Genetically enhanced, cord-blood derived immune cells strike B-cell cancers

Immune cells with a general knack for recognizing and killing many types of infected or abnormal cells also can be engineered to hunt down cells with specific targets on them to treat cancer, researchers at The University of Texas MD Anderson Cancer Center report in the journal Leukemia.

The team’s preclinical research shows that natural killer cells derived from donated umbilical cords can be modified to seek and destroy some types of leukemia and lymphoma. Genetic engineering also boosts their persistence and embeds a suicide gene that allows the modified cells to be shut down if they cause a severe inflammatory response.

A first-in-human phase I/II clinical trial of these cord-blood-derived, chimeric antigen receptor-equipped natural killer cells opened at MD Anderson in June for patients with relapsed or resistant chronic lymphocytic leukemia (CLL), acute lymphocytic leukemia (ALL), or non-Hodgkin lymphoma. All are cancers of the B cells, another white blood cell involved in immune response.

“Natural killer cells are the immune system’s most potent killers, but they are short-lived and cancers manage to evade a patient’s own NK cells to progress,” said Katy Rezvani, M.D., Ph.D., professor of Stem Cell Transplantation and Cellular Therapy.

“Our cord-blood derived NK cells, genetically equipped with a receptor that focuses them on B-cell malignancies and with interleukin-15 to help them persist longer — potentially for months instead of two or three weeks — are designed to address these challenges,” Rezvani said.

Moon Shots Program funds project

The clinical trial is funded by MD Anderson’s Moon Shots Program™, designed to more rapidly develop life-saving advances based on scientific discoveries.

The chimeric antigen receptor (CAR), so-called because it’s added to the cells, targets CD19, a surface protein found on B cells.

In cell lines and mouse models of lymphoma and CLL, CD19-targeted NK cells killed cancer cells and extended survival of animals compared to simply giving NK cells alone. Addition of IL-15 to the CD19 receptor was crucial for the longer persistence and enhanced activity of the NK cells against tumor cells.

NK cells are a different breed of killer from their more famous immune system cousins, the T cells. Both are white blood cells, but T cells are highly specialized hunters that look for invaders or abnormal cells that bear a specific antigen target, kill them and then remember the antigen target forever.

Natural killers have an array of inhibitory and activating receptors that work together to allow them to detect a wider variety of infected, stressed or abnormal cells.

“By adding the CD19 CAR, we’re also turning them into guided missiles,” said Elizabeth Shpall, M.D., professor of Stem Cell Transplantation and Cell Therapy.

Using a viral vector, the researchers transduce NK cells taken from cord blood with the CD19 CAR, the IL-15 gene, and an inducible caspase-9-based suicide gene.

Cell line tests found the engineered NK cells to be more efficient killers of lymphoma and CLL cells, compared to unmodified NK cells, indicating the engineered cells’ killing was not related to non-specific natural killer cell cytotoxicity.

Another experiment showed the engineered cord blood NK cells killed CLL cells much more efficiently than NK cells taken from CLL patients and engineered, highlighting the need to transplant CAR-engineered NK cells from healthy cord blood rather than use a patient’s own cells.

Suicide gene to counter cytokine release syndrome

Mouse model lymphoma experiments using a single infusion of low dose NK cells resulted in prolongation of survival. At a higher, double dose, none of the mice treated with the CD19/IL-15 NK cells died of lymphoma, with half surviving for 100 days and beyond. All mice treated with other types of NK cells died by day 41.

A proportion of mice treated with the higher dose of engineered NK cells died of cytokine release syndrome, a severe inflammatory response that also occurs in people treated with CAR T cells.

To counteract this toxicity, the researchers incorporated a suicide gene (iC9) that can be activated to kill the NK cells by treatment with a small-molecule dimerizer. This combination worked to swiftly reduce the engineered NK cells in the mouse model.

Subsequent safety experiments were conducted in preparation for the clinical trial. Rezvani, the principal investigator of the clinical trial, says the protocol calls for vigilance for signs of cytokine release syndrome, treatment with steroids and tocilizumab for low-grade CRS with AP1903 added to activate the suicide gene for grade 3 or 4 CRS.

NK CARs available off the shelf

T cells modified with chimeric antigen receptors against CD19 have shown efficacy in clinical trials. In these therapies, a patient’s own T cells are modified, expanded, and given back to the patient, a process that takes weeks. Finding a matched donor for T cells would be a challenge, but would be necessary because unmatched T cells could attack the recipient’s normal tissue – graft vs. host disease.

Rezvani and Shpall have given patients cord-blood derived NK cells in a variety of clinical trials and found that they do not cause graft vs. host disease, therefore don’t have to be matched. NK cells can be an off-the-shelf product, prepared in advance with the necessary receptor and given promptly to patients.

“CAR NK cells are scalable in a way that CAR T cells are not,” Rezvani noted.

A strength of T cells is the development of memory cells that persist and repeatedly attack cells bearing the specific antigen that return. NK cells do not seem to have a memory function, but Rezvani says the experience of the longer-lived mice, which are now more than a year old, raises the possibility that a prolonged NK cell attack will suffice.

Shpall, Rezvani and colleagues are developing cord blood NK CARs for other targets in a variety of blood cancers and solid tumors.

MD Anderson and the researchers have intellectual property related to the engineered NK cells, which is being managed in accordance with the institution’s conflict-of-interest rules.

Shpall founded and directs MD Anderson’s Cord Blood Bank, originally established to provide umbilical cord blood stem cells for patients who need them but cannot get a precise donor match. Donated by mothers who deliver babies at seven Houston hospitals and two others from California and Michigan, the bank now has 26,000 cords stored. MD Anderson researchers pioneered the extraction and expansion of NK cells from umbilical cords.

Promising new therapeutic approach for debilitating bone disease

An Australian-led research team has demonstrated a new therapeutic approach that can re-build and strengthen bone, offering hope for individuals with the debilitating bone cancer, multiple myeloma.

The findings were published today in the medical journal Blood, and were presented at an international meeting of bone biology experts in Brisbane earlier this month.

The researchers tested a new type of treatment that specifically targets a protein called sclerostin, which in healthy bones is an important regulator of bone formation. Sclerostin halts bone formation, and the researchers speculated that if they could inhibit the action of sclerostin, they could reverse the devastating bone disease that occurs with multiple myeloma.

Dr Michelle McDonald and Professor Peter Croucher, of the Bone Biology Division of the Garvan Institute of Medical Research in Sydney, led the study.

“Multiple myeloma is a cancer that grows in bone, and in most patients it is associated with widespread bone loss, and recurrent bone fractures, which can be extremely painful and debilitating,” says Dr McDonald.

“The current treatment for myeloma-associated bone disease with bisphosphonate drugs prevents further bone loss, but it doesn’t fix damaged bones, so patients continue to fracture. We wanted to re-stimulate bone formation, and increase bone strength and resistance to fracture.”

The new therapeutic approach is an antibody that targets and neutralises sclerostin, and in previous clinical studies of osteoporosis, such antibodies have been shown to increase bone mass and reduce fracture incidence in patients.

The researchers tested the anti-sclerostin antibody in mouse models of multiple myeloma, and found that not only did it prevent further bone loss, it doubled bone volume in some of the mice.

Dr McDonald says, “When we looked at the bones before and after treatment, the difference was remarkable – we saw less lesions or ‘holes’ in the bones after anti-sclerostin treatment.

“These lesions are the primary cause of bone pain, so this is an extremely important result.”

The researchers have a biomechanical method to test bone strength and resistance to fracture, and found that the treatment also made the bones substantially stronger, with more than double the resistance to fracture observed in many of the tests.

They then combined the new antibody with zoledronic acid, a type of bisphosphonate drug, the current standard therapy for myeloma bone disease.

“Bisphosphonates work by preventing bone breakdown, so we combined zoledronic acid with the new anti-sclerostin antibody, that re-builds bone. Together, the impact on bone thickness, strength and resistance to fracture was greater than either treatment alone,” says Dr McDonald.

The findings provide a potential new clinical strategy for myeloma. While this disease is relatively rare, with approximately 1700 Australians diagnosed every year, the prognosis is extremely poor, with less than half of those diagnosed expected to survive for more than five years.

Prof Croucher, Head of the Bone Biology Division at Garvan, says that preventing the devastating bone disease of myeloma is critical to improve the prognosis for these people.

“Importantly, myelomas, like other cancers, vary from individual to individual and can therefore be difficult to target. By targeting sclerostin, we are blocking a protein that is active in every person’s bones, and not something unique to a person’s cancer. Therefore, in the future, when we test this antibody in humans, we are hopeful to see a response in most, if not all, patients,” Prof Croucher says.

“We are now looking towards clinical trials for this antibody, and in the future, development of this type of therapy for the clinical treatment of multiple myeloma.

“This therapeutic approach has the potential to transform the prognosis for myeloma patients, enhancing quality of life, and ultimately reducing mortality.

“It also has clinical implications for the treatment of other cancers that develop in the skeleton.”

Study shows biomarkers can predict which ER-positive breast cancer patients respond best to first-line therapy

Two challenges in treating patients with estrogen-positive breast cancer (ER+) have been an inability to predict who will respond to standard therapies and adverse events leading to therapy discontinuation. A study at The University of Texas MD Anderson Cancer Center revealed new information about how the biomarkers retinoblastoma protein (Rb) and cytoplasmic cyclin E could indicate which patients will respond best to current first-line therapies.

The study also discovered that combining the current therapy with autophagy inhibitors will result in using one-fifth of the dosage of the standard treatment, which could significantly reduce side effects associated with this therapy. Findings were published in the June 27 issue of Nature Communications.

Standard treatment, consisting of palbociclib, often has adverse side effects and not all ER+ patients respond to the therapy. Palbociclib inhibits proteins called CDK4 and CDK6 (CDK4/6) and tumor cells escape this inhibition by activating autophagy, a process allowing cancer cells to thrive even when starved of nutrients. By combining palbocicilb with autophagy inhibitors in cells that express normal Rb and nuclear cyclin E, the dose of palbociclib was significantly reduced.

Khandan Keyomarsi, Ph.D., professor of Experimental Radiation Oncology, led a team that demonstrated how CDK4/6 and autophagy inhibitors synergistically induce cell senescence in Rb-positive cytoplasmic cyclin E-negative cancers. CDK4/6 inhibitors are approved by the Food and Drug Administration (FDA).

“Our findings could impact the majority of ER+ and HER2-negative breast cancers accounting for about 60 percent of advanced breast cancers,” said Keyomarsi. “We demonstrated for the first time evidence that Rb and cytoplasmic cyclin E status have a very strong effect on predicting response to the current standard first-line therapy for this population of patients, hormonal therapy plus palbociclib.We also discovered that by inhibiting the pathway such as autophagy that causes tumor cells to escape palbociclib growth inhibition, CDK4/6 inhibitor was more effective.”

Deregulation of cell cycle checkpoint proteins, such as CDK4/6, is a key hallmark of cancer, resulting in uncontrolled cellular growth and tumor formation. Some CDK4/6 inhibitors, including palbociclib, ribociclib and abemaciclib, have shown potential in pre-clinical and clinical studies in numerous solid tumors. Palbociclib has demonstrated benefits in Phase II and III trials in advanced ER+ breast cancers, doubling progression-free survival compared to drugs such as letrozole or fulvestrant, and is currently being evaluated clinically in other solid tumors.

“Data provided through The Cancer Genome Atlas revealed alterations in the CDK4/6/cyclin D pathway in about 35 percent of the patients, making them an ideal population for targeting CDK4/6,” said Keyomarsi. “Our study revealed that inhibition of CDK4/6 and autophagy pathways cooperate to induce sustained growth inhibition and senescence in vitro and in vivo, in breast and other solid tumors and showed how autophagy inhibition can significantly decrease the dose of palbociclib required to treat breast cancer patients. We believe this new strategy can improve the efficacy of other CDK4/6 inhibitor treatments like ribociclib and abemaciclib.”

The team’s findings indicated how Rb and cyclin E status predicts response to a combination of CDK4/6 and autophagy inhibition in pre-clinical models and that autophagy blockade is successful in reversing resistance to palbociclib.

“Palbociclib resistance is a significant limitation of this treatment which is not curative and does not prolong survival even though transient responses and prolongation of response have formed the basis of FDA approval,” said Keyomarsi. “Our study provides evidence that models of hormone receptor-negative cancer and even non-breast cancer malignancies can respond to the combination of palbociclib and autophagy inhibition, when selected based on Rb and cyclin E isoform status, representing a completely new therapeutic opportunity for these cancers.”

Keyomarsi and colleagues anticipate future clinical studies based on this pre-clinical and clinical evidence with the aim of developing translational and clinical applications.

Researching Radiosensitizers, a New Class of Drugs That Would Make Tumors More Vulnerable to Radiation Therapy

Two out of three cancer patients are treated with radiation, but the therapy often fails to wipe out the tumor or slow its growth. Southern Research is working to develop a new class of drugs that will help the radiation deliver a more powerful punch to the disease.

Dr. Bo Xu, M.D., Ph.D., Distinguished Fellow and Chair of Southern Research’s Oncology Department, said a radiosensitizer would greatly benefit cancer patients by improving the success rate of radiation by reducing resistance to the treatment.

“Our project focuses on making those tumor cells more vulnerable to radiation by targeting a critical survival mechanism that allows them to recover from the effects of radiation,” Xu said.

It’s a challenging project, in the works for almost a decade. It got started when Southern Research scientists began looking at fundamental biology concepts to identify a pathway that could play a role in the ability of cancer cells to survive radiation.

They discovered that disrupting the tumor’s self-protection mechanism – in this case, an interaction between two specific proteins – makes the cancer more sensitive to radiation treatment, Xu said.

“The whole idea is to use this strategy to find a new drug that can be used by patients who receive radiation. This drug wouldn’t have toxicity because if it got into the cell it wouldn’t mess up the major functions of the protein network,” he said.

“It would only work when radiation is delivered, and that radiation would be more effective. It’s like a catalyst.”

Using funding from the Alabama Drug Discovery Alliance (ADDA), a partnership with the University of Alabama at Birmingham, Southern Research scientists recently scanned thousands of compounds to identify potential drug candidates. The focus now is to validate the results of those scans and to identify lead compounds for more testing.

“Our hope is that in three years, we can identify a novel class of radiosensitizers that can help the approximately two-thirds of cancer patients who will eventually receive radiotherapy,” Xu said.

While some forms of cancer, such as lymphoma, are sensitive to radiation therapy, many others are not. Solid tumors with a low supply of oxygen, called hypoxic tumors, are tough to treat with radiation. So are cancer cells with a high DNA-repair capability.

To develop a radiosensitizer, Xu is taking aim at a protein that binds to DNA and recognizes the damage being done by radiation. The protein then joins forces with an enzyme to initiate a molecular repair job.

“If that recruitment is successful, then the DNA damage will be repaired, and the cancer cell will survive,” Xu said. “What we’re trying to do is to block this protein from finding the other one, so that the repair process will be diminished or affected. That way, the tumor cells will die.”

To prevent the DNA repair job from getting started, Xu is investigating a small peptide mimic, a small sequence of amino acids that is similar to a human protein but just a fraction of its size. These strands get to the site to block the interaction of the two natural, full-size proteins.

“This interference makes the cancer cell more vulnerable to radiation treatment,” he said.

Radiosensitizers are in demand, but they have proved difficult to develop. While the concept has been around for half a century, very few radiosensitizers have actually become available, according to Xu.

“While there are compounds that work synergistically with radiation, there are few drugs that were developed as a pure radiosensitizer,” he said.

In addition to the ADDA, the National Institutes of Health and the Department of Defense prostate cancer program have provided Southern Research with funding for this research over the years.

Cancer therapy shows promise for psoriasis treatment

HDAC inhibitors, already widely used to treat cancer, may be an effective therapy for psoriasis as well, scientists report.

They have shown that HDAC3 inhibitors are particularly adept at increasing expression of aquaporin-3, or AQP3, a channel that transports glycerin, a natural alcohol and water attractor, which helps skin look better and aids healthy production and maturation of high-turnover skin cells.

“We’ve found that HDAC3 normally inhibits expression of AQP3 and we think we can use this knowledge to treat patients with psoriasis,” said Dr. Vivek Choudhary, molecular biologist and physiologist in the Department of Physiology at the Medical College of Georgia at Augusta University.

MCG scientists knew that AQP3 levels were lower in psoriasis than in healthy skin, said Choudhary, corresponding author of the study in the Journal of Investigative Dermatology. The protein helps skin cells proliferate, differentiate into the right kind of cells and get to the right location in the body. It also aids the skin’s hydration, wound recovery and elasticity. Histone deacetylase, which they found suppresses AQP3, helps regulate gene expression and protein function.

Since the immune system is believed to play a key role in psoriasis, many current treatments generally suppress the immune response, which increases the risk of infections, even cancer. MCG scientists hope they can one day instead directly enhance the presence of AQP3 or maybe its key cargo glycerin.

Psoriasis is one of the most common skin disorders, with red, flaky patches most often erupting on the elbows, knees, scalp and back, said Dr. Wendy B. Bollag, cell physiologist in the MCG Department of Physiology and the study’s senior author.

Like cancer, inflammation and excessive proliferation of cells are a psoriasis hallmark. That common ground and other emerging clues got the scientists thinking about the treatment potential of HDAC inhibitors. But first they had to establish a relationship.

When they introduced a broad-acting HDAC inhibitor to normal skin cells, or keratinocytes, – both mouse and human – they found expression of AQP3 went up within 24 hours, the first time the relationship had been noted.

They reiterated that AQP3 was critical because when it was missing, there was no commensurate increase in glycerin. AQP3 knockout mice also further clarified AQP3’s role in skin hydration, elasticity and wound healing and that it is glycerin – rather than water – that is most key to these healthy functions.

They also found that p53, a known, natural tumor suppressor that also supports cell differentiation, helps the HDAC inhibitors enable more AQP3 and ultimately more glycerin, Choudhary said. HDACs also are known to inhibit p53 activity. However overexpressing p53 by itself did not result in increased functional levels of AQP3, the scientists found.

The MCG scientists first used the HDAC inhibitor, suberoylanilide hydroxamic acid, or SAHA, which was approved by the Food and Drug Administration more than a decade ago to treat cutaneous T cell lymphoma, which has symptoms that can include dry, itchy skin as well as enlarged lymph nodes.

“We think this is one of the ways it works,” Bollag said of SAHA and their new findings. They also used several other HDAC inhibitors and found the ones that suppressed HDAC3 were also most effective at increasing AQP3.

AQP3 is adept at hauling glycerin, the backbone of many lipids and typically a key ingredient in skin lotion. Bollag’s lab reported in the Journal of Investigative Dermatology in 2003 that glycerin helps skin cells mature properly. Inside skin cells, phospholipase D – an enzyme that converts fats or lipids in the external protective cell membrane into cell signals – and AQP3 interact. AQP3 hands off glycerin, which produces phosphatidylglycerol, which, in turn, aids skin cell differentiation.

“We think phosphatidylglycerol is the key,” Bollag said of the positive results. “If you don’t have enough, you may have psoriasis.”

The Bollag lab and others also had found that AQP3, which is present in psoriasis, appears rather immature and out of place, largely inside the cell cytoplasm instead of on the protective, outer cell membrane. The inner location puts quite a damper on its normal mature function of transporting glycerin, water and other substances through the membrane.

“If you use antibodies to visualize where AQP3 is in the keratinocytes, you will see it nicely outlining the cells because it’s right there on the plasma membrane,” Bollag said. “So clearly it’s normally expressed in keratinocytes but the fact that we can upregulate it even more with an HDAC3 inhibitor suggests that normally HDAC3 keeps it in check.”

Cambridge, Massachusetts-based biotech company Shape Pharmaceuticals Inc., currently has a topical version of an HDAC inhibitor in clinical trials for cutaneous T cell lymphoma. If psoriasis patients end up taking HDAC inhibitors, low doses or a topical application likely would help avoid some side effects, including nausea, Bollag said.

One way HDAC inhibitors help fight cancer is by temporarily loosening DNA, increasing the expression of tumor-suppressing genes and making the tumor more vulnerable. HDAC inhibitors also are being explored for their potential in treating neurological diseases such as Huntington’s.

Others have provided evidence that dysregulation of AQP3 contributes to psoriasis and AQP3 is linked to other skin diseases as well like atopic dermatitis – the most common type of eczema and vitiligo, which results in white patches on the skin.

Interestingly, even though psoriatic cells are known for their propensity to replicate, it’s hard to grow an adequate number of cells for scientific study: they increase a certain amount then go quiet. There also is no real animal model of psoriasis. Moving forward, the MCG scientists may try developing a model using a topical drug for genital warts since some patients who take it develop psoriasis.

Better treatment for kidney cancer thanks to new mouse model

Roughly 2-3 percent of all people suffering from cancer have kidney cancer. The most common form of this disease is called clear cell renal cell carcinoma (ccRCC). In roughly half of all patients with this disease, the tumor develops metastases and generally cannot be cured.

New Mouse Model for Investigating Kidney Cancer

The research of different types of cancer and the testing of new treatments depends on accurate mouse models. This is because the tumors in mice mirror the genetics as well as the molecular and cellular properties of tumors in humans. Despite decades of effort, however, researchers were unable to develop a mouse model of renal cell carcinoma – until now. Scientists conducting a long-term research project at the University of Zurich were able to develop a mouse model. The study was led by Sabine Harlander and her colleagues at the Institute of Physiology of the University of Zurich in the lab of Professor Ian Frew, who has recently joined the University of Freiburg in Germany. The researchers began by identifying the genes that often mutate in human renal cell carcinomas. They then mutated three of these genes simultaneously in renal cells of mice, which then developed renal cancer.

Gene Mutations Promote Uncontrolled Cell Division

The progression from gene mutation in the renal cells to the development of a tumor took eight to twelve months. This lengthy period of time, compared to a mouse’s lifetime, indicates that additional factors play a role in tumor development. The researchers therefore decided to take a closer look at the protein-encoding genes in the mouse tumors. They discovered that in all of the tumors at least one of the many genes responsible for the correct functioning of the primary cilium had mutated. The primary cilium is a hair-like structure found on the cell’s surface and is responsible for coordinating cell signaling, among other things.

Based on this finding, the researchers found that similar mutations also occur in renal cell carcinomas in humans. The scientists now believe that the loss of normal function in the primary cilium leads to the uncontrollable division of renal epithelial cells, which contributes to the formation of ccRCC. “This research project is a prime example of how mouse models can help us to better understand cancer diseases in human beings,” says Sabine Harlander.

Mouse Model Enables Development of Better Treatments

The new mouse model will make it possible to develop better therapies for renal cancer. For example, in the case of patients with renal carcinoma metastasis who are given different medications, some patients respond to the medications, while others do not. The same phenomenon can be observed when mice with renal cancer are treated with the same drugs as the humans. Some tumors shrink, while others do not. Now researchers can investigate the factors that contribute to why certain tumours respond to certain medications and not to others. “We hope that our mouse model, which allows us to combine drug testing and genetic analysis, will provide a deeper understanding of why tumors are sensitive or resistant to drugs,” states Ian Frew. Such vital information could be used to better adjust treatments to the characteristics of each patient.

The mouse model could also contribute to the further development of immunotherapies – a method in which the body’s immune system is stimulated, so that it intensifies its fight against tumor cells. In the last few years, much progress has been made in this field of cancer research, also for the treatment of renal cell carcinomas. Now, thanks to the new mouse model, it will be possible to study how renal tumors are able to develop in an environment with a normal immune system, and how cancer cells manage to evade the immune system’s attacks. Ultimately, the researchers’ goal is to use these new findings to improve the effectiveness of immunomodulatory treatments.

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.”