June 26, 2017 In this image of neurons in the cerebellum of the brain, the yellow cells are Purkinje cells in which the channelrhodopsin-2 gene is being produced. Credit: Horwitz Lab/UW Medicine Seattle

UW Medicine researchers have developed a technique for inserting a gene into specific cell types in the adult brain in an animal model.

Recent work shows that the approach can be used to alter the function of brain circuits and change behavior. The study appears in the journal Neuron in the NeuroResources section.

Gregory Horwitz, associate professor of physiology and biophysics at the University of Washington School of Medicine in Seattle, led the research team. He said that the approach will allow scientists to better understand what roles select cell types play in the brain's complex circuitry.

Researchers hope that the approach might someday lead to developing treatments for conditions, such as epilepsy, that might be curable by activating a small group of cells.

"The brain is made up of a mix of many cell types performing different functions. One of the big challenges for neuroscience is finding ways to study the function of specific cell types selectively without affecting the function of other cell types nearby," Horwitz said. "Our study shows it is possible to selectively target a specific cell type in an adult brain using this technique and affect behavior nearly instantly."

In their study, Horowitz and his colleagues at the Washington National Primate Research Center in Seattle inserted a gene into cells in the cerebellum, a small structure located at the back of the brain and tucked under the brain's larger cerebrum.

The cerebellum's primary function is controlling motor movements. Disorders of the cerebellum generally lead to often disabling loss of coordination. Recent research suggests the cerebellum may also be important in learning and may be involved in such conditions as autism and schizophrenia.

The cells the scientists selected to study are called Purkinje cells. These cells, named after their discoverer, Czech anatomist Jan Evangelista Purkinje, are some of the largest in the human brain. They typically make connections with hundreds of other brain cells.

"The Purkinje cell is a mysterious cell," said Horwitz. "It's one of the biggest and most elaborate neurons and it processes signals from hundreds of thousands of other brain cells. We know it plays a critical role in movement and coordination. We just don't know how."

The gene they inserted, called channelrhodopsin-2, encodes for a light-sensitive protein that inserts itself into the brain cell's membrane. When exposed to light, it allows ions - tiny charged particles - to pass through the membrane. This triggers the brain cell to fire.

The technique, called optogenetics, is commonly used to study brain function in mice. But in these studies, the gene must be introduced into the embryonic mouse cell.

"This 'transgenic' approach has proved invaluable in the study of the brain," Horwitz said. "But if we are someday going to use it to treat disease, we need to find a way to introduce the gene later in life, when most neurological disorders appear."

The challenge for his research team was how to introduce channelrhodopsin-2 into a specific cell type in an adult animal. To achieve this, they used a modified virus that carried the gene for channelrhodopsin-2 along with segment of DNA called a promoter. The promoter stimulates the cell to start expressing the gene and make the channelrhodopsin-2 membrane protein. To make sure the gene was expressed only by Purkinje cells, the researchers used a promoter that is strongly active in Purkinje cells, called L7/Pcp2."

In their paper, the researchers reported that by painlessly injecting the modified virus into a small area of the cerebellum of rhesus macaque monkeys, the channelrhodopsin-2 was taken up exclusively by the targeted Purkinje cells. The researchers then showed that when they exposed the treated cells to light through a fine optical fiber, they were able stimulate the cells to fire at different rates and affect the animals' motor control.

Horwitz said that it was the fact that Purkinje cells express L7/Pcp2 promoter at a higher rate than other cells that made them more likely to produce the channelrhodopsin-2 membrane protein.

"This experiment demonstrates that you can engineer a viral vector with this specific promoter sequence and target a specific cell type," he said. "The promoter is the magic. Next, we want to use other promoters to target other cell types involved in other types of behaviors."

Explore further: New insights into control of neuronal circuitry could lead to treatments for an inherited motor disorder

More information: Yasmine El-Shamayleh et al, Selective Optogenetic Control of Purkinje Cells in Monkey Cerebellum, Neuron (2017). DOI: 10.1016/j.neuron.2017.06.002

Journal reference: Neuron

Provided by: University of Washington

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Epilepsy's cause is doubtless pheromonal.

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Study shines light on brain cells that coordinate movement - Medical Xpress

June 26, 2017 Credit: CC0 Public Domain

Cancer tumours manipulate a natural cell process to promote their survival suggesting that controlling this mechanism could stop progress of the disease, according to new research led by the University of Oxford.

Non-sense mediated decay (NMD) is a natural physiological process that provides cells with the ability to detect DNA errors called nonsense mutations. It also enables these cells to eliminate the mutated message (decay) that comes from these faulty genes, before they can be translated into proteins that can cause disease formation. NMD is known among the medical community for the role it plays in the development of genetic diseases such as Cystic Fibrosis and some hereditary forms of cancers. But not all nonsense mutations can elicit NMD, so until now, it's wider impact on cancer was largely unknown.

Biomedical researchers and computer scientists from the University of Oxford Medical Sciences Division and the University of Birmingham developed a computer algorithm to mine DNA sequences from cancer to accurately predict whether or not an NMD would eliminate genes that had nonsense mutations. The work originally focused on ovarian cancers, and found that about a fifth of these cancers use NMD, to become stronger. This is because NMD ensures that the message from a gene called TP53, which ordinarily protects cells from developing cancer is almost completely eliminated. In the absence of NMD, a mutated TP53 might still retain some activity but NMD ensures that this is not the case.

Based on this research, the team predicts that because cancers essentially feed on NMD, they become dependent on it in some cases. If scientists were therefore able to inhibit or control the process, it is possible that they could also control cancer and prevent the progression of the disease.

Dr Ahmed Ahmed, Co-author and Professor of Gynaecology Oncology at the Nuffield Department of Obstetrics & Gynaecology and the head of the Ovarian Cancer Cell Laboratory, at the Weatherall Institute of Molecular Medicine at the University of Oxford, said: "Our first observations of evidence of the role of NMD in ovarian cancer were tantalizing. We found that NMD precisely explained why there was almost no expression of TP53 in certain ovarian cancers. We went on to test the role of NMD in other cancer types and the evidence of the role of NMD was compelling. This opens the door for exciting possibilities for customised treatments including individualized immunotherapies for patients in the future."

Following the ovarian cancer analysis, the team expanded the study to include other cancer types. They analysed about a million different cell mutations in more than 7,000 tumours from the Cancer Genome Atlas covering 24 types of cancer. The team was able to map how each cancer type used NMD revealing the remarkable extent to which NMD helps cancer to survive.

Katherine Taylor, CEO of Ovarian Cancer Action, who part-funded the research, said: "This is very exciting news. Professor Ahmed and his team have identified how cancer cells rely on a process called NMD for their survival. This discovery could help clinicians identify and inhibit the process, giving them much better control of a person's cancer.

"Ovarian cancer is a very complicated disease and survival rates are low, with only 46% of women living beyond five years after diagnosis. So understanding how we can prevent the disease from thriving is imperative if we are to improve the outcome for more women.

"It's fantastic to see how our funding is helping make real progress and we couldn't do this without the generosity of our supporters. We look forward to seeing where Professor Ahmed takes his research next."

Moving forward the team will focus on testing their theory and understanding to what degree stopping the NMD process allows them to control tumours.

Co-author, Dr Christopher Yau, a computational scientist at the Institute of Cancer and Genomic Sciences, University of Birmingham said: "As a result of these findings, we now plan to apply the same computer algorithm to determine if NMD affects cancer patients in The 100,000 Genomes Project. These investigations may pave the way to new treatment possibilities for NHS patients in the future."

Explore further: Two Oxford research discoveries offer hope for managing ovarian cancer

More information: The full paper citation is 'A pan-cancer genome-wide analysis reveals tumour dependencies by induction of nonsense-mediated decay,' and it will be published in Nature Communications on Monday 26 June 2017.

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Cancer hijacks natural cell process to survive - Medical Xpress - Medical Xpress

Dr. Purita has been a pioneer in the Regenerative Medicine space for over a decade.

Boca Raton, Florida (PRWEB) June 27, 2017

Robert J. Colucci CEO of PuRxCell, LLC announced today that PuRxCell, a Florida based company, has been established to train physicians at all levels of experience and their staffs in Dr. Puritas unique Stem Cell treatment and processing protocols. PuRxCell also provides a comprehensive line of products to support these cutting edge protocols.

Dr. Purita has been a pioneer in the Regenerative Medicine space for over a decade, said CEO R.J. Colucci. He continued, He has performed more than 8500 treatment procedures without serious adverse effect. These unique protocols not only improve treatment outcomes but also significantly reduce the cost of the treatment and increase margins, which is important in the world of increasing competition and reduced insurance reimbursements. Colucci said, Our mission at Purxcell is simple: Deliver individualized training to physicians - offer cutting edge treatment and processing protocols and products - lower the cost of treatment and provide physicians and their staffs a continuum of training and support."

In my 35 years as an Orthopedic Surgeon, I have never been as excited as I am over the advancements in Regenerative Medicine. Over the last 10 years I have focused my efforts on developing and refining cutting edge PRP and Stem Cell procedures and adjunct therapies, said Joseph Purita, PuRxCells Chief Medical Officer.

Purita continued, I have been hesitant to pass on specific details about my experiences and unique protocols. However, I have come to the conclusion that I want some of my colleagues and ultimately their patients to benefit from what I have learned. As such, PuRxCell is offering direct physician training as well as access to my proprietary treatment protocols and products. Purita noted, I was also unhappy with the current costs of disposable products for Stem Cell treatment protocols. I have expended considerable effort to finding ways to lower procedure and disposable costs while at the same time improving the quality of patient outcomes. All this eventually led me to form a dedicated, full service Regenerative Medicine company, PuRxCell. We have developed blood, adipose and bone marrow processing products designed to help customers dramatically reduce cost without sacrificing treatment outcomes.

Founded in 2016, Purxcell has multiple level of training programs tailored to individual physicians needs and experience levels. Physician training ranges from 2-5 days, while training for lab staff typically ranges from 1-2 days. PuRxCell also offers Regenerative Medicine practice and marketing support in line with the mission of providing full-service Regenerative Medicine products and services.

PuRxCell has its corporate headquarters office in Boca Raton, Florida, with satellite offices in Colorado and Coconut Creek, FL. More information about PuRxCell can be found at http://www.purxcell.com.

For additional information contact: Robert Colucci at r.colucci(at)purxcell(dot)com or at 877-498-5500 ext. 1

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Regenerative Medicine Pioneer Offers Comprehensive Stem Cell Training - PR Web (press release)

June 27, 2017 by Heather Lindsay Irradiated cancer cell with a small amount of DNA (green) in the cytoplasm. Credit: Dr. Claire Vanpouille-Box/Weill Cornell Medicine

An enzyme implicated in autoimmune diseases and viral infections also regulates radiation therapy's ability to trigger an immune response against cancer, Weill Cornell Medicine scientists found in a new study. Their discovery can help to better tailor treatment for patients.

Immunotherapy is an innovative approach to cancer treatment that unleashes the power of the immune system to fight the disease. The method has revolutionized treatment for several cancers. However, only a minority of patients responds to the treatment. Radiation therapy may boost patients' responses to immunotherapy, but the best way to achieve this effect has remained unclear.

In a study published June 9 in Nature Communications, investigators discovered that radiation in mice elicits the accumulation of DNA in a cellular compartment where it mimics the presence of a virus, generating molecular signals that are normally triggered by infection. The most important signal, known as interferon-beta, is required to activate immune cells that can kill virally infected cells. Thus, radiation therapy tricks the immune system to see the cancer cells as if they were infected by a virus, and by doing so, it activates the immune system against the tumor.

However, not all ways used to irradiate the tumor achieve this effect, due to the induction of an enzyme that clears the DNA, called TREX1.

"We found that the induction of interferon-beta by radiotherapy is under the control of TREX1," said senior study author Dr. Sandra Demaria, professor of radiation oncology and of pathology and laboratory medicine at Weill Cornell Medicine and member of the Sandra and Edward Meyer Cancer Center at Weill Cornell Medicine.

"When the radiation dose used is increased above a certain threshold, TREX1 levels increase in the cancer cells, blocking the production of interferon. Intriguingly, this mechanism of escape from immune detection by cancer cells irradiated with certain doses mimics one of the ways HIV evades the immune system."

For their study, the researchers treated mice with breast or colorectal cancer with different doses of radiation therapy, and also studied these treatments in human lung and breast cancer cells. They then evaluated mouse tumors and human cells in the laboratory to determine what cellular changes occurred. Importantly, they demonstrated in the mice that blocking the induction of TREX1 restored radiation's ability to induce effective anti-tumor immune responses when used with immunotherapy.

"The findings that TREX1 is a regulator of radiation therapy ability to activate the immune system and that its induction depends on the radiation dose used is potentially practice-changing, if these results are verified in patients," said study co-author Dr. Silvia Formenti, chairman of the Department of Radiation Oncology, the Sandra and Edward Meyer Professor of Cancer Research and associate director of the Sandra and Edward Meyer Cancer Center at Weill Cornell Medicine, and radiation oncologist-in-chief at NewYork-Presbyterian/Weill Cornell Medical Center.

The team is now studying whether the doses of radiation that induce TREX1 in carcinomas are similar in other types of cancers.

"This is important to help guide the choice of radiation to be used in clinical trials that test combinations of radiotherapy with immunotherapy in different cancers," Demaria said.

Formenti is leading several studies testing these combinations in patients with lung and other cancers.

As research and treatment progresses, "we need to be mindful when we use radiation to elicit an immune response," Formenti said. "It's important to use the dose per fraction that is likely to work the best to elicit an immune response, and we have now a way to determine what it is by measuring the levels of interferon-beta and TREX1 in each individual patient."

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Enzyme key to triggering anti-cancer immune response - Medical Xpress

The single-cell analysis market is expected to reach USD 3.59 billion by 2022 from USD 1.67 billion in 2017, at a CAGR of 16.5%.

Technological advancements in single-cell analysis products, increasing government funding for cell-based research, growing biotechnology and biopharmaceutical industries, wide applications of single-cell analysis in cancer research, growing focus on personalized medicine, and increasing incidence and prevalence of chronic and infectious diseases are driving the growth of the single-cell analysis market.

On the other hand, the high cost of single-cell analysis instruments may hinder the growth of the market in the coming years.

Consumables segment are expected to witness high growth during forecast period

Based on product, the single-cell analysis market is segmented into consumables and instruments.

Consumables are expected to register the highest CAGR during the forecast period.

Consumables are further segmented into beads, microplates, reagents, assay kits, and other consumables, whereas, instruments are further segmented into flow cytometers, NGS systems, PCR instruments, HCS systems, microscopes, cell counters, spectrophotometers, cell microarrays, and other instruments.

Players from the market are increasingly focusing on technological advancements and product development to launch efficient cytometry instruments which is driving the growth of the segment.

Human cells to dominate the single-cell analysis market

Based on the type of cells, the single-cell analysis market is segmented into human, animal, and microbial cells.

The human cells segment is expected to dominate this market with the largest share in 2017.

The large share of this segment can be attributed to the growing application areas of human stem cells and rising incidence of diseases such as cancer.

"Single-cell analysis market projected to register a CAGR of 16.5%" The single-cell analysis market is expected to reach USD 3.59 billion by 2022 from USD 1.67 billion in 2017, at a CAGR of 16.5%. Technological advancements in single-cell analysis products, increasing government funding for cell-based research, growing biotechnology and biopharmaceutical industries, wide applications of single-cell analysis in cancer research, growing focus on personalized medicine, and increasing incidence and prevalence of chronic and infectious diseases are driving the growth of the single-cell analysis market. On the other hand, the high cost of single-cell analysis instruments may hinder the growth of the market in the coming years.

"Consumables segment are expected to witness high growth during forecast period" Based on product, the single-cell analysis market is segmented into consumables and instruments. Consumables are expected to register the highest CAGR during the forecast period. Consumables are further segmented into beads, microplates, reagents, assay kits, and other consumables, whereas, instruments are further segmented into flow cytometers, NGS systems, PCR instruments, HCS systems, microscopes, cell counters, spectrophotometers, cell microarrays, and other instruments. Players from the market are increasingly focusing on technological advancements and product development to launch efficient cytometry instruments which is driving the growth of the segment.

"Human cells to dominate the single-cell analysis market" Based on the type of cells, the single-cell analysis market is segmented into human, animal, and microbial cells. The human cells segment is expected to dominate this market with the largest share in 2017. The large share of this segment can be attributed to the growing application areas of human stem cells and rising incidence of diseases such as cancer.

"NGS to account with the highest growth rate in the single-cell analysis market" Based on technique, the single-cell analysis market is segmented into flow cytometry, NGS, PCR, microscopy, mass spectrometry, and other techniques (including single-molecule fluorescence in situ hybridization, micromanipulation, and automated capillary electrophoresis). NGS segment is projected to grow at the highest rate during the forecast period.

"North America to dominate the market during forecast period" In 2017, North America is expected to account for the largest share of the single-cell analysis market. Factors such as increasing collaborations among prominent players, technological advancements and expanding biotechnology and pharmaceutical industries are supporting the growth of the single-cell analysis market in this region. Asian region is expected to register the highest CAGR during the forecast period. The large population in China and India, rising geriatric population, and increasing incidence of chronic diseases are the major factors driving the growth of the Asian market.

The primary interviews conducted for this report can be categorized as follows:

By Company Type: Tier 1 35%; Tier 2, 40%; Tier 3, 25%. By Designation: C- level- 35%; D-level- 25%; others- 40%. By Region: North America 43%; Europe- 19%; Asia- 29%; and Rest of the World (RoW) - 9%

*Note: Others include sales managers, marketing managers, and product managers Tiers of the companies are defined by their total revenue. As of 2016: Tier 1 = > USD 1 billion, Tier 2 = USD 100 million to USD 1 billion, and Tier 3 = < USD 100 million

List of Companies Benchmarked in the report

Merck KGaA (Germany) Becton, Dickinson and Company (U.S.) Promega Corporation (U.S.) Danaher Corporation (U.S.) General Electric Company (U.K.) Thermo Fisher Scientific Inc. (U.S.) Miltenyi Biotec (Germany) Illumina, Inc. (U.S.) Bio-Rad Laboratories, Inc.(U.S.) Fluidigm Corporation (U.S.) NanoString Technologies, Inc. (U.S.) Agilent Technologies (U.S.) Abcam Plc (U.S.) NuGEN Technologies Inc.(U.S.) LumaCyte (U.S.) PluriSelect Life Science UG & Co. KG (Germany) Sysmex Partec (U.S.) Bio-Techne Corporation (U.S.) Promega Corporation (U.S.) 10x Genomics (U.S.) WaferGen Bio-systems, Inc. (U.S.) Bruker (U.S.) Fluxion Biosciences (U.S.).

Research Coverage: The report provides a picture on global single-cell analysis across various medical devices. It aims at estimating the market size and future growth potential of this market across different segments such as products, cell type, technique, application, end user, and regions. Furthermore, the report also includes an in-depth competitive analysis of the key players in the market along with their company profiles, recent developments, and key market strategies.

Key Benefits of Buying the Report: The report will help the market leaders/new entrants in this market by providing them the closest approximations of the revenue numbers for the overall single-cell analysis market and the subsegments. This report will help stakeholders to better understand the competitor landscape and gain more insights to better position their businesses and make suitable go-to-market strategies. The report also helps the stakeholders to understand the pulse of the market and provides them information on key market drivers, restraints, challenges, and opportunities. Read the full report: http://www.reportlinker.com/p04579530/Single-Cell-Analysis-Market-by-Product-Cell-Type-Technique-Application-End-User-Global-Forecasts-to.html

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Single-cell Analysis Market is expected to reach USD 3.59 billion by 2022 - PR Newswire (press release)