Tag Archives: mouse

Research Roundup: Lupus protein identified, vaccine for Type 1 diabetes, new chronic pain treatment, and more!

Welcome to this week’s Research Roundup. These Friday posts aim to inform our readers about the many stories that relate to animal research each week. Do you have an animal research story we should include in next week’s Research Roundup? You can send it to us via our Facebook page or through the contact form on the website.

  • A protein that may cause Lupus has been identified.Lupus is a chronic inflammatory disease that occurs when your body’s immune system attacks its own tissues and organs. An estimated 1.5 million Americans, and at least 5 million people worldwide, have a form of lupus. Previous research has implicated the gene PRDM1 as a risk factor for lupus. Scientists looking at Blimp-1, a protein that is encoded by the PRDM1 gene, have found in mice thatthat a low level of or no Blimp-1 in a particular cell type led to an increase in the protein cathepsin S (CTSS) which caused the immune system to identify healthy cells as something to attack — particularly in females.” These results are particularly striking as women have an increased risk for lupus compared to men. While this work needs to be replicated and validated, this research provides some valuable insight into the etiology and treatment of lupus. This research was published in the journal Nature Immunology.
lupus mice

Mice from the Lupus study. Source: AJP Renal Physiology.

  • Vaccine for virus induced Type 1 diabetes successful in mice. Coxsackie B viruses are the most common enteroviruses and are believed to be associated with the development of Type 1 diabetes. Type 1 diabetes is  a common human disease defined by a decrease in the production of insulin, which is a hormone that allows blood glucose (sugar) to enter energy producing cells. Thus, without insulin your body cannot effectively produce energy. This week, a team of Finnish researchers published a preclinical evaluation of a Coxsackie B1 vaccine using mice and found that the vaccine successfully protected the mouse after administering the Coxsackie B1 virus. Pre-clinical trials in humans are the next logical step for this vaccine, and the researchers believe that this research will aid in the development of vaccines for other disease caused by enteroviruses such as; hand-foot-and-mouth disease, meningitis, and myocarditis. This research was published in the journal Vaccine.
  • High iron levels in brain linked to progression of Alzheimer’s. Alzheimer’s is a degenerative neurological disease that causes dementia in humans. Previous research has linked Alzheimer’s to the buildup of amyloid protein in the brain, but research on drugs that reduce amyloid levels have not successfully slowed the progression of the disease. New research from the University of Melbourne however, discovered that humans with high levels of iron and amyloid were suffering from rapid dementia, while those with just high levels of amyloid protein were stable. This finding will fuel a five year trial on whether an anti-iron drug can slow the progression of the disease. This research was published the journal Brain.
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Brain MRI using Quantitative Susceptibility Mapping (QSM). Source: University of Melbourne.

  • Compound protects macaques from simian HIV. Research being presented this week at the National AIDS Treatment Advocacy Project (NATAP) Conference on HIV Pathogenesis Treatment and Prevention in Paris shows that weekly administration of a compound called MK-8591 repeatedly protected 8 of 8 macaques from simian HIV (SHIV) 6 days after treatment. Researchers from the Aaron Diamond AIDS Research Center in New York, Merck, and the Tulane National Primate Research Center studies 16 male macaques, 8 of which received weekly treatments of MK-8591 for up to 14 weeks, the other 8 of which received a placebo. MK-8591 is a nucleoside reverse transcriptase translocation inhibitor (NRTTI) that thwarts HIV. All 8 monkeys treated with MK-8591 remained SHIV-free even after 12 challenges with SHIV, to the end of the 168-day study. In contrast, the monkeys not treated with the drug all became infected with SHIV. The researchers noted that protective intracellular active MK-8591 concentrations can be attained in humans at low drug doses. These new findings support the potential use of MK-8591 as a prophylactic treatment for high-risk individuals.
  • New drug acting at two opioid receptors shows promise to treat chronic pain without the adverse effects of morphine. An epidemic of opioid abuse is killing people by the hundred of thousands in the United States, becoming the main cause of death for people under 40. The epidemic has been traced to the prescription of new opioid analgesics like Oxycontin over the last years, which has led people to become addicted to it and then to harder drugs like heroin. A group of scientist followed the strategy of creating drugs that bind not only to the receptor for morphine in the brain, the mu-opioid receptor, but also to a new opioid receptor called the nociceptin FQ receptor. The new drug, BU08028, was shown to reduce pain responses in rats as effectively as morphine. Now the drug is being tested in rhesus monkeys, where it also decreased pain. Importantly, the monkeys showed no desire to self-administer BU08028, an indication that the drug is not addictive.
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Male rhesus macaque. Source: Kathy West.

  • Computer-designed opioid drug may decrease pain without producing respiratory arrest. People dying during the current opioid epidemic do so because the drugs inhibit the centers in the brain that drive breathing, leading to suffocation. Scientists proposed that the pain-relieving and breathing-suppressive effects of the opioids depend on different interactions of these drugs with the mu-opioid receptor, and set up to produce a compound that would suppress pain but not breathing. To do that, they modeled the drug binding site at the receptor using computers. After testing millions of compounds in the computer, they found a drug, PZM21, that showed promise. Then they tested PZM21 in mice and found that it suppressed pain but not breathing. They also found that PZM21 did not show the rewarding effect that typically lead to addiction. This demonstrates how even when new drugs are designed using computer models, animal studies are still needed to evaluate their effects.

Research Roundup: Malaria vaccine, mouse sperm in space, animal welfare prizes, and more!

Welcome to this week’s Research Roundup. These Friday posts aim to inform our readers about the many stories that relate to animal research each week. Do you have an animal research story we should include in next week’s Research Roundup? You can send it to us via our Facebook page or through the contact form on the website.

  • New study finds that mouse sperm stored in space still functions on Earth. Increasingly in the news we read about the upcoming reality of commercial space travel (for example, here and here). Of course, with such advances there is caution with respect to feasibility — and of course imagination with respect to possibilities (e.g., colonizing Mars). With such goals on the horizons, these researchers investigated whether sperm that had been freeze dried, and transported to the International Space Station (ISS) and then back to Earth would be able to produce viable offspring. To accomplish this they used freeze dried mouse spermatozoa — which provided a unique advantage, as the addition of water — maintains the viability of the sperm to fertilize an egg and allows for the sperm to be stored at room temperature. Other sperm when freeze dried do not survive. Microinjection  of these “space” sperm into an egg on Earth — produced healthy viable  “space offspring”. Moreover, these offspring all grew to healthy adults and were able to produce offspring of their own. This study was published in the Proceedings of the National Academy of Sciences of the USA.

Space mouse and pups. Source: PNAS

Laboratory frogs. Source: University of Portsmouth

  • Modified experimental vaccine protects monkeys from deadly malaria. Researchers at the National Institute of Allergy and Infectious Diseases (NIAID), part of the National Institutes of Health, discovered that a modified version of an experimental malaria vaccine completely protected 4 of 8 monkeys from a malaria parasite, and delayed the first appearance of the parasites in 3 more monkeys. Scientists modified an existing malaria vaccine by including a particular protein, RON2L, so that it closely mimicked the protein complex used by the parasite to infect blood cells. Vaccination with the modified vaccine resulted in more neutralizing antibody, indicating a better quality response to parasitic infection. Additionally, the modified vaccine seemed to protect against other parasite strains that differed from those used to create the vaccine, suggesting that this new modified vaccine may protect against multiple parasite strains. This research will pave the way toward eventual human trials. The study was published in NPJ Vaccines.

A female Aedes mosquito. Source: NIAID.

 

 

 

 

 

 

 

 

 

 

 

  • Researchers at the University of Helsinki has found the lymphatic vessels extend into the brain – overturning 300 years of accepted wisdom. By genetically altering mice using the luminescent GFP gene, so that lymphatic vessels glowed under light, Aleksanteri Aspelund found that there were lymphatic vessels in the brain. The research was repeated by Karl Alitalo with the same results.  Other researchers have found evidence linking problems with the lymphatic and glymphatic systems to Alzheimer’s; one study in mice showed it could lead to the buildup of amyloid beta in the brain – a key sign of the Alzheimer’s. The study was published in the Journal of Experimental Medicine.

    Red fluorescence of the membrane protein aquaporin-4 in an individual with Alzheimer’s (left) and a healthy individual (right). Source: OHSU

  • Mice help researchers identify genes responsible for a severe congenital heart defect.  Congenital heart disease affect up to 1 percent of all live births. Hypoplastic Left Heart Syndrome (HLHS) is a rare congenital heart disease resulting in an inability to effectively pump blood  throughout the body.  Current treatment involves multiple complex surgeries during the first few years of a child’s life. For some, the surgical interventions improve heart function.  For others, it does not,  leading to heart failure and the need for heart transplants. It has been known that genetic risk factors play a role in HLHS but specific genes have been hard to identify.  Researchers at the University of Pittsburgh Schools of the Health Sciences used fetal ultrasound imaging to look for structural heart defects in genetically modified mice to identify HLHS.  Then by comparing the genomes of affected and non affected mice, and confirming using CRISPR technology they found that mutations in two specific genes that interact were required for HLHS.,   Dr. Cecilia Lo, a professor and the F. Sargent Cheever Chair in Developmental Biology at Pitt says, “Studying diseases with complex genetics is extremely challenging…By understanding the genetics and biology of HLHS, this can facilitate development of new therapies to improve the prognosis for these patients.” This study was published in the journal Nature Genetics.
  • The University of Bristol has awarded prizes in its first Animal welfare and 3Rs Symposium. The 3Rs, developed by Russel and Burch in 1954, have advanced the humane treatment of animals used in research by advocating for replacement (aiming to replace animals where possible, with alternatives), to reduce the number of animals used to the minimum required to answer and experimental question and and to refine their experiments to minimise any adverse effects experienced by the animals.These awards went to three research projects that have advanced the 3Rs in their various lines of research.

“The research project that won first prize has developed a refined method for producing aortic aneurysms in mice.  An aortic aneurysm is a bulge in a section of the aorta, which is the body’s main artery, and if the bulge ruptures it can cause sudden death. The research team has also developed a new human aortic aneurysm model in the laboratory, potentially replacing the need for animal models, using arteries taken from the discarded umbilical cord of newly born babies.

The second prize was awarded to a research team who has developed a method for giving oral drugs using solutions that mice and rats both like and which avoids the need for restraint and reduces stress in the animals. The research team found that liquid foods such as condensed milk, milkshake and fruit puree baby food are good solutions to use for giving a wide range of drugs.
The final prize was awarded to a research team who has developed photographic techniques that can be used in conscious animals.  This new technique has revolutionised preclinical eye research and has markedly reduced the number of animals needed for research studies.”

The 3Rs. Source: Bayer

SR-Research Roundup

July 21st-July 28th

Welcome to this week’s Research Roundup. These Friday posts aim to inform our readers about the many stories that relate to animal research each week. Do you have an animal research story we should include in next week’s Research Roundup? You can send it to us via our Facebook page or through the contact form on the website.

  • A protein that may cause Lupus has been identified. Lupus is a chronic inflammatory disease that occurs when your body’s immune system attacks its own tissues and organs. An estimated 1.5 million Americans, and at least 5 million people worldwide, have a form of lupus. Previous research has implicated the gene PRDM1 as a risk factor for lupus. Scientists looking at Blimp-1, a protein that is encoded by the PRDM1 gene, have found in mice thatthat a low level of or no Blimp-1 in a particular cell type led to an increase in the protein cathepsin S (CTSS) which caused the immune system to identify healthy cells as something to attack — particularly in females.” These results are particularly striking as women have an increased risk for lupus compared to men. While this work needs to be replicated and validated, this research provides some valuable insight into the etiology and treatment of lupus. This research was published in the journal Nature Immunology.

Source: http://ajprenal.physiology.org/content/308/10/F1146

  • Vaccine for virus induced Type 1 diabetes successful in mice. Coxsackie B viruses are the most common enteroviruses and are believed to be associated with the development of Type 1 diabetes. Type 1 diabetes is a common human disease defined by a decrease in the production of insulin, which is a hormone that allows blood glucose (sugar) to enter energy producing cells. Thus, without insulin your body cannot effectively produce energy. This week, a team of Finnish researchers published a preclinical evaluation of a Coxsackie B1 vaccine using mice and found that the vaccine successfully protected the mouse after administering the Coxsackie B1 virus. Pre-clinical trials in humans are the next logical step for this vaccine, and the researchers believe that this research will aid in the development of vaccines for other disease caused by enteroviruses such as; hand-foot-and-mouth disease, meningitis, and myocarditis. This research was published in the journal Vaccine.
  • High iron levels in brain linked to progression of Alzheimer’s. Alzheimer’s is a degenerative neurological disease that causes dementia in humans. Previous research has linked Alzheimer’s to the buildup of amyloid protein in the brain, but research on drugs that reduce amyloid levels have not successfully slowed the progression of the disease. New research from the University of Melbourne however, discovered that humans with high levels of iron and amyloid were suffering from rapid dementia, while those with just high levels of amyloid protein were stable. This finding will fuel a five year trial on whether an anti-iron drug can slow the progression of the disease. This research was published the journal Brain.

Source:https://pursuit.unimelb.edu.au/articles/rusty-brains-linked-to-alzheimer-s?utm_source=twiter&utm_medium=social&utm_content=story

  • Compound protects macaques from simian HIV. Research being presented this week at the National AIDS Treatment Advocacy Project (NATAP) Conference on HIV Pathogenesis Treatment and Prevention in Paris shows that weekly administration of a compound called MK-8591 repeatedly protected 8 of 8 macaques from simian HIV (SHIV) 6 days after treatment. Researchers from the Aaron Diamond AIDS Research Center in New York, Merck, and the Tulane National Primate Research Center studies 16 male macaques, 8 of which received weekly treatments of MK-8591 for up to 14 weeks, the other 8 of which received a placebo. MK-8591 is a nucleoside reverse transcriptase translocation inhibitor (NRTTI) that thwarts HIV. All 8 monkeys treated with MK-8591 remained SHIV-free even after 12 challenges with SHIV, to the end of the 168-day study. In contrast, the monkeys not treated with the drug all became infected with SHIV. The researchers noted that protective intracellular active MK-8591 concentrations can be attained in humans at low drug doses. These new findings support the potential use of MK-8591 as a prophylactic treatment for high-risk individuals.
  • New drug acting at two opioid receptors shows promise to treat chronic pain without the adverse effects of morphine. An epidemic of opioid abuse is killing people by the hundred of thousands in the United States, becoming the main cause of death for people under 40. The epidemic has been traced to the prescription of new opioid analgesics like Oxycontin over the last years, which has led people to become addicted to it and then to harder drugs like heroin. A group of scientist followed the strategy of creating drugs that bind not only to the receptor for morphine in the brain, the mu-opioid receptor, but also to a new opioid receptor called the nociceptin FQ receptor. The new drug, BU08028, was shown to reduce pain responses in rats as effectively as morphine. Now the drug is being tested in rhesus monkeys, where it also decreased pain. Importantly, the monkeys showed no desire to self-administer BU08028, an indication that the drug is not addictive.
  • Computer-designed opioid drug may decrease pain without producing respiratory arrest. People dying during the current opioid epidemic do so because the drugs inhibit the centers in the brain that drive breathing, leading to suffocation. Scientists proposed that the pain-relieving and breathing-suppressive effects of the opioids depend on different interactions of these drugs with the mu-opioid receptor, and set up to produce a compound that would suppress pain but not breathing. To do that, they modelled the drug binding site at the receptor using computers. After testing millions of compounds in the computer, they found a drug, PZM21, that showed promise. Then they tested PZM21 in mice and found that it suppressed pain but not breathing. They also found that PZM21 did not show the rewarding effect that typically lead to addiction. This demonstrates how even when new drugs are designed using computer models, animal studies are still needed to evaluate their effects.

Nobel Prize 2016 – how yeast and mouse studies uncovered autophagy

Congratulations to Professor Yoshinori Ohsumi Tokyo Institute of Technology on being awarded the 2016 Nobel Prize in Physiology or Medicine for “for his discoveries of mechanisms for autophagy“!

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Yoshinori Ohsumi. Image: Tokyo Institute of Technology

The process of autophagy is hardly one familiar to most people, but is is absolutely crucial to all complex life on out planet, including ourselves. The name autophagy comes from the Greek words for “self” and “eating” and describes the ordered process through which cells break down and recycle unnecessary or damaged structures or proteins, and allows the cell to reach an equilibrium between the synthesis and degradation of proteins.

The discovery of autophagy

The process itself was identified through studies in tissues of mice and rats back in the 1950’s and 1960’s, by scientists including Christian de Duve, who was subsequently awarded the Nobel Prize in Physiology or Medicine in 1974 for this and other work. They first discovered that mammalian cells contain a compartment which they termed the lysosome where proteins are broken down, and then that proteins and other molecules that were to be degraded were first isolated from the rest of the cell by the formation of a membrane sac around the protein in question  (later called the autophagosome). The process through which the autophagosome fused with the lysosome to deliver its protein cargo for degradation was given the name autophagy by Christian de Duve.

Image: NobelPrize.org

Image: NobelPrize.org

Progress in understanding how autophagy worked was slow, as at the time the genes or proteins involved in regulating the process had been identified. With the research methods available at the time it was difficult to measure autophagy as it happened in mammalian cells, and hence difficult to determine how altering different components affected the overall process, a key step towards understanding their role. It may have seemed an unpromising field to join, but Yoshinori Ohsumi had a different career philosophy to most researchers, which he described in an interview given in 2012:

I am not very competitive, so I always look for a new subject to study, even if it is not so popular. If you start from some sort of basic, new observation, you will have plenty to work on.

From cells to genes

What was needed was a simple experimental system in which to study the process, and the bakers yeast Saccharomyces cerevisiae  – a simple single celled organism separated from us by hundreds of millions of years of evolution, but sharing many of our key biological processes – was one candidate. Yoshinori Ohsumi had worked with yeast, and in particular had identified many proteins in a subcellular component of the yeast cell known as the vacuole, which was important as there was evidence that the vacuole performed the same role in yeast cells as the lysosome in mammalian cells. Still, as the Nobel Prize website highlights there were still hurdles to overcome as he began his study of autophagy in yeast at the end of the 1980’s:

But Ohsumi faced a major challenge; yeast cells are small and their inner structures are not easily distinguished under the microscope and thus he was uncertain whether autophagy even existed in this organism. Ohsumi reasoned that if he could disrupt the degradation process in the vacuole while the process of autophagy was active, then autophagosomes should accumulate within the vacuole and become visible under the microscope. He therefore cultured mutated yeast lacking vacuolar degradation enzymes and simultaneously stimulated autophagy by starving the cells. The results were striking! Within hours, the vacuoles were filled with small vesicles that had not been degraded (Figure 2). The vesicles were autophagosomes and Ohsumi’s experiment proved that authophagy exists in yeast cells. But even more importantly, he now had a method to identify and characterize key genes involved this process.

With an experimental system available Yoshinori Ohsumi and his team studied the process of autophagy in thousands of mutant strains of yeast, and identified 15 individual genes (most of them of previously unknown function) that are essential for the process in yeast, tho order in which the key events in autophagy take place, and the roles of the individual genes in them. This was the work for which he was awarded the Nobel Prize.

From yeast genes to us!

But it is not the end of the story! Identifying the genes essential for autophagy in yeast, and their roles in the process, was a major breakthrough, but what about humans and other mammals?

It turns out that that in humans and other mammals there are counterparts to almost all the yeast autophagy genes, though the situation is made a lot more complicated by the face that mammals have more than one copy for each of the genes…starting with yeast was a wise move! Professor Noboru Mizushima of the University of Tokyo made an important advance when, working with Yoshinori Ohsumi,  he developed a transgenic mouse in which a protein called LC3 that is found in the autophagosome membrane is fused to Green Fluorescent Protein (GFP – see Nobel Prize for Chemistry 2008) which allowed him and his colleagues to observe and monitor the process of autophage in vivo in mice for the first time.

Laboratory Mice are the most common species used in research

This LC3-GFP transgenic mouse proved to be a very powerful research tool for studying mammalian autophagy, allowing not only the role of indicudual genes in the process to be determined, but also the role of autophagy itself in processes as diverse as early embryonic development, tumor suppression, nerve cell survival and function, and protection against infection.

This research is still at a relatively early stage, but techniques such as the LC3-GFP system in mice – and others used in organisms such as fruit flies, are showing us how defects in autophagy contribute to many diseases, including neurodegenerative disorders such as Parkinson’s Disease, and metabolic disorders such as type 2 Diabetes. While the development of specific therapies to correct these defects in autophagy is still some way off, it is already clear that understanding autophagy has the potential to improve the treatment of a wide range of illnesses.

What the work of Yoshinori Ohsumi demonstrates once again is the crucial contribution of basic biological research in model organisms that may at first glance appear to share little with us to the advancement of medicine.

Speaking of Research

 

 

Clinical trial success for Cystic Fibrosis gene therapy: built on animal research

This morning the Cystic Fibrosis Gene Therapy Consortium (GTC) announced the results of clinical trial in 140 patients with cystic fibrosis, which demonstrate the potential for gene therapy to slow – and potentially halt – the decline of lung function in people with the disorder. It is a success that is built on 25 years of research, in which studies in animals have played a crucial role.

Cystic fibrosis is one of the most commonly inherited diseases, affecting about one in every four thousand children born in the USA, and is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The CFTR gene produces a channel that allows the transport of chloride ions across membranes in the body, and the many mutations identified in cystic fibrosis sufferers either reduce the activity of the channel or eliminate it entirely. This defect in chloride ion transport leads to defects in several major organs including the lungs, digestive system, pancreas, and liver. While the severity of the disease and the number of organs affected varies considerably, cystic fibrosis patients often ultimately require lung transplant s, and too many still die early in their 20’s and 30’s as the disease progresses.

In a paper published in Lancet Respiratory Medicine today (1), the GTG members led by Professor Eric Alton of Imperial College London compared monthly delivery to the airway of a non-viral plasmid vector containing the CFTR gene in the liposome complex pGM160/GL67A using a nebuliser with a placebo group who received saline solution via the nebuliser. They reported stabilisation of lung function in the pGM169/GL67A group compared with a decline in the placebo group after a year. This is the first time that gene therapy has been shown to safely stabilise the disease, and while the difference between the treated and control group was modest, and the therapy is not yet ready to go into clinical use, it provides a sound bases for further development and improvement.

Blausen_0286_CysticFibrosis

The Chief Executive of the Cystic Fibrosis Trust, which is one of the main funders of the GTC, has welcomed the results, saying:

Further clinical studies are needed before we can say that gene therapy is a viable clinical treatment. But this is an encouraging development which demonstrates proof of concept.

“We continue to support the GTC’s ground-breaking work as well as research in other areas of transformational activity as part of our mission to fight for a life unlimited by cystic fibrosis.”

So how did animal research pave the way for this trial?

Following the identification of the CFTR gene in 1989 scientists sought to create animal models of cystic fibrosis with which to study the disease, and since the early 1990’s more than a dozen mouse models of cystic fibrosis have been created. In some of these the CFTR gene has been “knocked out”, in other words completely removed, but in others the mutations found in human cystic fibrosis that result in a defective channel have been introduced. These mouse models show many of the defects seen in human cystic fibrosis patients and over the past few years have yielded important new information about cystic fibrosis, and in 1993 Professor Alton and colleagues demonstrated that it is possible to deliver a working copy of the CFTR gene using liposomes to the lungs of CFTR knockout mice and correct some of the deficiencies observed.

To get a working copy of the CFTR gene to the lungs of cystic fibrosis patients Professor Alton and colleagues needed three things:

• A DNA vector containing the working CFTR gene that is safe and  can express sufficient amounts of the CFTR channel protein in the lungs to correct the disease

• A lipid-like carrier that can form a fatty sphere around the DNA vector to so that it can cross the lipid membrane of cells in the lung, as “naked” DNA will not do this efficiently.

• A nebuliser device that produces an aerosol of the gene transfer agent so that it can be inhaled into the lungs of the patient.

Several early attempts to use gene therapy using viral vectors to deliver the working copy of the CFTR gene to patients failed because the immune response rapidly neutralised the adenoviral vector (see this post for more information on challenges using adenoviral vectors), and while attempts to use non-viral vectors were more promising, it was found that they caused a mild inflammation in most patients, which would make then unsuitable for long term use. As reported in a paper published in 2008 the GTC members developed and assessed in mice a series of non-viral DNA vectors, repeatedly modifying them and testing their ability to both drive CFTR gene expression in the lungs and avoid inducing inflammation. They finally hit on a vector – named pGM169 – which fulfilled both key criteria.

Earlier the consortium had undertaken a study to determine which carrier molecule to use in their non-viral gene transfer agent (GTA). To do this they assessed 3 GTA’s, each consisting of a lipid like molecule that could form a sphere around the non-viral DNA vector; either the 25 kDa-branched polyethyleneimine (PEI), the cationic liposome GL67A, or as a compacted DNA nanoparticle formulated with polyethylene glycol-substituted lysine 30-mer. Because there are significant differences in airway physiology between mouse and human they carried out this study in sheep, whose lung physiology more closely matches that of humans. The study identified the cationic liposome GL67A as the most promising candidate, resulting in robust expression of the CFTR transgene in the sheep lungs.

Studies in sheep play a key role in the development of gene therapy for cystic fibrosis

Studies in sheep play a key role in the development of gene therapy for cystic fibrosis

It now remained to bring the DNA vector and carrier together. In a 2013 publication the consortium reported that repeated aerosol doses of pGM169/GL67A to sheep over a 32 week period were safe and induced expression of the CFTR transgene in the sheep lungs, although the level of expression varied between individuals (this variation was also observed in human CF patients in the clinical trial reported today). A final study, this time in mice, assessed the suitability of the Trudell AeroEclipse II nebuliser as a device to create stable pGM169/GL67A aerosols, finding that it did so in a reproducible fashion. When aerosolized to the mouse lung, the new pGM169/GL67A formulation was capable of directing persistent CFTR transgene expression for at least 2 months, with minimal inflammation. These studies provided the evidence to support the gene delivery system and dosage strategy used in the clinical trial reported today.

The trial results announced today are an important accomplishment, but they mark a beginning rather than the end for Cystic Fibrosis gene therapy. It will be necessary to improve the efficiency of the therapy before it can enter widespread clinical use. Animal research will certainly play an important part in this work, notably the observation that the efficiency of CFTR gene delivery using this strategy was varied between individuals in both sheep and humans indicates that sheep are a good model in which to assess changes to improve the consistency and effectiveness of the gene therapy.

If you would like to know more about this cystic fibrosis gene therapy clinical trial you can watch two videos recorded at a meeting for cystic fibrosis patients at ICL on the  Cystic Fibrosis Trust website.

Paul Browne

1) Alton E.W.F.W. et al. “Repeated nebulisation of non-viral CFTR gene therapy in patients with cystic fibrosis: a randomised, double-blind, placebo-controlled, phase 2b trial” Lancet Respiratory Medicine Published online July 3, 2015

Animal research successes spur growth in science…but PeTA can only complain

What do multiple myeloma, influenza, advanced breast cancer, atrial fibrillation, thyroid cancer, ear infection, advanced ovarian cancer and obesity all have in common? One commonality is obvious – they cause suffering, sickness and sometimes death in people around the world. Another commonality is less obvious – these are each conditions that are now being treated with new drugs just approved by the U.S. Food and Drug Administration (FDA) in the past three months alone. That’s right… in the period from Thanksgiving 2014 until now, new drugs that treat each of these conditions have become available, and these agents will be used to treat the illnesses that may affect millions of Americans. Eventually, they will likely have enormous worldwide impacts on these diseases. That’s something to be thankful for.

While some are thankful that the scientific progress is successfully tackling human suffering and disease, others cast doubt on the way that progress is achieved. In a newly published analysis entitled “Trends in animal use at US research facilities” [1], employees of People for the Ethical Treatment of Animals (PeTA) – a self-avowed animal rights organization – report that, amongst the largest research universities in the United States, the number of animals involved in research has grown by over 70% during the past 15 years. In their publication, the authors express alarm over the growing use of animals not covered by the Animal Welfare Act (AWA), mostly mice and fish, in biomedical research, without making any mention of the impact of this research growth.

This growth in animal research in the US is directly linked to an accelerating pace of scientific study and its benefits. A brief visit to the FDA’s “New Drugs at FDA page” makes it quickly apparent that the rate of approval of new medications is astounding. Where is this progress coming from? At least in part, it’s coming from the scientific discoveries that are pouring out of the research laboratories located in colleges and universities, institutes and pharmaceutical and biotechnology companies around the globe. A good example is the innovative BiTE antibody Blincyto (blinatumomab) which was approved for use in treating B-cell acute lymphoblastic leukemia in December 2014 (clinical evaluation against other cancers is ongoing); as we discussed in a blog post in 2008, animal research – particularly studies in mice – played a key role in its development and early evaluation.

Thanks to the researchers that occupy laboratories around the world, scientific discoveries are coming faster than ever, and all of us benefit. It’s not just that there is more research being done – it’s that the impact of the science is better than ever thanks to more advanced technologies, accumulating knowledge of how the body works and more advanced animals models, including ones that mimic human disease processes in increasingly sophisticated ways that promote new discoveries and new opportunities to develop novel drugs.

Why is the scale of animal research growing in the US? The answer is clear: scientific progress is cumulative. One discovery often enables multiple other lines of work. The discovery of the structure of DNA, for example, enabled thousands of efforts to find the genetic causes of disease. Because of this, successes build on successes and research grows.

What is the consequence of the growth in animal research? The answer is: new treatments, new cures, less sickness and longer, healthier lives.

In their paper, the PeTA employees fail to mention any of the following accomplishments, allow of which resulted from the growing scientific research efforts around the world:

But this isn’t the end. To these existing accomplishments, add the work that was started in the past 15 years and will yet unfold in the forthcoming decade AND the overwhelming progress in basic/fundamental research that will lead to new treatments and cures throughout the first half of the 21st century, and you have the recipe for a growing animal research infrastructure in this country.

As recent statistics from the UK indicate, the increase in the use of mice and fish in research is driven almost entirely by the increasing number of studies that involve the use of genetically-modified (GM) animals. In other words, the increase is driven by scientific and technological advances that had a profound impact on biomedical research over the past 15 years, rather than any desire to avoid using species regulated by the AWA (while mice and fish studied in Universities are not covered by the AWA, research involving them is regulated in multiple ways, including through the federal Office of Laboratory Animal Welfare which issues the PHS Guide for the Care and Use of Laboratory Animals).

“Recent statistics from the UK indicate, the increase in the use of mice and fish in research is driven almost entirely by the increasing number of studies that involve the use of genetically-modified (GM) animals.”

Growing study of GM animals has occurred because these models are enormously useful. To take just one example, the National Institute of Child Health and Development recently published an online article entitled “It’s in the DNA: Animal Models Offer Clues to Human Development”, discussing the role of animal models in helping to understand human development and developmental disorders. But this is far from the only example, studies in GM mice are key to many of the state-of-the-art emerging fields in biomedical research. These range from the very new areas of optogenetics – which uses light to control activation of individual cells – and gene editing techniques such as CRISPR that have the potential to cure genetic disorders, to new therapies such as cancer immunotherapy and treatments for rare genetic disorders such as progeria and Pompe disease which are being used to successfully treat patients for whom effective therapies were previously unavailable.

The rise in the numbers of zebra fish is also driven by their value as research models. As vertebrates they share over 84% of the genes that cause disease when defective in humans, while their rapid reproduction and transparent eggs make them ideal subjects for genetic and developmental studies. It’s not surprising that they are both an increasingly popular species in basic biomedical research, and in the preclinical evaluation of potential new therapies and of the environmental safety of chemicals.

In recent years zebra fish have become an increasingly popular species in biomedical research.

What the statistics presented by PeTA in their article don’t tell you is that, while the number of experiments and studies have increased, animal research increasingly involves Refined techniques that produce minimized harm to the subjects and Reduced numbers of animals per study. And of course, animal research directly led to the ability to Replace animals in some types of studies, altogether. The efficacy and efficiency of animal research is advancing, and individual discoveries are, on average, being made with fewer animals. That is a fact missed entirely by the PeTA article.

Furthermore, within the concept of refinement is the idea that researchers should use animals that will suffer less in a laboratory setting wherever possible [2]. So replacing a small number of “higher” mammals with a high number of “lower” animals is consistent with the 3Rs principles of animal welfare. PeTA neglect to mention that USDA statistics show a 40% fall in the use of AWA-covered species over the last 15 years, and it is likely that a small proportion of the rise in use of non-AWA covered species is due to technological advances that have allowed non-AWA species (e.g. GM mice) to replace AWA species (e.g. monkeys) in some studies, for example to develop new treatments for HIV/AIDS, in line with the principle of Refinement we have outlined.

Number of animals used annually for research in the US

“PeTA neglect to mention that USDA statistics show a 40% fall in the use of AWA-covered species over the last 15 years”

Through the implementation of these 3Rs, scientists ensure that they engage in socially-responsible and ethical work. What the authors of the PeTA study should do is to explain how achieving their end goal of a virtual end to animal research, which will reverse the trend of accelerating discovery and medical progress upon which it depends, is ethical or defensible.

  1. Goodman, J., Chandna A., and Roe K. 2015. Trends in animal use at US research facilities in: J Med Ethics. 0:1-3
  2. Richmond, J., 2014. Refinement Alternatives: Minimizing Pain and Distress in Allen, D. and Waters M. ed. In Vivo Toxicity Testing” in: Reducing, Refining and Replacing the Use of Animals in Toxicity Testing. Cambridge: RSC. pp. 133

David Jentsch

Primate research and twenty years of stem cell firsts

This guest post is by Jordana Lenon, B.S., B.A., Senior Editor, Wisconsin National Primate Research Center and University of Wisconsin-Madison Stem Cell and Regenerative Medicine Center. The research will also be featured this evening in a public talk at UW-Madison’s Wednesday Nite at the Lab. WN@tL: “Twenty Years of Stem Cell Milestones at the UW.”  Details and link are below. Update 1/8/15:  Dr. William Murphy’s talk  can now be viewed at:  http://www.biotech.wisc.edu/webcams?lecture=20150107_1900

As we enter 2015, the 20th anniversary of the first successful isolation and culture of primate pluripotent stem cells in the world, it’s time to look back and see how far we’ve come. Thanks to a young reproductive biologist who came from the University of Pennsylvania’s VMD/PhD program to the Wisconsin National Primate Research Center at the University of Wisconsin-Madison in 1991, and to those whose research his groundbreaking discoveries informed, the fields of cell biology and regenerative medicine will never be the same.

stem cell colonies

Pluripotent stem cells are right now being used around the world to grow different types of cells—heart muscle cells, brain cells, pancreatic cells, liver cells, retinal cells, blood cells, bone cells, immune cells and much more.

Cultures of these cells are right now being used to test new drugs for toxicity and effectiveness.

More and more of these powerful cells are right now moving out of the lab and into preclinical (animal) trials and early human clinical trials to treat disease. The results are being published in peer-reviewed scientific journal articles on stem cell transplant, injection and infusion, reprogramming, immunology, virology and tissue engineering.

Pluripotent stem cells and their derivatives are right now being studied to learn more about reproduction and development, birth defects, and the genetic origins of disease.

Embryonic, induced pluripotent, tissue specific (adult), and other types of stem cells and genetically reprogrammed cells are all being used by researchers due to the open and collaborative environment of scientific and medical enterprises in the U.S. and around the world.

All of this is happening right now because of discoveries made 20 years ago by researchers at the Wisconsin National Primate Research Center.

Here is a brief timeline of stem cell breakthroughs by WNPRC scientists:

  • 1995-James Thomson becomes the first to successfully isolate and culture rhesus monkey embyronic stem cells (ES cells) at the Wisconsin Regional Primate Research Center (PNAS)
  • 1996-Thomson repeats this feat with common marmoset ES cells (Biol Reprod).
  • 1998-Thomson publishes the neural differentiation of rhesus ES cells (APMIS).
  • 1998-Thomson’s famous breakthrough growing human ES (hES) cells is published in Science. (This research occurred off campus, with private funding.)

Many subsequent stem cell “firsts” were accomplished by scientists who conducted lengthy training with James Thomson or Ted Golos, reproduction and development scientists at the Wisconsin National Primate Research Center. These highlights include the following accomplishments by Primate Center researchers:

  • 2003-WNPRC Post-doctoral trainee Thomas Zwaka achieves homologous recombination with hES cells. A method for recombining segments of DNA within stem cells, the technique makes it possible to manipulate any part of the human genome to study gene function and mimic human disease in the laboratory dish (Nature Biotechnology).
  • 2004-WNPRC Post-doctoral trainee Behzad Gerami-Naini develops an hES model that mimics the formation of the placenta, giving researchers a new window on early development (Endocrinology).
  • 2005- WNPRC scientist Igor Slukvin and post-doc Maxim Vodyanik become the first to culture lymphocytes and dendritic cells from human ES cells (Blood, J Immunol).
  • 2005-WiCell’s Ren-He Xu, who completed his post-doctoral research at the WNPRC, grows hES cells in the absence of mouse-derived feeder cells (Nature Methods).
  • 2006-WiCell’s Tenneille Ludwig, a graduate student/post-doc/assistant scientist through the Primate Center with Barry Bavister, then James Thomson, formulates a media that supports hES cells without the need for contaminating animal products (Nature Biotechnology). Co-authoring the work is another former Primate Center post-doc, Mark Levenstein.
  • 2007-Junying Yu, WNPRC and Genome Center, in Jamie Thomson’s lab, grows induced pluripotent stem cells, or iPS cells. (Science). These are genetically reprogrammed mature cells that act like embryonic stem cells, but without the need to destroy the embryo.

Researchers at all of the National Primate Research Centers continue to make advances in this remarkable field of research and medicine. A few more milestones include the following:

  • 2007- Shoukhrat Mitalipov at the Oregon National Primate Research Center successfully converted adult rhesus monkey skin cells to embryonic stem cells using somatic cell nuclear transfer (Nature)
  • 2012- Shoukhrat Mitalipov at the Oregon National Primate Research Center generation chimeric rhesus monkeys using embryonic cells (Cell)
  • 2012-Alice Tarantal at the California NPRC successfully transplants human embryonic stem cells differentiated toward kidney lineages into fetal rhesus macaques.
  • 2013-Qiang Shi at the Texas Biomedical Research Institute and Gerald Shatten at the University of Pittsburgh – and previously with the Oregon National Primate Research Center and Wisconsin National Primate Research Center – genetically programs baboon embryonic stem cells to restore a severely damaged artery.
  • 2013-Shoukhrat Mitalipov at the Oregon National Primate Research Center produces human embryonic stem cells through therapeutic cloning, or somatic cell nuclear transfer (Cell)

NPRC Stem Cell Timeline 01.06.15

Before all of this happened, we must note that non-primate mammalian embryonic stem cells were first successfully isolated and cultured in 1981, by Martin Evans and Matthew Kaufman at the University of Cambridge, England. That breakthrough occurred almost 35 years ago. Jamie Thomson studied mouse embryonic stem cells in Pennsylvania before working on primate cells.

Even before that, in 1961, Ernest McCulloch and James Till at the Ontario Cancer Institute in Canada discovered the first adult stem cells, also called somatic stem cells or tissue-specific stem cells, in human bone marrow. That was 55 years ago.

So first it was human stem cells, then mouse, then monkey, then back to humans again. Science speaks back and forth. It reaches into the past, makes promises in the present, and comes to fruition in the future.

In every early talk I saw Jamie Thomson give about his seminal stem cell discoveries in the late 1990s and early 2000s – to staff, scientists, to the public, to Congress, to the news media – he would explain why he came to UW-Madison in the early 1990s to try to advance embryonic stem cell research. In large part, he said, it was because we had a National Primate Research Center here at UW-Madison, and also that we had leading experts in transplant and surgery at our medical school. After he joined the WNPRC as a staff pathologist and set up his lab, first he used rhesus and then marmoset embryos before expanding to cultures using human IVF patient-donated embryos off campus with private funding from Geron Corporation in Menlo Park, California.

Human And Mouse EmbryoIn these early talks, Jamie included images (see above) showing how very differently the mouse blastocyst (a days-old embryo, before implantation stage) is structured from the nonhuman primate and human primate blastocysts concerning germ layer organization and early development (ectoderm, mesoderm and endoderm). He also was able to show for the first time how differently stem cells derived from these early embryos grow in culture. In contrast to the mouse ES cells, the monkey cells, especially those of the rhesus monkey, grow in culture almost identically to human cells.

At the time, Thomson predicted that more scientists would study human ES cells in their labs over monkey ES cells, if human ES cells could become more standardized and available. Yet he emphasized that the NPRCs and nonhuman primate models would continue to play a critical role in this research, especially when it would advance to the point when animal models would be needed for preclinical research before attempting to transplant cells and tissues grown from ES cells. Both predictions have come true.

Jamie closed his talks, and still does, with this quotation:

“In the long run, the greatest legacy for human ES cells may be not as a source of tissue for transplantation medicine, but as a basic research tool to understand the human body.”

This simply and elegantly reminds us how basic research works: Many medical advances another 20 years from now will have an important link to the discoveries of today, which have their underpinnings in that early research in Jamie Thomson’s lab 20 years ago. It will become easy to forget where it all started, when many diseases of today, if not completely cured, will become so preventable, treatable and manageable that those diagnosed with them will spend more time living their lives than thinking about how to survive another day.

Just as I did not have to worry about polio, and my children did not have to worry about chicken pox, my grandchildren will hopefully see a world where leukemia, blindness, diabetes and mental illness do not have the disabling effects or claim as many young lives as they do today.

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WN@tL “Twenty Years of Stem Cell Milestones at the UW”

http://www.uwalumni.com/event/wntl-twenty-years-of-stem-cell-milestones-at-the-uw/

January 7 – 7:00PM – 8:15PM CT
Location: UW Biotechnology Center 425 Henry Mall, Room 1111, Madison, WI 53706
Cost: Free

Speaker: William L. Murphy, Stem Cell and Regenerative Medicine Centerwnatl_williammurphy

Don’t miss this fascinating talk covering stem cell milestones at the UW. Professor Murphy will talk about the work of his team at the Stem Cell and Regenerative Medicine Center, where they are creating biological materials that could radically change how doctors treat a wide range of diseases.

Bio: Murphy is the Harvey D. Spangler Professor of Engineering and a co-director of the Stem Cell and Regenerative Medicine Center. His work includes developing biomaterials for stem cell research. Specifically, Murphy uses biomaterials to define stem cell microenvironments and develop new approaches for drug delivery and gene therapy. His lab also uses bio-inspired approaches to address a variety of regenerative medicine challenges, including stem-cell differentiation, tissue regeneration and controlled drug delivery. Murphy has published more than 100 scientific manuscripts and filed more than 20 patent applications.