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Tag: Lieberman

Wistar-Led Team Awarded More Than $12 Million Grant from the NCI to Investigate Link Between Epstein-Barr Virus and Carcinomas

PHILADELPHIA — (JULY 26, 2023) — It’s been known since the 1960s that Epstein-Barr Virus (EBV) causes a variety of cancers, but research has overwhelmingly focused on its connection to lymphomas. Now, a multidisciplinary team of scientists led by The Wistar Institute has been awarded a more than $12 million National Cancer Institute (NCI) Program Project Grant (P01), a highly competitive five-year grant that includes a cross section of researchers from various disciplines and institutions throughout the country. The multidisciplinary team led by Wistar scientists is exploring the role of Epstein-Barr Virus in epithelial cancers. Epithelial cells form functional structures in organ tissue throughout the human body; they are often the site for solid organ cancers, including the most common cancers, which are known as carcinomas.

The new research will focus on basic questions about how EBV infection of normal epithelial cells transforms them into cancer-cells. Scientists also intend to build on this research to identify better and more selective therapeutic targets.

“We are investigating unexplored aspects of EBV and malignancies, potentially uncovering unique characteristics or pathways that can be targeted for therapeutic intervention,” said Italo Tempera, Ph.D., associate professor of the Gene Expression & Regulation Program of the Ellen and Ronald Caplan Cancer Center at The Wistar Institute. “This fresh perspective could lead to groundbreaking discoveries and innovative treatment strategies for EBV and epithelial malignancies.”

The project brings together scientists from The Wistar Institute and Harvard University, including experts in epigenetics, metabolomics and drug discovery. It’s the first time researchers from this variety of disciplines have combined their efforts to focus entirely on the EBV-epithelial cancer link.

“We’ve put together a new strategy, a new way of attacking the problem,” said Paul Lieberman, Ph.D., Hilary Koprowski, M.D., Endowed Professor and director of the Center for Chemical Biology and Translational Medicine at Wistar. “By working together across different modalities, there’s an opportunity for each of us to learn from the synergy and expertise of the other investigators.”

EBV is one of the most common human viruses, infecting an estimated 95% of people by the time they reach adulthood. Symptoms are usually mild, and most people recover within a few weeks. However, the virus can remain latent in the human body for years or even decades, and it causes some people to develop cancer later in life.

While research has historically focused on lymphomas, EBV-linked epithelial cancers are both more common and more deadly. Epithelial cancers represent 75% of the 200,000 EBV-related cancer cases diagnosed each year, and these cancers also have higher mortality rates and treatment failures.

“This grant put together a team that is now focused on this type of cancer that has been neglected, even though it’s the most common form of EBV cancers,” Lieberman said. The grant will fund three main research projects. The first will look at how EBV establishes a long-term infection within epithelial cells. The second will study how it causes genetic and metabolic changes to trigger cancer growth. Finally, researchers will use these findings to investigate new therapeutic strategies.

The research builds on past work by Lieberman’s lab, which has focused on developing small molecule inhibitors targeting EBV. He said the new project would focus on studying drugs that are already in development, and looking for ways to make them more targeted or use them in combination with other therapies.

Tempera said the group’s integrated approach sets it apart.“Our project will study both metabolic and epigenetic vulnerabilities simultaneously,” he said. “Combining these two aspects can provide a comprehensive understanding of the role of EBV infection in cancer and its underlying mechanisms, leading to unique insights and therapeutic opportunities.”

Co-authors: Ben Gewurz of Harvard; Joseph Salvino, Samantha Soldan, Andrew Kossenkov, Louise Showe, and Qin Liu of Wistar.

Unraveling the Ties That Bind: Epstein-Barr Virus and Multiple Sclerosis

A conversation with the Lieberman Lab delves into how EBV can trigger MS and potential therapeutic solutions that can be developed with this knowledge.

Epstein-Barr virus (EBV) is ubiquitous, establishing lifelong infection found throughout the world. It targets the immune system’s B cells and typically remains silent in immune system memory cells. Though infection with the virus is largely asymptomatic, specific biological and environmental conditions can enable the virus to cause more serious diseases. For instance, the virus can cause rare cancers that occur at much higher rates in immunosuppressed individuals. More recent research has found a connection between EBV and the neurodegenerative disease, multiple sclerosis (MS).

In a recent article published in Nature Reviews Microbiology, Paul Lieberman, Ph.D., Hilary Koprowski, M.D., Endowed Professor; program leader, Gene Expression & Regulation Program, Ellen and Ronald Caplan Cancer Center; and director, Center for Chemical Biology & Translational Medicine; and Samantha Soldan, Ph.D., staff scientist in the Lieberman laboratory, review evidence of EBV as a cause of MS and the implications of this knowledge in research and clinical spheres.

“Eliminating EBV latent infection should be a safe and effective way to treat EBV cancers and autoimmune disease, especially multiple sclerosis,” Lieberman says.

We spoke with Soldan about the link between EBV and MS as well as how this knowledge can be harnessed into potential therapeutics for the disease.

Q: What is some of the background linking EBV to cancer?

A: EBV was the first virus to be implicated as a causative agent of human cancer. EBV has been linked to nasopharyngeal cancers, stomach cancers, primary CNS lymphomas, Hodgkin’s and non-Hodgkin’s lymphomas, NK/T-cell lymphomas, and leiomyosarcomas, as well as several autoimmune disorders, including MS. It is currently estimated that 1.5-2% of all human cancers are attributable to EBV infection. All EBV-related cancers are associated with latent infection, where no infectious virus is produced by the tumor cells. Different types of EBV-associated cancers express different EBV latency genes. Notably, all EBV cancers express the viral protein EBNA1—required to maintain the EBV genome in latently infected cells and a primary interest of the Lieberman lab.

Q: What inspired this review to investigate evidence behind EBV as a cause of MS?

A: The case for EBV as a causative agent in MS has been mounting over the last 40 years. This year, two landmark studies were published: one providing a strong epidemiologic link to MS and the other suggesting a mechanism by which EBV may drive disease pathogenesis. These two studies have intensified the MS community’s interest in the link between EBV and MS, both as a disease trigger and as a potential driver of disease pathogenesis.

Dr. Paul Lieberman is a leading expert in EBV and in the study of EBV EBNA1 specifically, and he was invited to write a review article focused on the role of EBV in MS for Nature Reviews Microbiology. Paul asked me to be a co-author for this manuscript and I was delighted to have this opportunity. My background is in neurovirology, and I have been involved in research investigating the relationship between viruses and neuroinflammatory diseases, including MS, since graduate school. I am deeply invested in trying to better understand the role that EBV plays in MS.

Q: Why is multiple sclerosis a complex disease to study?

A: Multiple sclerosis is the most common demyelinating disorder of young adults, effecting more than one million individuals in the U.S. alone. MS is a heterogenous and often disabling disease that develops because of the interplay between the immune system and the environment in genetically susceptible individuals. The clinical progression of MS is variable and unpredictable with several distinct disease courses. In addition, MS patients often transition from a relapsing-remitting to a progressive disease course over time and the mechanism of the disease and central nervous system damage also evolve, which are a hallmark of MS.

Further complicating matters, epidemiological studies have shown that environmental factors (including EBV infection) that contribute to one’s risk of developing MS often occur many years before clinical onset. Collectively, these complex interactions between genetic, immunologic, and environmental risk factors makes attributing disease-contributing agents and designing preventative measures and effective therapies for MS very challenging.

Q: How does EBV trigger MS? Can you explain these processes?

A: How a ubiquitous agent like EBV triggers disease in a small percentage of those who are infected is an enigma. We face this challenge in understanding the role of EBV in cancer as well as MS. For MS and many autoimmune diseases, we can identify inflammatory and autoreactive immune responses and characterize immune responses to infectious agents and antigens that trigger immune response in patients. However, understanding what set immune cells on an autoreactive and inflammatory path before the patient is symptomatic is a difficult task.

Nevertheless, there are several theories as to how EBV may be both a trigger and a driver of MS and we discuss these in the review. We believe that it is likely that EBV is involved in the pathogenesis of MS at many levels and in different anatomic compartments.

Q: What are some potential therapies that could arise out of understanding the role of EBV in MS?

A: In recent years, therapies depleting B-cells have proven to be tremendously beneficial in MS. While EBV primarily infects B-cells, these B-cell depletion therapies eliminate cells regardless of whether they are infected by EBV, making it difficult to determine if any of the clinical benefit derived from these drugs is related to their effects on the virus.

There are several EBV specific therapies in development that have the potential to present new, effective options for patients with MS. In addition, they may also help us better understand the role of EBV as a trigger and driver of disease pathogenesis. These include vaccines to prevent the development of infectious mononucleosis, MS, and EBV-associated cancers; cell-based immunotherapies, including EBV-specific cytotoxic T cell lines; and EBV specific antivirals.

Q: Why is it important to review existing literature and reveal new directions for research?

A: I find review articles to be incredibly helpful, both to the writer and the reader. When writing a review, you must commit time and energy to refamiliarizing yourself with the latest literature as well as the history of the field. The process helps you take the proverbial 30,000-foot view and see the whole picture, forcing you to get your head out of the specific aspect of research that you are generally focused on and enabling you to generate new ideas and consider new approaches to your work. Reviews are also very important for colleagues and especially trainees to get perspective on where the field is headed and where there are gaps in our knowledge.

Q: What do you currently work on with Dr. Lieberman regarding MS and EBV and where is your research headed?

A: We are working in several directions to better understand the role that that EBV plays in the pathogenesis of MS. Our current work focuses on characterizing virus-host interactions and maintenance of EBV latency in EBV infected B-cells from MS patients compared to healthy controls. Dr. Chenhe Su, a postdoctoral fellow in the Lieberman lab, is also working very hard on these studies.

I am especially interested in developing better animal models to understand how host factors like age of exposure, genetic background, metabolism, and sex influence host-virus interactions and EBV reprogramming of B-cells. We are also testing EBV-specific antiviral therapies, including the EBNA1 inhibitor developed in the Lieberman lab, to determine its potential as a therapeutic agent for use in MS.

Viruses and Cancer

Viruses impact human health in many ways and can be the underlying agents that cause cancer.

The pandemic has reminded us how pervasive viruses are in human life. We know they can cause infection, making us very sick, but did you know that about 15 percent of all human cancers worldwide may be attributed to viruses?

Drs. Paul Lieberman, professor, and Italo Tempera, associate professor, both in the Gene Expression & Regulation Program of The Wistar Institute Cancer Center, investigate the link between the Epstein-Barr Virus (EBV) and malignant transformation to find ways to treat EBV-related cancers.

EBV is a very common virus. You know it as mononucleosis, and it is spread through saliva, but also through sexual contact, blood transfusions and organ transplants. It mostly infects B cells, a type of white blood cells that produces antibodies. The majority of the world population is infected and carries the virus in a silent state for life. However, in people with compromised immune systems, EBV increases the risk of B-cell lymphomas including Burkitt’s lymphoma and Hodgkin’s lymphoma.

Since it can also infect other types of cells, this virus is not only associated with blood tumors, but also with ten percent of all gastric cancers and most cancers of the nasopharynx, the region behind the nose and above the back of the throat.

How are viruses implicated in cancer?

They are particles made up of DNA or RNA surrounded by a protein coat. With such frugality comes the lack of the necessary machinery to replicate their genome or build proteins, so viruses hijack a host cell and use its machinery to make copies of themselves. In the case of oncogenic or cancer-causing viruses, this process can meddle with the cell’s genes and derail the mechanisms that keep cell proliferation in check, leading to uncontrolled growth.

Oncogenic viruses establish chronic but latent infections that don’t cause obvious symptoms in healthy individuals, because their immune system keeps infection at bay. In immunocompromised people, however, viruses are more likely to cause malignancies.

One approach to control EBV infection is by interfering with the complex and dynamic patterns of how viral genes are switched on or off — a process known as gene expression. The Tempera lab has explored the role of gene expression in the regulation of EBV latency, unveiling some viral and host cell factors that play key roles in regulating these patterns.

An EBV protein called latent membrane protein 1 (LMP1) is essential for the virus’s ability to make B cells cancerous.

The Tempera lab studies just how this protein affects the host cells. They discovered that LMP1 affects the function of an enzyme called poly(ADP-ribose) polymerase 1 (PARP1) and that inhibiting this protein suppresses malignant transformation, uncovering an important role of PARP1 in EBV-induced oncogenesis.

In a recent paper1, Tempera and colleagues demonstrated that LMP1 causes a switch in how infected cells produce the building blocks of fat that support tumorigenesis and cancer progression. Researchers also showed that targeting these metabolic changes could be an effective therapeutic strategy to treat EBV-associated cancer and that PARP1 inhibitors offset the metabolic changes caused by LMP1 that drive tumorigenesis.

“We have uncovered a potential new use for PARP inhibitors, which are currently used to treat recurrent ovarian cancers in women who have defects in their DNA repair system,” said Dr. Tempera. “Repurposing existing drugs saves time and money in the process of creating new therapies.”

Dr. Tempera joined Wistar as an associate professor in 2020, but that was not his first time working at the Institute. He had trained as a postdoctoral fellow in the Lieberman lab, which helped him launch his career in cutting-edge research on the epigenetic mechanisms underlying EBV infection. Returning to Wistar with his lines of research as an established investigator, he strengthened our program on virology and cancer.

“The outstanding scientific environment and technological support that benefited me during my training are a strong asset to further expand my research program.”

The lab of his mentor Dr. Lieberman is a reference point for EBV research as they have made seminal discoveries in the field and described several mechanisms that control replication and gene expression in latent EBV infection. One focus of their research is the Epstein-Barr nuclear antigen 1 (EBNA1) protein that is essential for efficient viral DNA replication. The team has been pursuing the development of small molecule inhibitors of EBNA1 as potential treatment against EBV-associated malignancies.

One of the lab’s latest studies2 led to an important basic discovery in the field, expanding the understanding of EBNA1’s function and providing new possibilities for inhibiting EBNA1 activity as an anticancer strategy.

The link between virus infection and human cancer is a complicated matter to unravel and has been studied for more than 100 years. The research underway in the Tempera and Lieberman labs is crucial to understanding the role viruses play in malignant transformation and hopefully finding future treatments to halt this process.

1 Epstein-Barr Virus-Encoded Latent Membrane Protein 1 and B-Cell Growth Transformation Induce Lipogenesis through Fatty Acid Synthase, J Virol, 2021
2 Cell-cycle-dependent EBNA1-DNA crosslinking promotes replication termination at oriP and viral episome maintenance, Cell, 2021

Wistar Scientists Make Pivotal Discovery on the Mechanism of Epstein-Barr Virus Latent Infection

PHILADELPHIA — (Jan. 21, 2021) — Researchers at The Wistar Institute have discovered a new enzymatic function of the Epstein-Barr Virus (EBV) protein EBNA1, a critical factor in EBV’s ability to transform human cells and cause cancer. Published in Cell, this study provides new indications for inhibiting EBNA1 function, opening up fresh avenues for development of therapies to treat EBV-associated cancers.

EBV establishes life-long, latent infection in B lymphocytes, which can contribute to development of different cancer types, including Burkitt’s lymphoma, nasopharyngeal carcinoma (NPC) and Hodgkin’s lymphoma.

The Epstein-Barr Nuclear Antigen 1 (EBNA1) serves as an attractive therapeutic target for these cancers because it is expressed in all EBV-associated tumors, performs essential activities for tumorigenesis and there are no similar proteins in the human body.

“We discovered an enzymatic activity of EBNA1 that was never described before, despite the intense research efforts to characterize this protein,” said Paul M. Lieberman, Ph.D., Hilary Koprowski, M.D., Endowed Professor, leader of the Gene Expression & Regulation Program at Wistar, and corresponding author of the study. “We found that EBNA1 has the cryptic ability to cross-link and nick a single strand of DNA at the terminal stage of DNA replication. This may have important implications for other viral and cellular DNA binding proteins that have similar cryptic enzyme-like activities.”

Lieberman and colleagues also found that one specific EBNA1 amino acid called Y518 is essential for this process to occur and, consequently, for viral DNA persistence in the infected cells.

They created a mutant EBNA1 protein in which the critical amino acid was substituted with another and showed that this mutant could not form covalent binding with DNA and perform the endonuclease activity responsible for generating single strand cuts.

In latently infected cells, the EBV genome is maintained as a circular DNA molecule, or episome, that is replicated by cellular enzymes along with the cell’s chromosomes. When the cell divides, the two episomal genomes segregate into the two daughter cells.

While it was known that EBNA1 mediates replication and partitioning of the episome during division of the host cell, the exact mechanism was not clear. The new study sheds light on the process and describes how the newly discovered enzymatic activity of EBNA1 is required to complete replication of the viral genome and maintenance of the episomal form.

“Our findings suggest that one could create small molecules to ‘trap’ the protein bound to DNA and potentially block replication termination and episome maintenance, similar to known inhibitors of topoisomerases,” said Jayaraju Dheekollu, Ph.D., first author on the study and staff scientist in the Lieberman Lab. “Such inhibitors may be used to inhibit EBV-induced transformation and treat EBV-associated malignancies.”

Co-authors: Andreas Wiedmer, Kasirajan Ayyanathan, Julianna S. Deakyne, and Troy E. Messick from The Wistar Institute. K.A. is currently employed at University of Pennsylvania and J.S.D. is currently employed at GlaxoSmithKline.

Work supported by: National Institutes of Health (NIH) grants RO1 CA093606, RO1 423 DE017336, P30 CA010815, and T32 CA09171.

Publication information: Cell Cycle-Dependent EBNA1-DNA Cross-Linking Promotes Replication Termination at oriP and Viral Episome Maintenance, Cell (2021). Online publication.

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The Wistar Institute is an international leader in biomedical research with special expertise in cancer, immunology, infectious disease research, and vaccine development. Founded in 1892 as the first independent nonprofit biomedical research institute in the United States, Wistar has held the prestigious Cancer Center designation from the National Cancer Institute since 1972. The Institute works actively to ensure that research advances move from the laboratory to the clinic as quickly as possible. wistar.org.

Spotlight on Wistar COVID-19 Researcher: Paul Lieberman, Ph.D.

Creating a new drug from scratch takes many years and billions of dollars. While discovery of new molecules against SARS-CoV-2 and other emerging viruses is imperative, the world also needs solutions for COVID-19 now. This is why there has been wide interest in repurposing existing drugs, for which safety and pharmacologic profiles are already available, as a viable strategy to save critical time and resources and quickly discover potential candidate drugs to combat the pandemic. 

Dr. Paul Lieberman is a molecular virologist in Wistar’s Cancer Center, better known for his pioneering studies on how certain viruses persist in the body in a latent, long-term infection that can lead to cancer. He leads a talented  drug-discovery team that has created first-in-class molecules directed against  Epstein-Barr virus (EBV) as a novel therapeutic approach for potentially treating EBV-associated cancers including Burkitt’s lymphoma, nasopharyngeal carcinoma and Hodgkin’s lymphoma.

Applying their antiviral drug discovery expertise and innovative methods to “drug” the RNA component of viruses and cancer cells, the Lieberman lab is now working on a drug-repurposing project to quickly identify FDA-approved molecules that trap and inactivate SARS-CoV-2 RNA, the genetic material that carries the life information for the virus, just like DNA does in our cells. 

SARS-CoV-2 RNA forms unique three-dimensional structures known as pseudoknots that are essential for viral replication and for the ability of the SARS-CoV-2 virus to cause disease. RNA pseudoknots may be a promising new target for therapeutic intervention. 

Working closely with the Molecular Screening Facility, which provides state-of-the-art technologies and industry standard expertise, Dr. Lieberman and his team rapidly set up a high-throughput, novel screening assay based on identifying molecules that bind to the SARS-CoV-2 pseudoknot structures and disrupt their function. His team is in the process of evaluating thousands of FDA-approved small molecule drugs. 

The lead candidates identified in the screen will then be further tested in cells and in preclinical models for their ability to stop infection and disease progression, and eventually to advance the best molecules into the clinic.

“We are taking a new, very focused approach to evaluate existing molecules,” said Dr. Lieberman. “This allows us to look for a very specific activity to deploy against the virus in molecules that have already tested safe in humans.”

Because RNA pseudoknot structures are similar in other coronaviruses, the outcome of this project could also have broader applications against other respiratory diseases.