaths. 

For the purposes of this review, the focus will remain on regulatory activities related to existing prescription opioid medications – as well as new pain-management products that will come up for approval in the future. 

“It’s laudable for the government to seek external input on its approach, particularly for a problem this large and this complicated,” Zettler said. “Although there isn’t likely to be one solution, we’re looking forward to using our expertise to help serve the public health and help inform the agency’s regulatory decisions.”

In mouse models of numerous types of cancer, scientists boosted activation of T cells, important fighters in an immune response, inside tumors in a way that improved their interactions with an antibody therapy currently being tested in clinical trials.

The researchers injected nanobodies carrying messenger RNA, molecules that translate genetic information into functional proteins, directly into the tumor site to help T cells generate specific receptors on their surfaces. Experimental monoclonal antibodies delivered six hours later could then bind to those receptors to carry out their cancer cell-killing functions.

The technique left six of 10 mice with lymphoma tumor-free, and was effective in melanoma when combined with additional existing drugs that help amplify the immune response.

“T cells are very important for fighting a lot of diseases – not just cancer – and it’s really difficult to modulate their function,” said Yizhou Dong, senior author of the study and associate professor of pharmaceutics and pharmacology at The Ohio State University.

“After injections of therapeutically relevant mRNA, the T cells decorate their surfaces with receptors, and that enables their additional functions: proliferating, recruiting other immune cells and production of helpful proteins. And when T cells significantly increase those receptors, antibodies can react with the receptors and carry out all the functions we know that interaction can produce.”

The study is published today (Dec. 14, 2021) in the journal Nature Communications.

While increasing T cell activation was the end goal of the research, designing the most effective nanoparticle to carry the messenger RNA was equally important. Dong’s lab has long focused on nanoparticle delivery of messenger RNA as a therapeutic strategy, producing promising results in animal studies against sepsis, genetic disorders and COVID-19.

The team designed nanoparticles for this purpose using one of many compounds that make up cell membranes.

“The idea was inspired by natural components on a cell membrane – we designed compounds that can have nice interactions with the cell membrane and help deliver mRNA into cells. That’s the idea,” Dong said.

Researchers then loaded the nanoparticle cargo: messenger RNA carrying instructions for the production of molecules that T cells express as part of their immune system function. These nanoparticles were injected directly into tumors in mouse models of specific cancers, and entered tumor-infiltrating T cells to amplify their expression of the receptors.

“We waited six hours until the cells produced enough receptors, and then injected antibodies into the tumors. They found their receptors on T cells and that triggered their functions,” said Dong, also an investigator in the Ohio State University Comprehensive Cancer Center.

Tests of the combined treatment produced the best results in mouse models of melanoma and B cell lymphoma. The nanoparticle and antibody delivery completely eliminated tumors in 60% of the mice – a significantly better result than treatment with the antibody alone. The immune response enhancement had staying power, as well: Lymphoma cells injected later into the treated tumor-free mice were unable to survive long enough to form tumors.

“If we used untreated mice, the tumor size increased a lot, but for those mice that had received treatment, it killed the primary tumor and upon rechallenge, tumors cannot grow,” Dong said.

Melanoma proved to be a tougher fight. However, when researchers supplemented the combination treatment with the addition of two antibodies that disrupt cancer cells’ ability to block the immune response, this approach resulted in a 50% complete response in the mice and protection against a later tumor rechallenge. This multi-therapy approach also reduced cancer’s spread in a mouse model of metastasis to the lungs.

Focusing treatment directly at the tumor site is a way of training the immune system to recognize local and circulating cancer cells while lowering the chances for whole-body side effects, Dong said.

The study provided evidence that this technology platform could be used to enhance immunotherapy.

“We want to test more materials and see if we can deliver mRNA to even more T cells to further increase efficiency,” Dong said. “Ultimately, we hope that for certain cancers, this may help produce stronger immune system function by inducing anti-tumor immunity.”

This work was supported by grants from the National Institute of General Medical Sciences, an Ohio State College of Pharmacy startup fund and the Professor Sylvan G. Frank Graduate Fellowship.

Co-authors, all from Ohio State, include Wenqing Li, Xinfu Zhang, Chengxiang Zhang, Jingyue Yan, Xucheng Hou, Shi Du, Chunxi Zeng, Weiyu Zhao, Binbin Deng, David McComb, Yuebao Zhang, Diana Kang, Junan Li and William Carson. Dong is a scientific advisory board member of the biopharmaceutical company Oncorus, Inc. in Cambridge, Massachusetts.

Scientists found that this chemo drug, used to treat many types of solid tumors and blood cancers, is able to enter heart cells by hitchhiking on a specific type of protein that functions as a transporter to move a drug from the blood into heart cells.

By introducing another anti-cancer drug in advance of the chemo, the researchers were able to block the transporter protein, effectively stopping the delivery of doxorubicin to those cardiac cells. This added drug, nilotinib, has been previously found to inhibit activation of other, related transport proteins.

The current findings are based on lab experiments in cell cultures and mice. The researchers are continuing studies with hopes to start designing human trials of the drug intervention later in 2021.

“The proposed intervention strategy that we’d like to use in the clinic would be giving nilotinib before a chemotherapy treatment to restrict doxorubicin from accessing the heart,” said first author Kevin Huang, who graduated in December from The Ohio State University with a PhD in pharmaceutical sciences. “We have pretty solid preclinical evidence that this intervention strategy might work.”

The study is published today (Jan. 25, 2021) in Proceedings of the National Academy of Sciences.

Doxorubicin has long been known for its potential to increase patients’ risk for serious heart problems, with symptoms sometimes surfacing decades after chemo, but the mechanisms have been a mystery. The risk is dose-dependent – the more doses a patient receives, the higher the risk for cardiac dysfunction later in life that includes arrhythmia and a reduction in blood pumped with each contraction, a hallmark symptom of congestive heart failure.

Huang worked in the lab of senior study authors Shuiying Hu and Alex Sparreboom, faculty members in pharmaceutics and pharmacology and members of the Ohio State Comprehensive Cancer Center’s Translational Therapeutics program. This research and other studies targeting different transport proteins to prevent chemo-related nerve pain were also part of Huang’s dissertation.

“Our lab works on the belief that drugs don’t naturally or spontaneously diffuse into any cell they would like to. We hypothesize that there are specialized protein channels found on specific cells that will facilitate movement of internal or external compounds into the cell,” Huang said.

For this work, the team focused on cardiomyocytes, cells composing the muscle behind the heart contractions that pump blood to the rest of the body. The researchers examined cardiomyocytes that were reprogrammed from skin cells donated by two groups of cancer patients who had been treated with doxorubicin – some who suffered cardiac dysfunction after chemo, and others who did not.

The scientists found that the gene responsible for production of the transport protein in question, called OCT3, was highly expressed in the cells derived from cancer patients who had experienced heart problems after treatment with doxorubicin.

“We used mouse models and engineered cell models to demonstrate doxorubicin does transport through this protein channel, OCT3,” Huang said. “We then looked prospectively into what this means from a therapy perspective.”

Blocking OCT3 became the goal once researchers found that genetically modified mice lacking the OCT3 gene were protected from heart damage after receiving doxorubicin. Further studies showed that inhibiting OCT3 did not interfere with doxorubicin’s effectiveness against cancer.

Hu and Sparreboom have specialized in a class of drugs called tyrosine kinase inhibitors, which block specific enzymes related to many cell functions. Nilotinib, a chronic myeloid leukemia drug, is a tyrosine kinase inhibitor that is also known to act on OCT3.

Additional experiments showed that cardiac function was preserved in mice that were pretreated with nilotinib before receiving doxorubicin – and the pretreatment did not interfere with doxorubicin’s ability to kill cancer cells.

The researchers plan to gather additional supporting evidence before pursuing a Phase 1 clinical trial testing the safety of two components of the proposed drug intervention in humans: blocking the function of the OCT3 transporter protein and demonstrating that inhibiting OCT3 in patients treated with doxorubicin protects those patients’ hearts from chemo-induced injury.

This work was supported by the National Institutes of Health, the Robert Bosch Stiftung, the German Research Foundation and Pelotonia funds from Ohio State. Huang was named a Pelotonia Graduate Fellow in 2018.

Additional Ohio State co-authors include Megan Zavorka Thomas, Eric Eisenmann, Muhammad Erfan Uddin, Duncan DiGiacomo, Alexander Pan, Sherry Xia, Yang Li, Yan Jin, Qiang Fu, Alice Gibson, Ingrid Bonilla, Cynthia Carnes, Kara Corps, Vincenzo Coppola, Sakima Smith, Daniel Addison, Ralf Bundschuh, Maryam Lustberg, Moray Campbell, Pearlly Yan and Sharyn Baker.

Scientists manipulated a natural cellular process to ramp up levels of two proteins used by the virus to infect other cells, packaged the protein-boosting instructions in nanoparticles and injected them into mice. Within a month, the mice had developed antibodies against the SARS-CoV-2 virus.

The technique involves altering specific sequences of messenger RNA, molecules that translate genetic information into functional proteins. While these sequences are not translated to proteins, the researchers changed their structures to promote higher-than-usual levels of proteins. The sequences are known as untranslated regions, or UTRs.

“We’ve been engineering messenger RNA for four years, and earlier this year we made some progress identifying a role for UTRs – and then COVID-19 happened,” said Yizhou Dong, senior author of the study and associate professor of pharmaceutics and pharmacology at The Ohio State University.

Though Phase 3 clinical trials of fast-tracked COVID-19 vaccine candidates are in progress, Dong said his lab’s platform offers a potential alternative.

“If the current vaccines work well, that’s wonderful. In case the field needs this, then it’s an option. It worked as a vaccine is expected to, and we can scale this up very fast,” he said. “For now, it’s a proof of concept – we’ve demonstrated we can optimize a sequence of messenger RNA to improve protein production, produce antigens and induce antibodies against those specific antigens.”

The study is published today in the journal Advanced Materials.

The crux of the method is typical to vaccine development: using snippets of a pathogen’s structure to produce an antigen – the foreign substance that triggers an appropriate immune response – and finding a safe way to introduce it to the body.

But the engineering technique takes antigen design to a new level by making use of messenger RNA UTRs, Dong said.

His lab worked with the two UTRs that bookend the start and finish of protein assembly, functioning as regulators of that process and influencing how the resulting protein interacts with others. UTRs themselves are strings of nucleotides, the molecules that compose RNA and DNA.

“For our application we tried to optimize the UTRs to improve the protein production process. We wanted as much protein produced as possible – so we can give a small dose of messenger RNA that produces enough antigen to induce antibodies against the virus,” Dong said.

The team experimented with two potential antigens that the novel coronavirus is known to use to cause infection: a spike protein on its surface and a receptor binding domain, a component of the spike protein, that the virus uses to make its way into host cells – a necessary step to make copies of itself. Both are used in other SARS-CoV-2 vaccine candidates.

After manipulating the messenger RNA for these two proteins, the team encased them in lipid nanoparticles developed previously in Dong’s lab. They injected mice with the experimental vaccine and gave them a booster two weeks later. A month after the first injection, immune cells in the mice had taken up the antigens of the two proteins and developed antibodies against them.

“It takes some time for the immune system to process the antigens and have cells produce antibodies,” Dong said. “In this study, we detected antibodies after 30 days.”

And even if this vaccine candidate is not needed for COVID-19, he is continuing to refine this latest method of engineering messenger RNA.

“UTR is a platform that we can apply to any type of messenger RNA. We are exploring other therapeutics,” Dong said.

This work was funded by a National Institutes of Health Maximizing Investigators’ Research Award, the National Institute of General Medical Sciences and the Ohio State College of Pharmacy startup fund.

Co-authors, all from Ohio State, include Chunxi Zeng, Xucheng Hou, Jingyue Yan, Chengxiang Zhang, Wenqing Li, Weiyu Zhao and Shi Du.

Scientists report today (Aug. 21) in the journal Science Advances that the lipid-based nanoparticles they engineered, carrying two sets of protein-making instructions, showed in animal studies that they have the potential to function as therapies for two genetic disorders.

In one experiment, the payload-containing nanoparticles prompted the production of the missing clotting protein in mice that are models for hemophilia. In another test, the nanoparticles’ cargo reduced the activation level of a gene that, when overactive, interferes with clearance of cholesterol from the bloodstream.

Each nanoparticle contained an applicable messenger RNA – molecules that translate genetic information into functional proteins.

“We demonstrated two applications for lipid-like nanomaterials that effectively deliver their cargo, appropriately biodegrade and are well-tolerated,” said Yizhou Dong, senior author of the study and associate professor of pharmaceutics and pharmacology at The Ohio State University.

“With this work, we have lowered potential side effects and toxicity, and have broadened the therapeutic window. This gives us confidence to pursue studies in larger animal models and future clinical trials.”

This work builds upon a collection of lipid-like spherical compounds that Dong and colleagues had previously developed to deliver messenger RNA. This line of particles was designed to target disorders involving genes that are expressed in the liver.

The team experimented with various structural changes to those particles, effectively adding “tails” of different types of molecules to them, before landing on the structure that made the materials the most stable. The tiny compounds have a big job to do: embarking on a journey through the bloodstream, carrying molecules to the target location, releasing the ideal concentration of messenger RNA cargo at precisely the right time and safely degrading.

The tests in mice suggested these particles could do just that.

The researchers injected nanoparticles containing messenger RNA holding the instructions to produce a protein called human factor VIII into the bloodstream of normal mice and mouse models for hemophilia. A deficiency of this protein, which enables blood to clot, causes the bleeding disorder. Within 12 hours, the deficient mice produced enough human factor VIII to reach 90 percent of normal activity. A check of the organs of both protein-deficient mice and normal mice showed that the treatment caused no organ damage.

“It can be helpful to think of this as a protein-replacement therapy,” Dong said.

In the second experiment, nanomaterials were loaded with two types of instructions: messenger RNA carrying the genetic code for a DNA base editor, and a guide RNA to make sure the edits occurred in a specific gene in the liver called PCSK9. Dozens of mutations that increase this gene’s activity are known to cause high cholesterol by reducing clearance of cholesterol from the bloodstream.

Analyses showed that the treatment resulted in the intended mutation of about 60 percent of the target base pairs in the PCSK9 gene, and determined that only a low dose was needed to produce high editing effect.

Dong credited academic and industry partners for helping advance this work. Co-corresponding authors include Denise Sabatino of Children’s Hospital of Philadelphia and Delai Chen from Boston-based Beam Therapeutics, who provided expertise in hemophilia and DNA base editing, respectively.

Dong and first author Xinfu Zhang are inventors on patent applications filed by Ohio State related to the lipid-like nanoparticles. This technology has been licensed for further clinical development.

This work was supported by the National Institute of General Medical Sciences, the National Heart, Lung and Blood Institute, and a startup fund from Ohio State’s College of Pharmacy.

Additional co-authors are Giang N. Nguyen of Children’s Hospital of Philadelphia; Weiyu Zhao, Chengxiang Zhang, Chunxi Zeng, Jingyue Yan, Shi Du, Xucheng Hou, Wenqing Li, Justin Jiang, Binbin Deng and David McComb of Ohio State; and Robert Dorkin, Aalok Shah, Luis Barrera, Francine Gregoire and Manmohan Singh of Beam Therapeutics.

The scientists used nanotechnology to transform donated healthy immune cells into a drug with enhanced power to kill bacteria.

In experiments treating mice with sepsis, the engineered immune cells eliminated bacteria in blood and major organs, dramatically improving survival rates.

This work focuses on a treatment for late-stage sepsis, when the immune system is compromised and unable to clear invading bacteria. The scientists are collaborating with clinicians specializing in sepsis treatment to accelerate the drug-development process.

“Sepsis remains the leading cause of death in hospitals. There hasn’t been an effective treatment for late-stage sepsis for a long time. We’re thinking this cell therapy can help patients who get to the late stage of sepsis,” said Yizhou Dong, senior author and associate professor of pharmaceutics and pharmacology at The Ohio State University. “For translation in the clinic, we believe this could be used in combination with current intensive-care treatment for sepsis patients.”

The study is published today (Jan. 6, 2020) in Nature Nanotechnology.

Sepsis itself is not an infection – it’s a life-threatening systemic response to infection that can lead to tissue damage, organ failure and death, according to The Centers for Disease Control and Prevention. The CDC estimates that 1.7 million adults in the United States develop sepsis each year, and one in three patients who die in a hospital has sepsis.

This work combined two primary types of technology: using vitamins as the main component in making lipid nanoparticles, and using those nanoparticles to capitalize on natural cell processes in the creation of a new antibacterial drug.

Cells called macrophages are one of the first responders in the immune system, with the job of “eating” invading pathogens. However, in patients with sepsis, the number of macrophages and other immune cells are lower than normal and they don’t function as they should.

In this study, Dong and colleagues collected monocytes from the bone marrow of healthy mice and cultured them in conditions that transformed them into macrophages. (Monocytes are white blood cells that are able to differentiate into other types of immune cells.)

The lab also developed vitamin-based nanoparticles that were especially good at delivering messenger RNA, molecules that translate genetic information into functional proteins.

The scientists, who specialize in messenger RNA for therapeutic purposes, constructed a messenger RNA encoding an antimicrobial peptide and a signal protein. The signal protein enabled the specific accumulation of the antimicrobial peptide in internal macrophage structures called lysosomes, the key location for bacteria-killing activities.

From here, researchers delivered the nanoparticles loaded with that messenger RNA into the macrophages they had produced with donor monocytes, and let the cells take it from there to “manufacture” a new therapy.

“Macrophages have antibacterial activity naturally. So if we add the additional antibacterial peptide into the cell, those antibacterial peptides can further enhance the antibacterial activity and help the whole macrophage clear bacteria,” Dong said.

After seeing promising results in cell tests, the researchers administered the cell therapy to mice. The mouse models of sepsis in this study were infected with multidrug-resistant Staphylococcus aureus and E. coli and their immune systems were suppressed.

Each treatment consisted of about 4 million engineered macrophages. Controls for comparison included ordinary macrophages and a placebo. Compared to controls, the treatment resulted in a significant reduction in bacteria in the blood after 24 hours – and for those with lingering bacteria in the blood, a second treatment cleared them away.

Dong considers the lipid nanoparticle delivery of messenger RNA into certain kinds of immune cells applicable to other diseases, and his lab is currently working on development of cancer immunotherapy using this technology.

This work was supported by the Ohio State College of Pharmacy start-up fund, a National Institute of General Medical Sciences Maximizing Investigators’ Research Award and Fundamental Research Funds for Chinese Central Universities.

Co-authors, all from Ohio State, include Xucheng Hou, Xinfu Zhang, Weiyu Zhao, Chunxi Zeng, Binbin Deng, David McComb, Shi Du, Chengxiang Zhang and Wenqing Li.

Ohio State will seek approval for the $155.9 million health sciences center during next week’s Board of Trustees meeting. Funding sources are to be determined. The project is poised to benefit colleges, including medicine, dentistry, nursing, optometry, pharmacy, public health and veterinary medicine, and strengthen the interdisciplinary curriculum of Ohio State.

“A healthier future demands health care leaders who are prepared to work together to address health challenges,” said Bruce A. McPheron, Ohio State’s executive vice president and provost. “Ohio State’s new health sciences education building is the place where students will become those leaders.”

The multiphase project advances Framework 2.0, the university’s long-term planning vision. It includes a 150,000-square-foot renovation of Hamilton Hall and construction of a new 100,000-square-foot building featuring flexible facilities to serve multiple disciplines. Program space will include:

• 33 interdisciplinary, state-of-the-art classrooms, each with new teaching technology, including virtual reality and enhanced anatomy and surgical practice labs
• 6,000-square-foot informal learning space
• 125-seat grand reading room

“Ohio State is in the process of creating a new model to replace the 100-year-old-plus model of academic medicine in use today,” said Dr. Hal Paz, executive vice president and chancellor for health affairs at Ohio State and Wexner Medical Center CEO. “This new academic health model takes advantage of the latest technology and brightest minds to educate professionals prepared to address the changing health needs of diverse populations. This is the key to building healthier communities.”

The Interdisciplinary Health Sciences Center will focus on inclusive work environments to create:

• Spaces for students in the health sciences to convene conversations and build team approaches to health care — a way to quickly assess patients’ needs and work with the right colleagues to design the best care plans for each individual.
• Opportunities for students to be at the forefront of innovation through modern technology — whether by using realistic virtual reality models to simulate patient care or learning how video chats, texting and other emerging “telehealth” innovations can help health care providers and patients stay in regular communication between in-person office visits.
• Warm and welcoming environments that support Ohio State’s work to build diverse communities of medical students. Research has shown that diversity in medicine — including increasing representation of women and people of color — leads to better health outcomes.

Design for the new Interdisciplinary Health Sciences Center began in August 2018. Pending board approval, construction could begin in October 2019, with completion targeted for late summer 2023.

Progress continues on a separate Health Sciences Faculty Office and Optometry Clinic at 11th and Neil avenues. The 106,000-square-foot building will be home to optometry clinics, retail spaces and faculty offices. The $35.9 million project is 60% of the way through steel work and targeting an October 2020 opening.

For more information about these projects and other major initiatives advancing Framework 2.0, visit Time and Change: Building the Future.