I HAVE PARKINSON'S DISEASES AND THOUGHT IT WOULD BE NICE TO HAVE A PLACE WHERE THE CONTENTS OF UPDATED NEWS IS FOUND IN ONE PLACE. THAT IS WHY I BEGAN THIS BLOG.
I COPY NEWS ARTICLES PERTAINING TO RESEARCH, NEWS AND INFORMATION FOR PARKINSON'S DISEASE, DEMENTIA, THE BRAIN, DEPRESSION AND PARKINSON'S WITH DYSTONIA. I ALSO POST ABOUT FUNDRAISING FOR PARKINSON'S DISEASE AND EVENTS. I TRY TO BE UP-TO-DATE AS POSSIBLE.
I AM NOT RESPONSIBLE FOR IT'S CONTENTS. I AM JUST A COPIER OF INFORMATION SEARCHED ON THE COMPUTER. PLEASE UNDERSTAND THE COPIES ARE JUST THAT, COPIES AND AT TIMES, I AM UNABLE TO ENLARGE THE WORDING OR KEEP IT UNIFORMED AS I WISH.IT IS IMPORTANT TO UNDERSTAND I AM A PERSON WITH PARKINSON'S DISEASE. I HAVE NO MEDICAL EDUCATION,
I JUST WANT TO SHARE WITH YOU WHAT I READ ON THE INTERNET. IT IS UP TO YOU TO DECIDE WHETHER TO READ IT AND TALK IT OVER WITH YOUR DOCTOR. I AM JUST THE COPIER OF DOCUMENTS FROM THE COMPUTER. I DO NOT HAVE PROOF OF FACT OR FICTION OF THE ARTICLE. I ALSO TRY TO PLACE A LINK AT THE BOTTOM OF EACH ARTICLE TO SHOW WHERE I RECEIVED THE INFORMATION SO THAT YOU MAY WANT TO VISIT THEIR SITE.
THIS IS FOR YOU TO READ AND TO ALWAYS KEEP AN OPEN MIND.
PLEASE DISCUSS THIS WITH YOUR DOCTOR, SHOULD YOU HAVE ANY QUESTIONS, OR CONCERNS. NEVER DO ANYTHING WITHOUT TALKING TO YOUR DOCTOR FIRST..
I DO NOT MAKE ANY MONEY FROM THIS WEBSITE. I VOLUNTEER MY TIME TO HELP ALL OF US TO BE INFORMED.
I WILL NOT ACCEPT ANY ADVERTISEMENT OR HEALING POWERS, HEALING FROM HERBS AND ETC. UNLESS IT HAS GONE THROUGH TRIALS AND APPROVED BY FDA. IT WILL GO INTO SPAM.
THIS IS A FREE SITE FOR ALL WITH NO ADVERTISEMENTS
THANK YOU FOR VISITING! TOGETHER WE CAN MAKE A DIFFERENCE!
Targeting specific calcium channels in dopamine-producing neurons may be a therapeutic target for Parkinson’s disease, according to research involving cells derived from Parkinson’s patients.
For most Parkinson’s patients, symptoms are idiopathic, or of unknown cause, making it difficult to understand the relationship between various factors that may cause disease. However, roughly 10% of patients show the familial incidence of Parkinson’s disease, which allows researchers to study the underlying mechanisms of disease in these patients, helping them understand common causes in all patients, as well as unravel potential therapeutic targets.
Prior generation of induced pluripotent stem cells (iPSCs) from patients with familial Parkinson’s successfully replicated the disease processes. iPSCs are derived from either skin or blood cells that have been reprogrammed back into a stem cell-like state, which allows for the development of an unlimited source of any type of human cell needed for therapeutic purposes.
Aiming to provide iPSCs-based ways to treat neurological diseases, Keio University School of Medicine and Eisai started a collaboration in 2013. Their joint research led to the generation of dopamine-producing neurons — progressively degenerated in Parkinson’s, leading to its motor symptoms — from familial Parkinson’s patients’-derived iPSC.
They also established a drug library of more than 1,000 existing compounds to screen treatment candidates.
The study, partially funded by Eisai, used neural progenitor cells, which differentiate into brain cells, including neurons, derived from two familial Parkinson’s disease patients with mutations in the PRKN gene (PARK2 type). PRKN is the most commonly implicated gene in young-onset Parkinson’s disease.
“The current procedure is more advanced in terms of simplicity and robustness for neuronal differentiation, enabling us to perform screening without complicated work,” researchers noted.
The approach enabled the efficient generation of dopamine-producing neurons in vitro, which, compared to healthy neurons, had reduced size of projections, as well as increased oxidative stress and apoptosis, or “programmed” cell death, as opposed to cell death caused by injury.
Oxidative stress is an imbalance between the production of free radicals and the ability of cells to detoxify them. These free radicals, or reactive oxygen species, are harmful to cells and are associated with a number of diseases, including Parkinson’s.
Gene editing in iPSCs to create similar mutations led to the same disease-related alterations.
Researchers also found that Parkinson’s-derived neurons were more susceptible to mitochondrial stress induced by rotenone, an agrochemical that acts as a mitochondrial inhibitor. Mitochondria are small cellular organelles that provide energy and are known as cells’ “powerhouses”.
Screening their library enabled the identification of several compounds able to suppress rotenone-induced apoptosis, particularly T-type calcium channel blockers benidipine (used for the treatment of blood pressure) and ML218.
These particular calcium channels are present in many neuronal cells within the central nervous system. They help mediate calcium influx into neurons after passing electrical impulses to communicate among themselves.
These blockers also were able to lower stress-induced apoptosis in dopamine-producing neurons derived from patients with a different type of familial Parkinson’s (PARK6), caused by a mutation in the PINK1 gene. These neurons also had higher levels of T-type calcium channels.
“These findings suggest that calcium homeostasis in [dopamine-producing] neurons might be a useful target for developing new drugs for [Parkinson’s] patients,” researchers wrote.
“In summary, we have established a robust platform to model [Parkinson’s] in a dish and revealed an additional layer of the pathogenesis of PD, offering a potential therapeutic target,” they added.
Future work will use experimental approaches to model the brain in vivo and to have different types of brain cells in the same dish with the goal of validating these therapeutic targets.
Of note, six of the study’s authors are employees at Eisai.
Although no marked differences in the asymmetry of facial expressions distinguished Parkinson’s disease patients and healthy individuals in a study, patients most clearly expressed an emotional reaction on the side of the face corresponding to the side of the body less affected by Parkinson’s motor symptoms, a study found.
Its researchers concluded patients’ facial asymmetry when displaying emotions is a consequence of general motor symptom asymmetry, rather than difficulties or problems in processing emotions.
Parkinson’s is characterized by the gradual loss of muscle control and lack of facial expression, sometimes accompanied by cognitive deficits.
Previous studies have suggested that hemispheric dominance — a phenomenon in which one side of the brain is more important than the other for a given function — in emotional processing can lead to an asymmetric facial expression.
Although Parkinson’s motor symptoms tend to be rather asymmetric (i.e., more pronounced on one side of the body), studies had not addressed facial and emotional asymmetry in Parkinson’s patients.
A research team in London and Italy explored the relationship between motor symptom asymmetry and facial expressiveness in Parkinson’s disease.
The study enrolled 20 patients and 20 healthy people serving as controls, who were video-recorded while displaying facial expressions: one that was neutral, and six basic emotions (anger, disgust, fear, happiness, sadness, and surprise).
The most expressive pictures obtained from the video-recordings were then cut down the middle and put through a program to generate ‘chimeric’ faces that showed only the right side of the face (right side manipulated to make full face) and the left side (a left-left combination). Investigators then asked nine healthy people with no prior connection to the study to rate which one of the two chimeric faces looked more expressive. Raters’ choices, reaction times, and confidence levels were recorded.
To evaluate a possible link between facial expressiveness and motor symptom asymmetry, researchers performed correlation analysis between the global facial laterality index (pooling all emotions together) as well as for the indexes of each emotion separately, and the body laterality index (defined by the side of the body most affected by Parkinson’s symptoms).
No substantial differences were found in how the nine raters judged emotional expressiveness on the two chimeric faces (right-right and left-left), whether within the Parkinson’s and the control group, or between the two groups.
In Parkinson’s patients, however, investigators found a correlation between the global facial laterality index and the body laterality index, suggesting that each patient’s most expressive side of the face corresponded to the body side less affected by Parkinson’s symptom’s.
“Despite the lack of significant facial asymmetry in PD [Parkinson’s disease] and healthy subjects, the relationship we found between the intensity of facial expression and motor symptom lateralization supports the hypothesis that there is some facial asymmetry of emotional expression in PD which relates to the general lateralisation of the motor features of the disorder rather than a specific abnormality in emotional processing,” the researchers wrote.
October 25, 2018 by Serena Gordon, Healthday Reporter
(HealthDay)—A new gene therapy might help improve motor symptoms in people with Parkinson's disease who aren't responding to other therapies, an early study has found.
"This is not a cure of Parkinson's disease," said James Beck, chief scientific officer of the Parkinson's Foundation. "This is a potentially good treatment for symptom control. It provides an additional way of providing dopamine to the brain, but it doesn't stop the progression of Parkinson's disease."
The new treatment uses a virus to deliver gene therapy to a targeted area of the brain. The gene therapy affects an enzyme called AADC. This enzyme transforms levodopa into dopamine in the brain.
Cells that make the neurotransmitter dopamine—a chemical messenger in the brain—die off in Parkinson's disease, according to the U.S. National Institute on Aging. A loss of dopamine causes the symptoms of Parkinson's disease, such as tremor and slow movements.
Standard treatments attempt to replace the lost dopamine. For example, one current medication is levodopa, but the cells that transform levodopa into dopamine have to be functioning for this treatment to work. As the dopamine-producing cells die off, it becomes harder and harder for the brain to respond to medications like this, Beck explained.
And, that's where the new gene treatment may help.
Led by Dr. Chadwick Christine, at the University of California, San Francisco, researchers used MRI scanning to locate the right area of the brain. Then they infused the new gene therapy into a targeted area of the brain called the putamen. The study team chose this area because these brain cells aren't destroyed by Parkinson's disease.
The phase 1 trial included 15 people who were no longer responding to other Parkinson's treatments. They all received one infusion of the gene therapy—known as VY-AADC.
After the treatment, researchers followed the patients' health for up to 36 months and found that the treatment was well-tolerated. The most serious side effects—a blood clot and an irregular heart rhythm caused by the blood clot—were related to the surgery used to deliver the treatment, and not the treatment itself, the researchers said.
The therapy also showed a meaningful improvement in the time people spent without movement symptoms each day. And the effect appeared to be lasting, with some of the patients followed for as long as three years.
Dr. Alessandro Di Rocco, director of the Movement Disorders Program at Northwell Health in Great Neck, N.Y., reviewed the findings.
"This study seems to be a step forward in perfecting the [gene therapy] vector and they were able to give it safely," he said. Di Rocco added that the decrease in symptoms "is a real effect."
But, like Beck, he cautioned that this is an early study with only a small number of people in the trial. Di Rocco also noted that the trial wasn't "blinded," so it's possible there was a placebo effect for some.
Both Di Rocco and Beck also expressed concern about the possible cost of such therapy.
The University of California, San Francisco, and University of Pittsburgh Medical Center researchers aren't the only team working on gene therapy for Parkinson's symptoms. A group of British researchers is actively recruiting Parkinson's patients to take part in a gene therapy trial that's also aimed at increasing the availability of dopamine in the brain.
Their trial will look at up to 30 patients receiving treatment in London or Paris. For this study, researchers are targeting a part of the brain called the striatum.
Results of the American study were presented Sunday at the American Neurological Association meeting, in Atlanta. Findings presented at meetings should be viewed as preliminary until they've been published in a peer-reviewed journal.
More information: James Beck, Ph.D., chief scientific officer, Parkinson's Foundation; Alessandro Di Rocco, M.D., director, Movement Disorders Program, Northwell Health, Great Neck, N.Y., and professor of neurology, Zucker School of Medicine at Hofstra University; Oct. 21, 2018, presentation, American Neurological Association meeting, Atlanta
A surprising offender has been emerging to drive the progression of Parkinson's, Alzheimer's, Huntington's and other neurodegenerative diseases: calcium. Calcium controls the production of fuel in mitochondria, the cell's powerhouses. But too much calcium can lead to cellular damage and even cell death. These events can cascade into neurodegenerative diseases and causes injury to the brain and heart during strokes and heart attacks.
Now, researchers at Jefferson (Philadelphia University + Thomas Jefferson University) have identified a molecular lock and key that control calcium's entry into mitochondria, and show how the key competes with a potent calcium-blocking compound. The finding reveals a new target for drug discovery.
"Our study gets directly to the pharmacologic and potential medical targeting of mitochondrial calcium uptake," said senior author Gyorgy Hajnoczky, MD, Ph.D., Director of the MitoCare Center, and Professor in the department of Pathology, Anatomy and Cell Biology at the Sidney Kimmel Medical College at Jefferson.
The work, another successful MitoCare Center collaboration, published October 25 in the journal Molecular Cell, with first author Melanie Paillard, Ph.D., a postdoctoral fellow in Hajnoczky's lab, and Gyorgy Csordas, MD and Suresh K Joseph, Ph.D., both professors at the MitoCare Center.
To control calcium's entry, mitochondria have specialized doors called mitochondrial calcium uniporter complexes. The doors have many parts including the pore through which calcium enters, called MCU, and a sort of gatekeeper protein that detects when calcium is at the door, called MICU1. When calcium levels are low, MICU1 keeps the doors closed. But when a surge of calcium arrives—as it does with each heartbeat, for example—MICU1 opens the door to the mitochondria. Dr. Hajnoczky and his colleagues wanted to understand how the MICU1 gatekeeper keeps the MCU pore closed.
Because calcium overload in mitochondria is implicated in multiple diseases, such as strokes and heart attacks and MCU controls calcium's entry, researchers are on the look out for compounds that block MCU's opening. The most effective compounds used in laboratories are ruthenium red and ruthenium 360. The drugs lock onto a section of the MCU pore to prevent calcium entry.
In a surprise finding, Dr. Hajnoczky's team discovered that the MCU pore has a deadbolt that can lock/open the door when a small part of the gatekeeper MICU1 interacts with it as a key. Ruthenium red/360 works as an alternative key for the same deadbolt to lock the door; thus competing with the gatekeeper MICU1. Ruthenium red/360 was able to block calcium much more effectively in liver cellsthat lacked the MICU1 gatekeeper protein, than in cells whose MICU1 was present.
Human heart tissue contains far less MICU1 than other tissue, suggesting that compounds like ruthenium red/360 might be more selectively in blocking calcium there. "We found that the dose response for the drugs totally depends on the presence of MICU1," said Dr. Hajnoczky. The researchers then confirmed the results in two other kinds of cells. The discovery suggested MICU1 attaches to the same location on MCU as the drugs.
Because ruthenium compounds attached to an area of the MCU known as DIME, the researchers looked for a similar area of attachment on the MICU1. They found a section of MICU1 that fits with MCU like a key to a lock. Mutating this area of MICU1, which the researchers named DIME Interacting Domain, or DID, reduced its ability to connect with MCU and prevented MICU1 from regulating calcium entry into mitochondria. Cells with mutant MICU1 were unable to manage the calcium levels in mitochondria, which led to oxidative stress and cellular damage.
The results indicate MICU1 attaches directly to the MCU and mitochondria need the DID region of MICU1 to control calcium levels and thus cell survival.
The team's results open a possible new avenue for therapies that control mitochondrial calcium levels. "Targeting the interaction of MICU1 with MCU is highly relevant for drug designing efforts," said Dr. Hajnoczky. "For diseases where mitochondria malfunction and cause cell death, a drug that blocks calcium entry might improve the patient outcomes."
More information: Melanie Paillard, Gyorgy Csordas, Kai-Ting Huang, Peter Varnai, Suresh K. Joseph and Gyorgy Hajnoczky, "MICU1 interacts with the D-ring of the MCU pore to control its Ca2+ flux and sensitivity to Ru360," Molecular Cell, DOI: 10.1016/j.molcel.2018.09.008 , https://www.cell.com/molecular-cell/fulltext/S1097-2765(18)30755-X
October 25, 2018 by Ariel Bleicher, University of California, San Francisco
Bruce Conklin (right), MD, is testing CRISPR-based surgeries on donated cells from Delaney Van Riper (left). These surgeries aim to cure patients like her who have rare genetic disorders. Credit: Steve Babuljak
One afternoon in July, deep within the labyrinthine halls of the Medical Sciences Building at UC San Francisco's hilltop campus on Parnassus Avenue, the laboratory of Alex Marson, MD, Ph.D., is buzzing. Doors clap. Gloves snap. Keyboards clack. Cells incubate in nutrient baths the color of Kool-Aid while machines resembling rice cookers spin mixtures of molecules, separating large from small. Every now and then, a printer whirs with notes for a new experiment, like a lunch order arriving in a restaurant kitchen.
Theo Roth, an MD-Ph.D. student, opens a deep freezer, releasing an icy cloud. Here, amid frosted boxes stacked on frosted shelves, is the impetus for all this activity – the reason Roth and Marson and their colleagues at UCSF and elsewhere have begun to suspect, with no small amount of excitement, that they are in the vanguard of a new era in medicine.
Roth pulls out a box and lifts from it a transparent plastic vial no taller than a toothpaste cap. Inside, he explains, are billions of intricately folded, ribbon-like molecules: proteins known as Cas9. When linked to other molecules called guide RNAs, the Cas9 proteins transform into …"… the magic CRISPR system," Roth says, holding the vial up to the light.
Its contents look like … well, nothing. "Just another clear liquid," Roth jests – because as he well knows, these molecules' humble appearance belies a singular and extraordinary power.
The Coming CRISPR Cures
If you've heard of CRISPR (pronounced "crisper"), a hot topic in science circles nowadays, you've likely encountered a dizzying array of definitions and divinations. Is CRISPR a therapy? A revolution? A pair of genetic scissors? A text editor? A genesis engine? A gateway to designer babies? And what does that catchy acronym – which stands for "clustered regularly interspaced short palindromic repeats" – even mean?
Put simply, CRISPR is a tool. In fact, it is many tools – more precisely described as CRISPR systems – exquisitely engineered for operating on life's tiniest anatomy: DNA, the substance of genes. These tools aren't the first of their kind, but they are by far the most exacting, the cheapest, and the easiest to use. Dispatched into living cells, they can be made to manipulate any gene in any tissue in any organism, whether microbe, mouse, or monkey.
Or human. Just six years after the discovery of CRISPR technology, hundreds of research labs around the world are now using it to study patients' cells and to create animal models of human diseases – from common illnesses to inherited disorders so rare that they may affect only a few families. This fast-growing body of research has proven a boon to medical science, showing how DNA – a spiraling chain of chemical bases strung together like rungs on a ladder – keeps us alive and healthy, and how even subtle changes in this code can make us sick.
But for physician-scientists like Marson and young trainees like Roth, the ascendancy of CRISPR systems raises an even grander hope: If these tools can illuminate the causes of disease in the laboratory, why not bring them into the clinic to treat patients?
What CRISPR scientists envision – the future they are now preparing for – is a whole new field of medicine. They even have a name for this nascent specialty: genome surgery. Just as today's surgeons use steel instruments to excise tumors, repair blood vessels, or transplant kidneys, tomorrow's genome surgeons could use CRISPR systems to remove one faulty gene, correct another, or replace a third – thereby curing genetic diseases at their source.
"Imagine a world where people go to the doctor, and they get their genome sequenced and learn they have a genetic disorder," says Jennifer Doudna, Ph.D., who famously pioneered the first CRISPR applications in 2012. "And instead of telling them they need to live with that disorder, we have the technology that can actually treat them – potentially even cure them."
It's a lofty vision but one that may be within reach. In 2014, Doudna – who is a professor of chemistry and of molecular and cell biology at UC Berkeley and a senior investigator at the Gladstone Institutes, an independent research institute affiliated with UCSF – helped found the Innovative Genomics Institute (IGI), a partnership between UCSF and UC Berkeley to apply CRISPR systems to improving human health. (Its mission later expanded to also encompass food security and environmental sustainability.) Since then, teams of researchers and clinicians in the Bay Area alone – many of them funded through the IGI – have begun to work toward new CRISPR surgeries that could treat an array of diseases, including genetic disorders of the eye, nerves, kidneys, blood, and immune system.
"The possibilities are mind-boggling," says Marson, an associate professor of microbiology and immunology and the scientific director of biomedicine at the IGI. He, together with Roth and others, is developing CRISPR-based techniques aimed at reprogramming patients' own immune cells to kill cancer and to ward off HIV. The day when treatments like these enter the clinic may not be so far off.
"Genome surgery is not science fiction anymore," Doudna says. "It's really coming down the pike."
New Hope for Rare Disorders
While Marson's team busies itself remodeling immune cells, a few miles away at the Gladstone Institutes, on UCSF's Mission Bay campus, a different sort of genome surgery is underway. There, in the laboratory of senior investigator Bruce Conklin, MD – a UCSF professor of medicine and IGI's deputy director – the cells of 19-year-old Delaney Van Riper are undergoing experimental procedures that could one day cure her of a worsening disability.
Van Riper was born with a rare disease called Charcot-Marie-Tooth (CMT), one of more than 6,000 known genetic disorders, which arise from specific variations in DNA. Such variations – called mutations – throw a wrench in a cell's protein production line, thus creating deviant or defunct molecules, like Ikea furniture assembled from garbled instructions. In some cases, a mutation in just one DNA base – out of the total 3 billion pairs of bases in the human genome – can wreak severe havoc.
Van Riper's mutation produces a miscreant protein that degrades her nerve cells' ability to relay messages between her brain and her muscles, causing her to slowly lose control of her limbs. She was diagnosed at age 7, after her father, a genetic counselor, noticed that she wasn't walking normally. By age 8, she wore leg braces, laughing along with the kids who called her Forrest Gump, "so they didn't see me as a cripple." By age 13, she struggled to hold a pencil.
"There are certain muscles I just don't have anymore," she says during a recent visit to the lab. She is seated at a conference table, where a dozen or so researchers from Conklin's group have gathered to meet her, many of them for the first time.
The researchers know her cells intimately, however. They have isolated them from samples of her blood and nurtured them in petri dishes. They have doused these blood cells with a cocktail of genes that turns them into stem cells, undifferentiated cells that can grow indefinitely.
Using another gene cocktail, they have coaxed the stem cells to become nerve cells like those at the root of Van Riper's disease. They have examined these diseased nerve cells through microscopes, studied their troublesome mutation, and sent in CRISPR systems to try to remove it.
All the while, Conklin and his team have dreamed about a day when a physician might inject CRISPR molecules directly into Van Riper's spine to heal the nerve cellsthere; a day when the success of this pilot surgery will lead to more CRISPR operations for more diseases; a day when patients who once had no hope will come to San Francisco from all over the world to seek these treatments out.
Now the researchers want to know all about this dark-haired teen who wears black skinny jeans, Converse sneakers, and a lip-ring; who has trouble using her hands and sometimes stumbles over her feet but sits with exquisite posture; who speaks eloquently and vulnerably about the disease that once made her question who she is and inspired her to become a writer and a medical trailblazer.
"How does it feel to be part of this project?" someone asks.
"It's nice to realize people are looking into a solution for people like me who don't have any solutions," Van Riper says. "I feel you really care." She flashes a grin and adds, "I like nerds."
"Do you worry about the risks?"
"I've lived long enough to have an experience of life with a disability. If something goes wrong, I don't think it would be as scary as some people think. We can't know until we do it. I'm fine being that person doing it."
"You're really brave."
"I know it's not a for-sure fix. Secretly, though, I do think it will work."
So do many of Conklin's patient volunteers. Some, like Van Riper, have CMT; others have genetic mutations that cause BEST disease, an eye disorder that leads to blindness.
Conklin's team is starting with these two rare diseases for several reasons. First, they each arise from well-known mutations in a single gene, making the CRISPR surgeries relatively simple to design. Second, they affect tissues where CRISPR systems can be easily administered and their effects easily measured. Third – and perhaps most important – these diseases are currently untreatable; any relief from their devastation is, for most patients, worth the potential risks (which may include, for instance, cuts in undesired parts of the genome).
"Almost universally, the first targets of genome surgery will be incurable diseases, where there is truly no other option," Conklin says. "If we can treat these, it will open the door to a new type of medicine."
An Unexpected Windfall
It's easy to see, even for the researchers involved, how the promise of genome surgery can sound like magic. Of course, the process is not magic at all but a very real, albeit exceptional, molecular operation that traces its origin to an unassuming source.
Starting in the 1980s, biologists studying bacteria and other microorganisms noticed strange regions of DNA in their genomes. Surprisingly, the regions contained segments that were palindromes – they read the same forward as backward – and that repeated at regular intervals, like books in which every paragraph begins with the word "RACECAR." Those oddities gave the segments their mouthful of a name: clustered regularly interspaced short palindromic repeats, soon shortened to CRISPRs.
Eventually, researchers determined that CRISPRs bookend pieces of DNA stolen from invading viruses, like frames around criminal mug shots. The whole DNA region serves as a kind of microbial defense force: Genes near the CRISPRs code for defender molecules, called CRISPR-associated (Cas) proteins, that execute viruses by chopping up their DNA; the viral mug shots, copied into RNA molecules that stick to the Cas proteins, serve as the defenders' guides.
For decades, CRISPR research remained a relatively obscure niche of biology. Then, in 2012, a team led by UC Berkeley's Doudna and Emmanuelle Charpentier, Ph.D., then of Sweden's Umeå University and now a director at the Max Planck Institute for Infection Biology in Berlin, published a paper that launched CRISPR to scientific fame.
The paper described how one particular Cas protein, Cas9, could be directed to cut not only bacteria-invading viruses but any piece of DNA, simply by changing Cas9's RNA guide. That ability – to create a specific DNA editor by supplying a specific RNA molecule – was revolutionary. RNA, after all, is easy to make in the lab. Scientists could therefore build a plethora of new Cas9-based tools in a fraction of the time and at a fraction of the cost of previous technologies. [See "Genome Editing Before CRISPR: A Brief History."]
This discovery ignited a CRISPR frenzy. Around the world, labs quickly embraced the so-called CRISPR-Cas9 system, using it to cut out and splice genes into bacteria, fungi, plants, animals, and, of course, human cells. "It was just remarkable how fast it spread," Doudna recalls. Soon, researchers were rejiggering Cas9 to create CRISPR tools with more diverse abilities, thus expanding Cas9's scalpel into an array of surgical instruments.
In 2013, for instance, Doudna teamed up with several UCSF researchers – including Stanley Qi, Ph.D. (now at Stanford University); Luke Gilbert, Ph.D. (the Goldberg-Benioff Professor); Jonathan Weissman, Ph.D.; and Wendell Lim, Ph.D. – to show that a mutated version of Cas9, called "dead" Cas9, or dCas9, could bind to a DNA target but not cut it. This insight proved incredibly fruitful: By fusing various other molecules to dCas9, the team could use the resulting systems to dial up or dial down gene expression without altering the underlying DNA. "Now we had a volume switch," says Weissman, a professor of cellular and molecular pharmacology and co-director of IGI.
Other labs soon found further add-ons: molecular tags to track genes' behavior; molecular proofreaders to edit single bases; molecular shields to stop rogue cuts; molecular switches to allow remote control. "It was kind of amazing," Weissman says. "In the course of just six years, we did everything we wanted and much more."
The challenge now for genome surgeons is to find which combinations of CRISPR systems, in what order and under what conditions, will treat a particular patient with a particular disease safely and effectively.
Back in the Marson lab, Roth has mixed the ingredients for his "magic" CRISPR system in a flask and left them under heat to allow them to assemble. Now, using a syringe-like pipette, he sucks up the CRISPR molecules and divvies them, squirt by tiny squirt, among the wells of a honeycombed plate. There, he will test the system on human T cells – a type of immune cell – to see how well his surgical procedure works.
"In a clinical setting, this would be done by a robot," he says, as if he's already envisioning a day when all this tinkering – and tedious pipetting – will not only satisfy scientific curiosities but also save lives.
Next, he adds to the wells another ingredient: genes. These particular genes code for a protein called a synthetic T-cell receptor, or TCR. Perched on the surface of T cells like border guards, receptors detect toxic particles or pathogens entering the body, thereby instigating an immune attack. A synthetic TCR is a lab-made receptor designed specifically to recognize cancer cells – in this case, some forms of melanoma. If all goes as Roth expects, the CRISPR system will splice the TCR genes into the T cells' DNA precisely where he wants them, turning the cells into cancer-killing agents. (In 2017, the U.S. Food and Drug Administration approved two older-generation T-cell therapies that use non-CRISPR technologies, one for acute lymphoblastic leukemia and the other for advanced lymphomas.)
"Genome surgery isn't just about repairing DNA," Roth says, now pipetting the human T-cells into the test wells to mingle with the CRISPR molecules and the TCR genes. "We also want to put new sequences into cells that impart new therapeutic functions."
Finally, he slides the entire well plate – with its motley crew of residents – into a breadbox-sized contraption: an electroporator. Click, click, click goes the electroporator, delivering a series of mild electric shocks. The shocks make the T cells' sack-like membranes permeable, letting the CRISPR molecules and the TCR genes slip through. When at last the electroporator ejects the cells, Roth sets them in an incubator to warm.
A couple of days later, after the CRISPR system has had time to perform its tricks, he will analyze the data. He will determine, to his satisfaction, that they are "somewhat as we expected." Then he will start preparing the next experimental run – one of hundreds he has done over the past year and will continue to do in the months to come – in the hope of making the procedure just a little easier, a little safer, a little more effective.
Like most genome surgery pioneers, he is cautiously optimistic that his efforts will pay off. The rapid rise of CRISPR technology, followed by early therapeutic progress, has given scientists and physicians alike reason to be hopeful – to "feel encouraged," as Doudna says, "that this is something that in the next few years will be increasingly available to patients."
At the same time, many important questions remain: How will physicians deliver CRISPR systems to hard-to-reach tissues such as the heart? How will they treat diseases with many underlying and interacting gene mutations? How will they educate patients about the risks and benefits? What exactly are the risks and benefits? What are the proper doses? How will these surgeries be regulated? Who will perform them? Who will pay for them? Who will have access to them?
"There's plenty of work still to be done," Roth says, speaking for the field as well as himself. He and his peers are like Apollo engineers – tweaking one more sensor, running one more simulation – before launching a space flight, with astronauts aboard, into the starry unknown.
Scientists began searching for ways to edit genomes in the 1960s. Working in test tubes, researchers at UCSF and Stanford bombarded DNA with various combinations of molecular widgets, all borrowed from bacteria. Some of these widgets slice apart DNA bases like miniature scythes; others fasten them together like glue.
In 1972, after a few years of trial and error – much of which took place in the UCSF laboratory of Herbert Boyer, Ph.D., then an assistant professor who would go on to co-found the biotechnology giant Genentech – the researchers eventually landed on a recipe for cutting and pasting DNA. For the first time, it was possible to mix and match genes to create hybrid sequences – called recombinant DNA – that had never before existed.
You could, let's say, take a virus like HIV, delete the genes that make it virulent, and splice in a gene from a human cell. You could then unleash copies of your recombinant virus in the cells of a patient with a diseased copy of this gene. The viruses would naturally insert the new gene into the cells' DNA, where it could compensate for its native, mutant twin and alleviate the disease's symptoms.
This scenario is the basis of gene therapy – imbuing cells with healthy genes to make up for sick ones. It's a promising approach. First tried in 1989, gene therapy progressed in fits and starts, plagued by unexpected setbacks – most notably the death of a patient in 1999. Those early setbacks, however, have been largely worked out, and "gene therapy 2.0" is now being tested in hundreds of clinical trials across the U.S., including several at UCSF clinics to treat sickle cell disease, beta thalassemia (a rare blood disorder), severe combined immunodeficiency syndrome (sometimes called "bubble boy disease"), and Parkinson's disease.
Still, the technology has its drawbacks. It's really more of a patch kit than a repair shop, and an imperfect one at that. Because gene therapy adds a new gene at an unpredictable spot in a cell's genome, the gene's fate isn't a sure thing. Genes, after all, don't work in isolation. They lie amid various DNA segments called regulatory DNA, which tell the cell how to read the code, much like notations on a music score. Consequently, a therapy gene – randomly inserted into the genome by a virus – might land near regulatory DNA that silences it, rendering it useless. Worse, it might disrupt a healthy gene or turn on a gene that causes cancer.
In the early 2000s, scientists went searching for tools they could better control. By cobbling together parts of natural proteins, they found they could synthesize artificial proteins able to target mutations at desired locations in a genome. One of the more capable creations, called a zinc-finger nuclease (ZFN), has already made its way into clinical trials. The first test in a human patient was led by Paul Harmatz, MD, at UCSF Benioff Children's Hospital Oakland – in partnership with Richmond, Calif.-based Sangamo Therapeutics – in 2017.
Engineering proteins, however, is no small feat. It takes months or even years to adapt a ZFN to target just one of the many thousands of known disease-causing mutations. The process is simply too time-consuming and costly to be of practical use in treating the vast majority of genetic diseases.
For nearly a decade, researchers struggled to find a better way – until, in 2012, CRISPR came along.
Editor's Note: Over the next few weeks, Medicare beneficiaries are allowed to make changes to their health plans. This period, called Medicare Open Enrollment, is an ideal time to explore different coverage options to see if they better meet your needs. Review the below checklist, originally shared in 2016, for tips on how to make the most out of Open Enrollment.
2018 Medicare Open Enrollment began Monday, October 15, and runs through Friday, December 7. During this period, all Medicare beneficiaries may make changes to their health care and prescription drug plans, which will take effect January 1, 2018. This allows individuals to compare coverage and find the plan that works best for their needs.
The Centers for Medicare and Medicaid Services has prepared a checklist for beneficiaries to consult during the Open Enrollment period.
Review your plan. Be sure to read any notices from your Medicare and/or prescription drug plan about changes for next year, especially your "Annual Notice of Change" letter. Look at your plan's information to make sure your drugs are still covered and your doctors are still in network.
Think about what matters most to you. Medicare health and drug plans change each year and so can your health needs. Do you need a new primary care doctor? Does your network include the specialist you want? Is your new medication covered by your current prescription drug plan? Does another plan offer the same value at a lower cost? Take stock of your health status and determine if you need to make a change.
Find out if you qualify for help paying for your Medicare. Learn about programs in your state to help with the costs of Medicare premiums, your Medicare Part A (hospital insurance) and Medicare Part B (medical insurance) deductibles, coinsurance and copayments, and Medicare prescription drug coverage costs. You can do this by visiting Medicare.gov or making an appointment with a local State Health Insurance Assistance Program (SHIP) counselor.
Shop for plans that meet your needs and fit your budget. You can use Medicare's plan finder tool at Medicare.gov/find-a-planto see what other plans are offered in your area. A new plan may:
Cost less
Cover your drugs
Allow you go to the providers you want, like your doctor or pharmacy
If you find that your current coverage still meets your needs, then you're done. Remember, during Medicare Open Enrollment, you can decide to stay in Original Medicare or join a Medicare Advantage plan. If you're already in a Medicare Advantage plan, you can switch back to Original Medicare or to a different Medicare Advantage plan.
Check your plan's star rating before you enroll. The Medicare Plan Finder includes star ratings for Medicare health and prescription drug plans. Plans are given an overall quality rating on a one to five star scale, with one star representing poor performance and five stars representing excellent performance. Use the star ratings to compare the quality of health and drug plans being offered.
Still have questions? MAPRx, a coalition of patient, care partner and health care organizations committed to helping people access the prescriptions drugs they need, has released a comprehensive brochure that answers frequently asked questions about open enrollment.
Then click on the original page showing each item you are interested in (showing on that page in orangish yellow,) another page will pop up for you to read.
