Stem Cells Do Geometry

Human embryonic cells
As seen under a microscope, human embryonic cells (colored dots) confined to circles measuring 1 millimeter across start to specialize and form distinct layers similar to those seen in early development. Credit: Aryeh Warmflash, Rockefeller University. View larger image

Each fluorescent point of light making up the multicolored rings in this image is an individual human embryonic cell in the early stages of development. Scientists seeking to understand the molecular cues responsible for early embryonic patterning found that human embryonic cells confined to areas of precisely controlled size and shape begin to specialize, migrate and organize into distinct layers just as they would under natural conditions.

Read the Inside Life Science article to learn more about this research, which has opened a new window for studying early development and could advance efforts aimed at using human stem cells to replace diseased cells and regenerate lost or injured body parts.

 

Meet Jennifer Doudna

Jennifer Doudna
Credit: Jennifer Doudna
Jennifer Doudna
Fields: Biochemistry and structural biology
Studies: New genome editing tool called CRISPR
Works at: University of California, Berkeley
Raised in: Hilo, Hawaii
Studied at: Pomona College, Harvard University
Recent honors: Winner of the Lurie Prize in the Biomedical Sciences Exit icon, an annual award that recognizes outstanding achievement by promising scientists age 52 or younger
If she couldn’t be a scientist, she’d like to be: A papaya farmer or an architect

Jennifer Doudna likes to get her hands dirty. Literally. When she’s not in her laboratory, she can often be found amid glossy green leaves and brightly colored fruit in her Berkeley garden. She recently harvested her first three strawberry guavas.

Coaxing tropical fruit plants from her childhood home in Hawaii to grow in Northern California is more than just a hobby—it’s an intellectual challenge.

“I like solving puzzles, I like the process of figuring things out, and I enjoy working with my hands,” says Doudna. “Those things were what really drew me to science in the beginning.”

Since she was a graduate student, Doudna’s professional puzzle has been RNA, a type of genetic material inside our cells. Recently, there has been an explosion of discoveries about the many roles RNA molecules play in the body. Doudna’s work probes into how RNA molecules work, what 3-D shapes they form and how their structures drive their functions.

“I’ve been fascinated by understanding RNA at a mechanistic level,” Doudna says.

While teasing out answers to these fundamental questions, Doudna’s lab has played a leading role in a discovery that is upending the field of genetic engineering, with exciting implications for human health.

Her Findings

The discovery started with bacteriophages—viruses that infect bacteria, just like the common cold infects humans. About 10 years ago, researchers using high-powered computing to sift through bacterial genomes began to find mysterious repetitive gene sequences that matched those from viruses known to infect the bacteria. The researchers named these sequences “clustered regularly interspaced short palindromic repeats,” or CRISPRs for short.

Over the next few years, scientists came to understand that these CRISPR sequences are part of something not previously thought to exist—an adaptive bacterial immune system, which remembers viruses fought off before and raises a response to fight them when exposed again. CRISPRs were this immune system’s reference library, holding records of viral exposure.

Somehow, bacteria with a CRISPR-based immune system (there are three types now known to scientists) use these records to command certain proteins to recognize and chop up DNA from returning viruses.

Wanting to know more about this process, Doudna’s team picked one protein in a CRISPR-based defense system to study. This protein, called Cas9, had been identified by other researchers as being essential for protection against viral invasion.

To their delight, Doudna’s group had hit the jackpot. Cas9 turned out to be the system’s scalpel. Once CRISPR identifies a DNA sequence from the invading virus, Cas9 slices the sequence out of the viral genome, destroying the virus’s ability to copy itself.

Doudna’s lab and their European collaborators also identified the other key components of the CRISPR-Cas9 system—two RNA molecules that guide Cas9 to the piece of viral DNA identified by CRISPR.

Even more importantly, the researchers showed that the two guide RNAs could be manipulated in the lab to create a tool that both recognizes any specified DNA sequence and carries Cas9 there to make its cut.

“That was really where we made the connection between the basic, curiosity-driven research that we were doing and recognizing that we had in our hands something that could be a very powerful technology for genome editing,” remembers Doudna.

She was right. After publication of their 2012 paper, the field of CRISPR-guided genetic manipulation exploded. Labs around the world now use the tool Doudna’s team developed to cut target gene sequences in organisms ranging from plants to humans. The technique is already replacing more time-consuming, less-reliable methods of creating ‘knock-out’ model organisms (those missing a specific gene) for laboratory research. CRISPR-based editing even allows more than one gene to be knocked out at the same time, something that was not possible with previous genome-editing techniques.

The ability of CRISPR systems to recognize DNA sequences with extraordinary precision also holds potential for human therapeutics. For example, a paper from another laboratory published early this year showed that, in a mouse model, CRISPR-based editing could cut out and replace a defective gene responsible for a type of muscular dystrophy. Researchers are testing similar CRISPR-based techniques in models of human diseases ranging from cystic fibrosis to blood disorders.

Doudna is a co-founder of two biotechnology companies hoping to harness the potential of CRISPR-based genome editing. Although the technology holds great promise, she acknowledges that much work needs to be done before CRISPR can be considered safe for human trials. Major challenges include assuring that no off-target cuts are made in the genome and finding a safe way to deliver the editing system to living tissues.

She is also excited to continue working with her research team, advancing the basic understanding of the CRISPR-based system.

“I’m very interested in seeing what we can contribute to the whole question about how you deliver a technology like this, how you can use it therapeutically in an organism. That’s an area where we hope that our biochemical understanding of this system will be able to contribute,” she concludes.

Modifying Bacterial Behavior

Biofilm
Communication through quorum sensing is key to the formation of biofilms, slimy bacterial communities that can cause infections and are often stubbornly resistant to antibiotics. Credit: P. Singh and E. Peter Greenberg.

Like a person trailing the aroma of perfume or cologne, bacteria emit chemical signals that let other bacteria of the same species know they’re there. Bacteria use this chemical communication system, called quorum sensing, to assess their own population size. When they sense a large enough group, or quorum, the microbes modify their behavior accordingly. Many disease-causing bacteria use quorum sensing to launch a coordinated attack when they’ve amassed in sufficient numbers to overwhelm the host’s immune response.

Chemist Helen Blackwell of the University of Wisconsin-Madison has been making artificial compounds that mimic natural quorum-sensing signals, as well as some that block a natural signal from binding to its protein target—a step needed to produce a change in bacterial behavior. By altering key building blocks in these protein targets one by one, Blackwell’s team found that small changes could convert an activation signal into an inhibitory signal, or vice versa, indicating that small-molecule control of quorum sensing is very finely tuned.

Improved understanding of the molecular basis of quorum sensing could help scientists design more potent compounds to disrupt these signals. Using such compounds to quiet quorum sensing may provide a new way to control disease-causing bacteria that reduces the chances an infection will become resistant to treatment.

This work was funded in part by NIH under grants R01GM109403 and T32GM008505.

Learn more:
University of Wisconsin-Madison News Release Exit icon
Blackwell Lab Exit icon
Learning From Bacterial Chatter Article from Inside Life Science
Bugging the Bugs Article from Findings Magazine

4 Timely Facts About Our Biological Clocks

Illustration of circadian rhythm.
Genes and proteins run biological clocks that help keep daily rhythms in synch. Credit: Wikimedia Commons. View larger image

After you roll your clocks back by an hour this Sunday, you may feel tired. That’s because our bodies—more specifically, our circadian rhythms—need a little time to adjust. These daily cycles are run by a network of tiny, coordinated biological clocks.

NIGMS’ Mike Sesma tracks circadian rhythm research being conducted in labs across the country, and he shares a few timely details about our internal clocks:

1. They’re incredibly intricate.

Biological clocks are composed of genes and proteins that operate in a feedback loop. Clock genes contain instructions for making clock proteins, whose levels rise and fall in a regular cyclic pattern. This pattern in turn regulates the activity of the genes. Many of the results from circadian rhythm research this year have uncovered more parts of the molecular machinery that fine-tune the clock. Earlier in the month, we blogged about an RNA molecule that cues the internal clock.

2. Every organism has them—from algae to zebras.

Many of the clock genes and proteins are similar across species, allowing researchers to make important findings about human circadian processes by studying the clock components of organisms like fruit flies, bread mold and plants.

3. Whether we’re awake or asleep, our clocks keep ticking.

While they might get temporarily thrown off by changes in light or temperature, our clocks usually can reset themselves.

4. Nearly everything about how our body works is tied to biological clocks.

Our clocks influence alertness, hunger, metabolism, fertility, mood and other physiological conditions. For this reason, clock dysfunction is associated with various disorders, including insomnia, diabetes and depression. Even drug efficacy has been linked to our clocks: Studies have shown that some drugs might be more effective if given earlier in the day.

Learn more:
Circadian Rhythms Fact Sheet
Resetting Our Clocks: New Details About How the Body Tells Time Article from Inside Life Science

Steering Cells Down the Right Path

In these time-lapse videos of 60 images taken over an hour, cell receptors move around the cell surface in search of the missing signal that will tell the cell where to go (top video). Once the receptors locate the signal (bottom video), they stay put in the region of the cell membrane that is closest to the signal. Credit: David Sherwood, Duke University.

Even traveling cells need help with directions. In fact, it’s crucial. For processes such as wound healing and organ development to take place, cells must be able to efficiently move throughout organisms. Receptor proteins on a cell’s surface rely on navigational signals from molecules called netrins to point them in the right direction.

The receptors don’t just sit around waiting for a signal. Studying the simple worm C. elegans, David Sherwood Exit icon and his research team at Duke University discovered that in the absence of netrin, the receptors rapidly cluster and reassemble in different areas of the cell’s membrane. When the receptors finally detect a netrin signal, they stabilize and correctly orient the cell. The finding might point to new ways to interfere with cells’ built-in navigation systems to hamper cell migration in metastatic cancer or encourage the regrowth of damaged cells in neurodegenerative diseases such as Parkinson’s.

This research was funded in part by NIH under grants R01GM100083 and P40OD010440.

Learn more:
Duke University News Release Exit icon

The Sweet Side of Chemistry

Glycans
Simple sugars connect in long, branched structures to create glycans. Credit: Stock image.

We’re in the middle of National Chemistry Week Exit icon, which this year focuses on “The Sweet Side of Chemistry—Candy.” Studying sugar chemistry is also relevant to our health.

The sugar in chocolate, taffy and other confections is a type of simple sugar called sucrose. In our bodies, sugars can exist in many forms, ranging from individual units like glucose to long, branched chains known as glycans containing thousands of individual sugar units linked together.

Glycans are involved in just about every aspect of how our cells work. They help make sure proteins are folded into the proper shape so they function correctly. They act as ZIP codes that direct newly made proteins to the right cellular locations. Some divert white blood cells to infection sites, and others serve as anchors for viruses to latch onto.

Because of the diverse and critical roles that glycans play in our bodies, chemists want to learn more about these molecules, with a long-term goal of harnessing them to treat or prevent disease. Read about some of their discoveries in the Why Sugars Might Surprise You article from Inside Life Science and the Life is Sweet article from Findings magazine. The You Are What You Eat chapter from ChemHealthWeb offers more details about the chemistry of sugar.

Cool Image: Of Surfaces and Stem Cells

Stem cells transform into neurons.
Stem cells grown on a soft surface begin to transform into neurons. Credit: Kiessling Lab, University of Wisconsin-Madison.

If you think this image looks like the fluorescent outline of a brain, you’re on the right track. The green threads show neurons that have just formed from unspecialized cells called stem cells.

Researchers led by Laura Kiessling Exit icon of the University of Wisconsin-Madison directed the stem cells to become neurons by changing the quality of the surface on which they grew. In experiments testing different gels used to grow stem cells in the lab, the scientists found that the stiffness of those gels influenced cell fate decisions.

When grown on a soft gel with a brain tissue-like surface, the stem cells began to transform into neurons. This happened without the addition of any of the proteins normally used to coax stem cells to specialize into different types of cells.

A better understanding of how stem cell fate is influenced by the mechanical properties of a surface could help researchers who are trying to harness the blank slate cells for tissue regeneration or other therapeutic uses.

This work also was funded by NIH’s National Institute of Biomedical Imaging and Bioengineering; National Heart, Lung, and Blood Institute; and National Institute of Neurological Disorders and Stroke.

Learn more:
University of Wisconsin-Madison News Release Exit icon

Nobel Prize for Powerful Microscopy Technology

Fibroblasts
The cells shown here are fibroblasts, one of the most common cells in mammalian connective tissue. These particular cells were taken from a mouse. Scientists used them to test the power of a new microscopy technique that offers vivid views of the inside of a cell. The DNA within the nucleus (blue), mitochondria (green) and cellular skeleton (red) is clearly visible. Credit: Dylan Burnette and Jennifer Lippincott-Schwartz, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health.

William E. Moerner was at a conference in Brazil when he learned he’d be getting a Nobel Prize in chemistry. “I was incredibly excited and thrilled,” he said of his initial reaction.

An NIGMS grantee at Stanford University, Moerner received the honor for his role in achieving what was once thought impossible—developing super-resolution fluorescence microscopy, which is so powerful it allows researchers to see and track individual molecules in living organisms in real time.

Nobel recipients usually learn of the prize via a phone call from Stockholm, Sweden, in early October. For those in the United States, the call typically comes between 2:30 a.m. and 5:45 a.m.

Every year, the NIGMS communications office prepares for the Nobel Prize announcements in physiology or medicine and chemistry, the categories in which our grantees are most likely to be recognized. If the Institute played a significant role in funding the prize-winning research, we work quickly to provide information and context to reporters covering the story on tight deadlines. We issue a statement, identify an in-house expert on the research and arrange interviews with reporters. It’s all to help get the word out about the research and the taxpayers’ role in supporting it.

This year’s in-house expert, Cathy Lewis, shared her thoughts on the prize to Moerner in an NIGMS Feedback Loop post. You can also read this year’s statement and see a full list of NIGMS-supported Nobel laureates.

Outwitting Antibiotic Resistance

Marine scene with fish and corals
The ocean is a rich source of microbes that could yield infection-fighting natural molecules. Credit: National Oceanic and Atmospheric Administration Exit icon.

Antibiotics save countless lives and are among the most commonly prescribed drugs. But the bacteria and other microbes they’re designed to eradicate can evolve ways to evade the drugs. This antibiotic resistance, which is on the rise due to an array of factors, can make certain infections difficult—and sometimes impossible—to treat.

Read the Inside Life Science article to learn how scientists are working to combat antibiotic resistance, from efforts to discover potential new antibiotics to studies seeking more effective ways of using existing ones.

 
 

An RNA Molecule That Cues the Internal Clock

Clock
Dysfunction in our internal clocks may lead to insufficient sleep, which has been linked to an increased risk for chronic diseases. Credit: Stock image.

Our internal clocks tell us when to sleep and when to eat. Because they are sensitive to changes in daytime and nighttime cues, they can get thrown off by activities like traveling across time zones or working the late shift. Dysfunction in our internal clocks may lead to insufficient sleep, which has been linked to an increased risk for chronic diseases like high blood pressure, diabetes, depression and cancer.

Researchers led by Yi Liu Exit icon of the University of Texas Southwestern Medical Center have uncovered a previously unknown mechanism by which internal clocks run and are tuned to light cues. Using the model organism Neurospora crassa (a.k.a., bread mold), the scientists identified a type of RNA molecule called long non-coding RNA (lncRNA) that helps wind the internal clock by regulating how genes are expressed. When it’s produced, the lncRNA identified by Liu and his colleagues blocks a gene that makes a specific clock protein.

This inhibition works the other way, too: The production of the clock protein blocks the production of the lncRNA. This rhythmic gene expression helps the body stay tuned to whether it’s day or night.

The researchers suggest that a similar mechanism likely exists in the internal clocks of other organisms, including mammals. They also think that lncRNA-protein pairs may contribute to the regulation of other biologic processes.

Learn more:
University of Texas Southwestern Medical Center News Release Exit icon
Circadian Rhythms Fact Sheet
Resetting Our Clocks: New Details About How the Body Tells Time Article from Inside Life Science
Remarkable RNAs Article from Inside Life Science