More than 70 years after the first sulfa drugs helped to revolutionize medical care and save millions of lives, St. Jude Children’s Research Hospital scientists have determined at an atomic level the mechanism these medications use to kill bacteria. The discovery provides the basis for a new generation of antibiotics that would likely be harder for bacteria to resist and cause fewer side effects.

The work focused on sulfa drugs and their target enzyme, dihydropteroate synthase (DHPS). Most disease-causing microorganisms need DHPS to help make the molecule folate, which is required for the production of DNA and some amino acids.

Working with enzymes from gram-negative and gram-positive bacteria, researchers used a variety of techniques to determine for the first time the key intermediate structure DHPS forms during the chemical reaction to advance folate production. The structure also explains at a molecular level how sulfa drugs function and how resistance causing mutations help bacteria withstand them.

The findings mark a major advance in both microbial biochemistry and anti-microbial drug discovery. The study is published in the March 2 issue of the journal Science.

“The structure we found was totally unexpected and really opens the door for us and others to design a new class of inhibitors targeting DHPS that will help us avoid side effects and other problems associated with sulfa drugs,” said Stephen White, Ph.D., chair of the St. Jude Department of Structural Biology and the paper’s corresponding author.

Co-author Richard Lee, Ph.D., a member of the St. Jude Department of Chemical Biology and Therapeutics, added: “Now we want to leverage this information to develop drugs against the opportunistic infections that threaten so many St. Jude patients.”

Sulfa drugs were discovered in the 1930s and became the first antibiotic in widespread use. Although the drugs were early victims of antibiotic resistance, they are still widely used against emerging infectious diseases and to prevent infections in patients with weakened immune systems, including St. Jude patients undergoing cancer chemotherapy. The growing problem of antibiotic resistance has prompted renewed interest in sulfa drugs as a possible source of new therapeutic targets, Lee said.

Previous work had shown that sulfa drugs target DHPS and work by mimicking a molecule called pABA. DHPS advances folate production by accelerating the fusion of pABA and another molecule called dihydropteridine pyrophosphate (DHPP). Until now, however, scientists did not know exactly how the DHPS reaction occurred or how sulfa drugs disrupted the process.

Working on enzymes from gram-positive Bacillus anthracis and gram-negative Yersinia pestis, the bacteria that cause anthrax and plague, researchers first used computational methods to predict the enzyme’s activity. Next they used a technique called X-ray crystallography to capture the unfolding chemical reaction and confirm the prediction. X-ray crystallography involves bombarding proteins trapped in crystals with X-rays to determine the protein structure.

Researchers showed that DHPP binds to a specific pocket in DHPS. Aided by magnesium, the binding promotes the break-up of DHPP and release of pyrophosphate. Two long flexible loops then create an intermediate structure that sets the stage for pABA to enter and bind in a second short-lived pocket, allowing pABA to fuse with the cleaved DHPP. Investigators captured all four actors in the drama in a single crystal structure, including the intermediate cleaved DHPP molecule whose existence was previously unknown.

The results showed that the mechanism involves a chemical reaction known as an Sn1 reaction rather than the anticipated Sn2 reaction. “This is a key finding for drug discovery because it reveals chemical features of the DHPS enzyme’s active site that we can exploit in developing new drugs,” said study co-author Donald Bashford, Ph.D., an associate member of the St. Jude Department of Structural Biology.

The study also provided insights into sulfa drug resistance. Investigators showed that the binding sites of pABA and the sulfa drugs overlap, but that sulfa drugs extend beyond the pocket in which pABA binds.

Mutations associated with drug resistance cluster around this extended region of the pABA pocket, which explains how mutations can prevent the drugs from binding without seriously affecting the binding of pABA. The work also highlights the transitory structure made by the two DHPS loops as a target for a new class of drugs that would be difficult for bacteria to develop resistance against.

“When we set out on this project eight years ago, a goal was to truly understand the catalytic mechanism of the DHPS protein and how the inhibitors targeting it work. I am ecstatic we’ve succeeded,” Lee said. The success grew out of an interdisciplinary effort and some luck, White said. The plague enzyme turned out to be well suited to this project.

Unlike the DHPS enzymes from other bacteria, the two extended loops are free to form the short-lived structure and the pABA pocket when the enzyme is immobilized in the crystal.

The study’s first authors are Mi-Kyung Yun of St. Jude and Yinan Wu, a University of Tennessee Health Sciences Center graduate student working in White’s laboratory. The other authors are Zhenmei Li, Ying Zhao, M. Brett Waddell and Antonio Ferreira, all of St. Jude. The research was supported in part by the National Institutes of Health and ALSAC.

The article Scientists Discover How Sulfa Drugs Kill Bacteria appeared first on The Toonari Post - News, Powered by the People!.

In 1962, St. Jude Children’s Research Hospital opened its doors amid an emotionally charged debate regarding how to treat childhood cancer. At that time, few children with the most common form of childhood cancer survived, and many physicians believed treatment was futile.

St. Jude physicians and researchers took a radically different approach, and these efforts proved pivotal in changing the way the world treats childhood cancer. St. Jude is recognized for playing a significant role in improving overall survival rates for childhood cancer, which have increased from 20 percent in 1962 to 80 percent today.

In recognition of this impact over the past 50 years, Tennessee Gov. Bill Haslam declared February “St. Jude Month” in the state of Tennessee. Founded by the late entertainer Danny Thomas, the hospital opened February 4, 1962.

“In the nearly four decades I’ve been at St. Jude, I’ve had the privilege of watching the organization grow from one star-shaped building to a sprawling campus of about 2.5 million square feet of research, clinical and administrative space,” said Dr. William E. Evans, St. Jude director and CEO.

“When I started, there were a few hundred people on staff. Now we have more than 3,700 employees. Driven by our patients, and thanks to our employees, our colleagues at ALSAC and the public support they generate, St. Jude will only continue to grow and flourish in the years to come.”

The history of St. Jude is marked with milestones in the treatment of pediatric cancer and other childhood illnesses. In 1971, St. Jude investigators showed that the combination of chemotherapy and radiation cured at least half of all children with acute lymphoblastic leukemia (ALL).

The most common form of childhood cancer, ALL, was previously considered almost universally fatal. Today, St. Jude patients with ALL have a 94 percent survival rate. In 1984, a St. Jude patient with sickle cell disease was the first to be cured with a bone marrow transplant.

St. Jude is currently engaged in the largest effort in the world to do whole genome sequencing of pediatric cancer tumors. The St. Jude Children’s Research Hospital – Washington University Pediatric Cancer Genome Project collaboration has already produced significant new findings related to aggressive forms of pediatric leukemia, eye tumors and brain tumors.

“St. Jude has a legacy of taking on the toughest of pediatric cancer questions, and that focus won’t change,” said James R. Downing, M.D., scientific director and deputy director at St. Jude. “We’re uniquely positioned as an institution to move research and treatment ahead. From the genetic data we collect from the genome project, we’re creating the foundation of knowledge to deliver the next decades’ childhood cancer discoveries and treatments.”

Throughout its five decades, St. Jude research has included work in cancer biology and genomics, pharmacogenomics, gene therapy, bone marrow transplant, drug discovery, radiation treatment, blood diseases and infectious diseases, integrated into a long series of innovative clinical trials.

According to Joseph Laver, M.D., St. Jude clinical director, “the unsurpassed family-centered care that is provided at St. Jude stems from the multidisciplinary team approach that has been a hallmark of St. Jude since the doors opened in 1962.”

“Looking toward the future, St. Jude is a national resource with a global mission and will continue to enhance its leadership as a resource for children with cancer and other catastrophic diseases,” Evans said. “Even though we’ve grown significantly, our mission has never wavered.

We’ve created a collaborative culture whose team members demonstrate unceasing compassion for our patients and families, innovation in our treatment and research, and quality in everything we do.”

Image Courtesy of   https://www.facebook.com/stjude

The article St. Jude Children’s Research Hospital Celebrates 50 Years appeared first on The Toonari Post - News, Powered by the People!.

St. Jude Children’s Research Hospital has been recognized by FORTUNE magazine as one of the “100 Best Companies to Work For” for the second consecutive year. Last year was the hospital’s first appearance in the publication’s annual list. Employees again ranked pride in the hospital’s mission—finding cures, saving children—as one of the top reasons St. Jude is a great place to work.

“Our employees look at the work they do at St. Jude as much more than a job,” said Dr. William E. Evans, St. Jude director and CEO. “As we celebrate our 50th anniversary this year, we are reminded that one of the keys to our success has been attracting outstanding people and giving them a place to do their best work. Employees embrace the mission and espouse the culture of St. Jude, which is one of collaboration, compassion, innovation and quality in all we do.”

St. Jude strives to continually improve in all areas, including work environment for employees. Based on employee ideas and needs, the hospital has created initiatives that have enhanced communication between senior leadership and staff; enhanced the Wellness Program with a comprehensive approach to fitness and health; and fostered programs and policies intended to promote diversity.

To select businesses for the “100 Best Companies to Work For” list, FORTUNE partners with the Great Place to Work Institute to conduct the most extensive survey of employees done in corporate America. More than 280 companies participated in this year’s survey. Two-thirds of a company’s score is based on the results of the Institute’s Trust Index survey, which is sent to a random sample of employees at each company. The survey asks questions that assess their attitudes about management’s credibility, job satisfaction and camaraderie.

The other third of the scoring is based on the company’s responses to the institute’s culture audit, which includes detailed questions about pay and benefit programs and a series of open-ended questions about hiring practices, internal communications, training, recognition programs and diversity efforts. Any company that is at least five years old with more than 1,000 U.S. employees is eligible.

Ranked No. 39 on this year’s list, St. Jude was No. 80 in 2011.

The article St. Jude Children’s Hospital One of the ’100 Best Companies to Work For’ appeared first on The Toonari Post - News, Powered by the People!.

Symptoms improved significantly in adults with the bleeding disorder hemophilia B following a single treatment with gene therapy developed by researchers at St. Jude Children’s Research Hospital in Memphis and demonstrated to be safe in a clinical trial conducted at the University College London (UCL) in the U.K.

The findings of the six-person study mark the first proof that gene therapy can reduce disabling, painful bleeding episodes in patients with the inherited blood disorder. Results of the Phase I study appear in the December 10 online edition of the New England Journal of Medicine. The research is also scheduled to be presented December 11 at the 53rd annual meeting of the American Society of Hematology in San Diego.

Four study participants stopped receiving protein injections to prevent bleeding episodes after undergoing the therapy and have not suffered spontaneous bleeding. Several have also participated in marathons and other activities that would have been difficult prior to gene therapy.

The study volunteers were all treated at the Royal Free Hospital in London under the care of Edward G.D. Tuddenham, M.D., Ph.D., a pioneer in the field of blood coagulation and a study co-author.

“This is a potentially life-changing treatment for patients with this disease and an important milestone for the field of gene therapy. It could have ramifications for the treatment of hemophilia A, other protein and liver disorders and chronic diseases such as cystic fibrosis,” said first author Amit Nathwani, M.D., Ph.D., a faculty member at the UCL Cancer Institute, Royal Free Hospital and NHS Blood and Transplant (NHSBT).

Hemophilia B is caused by an inherited mistake in the gene for making a protein called Factor IX, which is essential for normal blood clotting. The gene is carried on the X chromosome. As a result, hemophilia B is almost exclusively a disease of men. About 1 in 30,000 individuals inherit the mutation.

Previous efforts to ease hemophilia B symptoms by introducing a correct copy of the gene have been unsuccessful. The current study used adeno-associated virus (AAV) 8 as the vector to deliver the Factor IX gene along with additional genetic material into the patient’s liver. AAV8 was picked because the incidence of natural infection with AAV8 is low.

It belongs to a family of viruses that target liver cells but do not cause disease in humans or integrate into human DNA. Participants in the study received no immune suppressing drugs prior to gene therapy. This approach was jointly pioneered by St. Jude and UCL, initially in the laboratory of study co-author Arthur Nienhuis, M.D, a member of the St. Jude Department of Hematology.

For this study, each patient received a one-time infusion of the vector into a vein in the arm. Two patients each were treated with escalating doses of the vector. Following treatment, Factor IX levels rose in all six patients from less than 1 percent of normal levels prior to the gene therapy to between 2 and 12 percent.

Factor IX levels increased the most in the two study volunteers who received the highest dose of the experimental vector, researchers said. After treatment, levels of the protein ranged from 3 to 12 percent in those men.

Even modest increases that raise Factor IX production to more than 1 percent of normal levels have the potential to dramatically affect a patient’s quality of life and reduce bleeding episodes, said the study’s senior author Andrew Davidoff, M.D., chair of the St. Jude Department of Surgery.

“The first patient has been followed for the longest time, and his levels have remained at 2 percent for more than 18 months. These results are highly encouraging and support continued research. More patients are scheduled to be enrolled in future trials scheduled to begin later this year,” Davidoff said.

One of the participants who received the highest dose of the vector underwent successful, short-term steroid treatment after his liver enzymes rose slightly after the vector infusion. The rise signaled mild liver damage. The volunteer remained otherwise healthy, his Factor IX levels remain above pre-infusion levels, and his liver enzymes have returned to normal.

Liver enzymes also rose slightly, but remained in the normal range, for the other participant who received the highest dose of the vector. That participant also received a short course of steroids. Researchers believe an immune response targeting the vector triggered the elevated enzyme levels. A similar response was reported in earlier gene therapy trials conducted by other investigators using a different vector.

The vector used in this study was produced at the Good Manufacturing Practices (GMP) facility on the St. Jude campus. The GMP operates under U.S. government-approved manufacturing guidelines and produces highly specialized medicines, vaccines and other products that pharmaceutical companies are reluctant to pursue. The vector can also now be produced in a similar facility at UCL.

Image Courtesy of   https://www.facebook.com/stjude

The article Gene Therapy’s Success Against Hereditary Bleeding Disorder appeared first on The Toonari Post - News, Powered by the People!.