Sickle cell test sample (A. J. Kumar, Harvard University)
22 September 2014. A Cambridge, Massachusetts diagnostics company received a Small Business Innovation Research (SBIR) grant to develop a simple point-of-care test for sickle cell disease, a genetic blood disorder affecting a large percentage of people of African origin. The $225,000 grant from National Institutes of Health’s SBIR program to Daktari Diagnostics will support development and testing of a commercial prototype of the sickle cell diagnostic, in partnership with Harvard University, University of South Carolina, and University Teaching Hospital in Lusaka, Zambia.
Sickle cell disease is a genetic blood disorder affecting hemoglobin that delivers oxygen to cells in the body. People with sickle cell disease have hemoglobin molecules that cause blood cells to form in an atypical crescent or sickle shape. That abnormal shape causes the blood cells to break down, lose flexibility, and accumulate in tiny capillaries, leading to anemia and periodic painful episodes. The disease is prevalent worldwide, and affects 70,000 to 80,000 people in the U.S., including about 1 in 500 people of African descent.
Newborns in the U.S. are tested routinely for sickle cell disease, but not in many areas where resources are limited, including many places in Africa. Daktari Diagnostics develops medical tests for resource-limited regions, combining microfluidics (lab-on-a-chip) technologies with electrochemical sensing, which the company says are designed to be simple, portable, and inexpensive.
The grant — from NIH’s National Heart, Lung, and Blood Institute — supports development of a commercial prototype based on a device designed in the lab of Harvard chemistry professor George Whitesides by then-graduate student A. J. Kumar. The system devised by Kumar and colleagues harnesses a principle in chemistry that polymer chemicals separate into different layers when mixed with water. The team applied that idea to separating red blood cells of different densities in a mix of polymers and water.
Kumar, now a postdoctoral researcher in the Whitesides lab and the lead engineer on Daktari’s project, tested an early version of the device as a proof-of-concept exercise, with the results published earlier this month in the journal Proceedings of the National Academy of Sciences. The device basically consists of a narrow tube with a mix of water and polymers, where a drop of blood is drawn with a finger prick that wicks into the tube. Spinning the tube on a standard lab centrifuge separates the multiphase blood-polymer-water mixture into visually discernible layers, with the heavier sickle cells sinking to the bottom, all in about 12 minutes.
For the commercial prototype Daktari aims to develop a device with tubes designed for a simple, battery-powered centrifuge, pre-filled with the polymer-water mixture. Blood samples would be taken with a finger-prick or a heel-stick from newborns. If sickle cells are present in the blood, they will sink to a designated region in the tube, where technicians can easily identify them. As with the proof-of-concept, Daktari is aiming to have the commercial test return results in about 12 minutes.
A key part of the proposed commercial system is the simple battery-powered centrifuge to separate the blood-water-polymer mixture into layers. Daktari says it plans to apply its experience with a battery-powered, hand-held device to measure CD4 T-cells in blood as part of an HIV test.
Daktari says the test is called Mpana, an acronym for MultiPhase Analyzer, but also a Swahili word for ” a broad, wide, open channel.” The grant funding covers work on the device through May 2015, although the project is expected to continue through May 2016.
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E coli bacteria magnified (USDA Agricultural Research Service/Wikimedia Commons)
22 September 2014. Researchers at Massachusetts Institute of Technology are developing ways to fight antibiotic-resistant bacteria by modifying the genes that make the bacteria resistant to drugs. The team from MIT’s Synthetic Biology Group, led by engineering professor Timothy Lu, published its findings yesterday in the journal Nature Biotechnology (paid subscription required).
Antibiotic resistance is a continuing problem affecting some 2 million people in the U.S. each year, causing conditions such as multidrug-resistant tuberculosis and methicillin-resistant Staphylococcus aureus or MRSA infections. While infections from antibiotic resistant bacteria can occur anywhere, most deaths from these infections occur in health care facilities, including hospitals and nursing homes. According to the Centers for Disease Control and Prevention, at least 23,000 people die from antibiotic resistant infections each year, but many more deaths can be traced to other conditions complicated by these infections.
Bacteria become resistant to antibiotics when strains evolve to replicate in ways that avoid the toxic properties of current drugs designed to prevent replication. That resistance process is exacerbated by the overuse of antibiotics and poor infection control practices.
The team from Lu’s lab is taking a different approach from the traditional method of devising new compounds to prevent bacterial replication. The researchers instead target the genomes that give bacteria the ability to evolve into a resistant strain, with a technique known as Clustered Regularly Interspaced Short Palindromic Repeats, or CRISPR, in effect removing their ability to develop resistance.
CRISPR harnesses proteins the bacteria use to protect themselves against viruses that are natural enemies, specifically a protein called Cas9, with the ability to cut strands of DNA. Lu’s team is designing a form of RNA, a nucleic acid related to DNA, that guides the Cas9 proteins to cut specific genes, in this case the genes that enable bacteria to evolve and thus resist current antibiotics. The researchers use plasmids, circular DNA strands found in bacteria, and particles from the enemy viruses to deliver the Cas9 protein.
Among their first targets in the team’s tests was the gene expressing the NDM-1 enzyme that creates resistance to beta-lactam antibiotics, a class of drugs including penicillin and carbapenems, often used to treat infections in hospitals developing from catheters and ventilators. Their tests showed virtually all (99%) of the bacteria expressing NDM-1 are killed by antibiotics after being submitted to the CRISPR-Cas9 treatments, while untreated bacteria largely survive. The researchers also successfully tested the techniques with a mutated gene that causes resistance to quinolone antibiotics, used to treat hospital-acquired infections.
The tests included E. coli infections in waxworm larvae, where the CRISPR-Cas9 treatments enabled the larvae to survive the infections. The researchers are now conducting tests of the technology with lab mice, and eventually hope to advance the technology to human patients.
In August, Lu and colleagues reported an entirely different approach to combating antibacterial resistance, the application of high-throughput genomic sequencing to find combinations of genes that make them more susceptible to antibiotics. The result was Combinatorial Genetics En Masse, or CombiGEM, a library of 34,000 pairs of genes in bacteria coded for transcription factors — proteins that control the expression of other genes — and providing a genetic bar code for each pair.
The researchers then applied that library to identify combinations with the ability for antibiotics to kill up to 1 million times as many bacteria as they do now. Their tests with some of the more promising combinations showed they could kill highly resistant E. coli bacteria with NDM-1 enzymes, normally making them resistant to beta-lactam antibiotics.
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(National Cancer Institute)
19 September 2014. The pharmaceutical company Boehringer Ingelheim is licensing from biopharmaceutical enterprise CureVac a vaccine based on RNA to generate an immune response for treating lung cancer. The deal has a total potential value to CureVac of €465 million ($US 597 million).
CureVac, in Tübingen, Germany, develops vaccines and treatments based on messenger RNA, or mRNA, that translates the genetic code in DNA to proteins expressed through an individual’s genes. The company’s technology harnesses mRNA to generate an immune response, either as a vaccine to prevent infectious diseases or, in the case of cancer, to attack cancer cells. CureVac has immunotherapies in early- and intermediate-stage clinical trials to treat prostate and non-small cell lung cancer, the most common form of lung cancer.
Boehringer Ingelheim, in Ingelheim, Germany, offers afatinib as a treatment for non-small cell lung cancer on the market in the U.S., Europe, and Japan marketed under the brand names Giotrif or Gilotrif. The company also has drug candidates to treat colorectal and non-small cell lung cancer, as well as acute myeloid leukemia, in late-stage clinical trials.
Under the deal, Boehringer Ingelheim gains exclusive worldwide rights to develop and commercialize CureVac’s CV9202 vaccine now in an early-stage clinical trial as a treatment for non-small cell lung cancer. The underlying technology to design CV9202, known as RNActive, remains part of CureVac’s intellectual property.
Boehringer Ingelheim plans to test CV9202 as a supplement to its afatinib drug for treating some types of advanced or metastatic non-small cell lung cancer, or as a supplement to chemo- and radiation therapy to treat inoperable forms of the disease. The agreement gives CureVac a one-time payment of €35 million ($45 million), and eligibility for up to €430 million ($552 million) in milestone payments and royalties on sales.
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19 September 2014. Researchers with the companies Personal Genome Diagnostics and Blueprint Medicines identified genetic mutations associated with carcinosarcoma, a rare but deadly form of cancer affecting the female reproductive system also known as malignant mixed Mullerian tumors. The team that includes members from Johns Hopkins University and Oregon Health and Science University published its findings in today’s issue of the journal Nature Communications (paid subscription required).
Carcinosarcoma is an aggressive cancer made up of two forms of the disease, one affecting glandular cells and the other affecting soft tissue such as muscle or connective tissue. Tumors from carcinosarcoma generally appear in the uterus , but also in supporting organs and tissue. While carcinosarcoma accounts for 3 to 4 percent of uterine cancers, it has a higher mortality rate.
Personal Genome Diagnostics in Baltimore is a company founded by two of the paper’s authors, Victor Velculescu and Luis Diaz, both on the faculty of Johns Hopkins University’s medical school. The company conducts genomic sequencing of tumors and bioinformatics analyses to provide physicians with useful guidance to prescribe treatments for their patients.
Blueprint Medicines in Cambridge, Massachusetts develops cancer therapies that aim to limit the actions of kinases, enzymes supporting cancer growth resulting from genomic mutations. The company’s technology is designed to generate treatments for patients that address their precise genomic alterations. Christoph Lengauer, Blueprint’s chief scientist, was one of the paper’s senior authors, with Velculescu of Johns Hopkins and Personal Genome Diagnostics.
The research team analyzed the exomes of carcinosarcoma from 22 tumors with parallel high-throughput sequencing techniques to reveal the mutations in those tumors. The exome represents less than 2 percent of a whole genome, but accounts for about 85 percent genetic variations known to cause disease.
The findings revealed on average 43 mutations per tumor. In nearly two-thirds of the cases, the mutations affect genes regulating the chromosome structure and unpacking of the DNA to allow for transcription signals to reach their destinations, a process known as chromatin remodeling. About three-quarters of genetic alterations uncovered, say the authors, have potential as therapy targets, and in some cases were not previously associated with carcinosarcoma.
Personal Genome Diagnostics and Blueprint Medicines are already collaborating on identification of targets for Blueprint’s kinase inhibitor cancer treatments. Blueprint anticipates having 2 or 3 therapies in clinical testing by the end of 2015.
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18 September 2014. National Cancer Institute, part of National Institutes of Health, is funding development of lab tests using a patient’s own cancer cells to help determine the best treatments for the patient. The $1.975 million contract to biotechnology company Kiyatec Inc. in Greenville, South Carolina was awarded under the Small Business Innovation Research program, a government-wide vehicle for supported research and development by start-up enterprises.
Kiyatec’s technology provides personalized cancer testing using a patient’s own tumor cells, but cultured in the lab to allow for physicians to test potential therapies in a controlled setting before treating the patient. The company cites data from a Stanford University study, published last year in the journal Nature Medicine, showing three-dimensional cell models like those developed by Kiyatec offer a more realistic environment for testing outside the body than conventional lab cultures, including better representation of cancer’s genetic signatures.
The award, says Kiyatec, is a second-stage research project, building on development of a 3-D cell lab model for breast cancer, as a testing medium for potential treatments. That research, a nine-month proof-of-concept study, received a $295,000 award last September also from National Cancer Institute.
The new contract will support a two-year undertaking to adapt the company’s 3-D cell models to test for the cancer’s interaction with a patient’s immune system, as well as the patient’s blood supply, in this case formation of new blood vessels that support tumor growth. In addition, Kiyatec says the award will allow it to expand the company’s technology beyond its current focus on breast and ovarian cancer to glioblastoma multiforme, a highly malignant type of brain cancer with a low likelihood of survival.
Kiyatec expects the new funding will help it develop tools that give physicians drug profiles more personalized for their patients, particularly for predicting the ability of a treatment to invoke a patient’s immune system to fight the cancer. The technology, says the company, can also be applied in advance of clinical trials for identifying therapies with the best chance of succeeding with patients.
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3-D printed syringe pump (Emily Hunt, Michigan Technological University)
18 September 2014. Engineers at Michigan Technological University in Houghton produced a syringe pump, a common but often expensive piece of lab equipment, with three-dimensional printing that drastically cuts the cost of the device. The team led by Michigan Tech’s Joshua Pearce published its findings yesterday in the journal PLoS One, and makes the pump’s designs available through an open-source library.
Syringe pumps are computer-controlled devices for injecting liquids into solutions or cultures in precisely controlled quantities, as required in academic or corporate labs. Factory-manufactured syringe pumps, however, can be expensive, running into hundreds or thousands of dollars per unit, which often limits their use and number in research labs.
Pearce and colleagues created their syringe pump with a 3-D printer, using free and publicly-available components and software, which they say costs as little as 5 percent of the prices charged for factory-made devices. The team designed three basic configurations that requires inexpensive manufactured metal parts — e.g., bearings, rods, and stepper motors — while the casings, clamps, and wedges are 3-D printed from plastic filaments.The 3-D printing is done with a RepRap printer, itself the product of an open-source research and development project.
The software controlling the RepRap is written in OpenSCAD, an open-source 3-D solid modeling program that supports complex designs. The OpenSCAD design is translated using Cura, another open-source software, into a file format readable by the RepRap called g-code that controls actions of the print-head.
The devices are controlled by systems based on a Raspberry Pi, an open-source processing system about the size of a credit-card, that connects either through a wired or wireless link to the motor in the pump. While the researchers created three basic designs for the syringe pump, users can adjust the calibration, speed, and position of the pump through software. The controller can also drive more than one pump in parallel.
The Michigan Tech team tested the 3-D printed devices and found the pumps were able to deliver liquids within 1 to 5 percent of the desired amounts, depending on the size of pump. The tests included a configuration running two syringes simultaneously.
While the researchers designed three basic configurations, the use of 3-D printing makes it possible for labs to custom-design pumps for their own research needs. The low-price of the 3-D printed devices also enables labs to run more of their experiments in parallel rather than use a single pump sequentially.
“Not only have we designed a single syringe pump,” says Pearce in a university statement, “we’ve designed all future syringe pumps.”
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17 September 2014. The pharmaceutical company Sanofi and biopharmaceutical developer MyoKardia are collaborating on development and commercialization of three MyoKardia therapies for inherited heart diseases. The deal could earn MyoKardia as much as $200 million in milestone payments and equity investments from Sanofi.
MyoKardia, in South San Francisco, California, designs small-molecule therapies for two types of genetic heart disorders: hypertrophic cardiomyopathy and dilated cardiomyopathy that result from mutations in protein genes of muscles used in heart contractions. With hypertrophic cardiomyopathy, heart muscles become abnormally thick, making it more difficult for the heart to pump blood. In dilated cardiomyopathy, the heart’s left ventricle — the main pumping chamber — becomes enlarged, resulting in less pumping force than a healthy heart.
The company was started in 2012 by four academic researchers in heart muscle biology and cardiovascular genetics from Harvard Medical School, Brigham and Women’s Hospital, and University of Colorado, and received its initial financing of $38 million from Third Rock Ventures, a life sciences venture capital firm. In May, MyoKardia unveiled its Sarcomeric Human Cardiomyopathy Registry, or SHaRe, repository of de-identified clinical data on genetic heart disease, with data on some 5,800 patients, developed with researchers from 7 medical centers.
The agreement covers three MyoKardia therapies, two for hypertrophic cardiomyopathy and one addressing dilated cardiomyopathy. MyoKardia will be responsible for continued development of the three therapies through initial demonstrations of efficacy in patients, after which the two companies will share development costs of the hypertrophic cardiomyopathy drugs, while Sanofi covers further development costs of the dilated cardiomyopathy program.
MyoKardia will retain product product rights for the two hypertrophic cardiomyopathy therapies, while Sanofi has worldwide rights to develop and commercialize the dilated cardiomyopathy drug. The companies divide up geographic commercialization activities, with MyoKardia responsible for commercialization of the hypertrophic cardiomyopathy drugs in the U.S., and Sanofi responsible for areas outside the U.S. where it now operates. If further uses of the therapies emerge, Sanofi will have the option to co-promote either of the hypertrophic cardiomyopathy therapies in the U.S., while MyoKardia will have the option to co-promote the dilated cardiomyopathy drug in the U.S.
Also under the deal, Sanofi, based in Paris, paid MyoKardia an upfront payment of $45 million in licensing fees and equity investment. MyoKardia can qualify as well for up to $155 million in further payments for achieving milestones, research and development services, and further equity investments from Sanofi.
The agreement is part of a Sanofi initiative it calls Sunrise that seeks out partnerships with early-stage drug development projects that the company believes can benefit from investment and its commercialization expertise. In addition to MyoKardia, the Sunrise program has a collaboration with WarpDrive Bio, a developer of natural therapeutic products based on genomics.
Hat tip: FirstWord Pharma
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Chronic lymphocytic leukemia cells, stained purple (California Institute for Regenerative Medicine)
17 September 2014. An early stage clinical trial at University of California in San Diego is testing the safety of an antibody that in lab animals decreases the number of chronic lymphocytic leukemia cells, the most common form of leukemia among adults. The study is led by UC-San Diego medical school professor Thomas Kipps, who is also director of the Chronic Lymphocytic Leukemia Research Consortium.
Chronic lymphocytic leukemia is a disorder of the blood and bone marrow, where the bone marrow makes too many blood stem cells that fail to mature into healthy white blood cells. The overabundance of these leukemia cells crowd out the healthy blood cells, including other white blood cells that support the immune system as well as platelets and red blood cells. The disease causes individuals, primarily adults, to develop infections, anemia, and easier bleeding.
Kipps and colleagues at UC-San Diego designed an antibody called cirmtuzumab that targets a protein known as ROR1 found on the surface of chronic lymphocytic leukemia cells. The UC-San Diego team developed cirmtuzumab using antibody drug conjugate techniques devised by biotechnology company Concortis Biosystems. In June, Concortis Biosystems was acquired by Sorrento Therapeutics, also in San Diego, another biotechnology company designing antibodies as cancer therapies.
ROR1, the target for cirmtuzumab, is a protein that appears to occur only in cancer cells, not in healthy cells, thus the researchers believe this protein acts as a reliable biomarker of chronic lymphocytic leukemia. Previous research by Kipps’s lab found ROR1 to occur in a range of solid-tumor and blood-related cancers, where it combines with cancer-causing mutations early in the process to encourage the proliferation of cancer cells, as well as the spread of cancer or metastasizing to other parts of the body.
In tests of cirmtuzumab with lab cultures and animals, the UC-San Diego researchers found the antibody to target cells with ROR1 proteins, i.e. cancer cells, while avoiding healthy cells without ROR1. The findings also showed cirmtuzumab injections kill many more human chronic lymphocytic leukemia cells with ROR1 in lab animals than those not getting the injections, while cells without ROR1 proteins are unaffected. In addition, the results show the antibody has potential for killing pancreas and breast cancer cells expressing the ROR1 protein.
The clinical trial is enrolling 56 patients with chronic lymphocytic leukemia who have relapsed or advanced forms of the disease that do not respond to approved treatments. The patients will receive intravenous doses of cirmtuzumab at various levels, where clinicians will be watching for adverse reactions and overall tolerability of the treatments, although signs of clinical benefits to patients will also be noted. The treatments will be given every 14 days for 8 weeks.
The trial is expected to be completed in August 2016.
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3-D brain wiring illustration (NIH)
16 September 2014. The pharmaceutical companies Eli Lilly and Company and AstraZeneca will jointly develop a drug to treat Alzheimer’s disease, currently in early-stage clinical trials. In the $500 million deal, Lilly is licensing research and development conducted so far by AstraZeneca, with the companies dividing up further clinical development, commercialization, and manufacturing of the drug.
In Alzheimer’s disease, amyloid plaque develops in the brain, breaking down the ability of neurons or nerve cells to function efficiently, leading to death of neurons and shrinkage of brain tissue. Cognitive and memory loss are common symptoms of Alzheimer’s disease, which is the leading form of dementia, affecting more than 5 million people and the 6th leading cause of death in the U.S.
The companies plan to advance AstraZeneca’s drug, code-named AZD3293, into intermediate and late-stage clinical trials as a treatment for people with early symptoms of Alzheimer’s disease. AZD3293 is an oral beta secretase cleaving enzyme, or BACE, inhibitor that aims to prevent the build-up of amyloid plaque toxicity.
The small molecule drug acts by cleaving an amyloid precursor protein that releases and activates peptides inhibiting BACE, which in turn reduces toxicity of amyloid beta peptides making up the accumulating plaque. AstraZeneca tested AZD3293 in early-stage clinical trials, beginning in 2012, which show the drug can reduce levels of amyloid beta peptides in cerebro-spinal fluid of both patients with Alzheimer’s disease and healthy volunteers.
Under the new agreement, Lilly, in Indianapolis, will lead further clinical development of AZD3293 into intermediate and late-phase trials among patients in the early stages of Alzheimer’s disease. The two companies will share commercialization of the drug, while AstraZeneca will be responsible for manufacturing.
Lilly will pay AstraZeneca, based in London, up to $500 million in development and regulatory payments, with the first payment of $50 million expected in the 3rd quarter of 2014. The two companies plan to share equally future development and commercialization costs, as well as net revenues worldwide after launch of the drug.
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Science & Enterprise is a taking long weekend beginning tomorrow, 12 September through Monday, 15 September. We’ll resume posting on Tuesday 16 September.
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