The results come from a team of scientists investigating why a gene called ApoE4 makes people more prone to developing Alzheimer’s. People who carry two copies of the gene have roughly eight to 10 times the risk of getting Alzheimer’s disease than people who do not.

A team of scientists from the University of Rochester, the University of Southern California, and other institutions found that ApoE4 works through cyclophilin A, a well-known bad actor in the cardiovascular system, causing inflammation in atherosclerosis and other conditions. The team found that cyclophilin A opens the gates to the brain assault seen in Alzheimer’s.

“We are beginning to understand much more about how ApoE4 may be contributing to Alzheimer’s disease,” said Robert Bell, Ph.D., the post-doctoral associate at Rochester who is first author of the paper. “In the presence of ApoE4, increased cyclophilin A causes a breakdown of the cells lining the blood vessels in Alzheimer’s disease in the same way it does in cardiovascular disease or abdominal aneurysm. This establishes a new vascular target to fight Alzheimer’s disease.”

The team found that ApoE4 makes it more likely that cyclophilin A will accumulate in large amounts in cells that help maintain the blood-brain barrier, a network of tightly bound cells that line the insides of blood vessels in the brain and carefully regulates what substances are allowed to enter and exit brain tissue.

ApoE4 creates a cascade of molecular signaling that weakens the barrier, causing blood vessels to become leaky. This makes it more likely that toxic substances will leak from the vessels into the brain, damaging cells like neurons and reducing blood flow dramatically by choking off blood vessels.

Doctors have long known that the changes in the brain seen in Alzheimer’s patients – the death of crucial brain cells called neurons – begins happening years or even decades before symptoms appear. The steps described in Nature discuss events much earlier in the disease process.

The idea that vascular problems are at the heart of Alzheimer’s disease is one championed for more than two decades by Berislav Zlokovic, M.D., Ph.D., the leader of the team and a neuroscientist formerly with the University of Rochester Medical Center and now at USC. For 20 years, Zlokovic has investigated how blood flow in the brain is affected in people with the disease, and how the blood-brain barrier allows nutrients to pass into the brain, and harmful substances to exit the brain.

At Rochester, Zlokovic struck up a collaboration with Bradford Berk, M.D., Ph.D., a cardiologist and CEO of the Medical Center. For more than two decades Berk has studied cyclophilin A, showing how it promotes destructive forces in blood vessels and how it’s central to the forces that contribute to cardiovascular diseases like atherosclerosis and heart attack.

“As a cardiologist, I’ve been interested in understanding the role of cyclophilin A in patients who suffer from cardiovascular illness,” said Berk, a professor at the Aab Cardiovascular Research Institute. “Now our collaboration in Rochester has resulted in the discovery that it also has an important role in Alzheimer’s disease. The finding reinforces the basic research enterprise – you never know when knowledge gained in one area will turn out to be crucial in another.”

In studies of mice, the team found that mice carrying the ApoE4 gene had five times as much cyclophilin A compared to other mice in cells known as pericytes, which are crucial to maintaining the integrity of the blood-brain barrier. Blood vessels died, blood did not flow as completely through the brain as it did in other mice, and harmful substances like thrombin, fibrin, and hemosiderin, entered the brain tissue.

When the team blocked the action of cyclophilin A, either by knocking out its gene or by using the drug cyclosporine A to inhibit it, the damage in the mice was reversed. Blood flow resumed to normal, and unhealthy leakage of toxic substances from the blood vessels into the brain was slashed by 80 percent.

The team outlined the chain of events involved. Briefly:

• When ApoE4 is present, cyclophilin A is much more plentiful;
• Cyclophilin A causes an increase in the inflammatory molecule NF Kappa B;
• NF Kappa B boosts levels of certain types of molecules known as MMPs or matrix metalloproteinases that are known to damage blood vessels, reducing blood flow.

Altogether, the activity results in a dramatic boost in the amount of toxic substances in brain tissue. And when the cascade is interrupted at any of several points – when ApoE4 is not present, when cyclophilin A is blocked or shut off, or when NF Kappa B or the MMPs are inhibited – the blood-brain barrier is restored, blood flow returns to normal, and toxic substances do not leak into brain tissue.

For many years, researchers studying Alzheimer’s disease have been focused largely on amyloid beta, a protein structure that accumulates in the brains of patients with Alzheimer’s disease. The latest works points up the importance of other approaches, said Zlokovic, an adjunct professor at Rochester. At USC, Zlokovic is also deputy director of the Zilkha Neurogenetic Institute, director of the Center for Neurodegeneration and Regeneration, and professor and chair of the Department of Physiology and Biophysics.

“Our study has shown major neuronal injury resulting from vascular defects that are not related to amyloid beta,” said Zlokovic. “This damage results from a breakdown of the blood-brain barrier and a reduction in blood flow.

“Amyloid beta definitely has an important role in Alzheimer’s disease,” added Zlokovic. “But it’s very important to investigate other leads, perhaps where amyloid beta isn’t as centrally involved.”

In addition to Bell, Berk and Zlokovic, authors include, from Rochester, Ethan Winkler, Itender Singh, Abhay Sagare, Rashid Deane, Zhenhua Wu, and Jan Sallstrom. Additional authors include David Holtzman from Washington University School of Medicine, and Christopher Betsholtz and Annika Armulik of the Karolinska Institutet in Sweden.

At Rochester, Zlokovic’s team was anchored in the Center for Neurodegenerative and Brain Vascular Disorders, which was directed by Zlokovic, and in the Department of Neurosurgery. Bell was a graduate student with Zlokovic and now does research in the laboratory of Joseph Miano, Ph.D., at the Aab Cardiovascular Research Institute.

The work was funded by the National Institute of Neurological Disorders and Stroke and the National Institute on Aging.

Contact: Tom Rickey
tom_rickey@urmc.rochester.edu
585-275-7954
University of Rochester Medical Center

Published recently in Nature Cell Biology, the study reveals a pathway that, if deactivated, may help slow the development of the disease.

“We have the ability to target this pathway with drugs that already exist,” said lead investigator Michele Pagano, MD, the May Ellen and Gerald Jay Ritter Professor of Oncology in the Department of Pathology and a member of the NYU Cancer Institute at NYU Langone Medical Center, and a Howard Hughes Medical Institute investigator. “Many other lymphomas are also controlled by this pathway, so while we’re optimistic that this discovery will provide a way to kill multiple myeloma cells, we’re also very hopeful that this can be applied to other lymphomas and that it will have a major impact on these aggressive cancers.”

Pagano and colleagues put together several new pieces of the puzzle surrounding the survival and spread of multiple myeloma cells and the Nuclear Factor kappa B (NF-кB) pathway. This complicated pathway induces transcription of genes that control inflammation, immunity, and certain developmental processes. It is also known to frequently be involved in disease.

In normally functioning cells, the NF-кB pathway turns off and on, triggered by the accumulation and then degradation, or breakdown, of a protein called p100. When the pathway is “on,” p100 is degraded, allowing for pathway-dependent gene transcription. Several hours after the pathway’s activation, p100 begins to accumulate in the cell’s nucleus, naturally blocking the pathway, so that the gene transcription signal is temporarily blocked and transcription is halted.

In lymphomas, including multiple myeloma, however, the NF-кB pathway remains active, providing a refuge for lymphoma cells to hide. In fact, within this active pathway, the lymphoma cells are able to evade apoptosis, or cell death, allowing them to proliferate in an uncontrolled way.

“Activating mutations in the NF-кB pathway does not generally represent the initial oncogenic event,” Dr. Pagano said. “But they are necessary for the survival and spread of the cancer.”

Dr. Pagano explained the steps involved with pathway’s activation and deactivation in more scientific detail: The process begins in the pathway’s off state, with the accumulation of p100. To clear the pathway and naturally turn it back on, a sequence of events has to happen. First, a kinase, which the team identified as GSK3, phosphorylates p100. The phosphorylation draws the attention of Fbxw7α, a subunit of a ubiquitin ligase, which binds to the portion of p100 that has been phosphorylated by GSK3. The addition of Fbxw7α to the p100 protein then causes ubiquitin to seek out p100. Ubiquitin attaches to the protein and modifies it in a way such that it is recognized by a protease whose job it is to recognize and degrade any protein that has been modified by ubiquitin conjugation. As a result, p100 is degraded in the nucleus of the cell and the pathway is cleared and activated, turning on the gene transcription signal.

These new findings lead the researchers to conclude that the intersection of GSK3, Fbxw7α and p100 may serve as a potential intervention point for the treatment of multiple myeloma. Researchers believe if they can find a way to target the elimination of p100 they may be able to inactivate the pathway, which would eliminate the tumor cells’ safe haven so that they would be susceptible to apoptosis. This would in turn promote the death of multiple myeloma cells.

According to Dr. Pagano, this strategy may not be too far from becoming a reality. There are already drugs being tested in clinical trials for Alzheimer’s Disease that work by inhibiting GSK3. With the current study, the research team from NYU School of Medicine has shown that, by blocking GSK3 from phosphorylating p100, it is possible to prevent the degradation of p100, which then blocks the NF-кB pathway, thereby halting gene transcription and blocking tumor cells’ safe zone. Alternatively, pharmaceutical research may find a way to target Fbxw7α, which would keep the pathway turned off in the same way.

“Cancers are persistent and tenacious,” Dr. Pagano said. “There are millions of pieces to the puzzle of how they work and we’ve discovered a few more pieces of that puzzle. It is very possible that we can find an inhibitor of Fbxw7α since there are already drugs being tested that inhibit very similar enzymes.”

Moreover, Dr. Pagano explained, it is likely that the effects observed in multiple myeloma may be generalized to other B-cell neoplasms, types of lymphomas, especially those in which the tumor cells hide out in the NF-кB pathway.

“These new findings strongly suggest that by targeting this enzyme, we will kill multiple myeloma cells and other B-cell lymphomas,” he said. “And that, from a researcher’s perspective, is a very exciting prospect.”

The study was funded by the National Institutes of Health, the National Cancer Institute, the National Institute of General Medical Sciences, the Multiple Myeloma Research Foundation, the American Italian Cancer Foundation, and the Howard Hughes Medical Institute.

About NYU School of Medicine:

NYU School of Medicine is one of the nation’s preeminent academic institutions dedicated to achieving world class medical educational excellence. For 170 years, NYU School of Medicine has trained thousands of physicians and scientists who have helped to shape the course of medical history and enrich the lives of countless people. An integral part of NYU Langone Medical Center, the School of Medicine at its core is committed to improving the human condition through medical education, scientific research and direct patient care. The School also maintains academic affiliations with area hospitals, including Bellevue Hospital, one of the nation’s finest municipal hospitals where its students, residents and faculty provide the clinical and emergency care to New York City’s diverse population, which enhances the scope and quality of their medical education and training. Additional information about the NYU School of Medicine is available at http://school.med.nyu.edu/.

Contact: Jessica Guenzel
jessica.guenzel@nyumc.org
212-404-3591
NYU Langone Medical Center / New York University School of Medicine

The new research findings demonstrate that cardiac ischemia plays an important role in adenovector gene transfection (delivery) in mammalian hearts. Based on this understanding, using a standard balloon angioplasty catheter, researchers have developed and tested a new method to induce transient ischemia during a non-surgical interventional cardiac procedure, which when coupled with the infusion of nitroglycerin, boosts the delivery (cell transfection) of an adenovector gene construct into heart cells. The increase in adenovector-based gene transfection with the new technique is over two orders of magnitude (>100 fold).

Cardium’s new method of adenovector delivery takes advantage of the findings that transient ischemia appears to alter the permeability barrier of the vascular endothelium and may expose the blood to the coxsackie-adenovirus receptor mediating adenovector uptake by the heart. Balloon angioplasty catheters have been used for many years to dilate blocked coronary arteries, sometimes with use of a stent, and these catheters have also been used safely by cardiologists in patients with coronary artery disease to study the effects of brief ischemia. Cardium’s new technique inflates the balloon in non-narrowed areas, and only enough to briefly interrupt flow using inflation pressure that is less than that used for performing angioplasty.

Cardium’s recently initiated Russian-based ASPIRE Phase 3 / registration clinical study uses transient ischemia techniques during non-surgical percutaneous catheterization with a standard angioplasty catheter together with the intracoronary infusion of nitroglycerin with the Generx® [Ad5FGF-4] product candidate for the treatment of patients with myocardial ischemia and stable angina pectoris. These patients have atherosclerotic coronary artery disease, and the Company’s Generx product candidate is intended to stimulate the growth of new or additional collateral blood vessels to bypass blockages.

These studies were conducted at Emory University School of Medicine, led by Jakob Vinten-Johnasen, PhD., and co-sponsored by a Small Business Innovative Research grant from the National Institutes of Health (Cardium Therapeutics) and the Carlyle Fraser Heart Center (Emory). At the conference Gabor M. Rubanyi, MD, PhD, Cardium’s Chief Scientific Officer, will present the late-breaking poster entitled “Transient Ischemia is Necessary for Efficient Adenovector Gene Transfer in the Heart”, on May 17, 2012 from 3:00 to 5:30 p.m. in Exhibit Hall A. The poster presentation can be viewed at http://www.cardiumthx.com/pdf/Generx-ASGCT-Poster-Presentation-May-2012.pdf.

In addition, Dr. Rubanyi will also make an oral presentation titled: “New Perspectives for Angiogenic Gene Therapy to Treat Myocardial Ischemia in Patients with Coronary Disease” to attendees at the ASGCT Meeting today, May 16. The presentation will provide a historical overview of the Generx clinical development program and how these new and important preclinical findings have been incorporated into the protocol for the 100-patient Generx ASPIRE Phase 3 registration study which was recently initiated in the Russian Federation for patients with myocardial ischemia and stable angina pectoris. The presentation is now available for viewing at http://www.cardiumthx.com/pdf/Generx-ASGCT-May-2012-Rubanyi.pdf.

About Generx and the ASPIRE Study

Generx (Ad5FGF-4) is a disease-modifying regenerative medicine biologic that is being developed to offer a one-time, non-surgical option for the treatment of myocardial ischemia in patients with stable angina due to coronary artery disease, who might otherwise require surgical and mechanical interventions, such as coronary artery by-pass surgery or balloon angioplasty and stents. Similar to surgical/mechanical revascularization approaches, the goal of Cardium’s Generx product candidate is to improve blood flow to the heart muscle – but to do so non-surgically, following a single administration from a standard balloon angioplasty catheter. The video “Cardium Generx Cardio-Chant” provides an overview Generx and can be viewed at http://www.youtube.com/watch?v=pjUndFhJkjM.

In March 2012, Cardium reported on the ASPIRE Phase 3 registration study to evaluate the therapeutic effects of its lead product candidate, Generx in patients with myocardial ischemia due to coronary artery disease. The ASPIRE study, a 100-patient, randomized and controlled multi-center study to be conducted at up to eight leading cardiology centers in the Russian Federation, is designed to further evaluate the safety and effectiveness of Cardium’s Generx DNA-based angiogenic product candidate, which has already been tested in clinical studies involving 650 patients at more than one hundred medical centers in the U.S., Europe and elsewhere. The efficacy of Generx will be quantitatively assessed using rest and stress SPECT (Single-Photon Emission Computed Tomography) myocardial imaging to sensitively measure improvements in microvascular cardiac perfusion following a one-time, non-surgical, catheter-based administration of Generx.

The Cedars-Sinai Medical Center Nuclear Cardiology Core Laboratory in Los Angeles, California, will serve as the central core lab for the ASPIRE study and will be responsible for the analysis of SPECT myocardial imaging data electronically transmitted from the Russian medical centers participating in the ASPIRE study. Advanced Biosciences Research, an affiliate of bioRASI which is a global clinical research organization, is Cardium’s Russian sponsor and development partner and is responsible for the ASPIRE program management and regulatory compliance. The Russian Health Authority has assigned Generx the therapeutic drug trade name of Cardionovo™ for marketing and sales in Russia. Information about the ASPIRE study is available at http://clinicaltrials.gov/ct2/show/NCT01550614?term=cardium&rank=1.

About Cardium

Cardium is a health sciences and regenerative medicine company focused on the acquisition and strategic development of new and innovative bio-medical product opportunities and businesses with the potential to address significant unmet medical needs that have definable pathways to commercialization, partnering and other economic monetizations. Cardium’s current medical opportunities portfolio, which is focused on health sciences and regenerative medicine, includes the Tissue Repair Company, Cardium Biologics, and the Company’s in-house MedPodium Health Sciences healthy lifestyle product platform. The Company’s lead commercial product Excellagen™ topical gel for wound care management recently received FDA clearance for marketing and sale in the United States. Cardium’s lead clinical development product candidate Generx® is a DNA-based angiogenic biologic intended for the treatment of patients with myocardial ischemia due to coronary artery disease. In addition, consistent with its capital-efficient business model, Cardium continues to actively evaluate new technologies and business opportunities. In July 2009, Cardium completed the sale of its InnerCool Therapies medical device business to Royal Philips Electronics, the first asset monetization from the Company’s biomedical investment portfolio. News from Cardium is located at www.cardiumthx.com.

Forward-Looking Statements

Except for statements of historical fact, the matters discussed in this press release are forward looking and reflect numerous assumptions and involve a variety of risks and uncertainties, many of which are beyond our control and may cause actual results to differ materially from stated expectations. For example, there can be no assurance that enhancements in the uptake of adenovectors can be successfully applied to improve the uptake or therapeutic effects of Generx in human patients; that Generx can be successfully advanced in clinical studies outside of the U.S.; that results or trends observed in one clinical study or procedure will be reproduced in subsequent studies or procedures, or that clinical studies even if successful will lead to product advancement or partnering; that improvements in the formulation or use of Generx will be commercially practicable, or that Generx could be successfully advanced as a therapeutic in developing markets or that the results of studies in such markets could be used to advance or broaden the regulatory or commercialization activities of Generx in the U.S. or other markets; that the ASPIRE clinical study will be successful or will lead to approval of Generx by the Russian Health Authority for marketing and sales in Russia or lead to approvals in other countries of the Commonwealth of Independent States; that additional clinical evidence regarding the safety and effectiveness of Generx that might be obtained in Russia would be useful for optimizing and broadening commercial development pathways in other industrialized countries; that our products or product candidates will not be unfavorably compared to competitive products that may be regarded as safer, more effective, easier to use or less expensive; that FDA or other regulatory clearances or other certifications, or other commercialization efforts will be successful or will effectively enhance our businesses or their market value; that our products or product candidates will prove to be sufficiently safe and effective after introduction into a broader patient population; or that third parties on whom we depend will perform as anticipated.

Actual results may also differ substantially from those described in or contemplated by this press release due to risks and uncertainties that exist in our operations and business environment, including, without limitation, risks and uncertainties that are inherent in the development of complex biologics and in the conduct of human clinical trials, including the timing, costs and outcomes of such trials, our ability to obtain necessary funding, regulatory approvals and expected qualifications, our dependence upon proprietary technology, our history of operating losses and accumulated deficits, our reliance on collaborative relationships and critical personnel, and current and future competition, as well as other risks described from time to time in filings we make with the Securities and Exchange Commission. We undertake no obligation to release publicly the results of any revisions to these forward-looking statements to reflect events or circumstances arising after the date hereof.

Contact: Jessica Guenzel
jessica.guenzel@nyumc.org
212-404-3591
NYU Langone Medical Center / New York University School of Medicine

“This is the first study to show that apigenin, which can be extracted from celery, parsley and many other natural sources, is effective against human breast cancer cells that had been influenced by a certain chemical used in hormone replacement therapy,” said co-author Salman Hyder, the Zalk Endowed Professor in Tumor Angiogenesis and professor of biomedical sciences in the College of Veterinary Medicine and the Dalton Cardiovascular Research Center.

In the study, Hyder and his colleagues implanted cells of a deadly, fast-growing human breast cancer, known as BT-474, into a specialized breed of mouse. Some of the mice were then treated with medroxyprogesterone acetate (MPA), a type of progestin commonly given to post-menopausal women. A control group did not receive MPA.

Later one group of MPA-treated mice was treated with apigenin. Cancerous tumors grew rapidly in the mice which did not receive apigenin. In the apigenin-treated mice, breast cancer cell growth dropped to that of the control group, and the tumors shrank.

“We don’t know exactly how apigenin does this on a chemical level,” Hyder said. “We do know that apigenin slowed the progression of human breast cancer cells in three ways: by inducing cell death, by inhibiting cell proliferation, and by reducing expression of a gene associated with cancer growth. Blood vessels responsible for feeding cancer cells also had smaller diameters in apigenin-treated mice compared to untreated mice. Smaller vessels mean restricted nutrient flow to the tumors and may have served to starve the cancer as well as limiting its ability to spread.”

The mice in Hyder’s study were injected with apigenin. In the future, apigenin injections could be a safe alternative or supplement to the highly toxic chemotherapy drugs now in use.

“Chemotherapy drugs cause hair-loss, extreme fatigue and other side effects,” Hyder said. “Apigenin has shown no toxic side-effects even at high dosages. People have eaten it since pre-history in fruits and vegetables.”

Finding funding for clinical testing of apigenin in humans may be difficult, according to Hyder.

“Clinical trials of apigenin with humans could start tomorrow, but we have to wait for medical doctors to carry out that next step,” Hyder said. “One problem is, because apigenin doesn’t have a known specific target in the cancer cell, funding agencies have been reticent to support the research. Also, since apigenin is easily extracted from plants, pharmaceutical companies don’t stand to profit from the treatment; hence the industry won’t put money into studying something you can grow in your garden.”

The research team included Benford Mafuvadze, doctoral student in biomedical sciences; Yanyun Liang, research scientist Dalton Cardiovascular Research Center; Cynthia Besch-Williford, associate professor of veterinary pathobiology; and Xu Zhang, visiting researcher at the Dalton Cardiovascular Research Center.

The research was recently published online in the journal Hormones and Cancer.

Contact: Timothy Wall
walltj@missouri.edu
573-882-3346
University of Missouri-Columbia

Using a convergent functional genomics approach that incorporates a variety of experimental techniques, the scientists also were able to apply a panel of their top genes to data from other studies of schizophrenia and successfully identify which patients had been diagnosed with schizophrenia and which had not, according to a report published online today by the journal Molecular Psychiatry.

Evaluating the biological pathways in which the genes are active, the researchers also proposed a model of schizophrenia as a disease emerging from a mix of genetic variations affecting brain development and neuronal connections along with environmental factors, particularly stress.

“At its core, schizophrenia is a disease of decreased cellular connectivity in the brain, precipitated by environmental stress during brain development, among those with genetic vulnerability,” said principal investigator Alexander B. Niculescu III, M.D., Ph.D., associate professor of psychiatry and medical neuroscience at the IU School of Medicine and director of the Laboratory of Neurophenomics at the Institute of Psychiatric Research at the IU School of Medicine.

“For first time we have a comprehensive list of the genes that have the best evidence for involvement in schizophrenia,” said Niculescu, who is also staff psychiatrist and investigator at the Richard L. Roudebush Veterans Affairs Medical Center.

Schizophrenia is a relatively widespread psychiatric disease, affecting about 1 percent of the population, often with devastating impact. People with schizophrenia can have difficulty thinking logically and telling the difference between real and unreal experiences, and may engage in bizarre behavior.

When the test estimating the risk for schizophrenia is refined, it could provide guidance to caregivers and health care professionals about young people in families with a history of the disease, prompting early intervention and treatment when behavioral symptoms of schizophrenia occurred among those at higher risk, Dr. Niculescu said.

He stressed that a score indicating a higher risk of schizophrenia “doesn’t determine your destiny. It just means that your neuronal connectivity is different, which could make you more creative, or more prone to illness.”

“It’s all on a continuum; these genetic variants are present throughout the population. If you have too many of them, in the wrong combination, in an environment where you are exposed to stress, alcohol and drugs, and so on, that can lead to the development of the clinical illness,” he said.

The prototype test was able to predict whether a person was at a higher or lower risk of schizophrenia in about two-thirds of cases.

To identify and prioritize the genes reported Tuesday, the researchers combined data from several different types of studies. These included genome-wide association studies, gene expression data derived from human tissue samples, genetic linkage studies, genetic evidence from animal models, and other work. This approach, called convergent functional genomics, has been pioneered by Niculescu and colleagues, and relies on multiple independent lines of evidence to implicate genes in clinical disorders.

The authors noted that the results were stronger when analyses were performed using gene-level data, rather than analyses based on individual mutations — called single nucleotide polymorphisms, or SNPs — in those genes. Multiple different SNPs can spark a particular gene’s role in the development of schizophrenia, so evidence for the genes, and the biological mechanisms in which they play a role, was much stronger from study to study than was the evidence for individual SNPs.

Past research looking at individual mutations was difficult to replicate from study to study, Dr. Niculescu said. Tuesday’s paper, however, indicates that much of the research done in recent years has in fact produced consistent results at a gene and biological pathway level.

“There is a lot more reproducibility and concordance in the field than people realized,” he said.

“Finally now, by better understanding the genetic and biological basis of the illness, we can develop better tests for it, as well as better treatments. The future of medicine is not just treatment but prevention, so we hope this work will move things in the right direction.”

Additional authors of the paper are Mikias Ayalew, Helen Le-Niculescu, Daniel Levey, Nitika Jain, Bharathi Eddula-Changala, Sagar Patel, Evan Winiger, Alan Breier, Anantha Shekhar, John Nurnberger, and Daniel Koller from IU; Aiden Corvin from Trinity College; Mark Geyer and Ming Tsuang from UC San Diego; Daniel Salomon and Nicholas Schork from The Scripps Research Institute; Richard Amdur and Ayman Fanous from Washington DC VA Medical Center; and Michael O’ Donovan from Cardiff University.

Support for the research was provided by a National Institutes of Health Director’s New Innovator Award (1DP2OD007363) and a Veterans Administration Merit Award (1I01CX000139-01).

Within a few hours of injecting empty-handed DNA nanoparticles, Georgia Health Sciences University researchers were surprised to see increased expression of an enzyme that calms the immune response.

In an animal model of rheumatoid arthritis, the enhanced expression of indoleomine 2,3 dioxygenase, or IDO, significantly reduced the hallmark limb joint swelling and inflammation of this debilitating autoimmune disease, researchers report in the study featured on the cover of The Journal of Immunology.

“It’s like pouring water on a fire,” said Dr. Andrew L. Mellor, Director of the GHSU’s Medical College of Georgia Immunotherapy Center and the study’s corresponding author. “The fire is burning down the house, which in this case is the tissue normally required for your joints to work smoothly,” Mellor said of the immune system’s inexplicable attack on bone-cushioning cartilage. “When IDO levels are high, there is more water to control the fire.”

Several delivery systems are used for gene therapy, which is used to treat conditions including cancer, HIV infection and Parkinson’s disease. The new findings suggest the DNA nanoparticle technique has value as well for autoimmune diseases such as arthritis, type 1 diabetes and lupus. “We want to induce IDO because it protects healthy tissue from destruction by the immune system,” Mellor said.

The researchers were exploring IDO’s autoimmune treatment potential by inserting the human IDO gene into DNA nanoparticles. They hoped to enhance IDO expression in their arthritis model when Dr. Lei Huang, Assistant Research Scientist and the paper’s first author, serendipitously found that the DNA nanoparticle itself produced the desired result. Exactly how and why is still being pursued. Early evidence suggests that immune cells called phagocytes, white blood cells that gobble up undesirables like bacteria and dying cells, start making more IDO in response to the DNA nanoparticle’s arrival. “Phagocytes eat it and respond quickly to it and the effect we measure is IDO,” Mellor said.

Dr. Tracy L. McGaha, GHSU immunologist and a co-author on the current study, recently discovered that similar cells also prevented development of systemic lupus erythematosus in mice.

Follow-up studies include documenting all cells that respond by producing more IDO. GHSU researchers already are working with biopolymer experts at the Massachusetts Institute of Technology, the University of California, Berkeley and the Georgia Institute of Technology to identify the optimal polymer.

The polymer used in the study is not biodegradable so the researchers need one that will eventually safely degrade in the body. Ideally, they’d also like it to target specific cells, such as those near inflamed joints, to minimize any potential ill effects.

“It’s like a bead and you wrap the DNA around it,” Mellor said of the polymer. While the DNA does not have to carry anything to get the desired response in this case, DNA itself is essential to make cells express IDO. To ensure that IDO expression was responsible for the improvements, they also performed experiments in mice given an IDO inhibitor in their drinking water and in mice genetically altered to not express IDO. “Without access to the IDO pathway, the therapy no longer works,” Mellor said.

Drs. Andrew Mellor and David Munn reported in 1998 in the journal Science that the fetus expresses IDO to help avoid rejection by the mother’s immune system. Subsequent studies have shown tumors also use IDO for protection and clinical trials are studying the tumor-fighting potential of an IDO inhibitor. On the flip side, there is evidence that increasing IDO expression can protect transplanted organs and counter autoimmune disease.

Mellor is the Bradley-Turner and Georgia Research Alliance Eminent Scholar in Molecular Immunogenetics at MCG. The research was funded by the Carlos and Marguerite Mason Trust and the National Institutes of Health and a patent is pending on the findings.

Contact: Toni Baker
tbaker@georgiahealth.edu
706-721-4421
Georgia Health Sciences University

The findings provide valuable insight as to why some cancers metastasize to bone, and could eventually result in new metastasis-prevention drugs, said Laurie McCauley, professor in the Department of Periodontics and Oral Medicine at the University of Michigan School of Dentistry and principal investigator on the study.

The really good news is that researchers reversed the tumor-friendly effect of the drug, called cyclophosphamide, by inhibiting another cell-communicating protein in the bone marrow, called CCL2.

“This work is early and still at the pre-clinical level,” said McCauley, who also has an appointment in the Department of Pathology at the U-M Health System. “However, the biggest potential impact is in metastasis preventive strategies.

“If we better understood the specific mediators, or conditions, in the bone marrow that support tumors, we could develop more effective therapeutics to prevent local cancers from spreading and hence reduce metastasis to the bone.”

The study highlights the potential for the bone marrow to provide the right environment for tumors to metastasize, said Serk In Park, first author and a postdoctoral fellow in McCauley’s lab. Many cancers, such as prostate and breast cancer, are fond of spreading, or metastasizing, to bones.

Researchers administered the chemotherapy drug cyclophosphamide experimentally to manipulate the environment inside the bone marrow prior to exposing experimental tumors. Cyclophosphamide therapy is used in certain cancers to slow cell growth, and McCauley’s group experimented with its use in a pre-metastatic mode using a prostate cancer model.

While effective at attacking tumor cells, a side effect of cyclophosphamide (and many other chemotherapy drugs) is that it suppresses certain bone marrow cells that help the immune system and increases some harmful cells. Researchers hypothesized correctly that the drug would make the bone marrow more tumor-friendly.

The paper, “Cyclophosphamide Creates a Receptive Microenvironment for Prostate Cancer Skeletal Metastasis,” appears in the journal Cancer Research.

Contact: Laura Bailey
baileylm@umich.edu
734-647-1848
University of Michigan

The study is the first example of using bone cell progenitors derived from human embryonic stem cells to grow compact bone tissue in quantities large enough to repair centimeter-sized defects. When implanted in mice and studied over time, the implanted bone tissue supported blood vessel ingrowth, and continued development of normal bone structure, without demonstrating any incidence of tumor growth.

Dr. Marolt’s work is a significant step forward in using pluripotent stem cells to repair and replace bone tissue in patients. Bone replacement therapies are relevant in treating patients with a variety of conditions, including wounded military personnel, patients with birth defects, or patients who have suffered other traumatic injury.

Since conducting this work as proof of principle at Columbia University, Dr. Marolt has continued to build upon this research as an Investigator in the NYSCF Laboratory, developing bone grafts from induced pluripotent stem (iPS) cells. iPS cells are similar to embryonic stem cells in that they can also give rise to nearly any type of cell in the body, but iPS cells are produced from adult cells and as such are individualized to each patient. By using iPS cells rather than embryonic stem cells to engineer tissue, Dr. Marolt hopes to develop personalized bone grafts that will avoid immune rejection and other implant complications.

The New York Stem Cell Foundation has supported Dr. Marolt’s research throughout her career, first through a NYSCF – Druckenmiller Fellowship to fund her post-doctoral work at Columbia University, and now with a NYSCF – Helmsley Investigator Award at The New York Stem Cell Foundation Laboratory. “The continuity of funding provided by NYSCF has allowed me to continue my research uninterrupted, making progress more quickly than would have otherwise been possible,” Dr. Marolt said.

The New York Stem Cell Foundation (NYSCF) conducts cutting-edge translational stem cell research in its laboratory in New York City and supports research by stem cell scientists at other leading institutions around the world. More information is available at www.nyscf.org.

Contact: David McKeon
dmckeon@nyscf.org
212-365-7440
New York Stem Cell Foundation

Even though HMO are a major component of human milk, present in higher concentration than protein, many of their actions in the infant are not well understood. Furthermore, they’re virtually absent from infant formula. The scientists wanted to find out what formula-fed babies were missing.

“We refer to HMO as the fiber of human milk because we don’t have the enzymes to break down these compounds. They pass into the large intestine where the bacteria digest them.

“We’re curious about the role they play in the development of the breast-fed infant’s gut bacteria because the bacteria found in the guts of formula-fed infants is different,” said Sharon Donovan, the U of I’s Melissa M. Noel Endowed Professor in Nutrition and Health.

With this study, Donovan is gaining insight into the mystery. For the first time, scientists have shown that a complex mixture of HMO and a single HMO component produce patterns of short-chain fatty acids that change as the infant gets older.

A healthy microbiome has both short- and long-term effects on an infant’s health. In the short term, beneficial bacteria protect the infant from infection by harmful bacteria. In the long term, beneficial bacteria strengthen the immune system so that it can fend off chronic health problems like food allergies and asthma, she said.

In the study, breast milk was obtained from mothers of preterm infants at Chicago’s Rush University Medical Center, and the HMO were isolated and analyzed. The scientists tested bacteria from 9- and 17-day-old sow-reared and formula-fed piglets. Because piglets grow so rapidly, these ages reflect approximately three- and six-month-old human infants.

The colon bacteria were added to test tubes containing HMO and two prebiotics commonly used in infant formulas. These mixtures were allowed to ferment and then sampled to see how the bacterial population was changing over time and what products were being produced by the bacteria.

“When the HMOs were introduced, the bacteria produced short-chain fatty acids, at some cases at higher levels than other prebiotics now used in infant formula. The short-chain fatty acids can be used as a fuel source for beneficial bacteria and also affect gastrointestinal development and pH in the gut, which reduces the number of disease-causing pathogens,” she said.

Further, different HMOs produced different patterns of short-chain fatty acids, and the composition of bacteria in the gut changed over time. “It was distinctly different at 9 vs. 17 days, making it likely that the functions of HMO change as the human infant gets older,” she said.

According to Donovan, HMO are critically important in understanding how breastfeeding protects babies.

“Several companies are now able to synthesize HMO, and in the future, we may be able to use them to improve infant formula. There’s evidence that these compounds can bind to receptors on immune cells and, to our knowledge, no current prebiotic ingredient can do that,” she said.

The study was published in the April issue of the Journal of Nutrition. Co-authors are Min Li, Laura L. Bauer, Xin Chen, Mei Wang, Theresa B. Kuhlenschmidt, and George C. Fahey Jr., all of the U of I. Funding was provided by a grant from the National Institutes of Health.

Contact: Phyllis Picklesimer
p-pickle@illinois.edu
217-244-2827
University of Illinois College of Agricultural, Consumer and Environmental Sciences

Hamamy syndrome is a rare genetic disorder which is marked by abnormal facial features and defects in the heart, bone, blood and reproductive cells. Its exact cause was unknown until now. The international team, led by scientists at IMB, have pinpointed the genetic mistake to be a mutation in a single gene called IRX5.

This is the first time that a mutation in IRX5 (and the family of IRX genes) has ever been discovered in man. IRX5 is part of a family of transcription factors that is highly conserved in all animals, meaning that this gene is present not only in humans but also in mice, fish, frogs, flies and even worms. Using a frog model, the scientists demonstrated that Irx5 orchestrates cell movements in the developing foetus which underlie head and gonad formation.

Carine Bonnard, a final-year PhD student at IMB and the first author of the paper, said, “Because Hamamy syndrome causes a wide range of symptoms, not just in newborn babies but also in the adult, this implies that IRX5 is critical for development in the womb as well as for the function of many organs in our adult body. For example, patients with this disease cannot evacuate tears from their eyes, and they will also go on to experience repetitive bone fractures or progressive myopia as they age. This discovery of the causative gene is a significant finding that will catalyze research efforts into the role of the Irx gene family and greatly increase our understanding of human health, such as bone homeostasis, or gamete formation for instance.”

“We believe that this discovery could open up new therapeutic solutions to common diseases like osteoporosis, heart disease, anaemia which affect millions of people worldwide,” said Dr Bruno Reversade, Senior Principle Investigator at IMB. “The findings also provide a framework for understanding fascinating evolutionary questions, such as why humans of different ethnicities have distinct facial features and how these are embedded in our genome. IRX genes have been repeatedly co-opted during evolution, and small variation in their activity could underlie fine alterations in the way we look, or perhaps even drastic ones such as the traits seen in an elephant, whale, turtle or frog body pattern.”

Only a handful of people in the world have been identified with Hamamy Syndrome making it a very rare genetic disorder. Rare genetic diseases, usually caused by mutations in a single gene, provide a unique opportunity to better understand more common disease processes. These “natural” experiments are similar to carefully controlled knockout animal experiments in which the function of single genes are analyzed and often give major insights into general health issues.

Prof Birgitte Lane, Executive Director of IMB, said, “Understanding how various pathways in the human body function is the foundation for developing new therapeutic targets. This is an important piece of research that I believe will be of great interest to many scientists and clinicians around the world because of the clinical and genetic insights it brings to a large range of diseases.”

Notes for editor:

The research findings described in this news release can be found on Nature Genetics’s website under the title “Mutations in IRX5 impair craniofacial development and germ cell migration via SDF1″ by Carine Bonnard1, Anna C Strobl2, Mohammad Shboul1, Hane Lee3, Barry Merriman3, Stanley F Nelson3, Osama H Ababneh4, Elif Uz5, 6, Tülay Guran7, Hülya Kayserili8, Hanan Hamamy9, 10 & Bruno Reversade1, 11.

1 Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore
2 Division of Systems Biology, Medical Research Council National Institute for Medical Research, London, UK
3 Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, California, USA
4 Department of Opthalmology, Faculty of Medicine, University of Jordan, Amman, Jordan
5 Department of Biology, Faculty of Arts and Sciences, Duzce University, Duzce, Turkey
6 Gene Mapping Laboratory, Department of Medical Genetics, Hacettepe University Medical Faculty, Ankara, Turkey
7 Pediatric Endocrinology and Diabetes, Marmara University Hospital, Istanbul, Turkey
8 Medical Genetics Department, Istanbul Medical Faculty, Istanbul University, Istanbul, Turkey
9 Department of Genetic Medicine and Development, Geneva University Hospital, Geneva, Switzerland
10 National Center for Diabetes, Endocrinology and Genetics, Amman, Jordan
11 Department of Pediatrics, National University of Singapore, Singapore
Correspondence should be addressed to B.R. (Bruno@reversade.com)

The article can be accessed from http://www.nature.com/ng/journal/vaop/ncurrent/full/ng.2259.html.

Agency for Science, Technology and Research (A*STAR)

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About the Institute of Medical Biology (IMB)

IMB is one of the Biomedical Sciences Institutes of the Agency for Science, Technology and Research (A*STAR). It was formed in 2007, the 7th and youngest of the BMRC Research Institutes, with a mission to study mechanisms of human disease in order to discover new and effective therapeutic strategies for improved quality of life. From 2011, IMB also hosts the inter-research institute Skin Biology Cluster platform.

IMB has 20 research teams of international excellence in stem cells, genetic diseases, cancer and skin and epithelial biology, and works closely with clinical collaborators to target the challenging interface between basic science and clinical medicine. Its growing portfolio of strategic research topics is targeted at translational research on the mechanisms of human diseases, with a cell-to-tissue emphasis that can help identify new therapeutic strategies for disease amelioration, cure and eradication. For more information about IMB, please visit www.imb.a-star.edu.sg.

About the Reversade Laboratory

Dr. Reversade, a human geneticist and embryologist holds a Senior Principal Investigator position at IMB and an adjunct faculty position at the Department of Paediatrics in the National University of Singapore. He is a Fellow of the Branco Weiss Foundation based at ETH in Switzerland and also the first recipient of an A*STAR Investigatorship, a programme which provides competitive and prestigious fellowships to support the next generation of international scientific leaders, offering funding and access to state-of-the-art scientific equipment and facilities at A*STAR.

For more information about Dr. Reversade’s laboratory, please visit www.reversade.com.

About the Agency for Science, Technology and Research (A*STAR)

The Agency for Science, Technology and Research (A*STAR) is the lead agency for fostering world-class scientific research and talent for a vibrant knowledge-based and innovation-driven Singapore. A*STAR oversees 14 biomedical sciences and physical sciences and engineering research institutes, and six consortia & centres, located in Biopolis and Fusionopolis as well as their immediate vicinity.

A*STAR supports Singapore’s key economic clusters by providing intellectual, human and industrial capital to its partners in industry. It also supports extramural research in the universities, and with other local and international partners.

For more information about A*STAR, please visit www.a-star.edu.sg.

Contact: Ong Siok Ming
ong_siok_ming@a-star.edu.sg
65-682-66254
Agency for Science, Technology and Research (A*STAR), Singapore