Study redefines HPV-related head and neck cancers

Much of what we thought we knew about the human papilloma virus (HPV) in HPV-related head and neck cancers may be wrong, according to a newly published study by Virginia Commonwealth University (VCU) researchers that analyzed data from The Human Cancer Genome Atlas. Head and neck cancers involving HPV are on the rise, and many experts believe we are seeing the start of an epidemic that will only get worse in the coming years.

The Cancer Genome Atlas is a collaboration between the National Cancer Institute (NCI) and the National Human Genome Research (NHGR) Institute that makes publicly available genomic information on tumor samples from 33 different types of cancers. Its aim is to help the cancer research community improve the prevention, diagnosis and treatment of cancer.

It is thought that there are two main forms of HPV-related cancers, episomal and integrated. In episomal variants, the HPV genome replicates independently. Integrated HPV has become part of the DNA of the host cell and relies on it for replication. Previously, it was believed that most HPV-related head and neck cancers had integrated HPV, as is what is believed with HPV-related cervical cancers. However, Windle’s study, recently published in the journal Oncotarget, found that HPV DNA is maintained separate from the human genome in the majority of HPV-related head and neck cancers, though, in many cases, the HPV genome can acquire a small piece of human DNA making it look like integrated HPV. This viral-human hybrid represents a new category of episomal HPV in HPV-related cancers.

“Our work challenges the idea that finding HPV DNA joined to human DNA means that HPV is integrated. With this new view of the state of HPV, we conclude that episomal HPV is the predominant state in HPV-related head and neck cancers,” says Brad Windle, member of the Cancer Molecular Genetics research program at VCU Massey Cancer Center, professor at the Philips Institute for Oral Health Research at the VCU School of Dentistry and co-principle investigator on the study. “This is an important distinction because patients with episomal HPV cancer respond better to therapy than patients with integrated HPV cancer.”

Windle’s team analyzed the genomes of all 520 HNC samples in The Cancer Genome Atlas and found that 72 were HPV positive. The large majority of these cancers had a common type of the virus known as HPV16 present, so they focused on that virus type. The data showed that 75 percent of the HPV16 samples had the HPV genome in the episomal state, and about half of the genomes contained a piece of human DNA within their circular structure.

The researchers also found that 73 percent of the tumor samples were still dependent on proteins known as E1 and E2 for replication. This is important because when the HPV genome integrates with human DNA, expression of the HPV E2 protein–essential for independent replication–is lost. The presence of E2, or lack thereof, in tumor biopsies could be a reliable way for physicians to determine the cancer type and provide a more accurate prognosis.

“Perhaps our most striking outcome is the potential to target the E1 and E2 proteins for diagnosis and treatment,” says Windle. With nearly three quarters of these cancers dependent on E1 and E2 for replication, we could develop drugs that target these proteins and promote cell death.”

Windle’s team plans to continue studying the integration of HPV in HPV-related head and neck cancers, and suggests that viral-human DNA hybrid HPV should be further explored in HPV-related cervical cancers. His team is currently working with Massey clinicians in order to use this information to assess patients’ prognosis in the clinic.

Epilepsy drug therapies to be improved by new targeted approach

New research from the University of Liverpool, in collaboration with the Mario Negri Institute in Milan, published today in the Journal of Clinical Investigation, has identified a protein that could help patients with epilepsy respond more positively to drug therapies.

Epilepsy continues to be a serious health problem and is the most common serious neurological disease. Despite 30 years of drug development, approximately 30% of people with epilepsy do not become free of fits (also called seizures) with currently available drugs.

New, more effective drugs are therefore required for these individuals. We do not fully understand why some people develop seizures, why some go onto develop epilepsy (continuing seizures), and most importantly, why some patients cannot be controlled with current drugs.

Inflammation

There is now increasing body of evidence suggesting that local inflammation in the brain may be important in preventing control of seizures. Inflammation refers to the process by which the body reacts to insults such as having a fit. In most cases, the inflammation settles down, but in a small number of patients, the inflammation continues.

The aim of the research, undertaken by Dr Lauren Walker while she was a Medical Research Council (MRC) Clinical Training Fellow, was to address the important question of how can inflammation be detected by using blood samples, and whether this may provide us with new ways of treating patients in the future to reduce the inflammation and therefore improve seizure control.

The research focused on a protein called high mobility group box-1 (HMGB1), which exists in different forms in tissues and bloodstream (called isoforms), as it can provide a marker to gauge the level of inflammation present.

Predicting drug response

The results showed that there was a persistent increase in these isoforms in patients with newly-diagnosed epilepsy who had continuing seizure activity, despite anti-epileptic drug therapy, but not in those where the fits were controlled.

An accompanying drug study also found that HMGB1 isoforms may predict how an epilepsy patient’s seizures will respond to anti-inflammatory drugs.

Dr Lauren Walker, said: “Our data suggest that HMGB1 isoforms represent potential new drug targets, which could also identify which patients will respond to anti-inflammatory therapies. This will require evaluation in larger-scale prospective trials.”

Innovative scheme

Professor Sir Munir Pirmohamed, Director of the MRC Centre for Drug Safety Science and Programme lead for the MRC Clinical Pharmacology scheme, said: “The MRC Clinical Pharmacology scheme is a highly successful scheme to train “high flyers” who are likely to become future leaders in academia and industry.

“Dr Walker’s research is testament to this and shows how this innovative scheme, which was jointly funded by the MRC and Industry, can tackle areas of unmet clinical need, and identify new ways of treating patients with epilepsy using a personalised medicine approach”.

Stem Cell Trial for Stroke Patients Suffering Chronic Motor Deficits Begins at UTHealth

A clinical trial to evaluate the safety and efficacy of a stem cell product injected directly into the brain to treat chronic motor deficits from ischemic stroke has begun at McGovern Medical School at The University of Texas Health Science Center at Houston (UTHealth).

McGovern Medical School at UTHealth is the only site in Texas and the central south portion of the country to open enrollment for the multi-institutional, phase 2B study – the first in the U.S. for chronic stroke. Surgeries will be conducted at Memorial Hermann-Texas Medical Center.

“This trial is one of the first randomized, sham-controlled studies to test the efficacy of administering adult-derived stem cells in patients disabled with a chronic stroke,” said Sean I. Savitz, M.D., professor and the Frank M. Yatsu Chair in Neurology at McGovern Medical School and director of the UTHealth Institute for Stroke and Cerebrovascular Disease. “We were chosen as one of only a handful of referral centers in the nation and patients from all over the country will be referred to our center for this trial. Overall, the study adds to our growing regenerative medicine program for patients with neurological disorders.”

In the double-blind, sham-surgery controlled study, patients randomized to the study intervention will receive a stem cell product made by SanBio and patients must have chronic motor deficits from an ischemic stroke to be eligible for the study. The product, administered through tiny holes bored into the skull and placed near the site of the damage, came from the bone marrow of two healthy adult donors. Enrollment is limited to patients who are between six and 60 months post-stroke and have a chronic motor neurological deficit.

Results of a phase 1/2A study of the stem cell product, presented at the International Society of Stem Cell Research Meeting and published in the journal, Stroke, showed statistically significant improvements in motor function and no safety concerns.

The UTHealth Stroke Program at McGovern Medical School, led by Savitz, is one of the most active research and clinical programs in the country. It was one of the lead sites in the National Institute of Neurological Disease and Stroke’s (NINDS) tPA stroke study; was one of eight centers in the country funded by the NIH to conduct specialized translational research to develop novel acute stroke therapies; and receives NINDS fellowship funding to train the next generation of academic leaders in cerebrovascular disease.

Trigger for autoimmune disease identified

Researchers at National Jewish Health have identified a trigger for autoimmune diseases such as lupus, Crohn’s disease and multiple sclerosis. The findings, published in the April 2017 issue of Journal of Clinical Investigation, help explain why women suffer autoimmune disease more frequently than men, and suggest a therapeutic target to prevent autoimmune disease in humans.

“Our findings confirm that Age-associated B Cells (ABCs) drive autoimmune disease,” said Kira Rubtsova, PhD, an instructor in biomedical science at National Jewish Health. “We demonstrated that the transcription factor T-bet inside B cells causes ABCs to develop. When we deleted T-bet inside B cells, mice prone to develop autoimmune disease remained healthy. We believe the same process occurs in humans with autoimmune disease, more often in elderly women.”

Autoimmune diseases occur when the immune system attacks and destroys the organs and tissue of its own host. Dozens of autoimmune diseases afflict millions of people in the United States. Several autoimmune diseases, including lupus, rheumatoid arthritis and multiple sclerosis strike women two to 10 times as often as men. Overall, about 80 percent of autoimmune patients are women. There is no cure for autoimmune disease.

B cells are important players in autoimmune disease. The National Jewish Health research team, led by Chair of Biomedical Science Philippa Marrack, PhD, previously identified a subset of B cells that accumulate in autoimmune patients, autoimmune and elderly female mice. They named the cells Age-associated B cells, or ABCs. Subsequent research showed that the transcription factor T-bet plays a crucial role in the appearance of ABC.

Transcription factors bind to DNA inside cells and drive the expression of one or several genes. Researchers believe that T-bet appears inside cells when a combination of receptors on B-cell surfaces — TLR7, Interferon-gamma and the B-cell receptor — are stimulated.

Through breeding and genetic techniques the research team eliminated the ability of autoimmune-prone mice to express T-bet inside their B cells. As a result, ABCs did not appear and the mice remained healthy. Kidney damage appeared in 80 percent of mice with T-bet in the B cells and in only 20 percent of T-bet-deficient mice. Seventy-five percent of mice with T-bet in their B cells died by 12 months, while 90 percent of T-bet-deficient mice survived 12 months.

“Our findings for the first time show that ABCs are not only associated with autoimmune disease, but actually drive it,” said Dr. Rubtsova.

ABCs have attracted increasing interests since their discovery in 2011. Dr. Rubtsova and her colleagues at National Jewish Health have expanded their study of ABCs beyond autoimmune disease and are looking at their involvement in sarcoidosis, hypersensitivity pneumonitis and chronic beryllium disease.

New Progress Toward Finding Best Cells for Liver Therapy

Study shows transplanted fetal rat liver cells multiply and give rise to new cells in injured adult liver.

In a new study, researchers demonstrate successful transplantation of fetal rat liver cells to an injured adult rat liver. The work is an important step toward using transplanted cells to treat liver failure, which currently requires an organ transplant.

Jennifer Sanders, PhD, assistant professor of pediatrics at Brown University, will present the new research at the American Society for Investigative Pathology annual meeting during the Experimental Biology 2017 meeting, to be held April 22–26 in Chicago.

“There are too few donor livers, so many people die of liver diseases such as hepatitis and cirrhosis without ever getting a transplant,” said Sanders. “Understanding the behavior of fetal liver cells may lead to ways to select the best cells for transplantation into people whose livers are failing.”

In the new study, the researcher removed liver cells from a rat fetus near the end of gestation and transplanted them into an injured adult rat liver. In the new liver, the transplanted cells multiplied for a long period and gave rise to new hepatocytes—the main cell type found in the liver—as well as the cells that form the bile ducts and line the blood vessels. Adult rat liver cells cannot multiply and differentiate after transplantation.

“Most previous studies have used very immature fetal rat cells and have not attempted to characterize the cell population prior to transplantation,” said Sanders.  “We are using late-gestation fetal rat hepatocytes that can carry out many of the functions of adult liver cells, and we characterized the cells based on expression of markers on their surface.”

To better understand how fetal and adult hepatocytes differ, the researchers examined proteins called histones that regulate DNA structure. They identified histone differences that may allow the fetal cells to grow and survive when transplanted into an injured adult liver.

“Our prior studies have shown that fetal rat hepatocyte proliferation, growth and gene-expression regulation are different than in adult rat hepatocytes,” said Sanders. “This has led us to believe that maintenance of DNA structure is very important for the behavior of fetal rat hepatocytes and the ability of these cells to repopulate an injured adult liver.”

In addition to this work’s implications for cell-based liver therapies, better understanding of how DNA structure, gene expression and protein function are regulated together in the normal fetal liver cell could help scientists understand the events that lead to liver cancer.

As a next step, the researchers are working to determine how the adult liver environment affects transplanted fetal cells. They want to find out whether transplanted fetal cells differentiate in a way that makes them indistinguishable from normal adult hepatocytes.

Breast Cancer Drug Dampens Immune Response, Protecting Light-Sensing Cells of the Eye

Tamoxifen could be repurposed to treat degenerative diseases of the retina

The breast cancer drug tamoxifen appears to protect light-sensitive cells in the eye from degeneration, according to a new study in mice. The drug prevented immune cells from removing injured photoreceptors, the light-sensitive cells of the retina in the back of the eye. The study, recently reported in the Journal of Neuroscience, suggests tamoxifen might work for the treatment of age-related macular degeneration (AMD) and retinitis pigmentosa (RP), blinding diseases that lack good treatment options. The study was conducted by researchers at the National Eye Institute (NEI), part of the National Institutes of Health.

Although commonly used for cancer treatment, tamoxifen is used in the laboratory as a tool to activate specific genes in genetically engineered mice. The tool allows researchers to turn genes on and off in specific tissues at will. Wai Wong, M.D., Ph.D., chief of NEI’s Unit on Neuron-Glia Interactions in Retinal Disease, and his team were using tamoxifen for this purpose when they noticed something odd. Xu Wang, Ph.D., staff scientist in the Wong laboratory and lead author of the study, observed that mice treated with tamoxifen gained resistance to light-induced eye injuries. Light injury, induced by exposing mice to short-duration, high-intensity light, normally leads to degeneration of photoreceptors. But in the tamoxifen-treated mice, the team unexpectedly observed little to no photoreceptor degeneration.

The team investigated the effects of tamoxifen on light-induced photoreceptor degeneration in normal mice and mice with a disease similar to RP. Live retinal imaging and tissue analyses showed significantly lower levels of photoreceptor degeneration, compared to control mice that did not received tamoxifen. Tamoxifen-treated mice also demonstrated higher photoreceptor function, compared to controls.

How was tamoxifen exerting this protective effect? In an earlier study in 2015, Wong showed that light injury triggers a neurotoxic immune response in the retina. “The immune system becomes alerted to the stressed photoreceptors and goes into culling mode, clearing them out of the retina,” he explained. Wong and his team surmised that tamoxifen was inhibiting this immune response, rather than protecting the photoreceptors directly.

To investigate this hypothesis, Wong’s team cultured microglia — immune cells in the retina — and found that tamoxifen reduced their ability to remove and kill photoreceptor cells. Tamoxifen also reduced levels of inflammatory cytokines — signaling molecules that trigger inflammation — produced by the microglia.

Tamoxifen did not appear to directly influence the physiology of photoreceptors or protect photoreceptors in the absence of microglia, suggesting that the inhibition of microglia is a key mechanism underlying tamoxifen’s protective effect. The investigators are currently studying at molecular level how tamoxifen is able to inhibit the microglia.

In August 2016, Wong’s laboratory filed a patent for use of tamoxifen in retinal degenerative disorders. The new use of the drug is unexpected, as tamoxifen’s only previously known association with the retina had been a low risk of retinopathy among breast cancer patients.

RP is a group of rare genetic disorders affecting the retina. Worldwide, RP affects about 1 in 4,000 people. Symptoms typically appear during childhood and slowly progress over many years, often causing blindness. AMD is a leading cause of vision loss among people age 50 and older. About two million Americans have AMD, which affects central vision.

The tamoxifen dose used in Wong’s mouse study was equivalent to eight times the FDA-approved dose for breast cancer. The researchers are currently investigating whether the protective effects are retained at lower doses.

The work “sets us up for a clinical trial in the not-so-distant future,” said Wong. “Translation to the clinic can happen reasonably rapidly because tamoxifen, as an FDA-approved drug, already has a well-characterized safety profile,” he explained.

Stem Cell Treatment May Restore Vision to Patients with Damaged Corneas

Researchers working as part of the University of Georgia’s Regenerative Bioscience Center have developed a new way to identify and sort stem cells that may one day allow clinicians to restore vision to people with damaged corneas using the patient’s own eye tissue. They published their findings in Biophysical Journal.

The cornea is a transparent layer of tissue covering the front of the eye, and its health is maintained by a group of cells called limbal stem cells. But when these cells are damaged by trauma or disease, the cornea loses its ability to self-repair.

“Damage to the limbus, which is where the clear part of the eye meets the white part of the eye, can cause the cornea to break down very rapidly,” said James Lauderdale, an associate professor of cellular biology in UGA’s Franklin College of Arts and Sciences and paper co-author. “The only way to repair the cornea right now is do a limbal cell transplant from donated tissue.”

In their study, researchers used a new type of highly sensitive atomic force microscopy, or AFM, to analyze eye cell cultures. Created by Todd Sulchek, an associate professor of mechanical engineering at Georgia Tech, the technique allowed researchers to probe and exert force on individual cells to learn more about the cell’s overall health and its ability to turn into different types of mature cells.

They found that limbal stem cells were softer and more pliable than other cells, meaning they could use this simple measure as a rapid and cost-effective way to identify cells from a patient’s own tissue that are suitable for transplantation.

“Todd’s technology is unique in the tiniest and most sensitive detection to change,” said Lauderdale. “Just think about trying to gently dimple or prod the top of an individual cell without killing it; with conventional AFM it’s close to impossible.”

Building on their findings related to cell softness, the research team also developed a microfluidic cell sorting device capable of filtering out specific cells from a tissue sample.

With this device, the team can collect the patient’s own tissue, sort and culture the cells and then place them back into the patient all in one day, said Lauderdale. It can take weeks to perform this task using conventional methods.

The researchers are quick to caution that more tests must be done before this technique is used in human patients, but it may one day serve as a viable treatment for the more than 1 million Americans that lose their vision to damaged corneas every year.

The group first started this research with the hope of helping children with aniridia, an inherited malformation of the eye that leads to breakdown of the cornea at an early age.

Because aniridia affects only one in 60,000 children, few organizations are willing to commit the resources necessary to combat the disease, Lauderdale said.

“Our first goal in working with such a rare disease was to help this small population of children, because we feel a close connection to all of them,” says Lauderdale, who has worked with aniridia patients for many years. “However, at the end of the day this technology could help hundreds of thousands of people, like the military who are also interested in corneal damage, common in desert conditions.”

Steven Stice, a Georgia Research Alliance Eminent Scholar, who plays an important role in fostering cross-interdisciplinary collaboration as director of the RBC, initially brought the researchers together and encouraged a seed grant application through the center for Regenerative Engineering and Medicine, or REM, a joint collaboration between Emory University, Georgia Tech and UGA.

“A culture is developing around seed funding that is all about interdisciplinary collaboration, sharing of resources, and coming together to make things happen,” said Stice. “Government funding agencies place a high premium on combining skills and disciplines. We can no longer afford to work in an isolated laboratory using a singular approach.”

The REM seed funding program is intended to stimulate new, unconventional collaborative research and requires equal partnership of faculty from two of the participating institutions.

“We tend to get siloed experimentally,” says Lauderdale. “To a biologist like me, all cells are very different and all atomic force microscopes are the same. To an engineer like Todd it’s just the opposite.”

Study Identifies Common Gene Variants Associated with Gallbladder Cancer

By comparing the genetic code of gallbladder cancer patients with those of healthy volunteers at nearly 700,000 different locations in the genome, researchers say they have found several gene variants which may predispose individuals to develop the disease.

The findings, published March 5 in The Lancet Oncology, could lead to a better understanding of the causes of this highly fatal condition, which could in turn lead to better treatments for the disease. The work is a collaboration between the Johns Hopkins Bloomberg School of Public Health, the National Cancer Institute and Tata Memorial Cancer Centre in Mumbai, India.

Although gallbladder cancer is rare in most parts of the world, it is far more common among some ethnic groups, such as Native Americans in North America, and in certain geographic regions, including Central and South America and East and Southeast Asia. The 178,000 new cases diagnosed worldwide each year are centered primarily in these high-risk regions.

“Using the latest technologies to look at the causes – notably the genetic underpinnings – of this understudied disease just makes a lot of sense,” says study co-leader Nilanjan Chatterjee, PhD, Bloomberg Distinguished Professor in the Department of Biostatistics at the Bloomberg School and a professor of oncology at the Johns Hopkins Kimmel Cancer Center

The gallbladder is a tiny organ in the abdomen which stores bile, the digestive fluid produced by the liver. When gallbladder cancer is discovered early, the chances for survival are good, but most gallbladder cancers are discovered late as it is difficult to diagnose since it often causes no specific symptoms.

To search for which genes might be important in gallbladder cancer, investigators at the Tata Memorial Centre gathered blood samples from 1,042 patients who were treated at the Centre’s Hospital in Mumbai between Sept. 2010 and June 2015. The researchers also collected blood samples during this time from 1,709 healthy volunteers with no known cancers who were visiting patients at the hospital.

To make the groups comparable, they were matched by their ages, sex and geographic regions in India from which the patients came from.

The scientists then ran these blood samples through a whole genome analysis of common single nucleotide polymorphisms (SNPs), places where the genome between different individuals vary by changes in single nucleotides, the smallest units that make up the genome.

Through a series of biostatistical and bioinformatics analyses, they found highly significant association for multiple DNA variants near two genes — ABCB4 and ABCB1 — known to be involved in moving lipids through the liver, gallbladder and bile ducts. A previous study had associated ABCB4 with the formation of gallstones, a known risk factor for gallbladder cancer. But the new results show for the first time that common inherited variants in this region may predispose individuals to gallbladder cancer itself, independent of gallstone status, Chatterjee says.

The researchers later replicated these results using blood samples gathered from 447 more patients with gallbladder cancer and 470 healthy volunteers from Tata Memorial Hospital and Sanjay Gandhi Postgraduate Institute of Medical Science in Uttar Pradesh, India.

They also ran another analysis to estimate how much variation in gallbladder cancer risk can be explained by the discovery of additional common variants. They say they hope to conduct similar studies of larger groups of people in the future.

“Gallbladder cancer, like many other cancers and complex diseases, is likely to be associated with many genetic markers, each of which may have small effects, but in combination they can explain substantial variation in risk,” Chatterjee says.

The researchers estimate as much as 25 percent of gallbladder cancer risk could be explained by common genetic variants. Although the specific genetic variants the current study has identified explain a small fraction of this risk, the fact that they are in close proximity to genes known to be important for transporting a certain class of lipids from liver to gallbladder could provide an important clue to the cause of the disease.

The team is currently planning to investigate the ABCB4/ABCB1 region in more depth by fully sequencing this region in some of the current study participants to understand whether there are additional risk variants there. They also plan to conduct larger studies to look for additional genes associated with gallbladder cancer. By better understanding the function of the genetic risk variants, as well as by investigating environmental and lifestyle causes, Chatterjee says, researchers might eventually be able to develop new treatments or interventions to prevent this disease from occurring in patients at high risk.

New Assay May Lead to a Cure for Debilitating Inflammatory Joint Disease

Current treatments for rheumatoid arthritis relieve the inflammation that leads to joint destruction, but the immunologic defect that triggers the inflammation persists to cause relapses, according to research conducted at NYU Langone Medical Center and the University of Pittsburgh.

Known as autoantibodies and produced by the immune system’s B cells, these defective molecules mistakenly attack the body’s own proteins in an example of autoimmune disease. Now the results of a study just published in Arthritis & Rheumatology suggest that clinical trials for new rheumatoid arthritis (RA) drugs should shift from their sole focus on relieving inflammation to eliminating the B cells that produce these antibodies.

“We have developed a test for measuring the underlying autoimmunity in rheumatoid arthritis patients that should be used to evaluate new treatment regimens,” says senior author Gregg Silverman, MD, professor in the Departments of Medicine and Pathology at NYU Langone and co-director of its Musculoskeletal Center of Excellence. “We believe this provides a road to a cure for rheumatoid arthritis.”

Rheumatoid arthritis is a chronic inflammatory autoimmune disease that affects 1.5 million people in the United States. The current standard of care begins with methotrexate, a drug that reduces inflammation. It is often followed by drugs that block a molecule called tumor necrosis factor (TNF), which promotes inflammation. Both of these classes of drugs can blunt the swelling and inflammation associated with rheumatoid arthritis and at times even allow patients to go into clinical remission that requires continued treatment. But when patients halt these medications, symptoms generally flare up either sooner or later. According to Silverman, the reduction of inflammation does not directly reflect the autoimmune disease that causes rheumatoid arthritis.

In the study, researchers focused on “memory” B cells, immune system cells that remember the initial errant immune encounter that recognized the body’s own proteins as foreign. In rheumatoid arthritis, memory B cells secrete molecules called anti-citrullinated protein antibodies (ACPAs). Doctors currently confirm an RA diagnosis with a blood test that looks for ACPAs, which are present in 80 percent of RA patients.

Silverman and his colleagues developed sensitive assays to detect a range of different autoantibodies present in the disease. The researchers then established a cell culture system to stimulate memory B cells, and used the assays to test what kind of antibodies the B cells produced.

The researchers tested blood samples from RA patients and from healthy donors. They found high levels of APCA-secreting memory B cells in the blood of patients with these autoantibodies, but not in patients without autoantibodies or in the healthy volunteers.

They then looked at patients who had achieved remission with either methotrexate or a TNF inhibitor. The researchers found that APCA levels were directly proportional to the recirculating memory B cells in the blood stream, confirming that current drug treatments do not affect the underlying autoimmunity in rheumatoid arthritis.

The next step, Silverman says, is to conduct long-term prospective clinical trials of new RA drugs, using the team’s new test to determine each drug’s effect on autoimmunity. The current metrics for evaluating the effectiveness of new rheumatoid arthritis drugs remain focused on reducing inflammation but not curing the disease, he says.

“We need to develop longer-term vision of how to improve the treatment of rheumatoid arthritis,” Silverman says. “This new tool may show that agents that target other molecules or cells have advantages that were previously not considered now that we can better measure those effects.”

University Hospitals Seidman Cancer Center Enrolls First Patient in New National Head and Neck Cancer Study

Study tests safety of immunotherapy drug added to regimen of surgery, chemotherapy and radiation therapy; same drug used on President Jimmy Carter’s brain cancer

University Hospitals (UH) Seidman Cancer Center patient Richard Bartlett, 62, of Magnolia, Ohio, has become the first in the nation to enroll in a new study for very high risk head and neck cancer.

“The study is one of the first ever to use ‘quadra-modality’ therapy, or in other words, four different types of therapy for this cancer,” said Min Yao, MD, PhD, the UH principal investigator, a radiation oncologist at UH and a professor of radiation oncology at the Case Western Reserve University School of Medicine.

Standard treatment for this cancer is surgery, followed by radiation and chemotherapy. This study will add an immunotherapy drug called pembrolizumab to activate the body’s immune system in the fight against the cancer. The drug, originally developed to treat melanoma, made the news in 2015 when President Jimmy Carter was treated with it for his brain metastases from melanoma.

Mr. Bartlett volunteered for the study for several reasons. “I want to have the best outcome possible,” he said, “and I have a responsibility to my family. I also know that people in the past have made sacrifices for research and I know cancer research is in its infancy in many ways and I’d like to do what I can to help.”

Pembrolizumab is one of the first immunotherapy drugs. Instead of directly killing cancer cells, these drugs boost the immune system to do the job.

This phase I trial will study the side effects and best dose and schedule of pembrolizumab when given together with the chemotherapy drug called cisplatin and radiation therapy.

Despite advances in cancer detection and treatment, the five-year overall survival rate for high risk head and neck squamous cell cancer is only 40 to 60 percent.

“This is a four-pronged attack on the cancer,” said Dr. Yao. “With surgery, we remove as much of the tumor as we can. Chemotherapy works to stop the growth of tumor cells, either by killing the cells or by stopping them from dividing. Radiation therapy uses high-energy X-rays to kill tumor cells. And now with pembrolizumab, we trigger the immune system in the fight.”

This phase 1 study is for safety and will lay the groundwork for a future phase 3 study.

The study will enroll 56 patients nationally in seven study sites. Data collection is estimated to be completed by May 2018. UH Seidman Cancer Center and CWRU School of Medicine comprise the only Ohio site. The Cleveland site for the study is funded by a National Cancer Institute grant to the CWRU School of Medicine.

Mr. Bartlett’s head and neck cancer was discovered after he went to a dentist for a wisdom tooth extraction and a sore on the inside of his right cheek. He thought the sore was caused by the problem tooth rubbing against the cheek. The dentist recommended a biopsy on the sore.

It turned out to be cancerous and he was referred to Pierre Lavertu, MD, Director of Head and Neck Surgery and Oncology at UH Cleveland Medical Center. Dr. Lavertu and Chad Zender, MD, of the UH Department of Otolaryngology, did the surgery.

Mr. Bartlett had surgery on Dec. 22, 2016 and his cancer was more aggressive than originally thought. He has begun his chemo, radiation, and immunotherapy under the care of Michael Gibson, MD, PhD, Medical Director of the Head and Neck Oncology Team at UH Seidman Cancer Center, and Dr. Yao of the Radiation Oncology Team. All of his physicians are members of the faculty at the CWRU School of Medicine.

Mr. Bartlett and his wife Nancy have been together for 25 years and have been married the past 10 years. They met when she worked as a cashier in a store where he was shopping. He found a plastic flower on the floor and gave it to her. “There was no turning back after that,” laughed Mr. Bartlett.

Nancy said they are both pleased with the care that Richard has received at UH Seidman Cancer Center. “The doctors and the staff have been very nice. We came here because they have the latest in cancer care,” she said.

Scientists Identify Chain Reaction That Shields Breast Cancer Stem Cells From Chemotherapy

Working with human breast cancer cells and mice, researchers at Johns Hopkins say they have identified a biochemical pathway that triggers the regrowth of breast cancer stem cells after chemotherapy.

The regrowth of cancer stem cells is responsible for the drug resistance that develops in many breast tumors and the reason that for many patients, the benefits of chemo are short-lived. Cancer recurrence after chemotherapy is frequently fatal.

“Breast cancer stem cells pose a serious problem for therapy,” says lead study investigator Gregg Semenza, M.D., Ph.D., the C. Michael Armstrong Professor of Medicine, director of the Vascular Biology Program at the Johns Hopkins Institute for Cell Engineering and a member of the Johns Hopkins Kimmel Cancer Center. “These are the cells that can break away from a tumor and metastasize; these are the cells you most want to kill with chemotherapy. Paradoxically, though, cancer stem cells are quite resistant to chemotherapy.”

Semenza says previous studies have shown that resistance to chemotherapy arises from the hardy nature of cancer stem cells, which are often found in the centers of tumors, where oxygen levels are quite low. Their survival is made possible through proteins known as hypoxia-inducible factors (HIFs), which turn on genes that help the cells survive in a low-oxygen environment.

In this new study, described Feb. 21 in Cell Reports, Semenza and his colleagues conducted gene expression analysis of multiple human breast cancer cell lines grown in the laboratory after exposure to chemotherapy drugs, like carboplatin, which stops tumor growth by damaging cancer cell DNA. The team found that the cancer cells that survived tended to have higher levels of a protein known as glutathione-S-transferase O1, or GSTO1. Experiments showed that HIFs controlled the production of GSTO1 in breast cancer cells when they were exposed to chemotherapy; if HIF activity was blocked in these lab-grown cells, GSTO1 was not produced.

Semenza notes that GSTO1 and related GST proteins are antioxidant enzymes, but GSTO1’s role in chemotherapy resistance did not require its antioxidant activity. Instead, following exposure to chemotherapy, GSTO1 binds to a protein called the ryanodine receptor 1, or RYR1, that triggers the release of calcium, which causes a chain reaction that transforms ordinary breast cancer cells into cancer stem cells.

To more directly assess the role of GSTO1 and RYR1 in the breast tumor response to chemotherapy, the researchers injected human breast cancer cells into the mammary gland of mice and then treated the mice with carboplatin after tumors had formed. In addition to using normal breast cancer cells in the experiments, the team also used cancer cells that had been genetically engineered to lack either GSTO1 or RYR1. Loss of either GSTO1 or RYR1, the researchers report, decreased the number of cancer stem cells in the primary tumor, blocked metastasis of cancer cells from the primary tumor to the lungs, decreased the duration of chemotherapy required to induce remission and increased the duration of time after chemotherapy was stopped that the mice remained tumor-free.

Although the study showed that blocking the production of GSTO1 may improve the efficacy of chemotherapy drugs, such as carboplatin, GSTO1 is only one of many proteins that are produced under the control of HIFs in breast cancer cells that have been exposed to chemotherapy. The Semenza lab is working to develop drugs that can block the action of HIFs, with the hope that HIF inhibitors will make chemotherapy more effective.

Lysosomes in Healthy Neurons and in Neurons with Juvenile Batten Disease

Researchers at Baylor College of Medicine, Jan and Dan Duncan Neurological Research Institute at Texas Children’s Hospital and King’s College London have discovered a treatment that improves the neurological symptoms in a mouse model of juvenile Batten disease. This discovery brings hope to patients and families affected by the disease that a treatment might be available in the future. The study appears in Nature Communications.

“Patients with juvenile Batten disease are born healthy and reach the expected developmental milestones of the first 4 to 6 years of age,” said senior author Dr. Marco Sardiello, assistant professor of molecular and human genetics at Baylor. “Then, these children progressively regress their developmental achievements; they gradually lose their vision and develop intellectual and motor disabilities, changes in behavior and speech difficulties. Most people with this condition live into their 20s or 30s. This inherited, rare disease has no cure or treatment other than palliative care.”

“As we started this project, patients and families affected by this condition visited us in the laboratory,” said first author Dr. Michela Palmieri, who was a postdoctoral fellow in the Sardiello lab during this project and currently is at the San Raffaele Scientific Institute in Milan, Italy. “We were deeply affected by our interactions with the patients and their families and this further motivated us to pursue this research with the hope that maybe one day it will lead to a treatment that will improve the lives of people affected by this condition.”

Juvenile Batten disease, a problem with cellular waste management

Like a large dynamic city, a cell carries out many activities that generate waste. Waste needs to be disposed of properly in order for the city to continue its activities without interruption. If waste management fails, waste progressively accumulates and eventually leads to interruption and paralysis of the activities of the city. Something similar happens in cells when cellular waste is not discarded.

The lysosomes are the structures in charge of clearing the waste produced by the cell’s regular functions. Lysosomes are sacs inside all cells containing enzymes that degrade cellular waste into its constituent components, which the cell can recycle or discard. When lysosomes fail and cellular waste accumulates, disease follows. Although all types of cells can be affected by defects in lysosomal waste processing and cellular waste accumulation, brain cells – neurons – are particularly susceptible.

“In juvenile Batten disease, one of nearly 50 human lysosomal storage disorders, the function of brain cells is progressively affected by the accumulation of cellular waste,” Sardiello said. “This accumulation leads to perturbation of many cellular processes, cell death and progressive regression of motor, physical and intellectual abilities.”

A novel approach to finding a treatment

“A few years back we discovered a protein in cells called TFEB, a master transcription factor that stimulates the cell to produce more lysosomes and degrade cellular waste more effectively,” said Sardiello. “So we thought about counteracting the accumulation of cellular waste in Batten disease by acting on TFEB.”

“We and others had found that enhancing the activity of TFEB genetically can help counter the accumulation of cellular waste in different diseases,” Sardiello said. “What was missing was a way to activate TFEB with a drug that in the future could be put in a pill to treat the condition. We focused on investigating how to activate TFEB pharmacologically.”

“We discovered that TFEB is under the control of another molecule called Akt, which is a kinase, a protein that can modify other proteins,” said Palmieri. “Akt has been studied in detail. There are drugs available that can modulate the activity of Akt.”

The researchers discovered that Akt modifies TFEB by adding a chemical group, a phosphate, to it. This chemical modification inactivates TFEB.

“We wanted to inhibit Akt to keep TFEB more active,” said Palmieri. “We discovered that the sugar trehalose is able to do this job.”

Testing a treatment for juvenile Batten disease in a mouse model of the condition

The scientists tested the effect of trehalose in a mouse model of juvenile Batten disease.

“We dissolved trehalose in drinking water and gave it to mice that model juvenile Batten disease,” said Sardiello. “Then, over time we examined the mice’s brain cells under the microscope. We found that the continuous administration of trehalose inhibits Akt and activates TFEB in the brains of the mice. More active TFEB meant more lysosomes in the brain and increased lysosomal activity, followed by decreased accumulation of the storage material and reduced tissue inflammation, which is one of the main features of this disease in people, and reduced neurodegeneration. These changes resulted in the mice living significantly longer. This is a good start toward finding a treatment for people with this disease.”

“We are very excited that these findings put research a step closer to understanding the mechanisms that underlie human lysosomal storage diseases,” said Palmieri. “We hope that our research will help us design treatments to counteract this and other human diseases with a pathological storage component, such as Alzheimer’s, Huntington’s and Parkinson’s diseases, and hopefully ameliorate the symptoms or reduce the progression of the disease for those affected.”

Personalized Cancer Therapy on the Horizon Thanks to New Genomic Cancer Research Partnership

Gene Editing Institute at Christiana Care Health System partners with NovellusDx in BIRD Foundation Grant

Wilmington, Del, Jan. 30, 2017 – For its enormous potential to accelerate the development of personalized cancer therapies, the Gene Editing Institute of Christiana Care Health System’s Helen F. Graham Cancer Center & Research Institute has been awarded a grant of $900,000 from the U.S.-Israel Binational Industrial Research and Development (BIRD) Foundation in partnership with the biotechnology company NovellusDx.

The BIRD Foundation promotes collaboration between U.S. and Israeli companies in a wide range of technology fields for the purpose of joint product development. Projects submitted to the BIRD Foundation undergo evaluation by the U.S. National Institute of Standards and Technology of the U.S. Department of Commerce and by the Israel Innovation Authority.

The grant allows the Gene Editing Institute to partner with Jerusalem-based NovellusDx on a new series of state-of-the-art gene editing technologies that help identify the genetic mechanism responsible for both the onset and progression of many types of cancer. The two organizations are collaborating on a licensing agreement to commercialize the gene editing technologies that result from the research.

“Thanks to this generous BIRD Foundation grant, this partnership promises to be a catalyst that will speed progress in personalized medicine for many forms of cancer, accelerating the path to prevention, diagnosis, treatment, and ultimately, to a cure of cancer,” said Nicholas J. Petrelli, M.D., the Bank of America endowed medical director of the Helen F. Graham Cancer Center & Research Institute at Christiana Care Health System.

“We are honored to partner with the exceptional team at NovellusDx to advance genomic cancer research and to discover new gene editing techniques,” said Eric Kmiec, Ph.D., director of the Gene Editing Institute. “Our partnership is not only based on the skills of both organizations, but on the unique opportunity to license our gene editing technology with a company capable of commercializing it. The due diligence and peer review process for this award are extensive. I’m enormously grateful to the Research Institute at the Philadelphia-Israeli Chamber of Commerce for its invaluable support of our application.”

NovellusDx has established a unique approach to identify unknown “driver” gene mutations that often accelerate or facilitate cancer progression. With clinical partners throughout the world, including at MD Anderson Cancer Center and Massachusetts General Hospital in the U.S., NovellusDx obtains DNA sequence information and creates a personal profile of the genetic mutations from individual patients. The Gene Editing Institute will use its expertise in gene editing to re-create these mutations that allows NovellusDx and its partners to identify, design and implement the most effective therapy for each patient.

Cancer genomics plays a critical role in pharmacogenomics, or the study of how genes impact a patient’s response to drugs. “With our joint research, we hope to develop gene editing technologies that help develop effective, safe medications and doses that can be tailored to a person’s genetic profile,” Dr. Kmiec said. “This will lead to precision and personalized cancer therapy at its very best.”

“We have been working closely with Dr. Kmiec and the Gene Editing Institute for the last nine months to generate preliminary data to support this ground-breaking idea and grant application,” said Haim Gil-Ad, CEO of NovellusDx. “We are excited that the BIRD Foundation with its stringent review process found our application worthy of the generous funding, which also provides external validation. This work has the potential to change the way functional genomics is done. Once the genetic makeup is known, we will be immediately able to test and monitor the effect of the patient mutations in live cells.”

The BIRD Foundation grant recognizes the Gene Editing Institute’s pioneering work to advance gene editing toward clinical applications in cancer research. The Gene Editing Institute is partnering with The Wistar Institute to develop translational genetic approaches to melanoma cancer research, and with Bio-Rad Inc. to advance a gene editing educational curriculum. In addition, with funding from the U.S. National Institutes of Health, the Gene Editing Institute is developing a gene editing strategy for the treatment of sickle cell anemia.

The BIRD Foundation supports projects without receiving any equity or intellectual property rights in the participating companies or in the projects themselves. BIRD funding is repaid as royalties from sales of products that were commercialized as a result of BIRD support. The Foundation shares the risk and does not require repayment if the project fails to reach the sales stage.

The Gene Editing Institute at the Graham Cancer Center is a worldwide leader in personalized genetic medicine. Founded and led by Dr. Kmiec, the Gene Editing Institute is unlocking the genetic mechanisms that drive cancer and that can lead to new therapies and pharmaceuticals to revolutionize cancer treatment. The Gene Editing Institute also provides instruction in the design and implementation of these precise new genetic tools.

KU Reseachers Find Statins May Hold Keys to Future Cancer Treatment

Researchers at the University of Kansas Medical Center have found that high doses of drugs commonly used to fight high cholesterol can destroy a rogue protein produced by a damaged gene that is associated with nearly half of all human cancers.

Tomoo Iwakuma, M.D., Ph.D., an associate professor in the Department of Cancer Biology, and his team have published the first research showing how the use of statins, such as Lipitor (atorvastatin), Crestor (rosuvastatin) and Mevacor (lovastatin), can shut down structurally mutated p53 proteins that can accelerate cancer progression, while not harming proteins produced by healthy p53 genes. Although statins are not a cancer treatment per se, the understanding of how they affect mutated forms of p53 could lead to new medications designed specifically to knock out the damaged p53.

“I could have kept working for 20 years or longer without any big finding,” said Iwakuma, whose work appeared in the November 2016 issue of Nature Cell Biology and has been recommended for F1000Prime, a prestigious peer-review service that identifies research that is likely to influence biomedical and clinical knowledge. “This is the most exciting work of my science life, because it will contribute to treating cancer.”

P53 gene and cancer

Cancer is essentially caused by mutations to the genes that regulate cell growth or cell death. Of the hundreds of genetic culprits that have been implicated with causing various cancers, p53, dubbed the “guardian of the genome,” is the mightiest of them all. Mutant forms of p53 have been found in nearly half of all malignant tumors and nearly every type of human cancer.

When p53 works properly, it produces proteins that keep cells from growing and dividing too quickly. When p53 becomes mutated, either spontaneously or through heredity, its regulating abilities no longer work and cells can grow out of control, forming tumors and invading normal tissues – that’s cancer.

Compounding the problem that mutant p53 can no longer suppress the growth of tumors is that fact that it can also actually accelerate the progression of cancer and drug resistance.

The challenge for Iwakuma and his team was to find out how to eliminate the misbehaving protein, while leaving cells containing healthy p53 needed for normal cell growth unharmed.

Hunting for a weapon

Four years ago, Iwakuma and his lab team collaborated with the High Throughput Screening Laboratory (HTC) on the University of Kansas Lawrence campus to screen compounds to find out which ones might degrade mutant p53. Of the nearly 9,000 compounds they tested, about 2,400 were Food and Drug Administration (FDA)-approved drugs, while the others were non-FDA approved and uncharacterized compounds.

When Iwakuma got an email from the HTC listing the 10 compounds that the screenings had shown promise in reducing mutant p53 levels, he was shocked to see that some of them were statins.

“At first I thought, ‘What? This must be wrong,'” said Iwakuma, who first became interested in p53 as a post-doctoral student at the University of Texas MD Anderson Cancer Center.

Early screenings often produce false positives, so Iwakuma had to verify the lab results, first testing them in cells and then in mice. The KU researchers injected the mice with cells expressing mutant p53, waited for tumors to form, and then treated them with high doses of statins for 21 days. They found that tumors did not grow well in mice treated with statins compared to the controls, and they learned the statins worked only on structurally mutated (misfolded) p53, as opposed to p53 mutated at the spot where it binds to DNA. This was an important discovery, particularly since clinical research with statins had not considered the type of p53 mutation.

“We found that only the structural mutation is affected,” Iwakuma said. “Which explains why clinical studies with statins were inconclusive.”

Just the beginning

While the team was elated with its findings, the researchers knew their work was just beginning.

“Once we knew for sure statins degraded mutant p53, we still had to figure out how,” explained Atul Ranjan, Ph.D., a post-doctoral researcher in cancer biology at KU and co-author on the study. “We needed to find out exactly how the statins work for p53 degradation; which other proteins are involved in the mechanism.”

So Alejandro Parrales, Ph.D., another co-author on the study and a post-doc in Iwakuma’s lab, began looking at heat shock proteins, which are known for their efforts to correct misfolded proteins, as a possible piece to the puzzle. The researchers identified DNAJA1 as a heat shock protein that binds to misfolded mutant p53 and thus protects the mutant p53 from an enzyme that flags damaged or misshapen proteins for destruction.

It turned out that the same mechanisms that help statins reduce cholesterol are at work preventing mutant p53 from binding to DNAJA1, leaving these mutant proteins unprotected. As a result, mutant p53 is free to attach to the enzyme that leads to its degradation. And since mutant p53 is not usually present in normal cells, all this happens without affecting healthy cells.

Going forward, researchers know that many challenges await them, including finding ways to target DNAJA1 directly, now that they know its absence results in mutant p53 being degraded. Iwakuma also sees potential to use statins or another p53-degrading drug in conjunction with chemotherapy.

Mutant p53 makes human cancer cells more metastatic and resistant to chemotherapy,” he said. “That’s a primary reason to get rid of it — to improve survival in cancer patients.”

‘Collateral’ Lethality May Offer New Therapeutic Approach for Cancers of the Pancreas, Stomach and Colon

Cancer cells often delete genes that normally suppress tumor formation. These deletions also may extend to neighboring genes, an event known as “collateral lethality,” which may create new options for development of therapies for several cancers.

Scientists at The University of Texas MD Anderson Cancer Center have discovered that during early cancer development when a common tumor suppressor known as SMAD4 is deleted, a nearby metabolic enzyme gene called malic enzyme 2 (ME2) also is eradicated, suggesting the possibility of malic enzyme inhibitors as a novel therapy approach. Study findings were published in the Jan. 18 online issue of Nature.

“In an effort to expand therapeutic strategies beyond oncogenic targets to those not directly linked to cancer development, we have identified collateral lethal vulnerability in pancreatic cancers that can be targeted pharmacologically in certain patient populations,” said Prasenjit Dey, Ph.D., postdoctoral fellow in Cancer Biology and co-author of the Nature article. “Genomic data across several cancers further suggest this therapeutic strategy may aid many cancer patients, including those with stomach and colon cancers.”

Collateral lethality occurs when tumor suppressor genes are deleted, a nearly universal occurrence in cancer. Correspondingly, a large number of genes with no direct role in tumor progression also are deleted as a result of their proximity to tumor suppressor genes.

SMAD4 is deleted in one-third of pancreatic cancers. The research team found that when the SMAD4 gene is eradicated in mice, it also results in depletion of ME2 levels. The genetic depletion of ME3, a sister gene to ME2, sets off a complex chain of events that ultimately regulates an amino acid group called branched chain amino acid (BCAA), which are crucial to cancer’s ability to thrive. Thus, if a therapy could be developed that inhibits ME3, it might prevent ME2-deleted tumor growth.

“Our work suggests a mechanism for cell lethality involving the regulation of BCAAs as crucial elements in pancreatic cancer by regulating ME3,” said Ronald DePinho, M.D., professor of Cancer Biology, senior author of the Nature paper and president of MD Anderson. “We propose that highly specific ME3 inhibitors could provide an effective therapy for many cancer patients, but more research must be done.”

Moffitt Cancer Center Researchers Report Promising Clinical Activity and Minimal Toxicities for HER2-Targeted Dendritic Cell Vaccines in Early-Stage Breast Cancer Patients

HER2-Targeted Dendritic Cell Vaccines Stimulate Immune Responses and Regression of HER2-Expressing Early-Stage Breast Tumors

Deregulation and inhibition of the immune system contributes to cancer development. Many therapeutic strategies aim to restimulate the immune system to recognize cancer cells and target them for destruction. Researchers from Moffitt Cancer Center report that a dendritic cell vaccine that targets the HER2 protein on breast cancer cells is safe and effectively stimulates the immune system leading to regression of early-stage breast cancer.

The HER2 protein is overexpressed in 20-25% of all breast cancer tumors and is associated with aggressive disease and poor prognosis. Moffitt researchers have previously shown that immune cells are less able to recognize and target cancer cells that express HER2 as breast cancer progresses into a more advanced and invasive stage. This suggests that strategies that can restimulate the immune system to recognize and target HER2 early during cancer development may be effective treatment options.

The Moffitt researchers previously developed a vaccine that helps the immune system recognize the HER2 protein on breast cancer cells. Their approach involves creating the vaccine from immune cells called dendritic cells that are harvested from each individual patient to create a personalized vaccine.

In order to determine if the HER2-dendritic cell vaccine is safe and effective, the Moffitt researchers performed a clinical trial in 54 women who have HER2-expressing early-stage breast cancer. The dendritic cell vaccines were prepared by isolating dendritic cells from each patients’ blood and exposing them to fragments of the HER2 protein. Patients were injected with a dose of their personal dendritic cell vaccine once a week for 6 weeks into either a lymph node, the breast tumor, or into both sites.

The researchers report that the dendritic cell vaccines were well-tolerated and patients only experienced low-grade toxicities. The most common adverse events were fatigue, injection site reactions, and chills. They also show that the vaccine was able to stimulate an immune response in the majority of the patients. Approximately 80% of evaluable patients had a detectable immune response in their peripheral blood and/or in their sentinel lymph node wherein their cancer is most likely to spread to first. Importantly, the immune responses among the patients were similar, regardless of the route of vaccine administration.

The Moffitt researchers assessed the effectiveness of the vaccine by determining the percentage of patients who had detectable disease within surgical specimens after resection. The absence of disease is termed a pathological complete response (pCR). They report that 13 patients achieved a pCR and patients who had early non-invasive disease called ductal carcinoma in situ (DCIS) achieved a higher rate of pCR than patients who had early-stage invasive disease. Interestingly, patients who achieved a pCR had a higher immune response within their local sentinel lymph nodes.

“These results suggest that vaccines are more effective in DCIS, thereby warranting further evaluation in DCIS or other minimal disease settings, and the local regional sentinel lymph node may serve as a more meaningful immunologic endpoint,” said Brian J. Czerniecki, MD, PhD, Chair of the Department of Breast Oncology at Moffitt Cancer Center.

Scripps Florida Scientists Uncover New Way to Defeat Therapy-Resistant Prostate Cancer

A new study led by scientists from the Florida campus of The Scripps Research Institute (TSRI) sheds light on a signaling circuit in cells that drives therapy resistance in prostate cancer. The researchers found that targeting the components of this circuit suppresses advanced prostate cancer development.

The study, led by TSRI Associate Professor Jun-Li Luo, was published online ahead of print in the journal Molecular Cell.

A New Strategy to Fight Prostate Cancer

Prostate cancer—which, according to the American Cancer Society, affects one in six American men—is the second-leading cause of death after lung cancer in American men.
Currently, the most effective treatment of advanced prostate cancer is to deprive the cancer of what feeds it—androgen hormones, such as testosterone. Unfortunately, almost all patients eventually develop resistance to this therapy, leaving doctors with no options to counteract the inevitable.

The new study shows that a “constitutively active” signaling circuit can trigger cells to grow into tumors and drive therapy resistance in advanced prostate cancer. A cell signal pathway with constitutive activity requires no binding partner (ligand) to activate; instead, the signaling circuit continually activates itself.

This signaling circuit, which is composed of the protein complex IκBα/NF-κB (p65) and several other molecules, controls the expression of stem cell transcription factors (proteins that guide the conversion of genetic information from DNA to RNA) that fuel the aggressive growth of these resistant cancer cells.

“The fact that the constitutive activation of NF-kB in the circuit is independent of traditional activation opens the door for potential treatment options,” said Luo.

Targeting Other Signaling Components Shows Promise

NF-kB plays important roles in cancer development, and it is regarded as one of the most important targets for cancer therapy. However, the use of NF-kB inhibitors in treating cancer is complicated by severe side effects related to immunosuppression caused by indiscriminate inhibition of NF-kB in normal immune cells.

Luo noted that targeting the other non-IκBα/NF-κB components in this signaling circuit would avoid the suppression of NF-κB in normal immune cells while keeping the potent anti-cancer efficacy.

In addition to IκBα/NF-κB, the signaling circuit includes the microRNA miR-196b-3p, Meis2 and PPP3CC. While miR-196b-3p promotes tumor development, Meis2, which is an essential developmental gene in mammals, can disrupt the circuit when overexpressed. The protein PPP3CC can inhibit NF-κB activity in prostate cancer cells.

“Disrupting this circuit by targeting any of its individual components blocks the expression of these transcription factors and significantly impairs therapy-resistant prostate cancer,” said TSRI Research Associate Ji-Hak Jeong, the first author of the study.

Rare Obesity Syndrome Therapeutic Target Identified

Columbia University Medical Center (CUMC) researchers have discovered that a deficiency of the enzyme prohormone covertase (PC1) in the brain is linked to most of the neuro-hormonal abnormalities in Prader-Willi syndrome, a genetic condition that causes extreme hunger and severe obesity beginning in childhood. The discovery provides insight into the molecular mechanisms underlying the syndrome and highlights a novel target for drug therapy.

The findings were published online today in the Journal of Clinical Investigation.

“While we’ve known for some time which genes are implicated in Prader-Willi syndrome, it has not been clear how those mutations actually trigger the disease,” said lead author Lisa C. Burnett, PhD, a post-doctoral research scientist in pediatrics at CUMC. “Now that we have found a key link between these mutations and the syndrome’s major hormonal features, we can begin to search for new, more precisely targeted therapies.”

An estimated one in 15,000 people have Prader-Willi syndrome (PWS). The syndrome is caused by abnormalities in a small region of chromosome 15, which leads to dysfunction in the hypothalamus—which contains cells that regulate hunger and satiety—and other regions of the brain. A defining characteristic of PWS is insatiable hunger. People with PWS typically have extreme obesity, reduced growth hormone and insulin levels, excessive levels of ghrelin (a hormone that triggers hunger), and developmental disabilities. There is no cure and few effective treatments for PWS.

Dr. Burnett and her colleagues used stem cell techniques to convert skin cells from PWS patients and unaffected controls into brain cells. Analysis of the stem cell-derived neurons revealed significantly reduced levels of PC1 in the patients’ cells, compared to the controls. The cells from PWS patients also had abnormally low levels of a protein, NHLH2, which is made by NHLH2, a gene that also helps to produce PC1.

To confirm whether PC1 deficiency plays a role in PWS, the researchers examined transgenic mice that do not express Snord116, a gene that is deleted in the region of chromosome 15 that is associated with PWS. The mice were found to be deficient in NHLH2 and PC1 and displayed most of the hormone-related abnormalities seen in PWS, according to study leader Rudolph L. Leibel, MD, professor of pediatrics and medicine and co-director of the Naomi Berrie Diabetes Center at CUMC.

“The findings strongly suggest that PC1 is a good therapeutic target for PWS,” said Dr. Burnett. “There doesn’t seem to be anything wrong with the gene that makes PC1—it’s just not getting activated properly. If we could elevate levels of PC1 using drugs, we might be able to alleviate some of the symptoms of the syndrome.”

“This is an outstanding example how research on human stem cells can lead to novel insight into a disease and provide a platform for the testing of new therapies,” said Dieter Egli, PhD, a stem cell scientist who is an assistant professor of developmental cell biology (in Pediatrics) and a senior author on the paper.

“This study changes how we think about this devastating disorder,” said Theresa Strong, PhD, chair of the scientific advisory board of the Foundation for Prader-Willi Research and the mother of a child with PWS. “The symptoms of PWS have been very confusing and hard to reconcile. Now that we have an explanation for the wide array of symptoms, we can move forward with developing a drug that addresses their underlying cause, instead of treating each symptom individually.”

Following the findings reported in this paper, the Columbia research team began collaborating with Levo Therapeutics, a PWS-focused biotechnology company, to translate the current research into therapeutics.

‘Rewired’ Cells Show Promise for Targeted Cancer Therapy

Human immune cells rationally engineered to sense and respond to tumor signal

A major challenge in truly targeted cancer therapy is cancer’s suppression of the immune system. Northwestern University synthetic biologists now have developed a general method for “rewiring” immune cells to flip this action around.

“Right now, one of the most promising frontiers in cancer treatment is immunotherapy — harnessing the immune system to combat a wide range of cancers,” said Joshua N. Leonard, the senior author of the study. “The simple cell rewiring we’ve done ultimately could help overcome immunosuppression at the tumor site, one of the most intransigent barriers to making progress in this field.”

When cancer is present, molecules secreted at tumor sites render many immune cells inactive. The Northwestern researchers genetically engineered human immune cells to sense the tumor-derived molecules in the immediate environment and to respond by becoming more active, not less.

This customized function, which is not observed in nature, is clinically attractive and relevant to cancer immunotherapy. The general approach for rewiring cellular input and output functions should be useful in fighting other diseases, not just cancer.

“This work is motivated by clinical observations, in which we may know why something goes wrong in the body, and how this may be corrected, but we lack the tools to translate those insights into a therapy,” Leonard said. “With the technology we have developed, we can first imagine a cell function we wish existed, and then our approach enables us to build — by design — a cell that carries out that function.”

Currently, scientists and engineers lack the ability to program cells to exhibit all the functions that, from a clinical standpoint, physicians might wish them to exhibit, such as becoming active only when next to a tumor. This study addresses that gap, Leonard said.

Leonard, who focuses on integrating synthetic biology into medicine, is an associate professor of chemical and biological engineering at the McCormick School of Engineering. He is a member of the Robert H. Lurie Comprehensive Cancer Center of Northwestern University.

The research comes out of a rich collaboration that Leonard’s team has with clinical oncologists, immunologists and basic cancer researchers at Northwestern University Feinberg School of Medicine as well as other synthetic biologists.

The study, to be published Dec. 12 by the journal Nature Chemical Biology, provides details of the first synthetic biology technology enabling researchers to rewire how mammalian cells sense and respond to a broad class of physiologically relevant cues. Kelly A. Schwarz, a graduate student in Leonard’s research group, is the study’s first author.

“This work is exciting because it addresses a key technical gap in the field,” Schwarz said. “There is great promise for using engineered cells as programmable therapies, and it is going to take technologies such as this to truly realize that goal.”

Starting with human T cells in culture, the research team genetically engineered changes in the cells’ input and output, including adding a sensing mode, and built a cell that is relevant to cancer immunotherapy.

Specifically, the engineered cells sense vascular endothelial growth factor (VEGF), a protein found in tumors that directly manipulates and in some ways suppresses the immune response. When the rewired cells sense VEGF in their environment, these cells, instead of being suppressed, respond by secreting interleukin 2 (IL-2), a protein that stimulates nearby immune cells to become activated specifically at that site. Normal unmodified T cells do not produce IL-2 when exposed to VEGF, so the engineered behavior is both useful and novel.

This work was carried out in cells in culture, and the technology next will be tested in animal studies.

While Leonard’s team has initially focused on the application of this cell programming technology to enabling cancer immunotherapy, it can be readily extended to distinct cellular engineering goals and therapeutic applications. Leonard’s “parts” are also intentionally modular, such that they can be combined with other synthetic biology innovations to write more sophisticated cellular programs.

“To truly accelerate the rate at which we can translate scientific insights into treatments, we need technologies that let us rapidly try out new ideas, in this case by building living cells that manifest a desired biological function,” said Leonard, who also is a founding member of the Center for Synthetic Biology and a member of the Chemistry of Life Processes Institute.

“Our technology also provides a powerful new tool for fundamental research, enabling biologists to test otherwise untestable theories about how cells coordinate their functions in complex, multicellular organisms,” he said.

URI Scientist: Rare Childhood Disease Linked to Major Cancer Gene

Researcher discovers novel connection between Fanconi anemia, PTEN

A team of researchers led by a University of Rhode Island scientist has discovered an important molecular link between a rare  childhood genetic disease, Fanconi anemia, and a major cancer gene called PTEN. The discovery improves the understanding of the molecular basis of Fanconi anemia and could lead to improved treatment outcomes for some cancer patients.

According to Niall Howlett, URI associate professor of cell and molecular biology and Rhode Island’s leading expert on Fanconi anemia, the disease is characterized by birth defects, bone marrow failure and increased cancer risk. He said the genes that play a role in the development of the disease are also important in the development of hereditary breast and ovarian cancer.

Howlett’s new study now establishes a molecular link between Fanconi anemia and a gene strongly associated with uterine, prostate and brain cancer. This research was published this month in the journal Scientific Reports, with URI graduate student Elizabeth Vuono as lead author.

About 1 in 150,000 children in the United States is born with Fanconi anemia.

“People often ask why we study such a rare disease,” said Howlett, who has been studying Fanconi anemia for nearly 20 years. “First and foremost, there is no cure or effective treatments for it. So a greater understanding of the molecular basis of Fanconi anemia is critical to address this need.”

In addition, Howlett said there are countless examples of how the study of Fanconi anemia has greatly benefited the general population. The first umbilical cord blood transplant, for example, was performed with a Fanconi anemia patient. Bone marrow transplants have become much safer and more effective because of studies with Fanconi anemia patients. And new breast and ovarian cancer genes have been discovered as a result of studies on the molecular biology of Fanconi anemia.

Howlett’s current research is another example of the broader impact of Fanconi anemia studies.

The URI researcher speculated about the existence of a biochemical link between Fanconi anemia and PTEN. Mutations in PTEN occur frequently in uterine, prostate and brain cancer.

“The PTEN gene codes for a phosphatase – an enzyme that removes phosphate groups from proteins,” explained Howlett. “Many Fanconi anemia proteins have phosphate groups attached to them when they become activated. However, how these phosphate groups are removed is poorly understood.”

Howlett said that cells from Fanconi anemia patients are characteristically sensitive to a class of drugs widely used in cancer chemotherapy called DNA crosslinking agents.

“So we performed an experiment to determine if Fanconi anemia and PTEN were biochemically linked,” he said. “By testing if cells with mutations in the PTEN gene were also sensitive to DNA crosslinking agents, we discovered that Fanconi anemia patient cells and PTEN-deficient cells were practically indistinguishable in terms of sensitivity to these drugs. This strongly suggested that the Fanconi anemia proteins and PTEN might work together to repair the DNA damage caused by DNA crosslinking agents.”

By using epistasis analysis, a genetic method that determines if genes work together, Howlett and his research group found that the Fanconi anemia proteins and PTEN do indeed function together in this repair pathway.

“Before this work, Fanconi anemia and PTEN weren’t even on the same radar,” said Howlett. “This is really important to understanding how this disease arises and what its molecular underpinnings are. The more we can find out about its molecular basis, the more likely we are to come up with strategies to treat the disease.”

Howlett’s research is equally important to cancer patients who do not have Fanconi anemia. He said that since his study found that cells missing PTEN are highly sensitive to DNA crosslinking agents, it should be possible to predict whether a particular cancer patient will respond to this class of chemotherapy drug by conducting a simple DNA test.

“We can now predict that if a patient has cancer associated with mutations in PTEN, then it is likely that the cancer will be sensitive to DNA crosslinking agents,” he said. “This could lead to improved outcomes for patients with certain types of PTEN mutations.”