Category Archives: Cell Biology

Cell Biology

Researchers solve a long-held mystery of X chromosome inactivation – News-Medical.net

Researchers at Massachusetts General Hospital (MGH) have solved a mystery that has long puzzled scientists: How do the bodies of female humans and all other mammals decide which of the two X chromosomes it carries in each cell should be active and which one should be silent?

In a breakthrough study published in Nature Cell Biology, the MGH team discovered the role of a critical enzyme in the phenomenon known as X chromosome inactivation (XCI), which is essential for normal female development and also sets the stage for genetic disorders known as X-linked diseases (such as Rett Syndrome) to occur.

Scientists have known for over a half-century that female mammals undergo XCI during embryo formation. Females have two copies of the X chromosome, and each carries many genes.

Having genes expressed on both X chromosomes would be toxic to the cell, as would having both X chromosomes inactivated. To avoid these fates, females evolved with a mechanism that inactivates, or silences, one of the chromosomes.

Over the years, investigators have made strides in understanding how XCI occurs. In 2006, a team led by Jeannie Lee, MD, PhD, of the Department of Molecular Biology at MGH reported that during embryo development the two X chromosomes briefly come together, or pair.

She and her colleagues have since uncovered conclusive evidence that pairing is necessary for the body to decide which X chromosome to inactivate. "But until now, no one knew what one X chromosome was saying to the other to make the decision," says Lee, who is senior author of the Nature Cell Biology paper.

To find out, Lee and her colleagues had to develop sophisticated molecular tools that allow them to study key proteins involved in XCI, which were previously difficult to measure. It was already known that, prior to pairing, both X chromosomes are identical, or "symmetrical," meaning that they express the same genes.

Importantly, both express a form of noncoding RNA called Xist, which plays a vital role in inactivating the X chromosome. However, both X chromosomes also express another form of RNA, Tsix, which blocks Xist and prevents XCI.

In the Nature Cell Biology paper, Lee and her team show that an enzyme called DCP1A randomly chooses one X chromosome to bind to, and in doing so it cuts off, or "decaps," Tsix's protective cover, making the RNA unstable. However, because DCP1A exists in tiny quantities, there is only enough to bind to one X chromosome. "DCP1A flips the switch that starts the entire cascade of X chromosome inactivation," says Lee.

As a result, a protein called CTCF--the "glue" that holds X chromosomes together during pairing--binds to the unstable Tsix RNA and causes it to shut down permanently. Xist is then able to complete the silencing of that X chromosome.

DCP1A allows the two X chromosomes to have a fateful 'conversation', noting that there are many other instances where the body must choose which copy of a gene to express in order to maintain a healthy state. "This discovery, will help scientists understand how other molecular conversations take place in the cell."

Jeannie Lee, MD, PhD, Professor and Director, Department of Molecular Biology, Massachusetts General Hospital

Jeannie Lee, MD, Ph.D., of the Department of Molecular Biology at MGH, is also director of the Lee Laboratory and a professor of Genetics at Harvard Medical School.

Source:

Journal reference:

Aeby, E., et al. (2020) Decapping enzyme 1A breaks X-chromosome symmetry by controlling Tsix elongation and RNA turnover. Nature Cell Biology. doi.org/10.1038/s41556-020-0558-0.

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Researchers solve a long-held mystery of X chromosome inactivation - News-Medical.net

Cell Biology

Researchers find method to regrow cartilage in the joints – Stanford Medical Center Report

Damaged cartilage can be treated through a technique called microfracture, in which tiny holes are drilled in the surface of a joint. The microfracture technique prompts the body to create new tissue in the joint, but the new tissue is not much like cartilage.

Microfracture results in what is called fibrocartilage, which is really more like scar tissue than natural cartilage, said Chan. It covers the bone and is better than nothing, but it doesnt have the bounce and elasticity of natural cartilage, and it tends to degrade relatively quickly.

The most recent research arose, in part, through the work of surgeon Matthew Murphy, PhD, a visiting researcher at Stanford who is now at the University of Manchester. I never felt anyone really understood how microfracture really worked, Murphy said. I realized the only way to understand the process was to look at what stem cells are doing after microfracture. Murphy is the lead author on the paper. Chan and Longaker are co-senior authors.

For a long time, Chan said, people assumed that adult cartilage did not regenerate after injury because the tissue did not have many skeletal stem cells that could be activated. Working in a mouse model, the team documented that microfracture did activate skeletal stem cells. Left to their own devices, however, those activated skeletal stem cells regenerated fibrocartilage in the joint.

But what if the healing process after microfracture could be steered toward development of cartilage and away from fibrocartilage? The researchers knew that as bone develops, cells must first go through a cartilage stage before turning into bone. They had the idea that they might encourage the skeletal stem cells in the joint to start along a path toward becoming bone, but stop the process at the cartilage stage.

The researchers used a powerful molecule called bone morphogenetic protein 2 (BMP2) to initiate bone formation after microfracture, but then stopped the process midway with a molecule that blocked another signaling molecule important in bone formation, called vascular endothelial growth factor (VEGF).

What we ended up with was cartilage that is made of the same sort of cells as natural cartilage with comparable mechanical properties, unlike the fibrocartilage that we usually get, Chan said. It also restored mobility to osteoarthritic mice and significantly reduced their pain.

As a proof of principle that this might also work in humans, the researchers transferred human tissue into mice that were bred to not reject the tissue, and were able to show that human skeletal stem cells could be steered toward bone development but stopped at the cartilage stage.

The next stage of research is to conduct similar experiments in larger animals before starting human clinical trials. Murphy points out that because of the difficulty in working with very small mouse joints, there might be some improvements to the system they could make as they move into relatively larger joints.

The first human clinical trials might be for people who have arthritis in their fingers and toes. We might start with small joints, and if that works we would move up to larger joints like knees, Murphy says. Right now, one of the most common surgeries for arthritis in the fingers is to have the bone at the base of the thumb taken out. In such cases we might try this to save the joint, and if it doesnt work we just take out the bone as we would have anyway. Theres a big potential for improvement, and the downside is that we would be back to where we were before.

Longaker points out that one advantage of their discovery is that the main components of a potential therapy are approved as safe and effective by the FDA. BMP2 has already been approved for helping bone heal, and VEGF inhibitors are already used as anti-cancer therapies, Longaker said. This would help speed the approval of any therapy we develop.

Joint replacement surgery has revolutionized how doctors treat arthritis and is very common: By age 80, 1 in 10 people will have a hip replacement and 1 in 20 will have a knee replaced. But such joint replacement is extremely invasive, has a limited lifespan and is performed only after arthritis hits and patients endure lasting pain. The researchers say they can envision a time when people are able to avoid getting arthritis in the first place by rejuvenating their cartilage in their joints before it is badly degraded.

One idea is to follow a Jiffy Lube model of cartilage replenishment, Longaker said. You dont wait for damage to accumulate you go in periodically and use this technique to boost your articular cartilage before you have a problem.

Longaker is the Deane P. and Louise Mitchell Professor in the School of Medicine and co-director of the Institute for Stem Cell Biology and Regenerative Medicine. Chan is a member of the Institute for Stem Cell Biology and Regenerative Medicine and Stanford Immunology.

Other Stanford scientist taking part in the research were professor of pathology Irving Weissman, MD, the Virginia and D. K. Ludwig Professor in Clinical Investigation in Cancer Research; professor of surgery Stuart B. Goodman, MD, the Robert L. and Mary Ellenburg Professor in Surgery; associate professor of orthopaedic surgery Fan Yang, PhD; professor of surgery Derrick C. Wan, MD; instructor in orthopaedic surgery Xinming Tong, PhD; postdoctoral research fellow Thomas H. Ambrosi, PhD; visiting postdoctoral scholar Liming Zhao, MD; life science research professionals Lauren S. Koepke and Holly Steininger; MD/PhD student Gunsagar S. Gulati, PhD; graduate student Malachia Y. Hoover; former student Owen Marecic; former medical student Yuting Wang, MD; and scanning probe microscopy laboratory manager Marcin P. Walkiewicz, PhD.

The research was supported by the National Institutes of Health (grants R00AG049958, R01 DE027323, R56 DE025597, R01 DE026730, R01 DE021683, R21 DE024230, U01HL099776, U24DE026914, R21 DE019274, NIGMS K08GM109105, NIH R01GM123069 and NIH1R01AR071379), the California Institute for Regenerative Medicine, the Oak Foundation, the Pitch Johnson Fund, the Gunn/Olivier Research Fund, the Stinehart/Reed Foundation, The Siebel Foundation, the Howard Hughes Medical Institute, the German Research Foundation, the PSRF National Endowment, National Center for Research Resources, the Prostate Cancer Research Foundation, the American Federation of Aging Research and the Arthritis National Research Foundation.

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Researchers find method to regrow cartilage in the joints - Stanford Medical Center Report

Cell Biology

How We Could Control Cells in the Body to Treat Conditions – Lab Manager Magazine

WASHINGTON, DC Like electronic devices, biological cells send and receive messages, but they communicate through very different mechanisms. Now, scientists report progress on tiny communication networks that overcome this language barrier, allowing electronics to eavesdrop on cells and alter their behaviorand vice versa. These systems could enable applications including a wearable device that could diagnose and treat a bacterial infection or a capsule that could be swallowed to track blood sugar and make insulin when needed.

The researchers will present their results today at the American Chemical Society (ACS) Fall 2020 Virtual Meeting & Expo. ACS is holding the meeting through Thursday. It features more than 6,000 presentations on a wide range of science topics.

"We want to expand electronic information processing to include biology," says principal investigator William E. Bentley, PhD. "Our goal is to incorporate biological cells in the computational decision-making process."

The new technology Bentley's team developed relies on redox mediators, which move electrons around cells. These small molecules carry out cellular activities by accepting or giving up electrons through reduction or oxidation reactions. Because they can also exchange electrons with electrodes, thereby producing a current, redox mediators can bridge the gap between hardware and living tissue. In ongoing work, the team, which includes co-principal investigator Gregory F. Payne, PhD, is developing interfaces to enable this information exchange, opening the way for electronic control of cellular behavior, as well as cellular feedback that could operate electronics.

"In one project that we are reporting on at the meeting, we engineered cells to receive electronically generated information and transmit it as molecular cues," says Eric VanArsdale, a graduate student in Bentley's lab at the University of Maryland, who is presenting the latest results at the meeting. The cells were designed to detect and respond to hydrogen peroxide. When placed near a charged electrode that generated this redox mediator, the cells produced a corresponding amount of a quorum sensing molecule that bacteria use to signal to each other and modulate behavior by altering gene expression.

In another recent project, the team engineered two types of cells to receive molecular information from the pathogenic bacteriaPseudomonas aeruginosa and convert it into an electronic signal for diagnostic and other applications. One group of cells produced the amino acid tyrosine, and another group made tyrosinase, which converts tyrosine into a molecule called L-DOPA. The cells were engineered so this redox mediator would be produced only if the bacteria released both a quorum sensing molecule and a toxin associated with a virulent stage ofP. aeruginosa growth. The size of the resulting current generated by L-DOPA indicated the amount of bacteria and toxin present in a sample. If used in a blood test, the technique could reveal an infection and also gauge its severity. Because this information would be in electronic form, it could be wirelessly transmitted to a doctor's office and a patient's cell phone to inform them about the infection, Bentley says. "Ultimately, we could engineer it so that a wearable device would be triggered to give the patient a therapeutic after an infection is detected."

The researchers envision eventually integrating the communication networks into autonomous systems in the body. For instance, a diabetes patient could swallow a capsule containing cells that monitor blood sugar. The device would store this blood sugar data and periodically send it to a cell phone, which would interpret the data and send back an electronic signal directing other cells in the capsule to make insulin as needed. As a step toward this goal, VanArsdale developed a biological analog of computer memory that uses the natural pigment melanin to store information and direct cellular signaling.

In other work, Bentley's team and collaborators including Reza Ghodssi, PhD, recently designed a system to monitor conditions inside industrial bioreactors that hold thousands of gallons of cell culture for drug production. Currently, manufacturers track oxygen levels, which are vital to cells' productivity, with a single probe in the side of each vessel. That probe can't confirm conditions are uniform everywhere in the bioreactor, so the researchers developed "smart marbles" that will circulate throughout the vessel measuring oxygen. The marbles transmit data via Bluetooth to a cell phone that could adjust operating conditions. In the future, these smart marbles could serve as a communication interface to detect chemical signals within a bioreactor, send that information to a computer, and then transmit electronic signals to direct the behavior of engineered cells in the bioreactor. The team is working with instrument makers interested in commercializing the design, which could be adapted for environmental monitoring and other uses.

- This press release was originally published on theACS website

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How We Could Control Cells in the Body to Treat Conditions - Lab Manager Magazine

Cell Biology

Loss of Enzyme Increases Metabolism and Exercise Endurance in Mice – Technology Networks

Sugars and fats are the primary fuels that power every cell, tissue and organ. For most cells, sugar is the energy source of choice, but when nutrients are scarce, such as during starvation or extreme exertion, cells will switch to breaking down fats instead.

The mechanisms for how cells rewire their metabolism in response to changes in resource availability are not yet fully understood, but new research reveals a surprising consequence when one such mechanism is turned off: an increased capacity for endurance exercise.

In a study published in Cell Metabolism, Harvard Medical School researchers identifiedacritical role oftheenzyme, prolyl hydroxylase 3 (PHD3), in sensing nutrient availability and regulating the ability of muscle cells to break down fats. When nutrients are abundant, PHD3 acts as a brake that inhibits unnecessary fat metabolism. This brake is released when fuel is low and more energy is needed, such as during exercise.

Remarkably, blocking PHD3 production in miceleadsto dramatic improvements in certain measures of fitness, the research showed. Compared with their normal littermates, mice lacking the PHD3 enzyme ran 40 percent longer and 50 percent farther on treadmills andhadhigher VO2 max, a marker of aerobic endurance that measures maximum oxygen uptake during exercise.

The findings shed light on a key mechanism for how cells metabolize fuels and offer clues toward a better understanding of muscle function and fitness, the authors said.

Our results suggest that PHD3 inhibition in whole body or skeletal muscle is beneficial for fitness in terms of endurance exercise capacity, running time and running distance, said senior study authorMarcia Haigis, professor of cell biology in the Blavatnik Institute at HMS. Understanding this pathway and how our cells metabolize energy and fuels potentially has broad applications in biology, ranging from cancer control to exercise physiology.

However, further studies are needed to elucidate whetherthis pathway canbe manipulated in humans to improve muscle function in disease settings, the authors said.

Haigis and colleagues set out to investigate the function of PHD3, an enzyme that they had found to play a role regulating fat metabolism in certaincancersin previous studies. Their work showed that, under normal conditions, PHD3 chemically modifies another enzyme, ACC2, which in turn prevents fatty acids from entering mitochondria to be broken down into energy.

In the current study, the researchers experiments revealed that PHD3 and another enzyme called AMPK simultaneously control the activity of ACC2 to regulate fat metabolism, depending on energy availability.

In isolated mouse cells grown in sugar-rich conditions, the team found that PHD3 chemically modifies ACC2 to inhibit fat metabolism. Under low-sugar conditions, however, AMPK activates and places a different, opposing chemical modification on ACC2, which represses PHD3 activity and allows fatty acids to enter the mitochondria to be broken down for energy.

These observations were confirmed in live mice that were fasted to induce energy-deficient conditions. In fasted mice, the PHD3-dependent chemical modification to ACC2 was significantly reduced in skeletal and heart muscle, compared to fed mice. By contrast, the AMPK-dependent modification to ACC2 increased.

Longer and further

Next, the researchers explored the consequences when PHD3 activity was inhibited, using genetically modified mice that do not express PHD3. Because PHD3 is most highly expressed in skeletal muscle cells and AMPK has previously been shown to increase energy expenditure and exercise tolerance, the team carried out a series of endurance exercise experiments.

The question we asked was if we knock out PHD3, Haigis said, would that increase fat burning capacity and energy production and have a beneficial effect in skeletal muscle, which relies on energy for musclefunctionand exercisecapacity?

To investigate, the team trained young, PHD3-deficient mice to run on an inclined treadmill. They found that these mice ran significantly longer and further before reaching the point of exhaustion, compared to mice with normal PHD3. These PHD3-deficient mice also had higher oxygen consumption rates, as reflected by increased VO2 and VO2 max.

Aftertheendurance exercise, the muscles of PHD3-deficient mice had increased rates of fat metabolism and an altered fatty acid composition and metabolic profile. The PHD3-dependent modification to ACC2 was nearly undetectable, but the AMPK-dependent modification increased, suggesting that changes to fat metabolism play a role in improving exercise capacity.

These observations held true in mice genetically modified to specifically prevent PHD3 production in skeletal muscle, demonstrating that PHD3 loss in muscle tissues is sufficient to boost exercise capacity, according to the authors.

It was exciting to see this big, dramatic effect on exercise capacity, which could be recapitulated with a muscle-specific PHD3 knockout, Haigis said. The effect of PHD3 loss was very robust and reproducible.

The research team also performed a series of molecular analyses to detail the precise molecular interactions that allow PHD3 to modify ACC2, as well as how its activity is repressed by AMPK.

The study results suggest a new potential approach for enhancingexercise performance by inhibiting PHD3.While the findings are intriguing, the authors note that further studies are needed to better understand precisely how blocking PHD3 causes a beneficial effect on exercise capacity.

In addition, Haigis and colleagues found in previous studies that in certain cancers, such as some forms of leukemia, mutated cells express significantly lower levels of PHD3 and consume fats to fuel aberrant growth and proliferation. Efforts to control this pathway as a potential strategy for treating such cancers may help inform research in other areas, such as muscle disorders.

It remains unclear whether there are any negative effects of PHD3 loss. To know whether PHD3 can be manipulated in humansfor performance enhancement in athletic activities or as a treatment for certain diseases will require additional studies in a variety of contexts, the authors said.

It also remains unclear if PHD3 loss triggers other changes, such as weight loss, blood sugar and other metabolic markers, which are now being explored by the team.

A better understanding of these processes and the mechanisms underlying PHD3 function could someday help unlock new applications in humans, such as novel strategies for treating muscle disorders, Haigis said.

Reference: Yoon et al. (2020).PHD3 Loss Promotes Exercise Capacity and Fat Oxidation in Skeletal Muscle. Cell Metabolism.DOI: 10.1016/j.cmet.2020.06.017.

This article has been republished from the following materials. Note: material may have been edited for length and content. For further information, please contact the cited source.

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Loss of Enzyme Increases Metabolism and Exercise Endurance in Mice - Technology Networks

Cell Biology

Boundless Bio Announces Publication in Nature Genetics Detailing the Association Between Extrachromosomal DNA-Based Oncogene Amplification and Poor…

SAN DIEGO--(BUSINESS WIRE)--Boundless Bio, a company developing innovative new therapies directed to extrachromosomal DNA (ecDNA) in aggressive cancers, today announced research published in the journal Nature Genetics that demonstrates that ecDNA-based oncogene amplification drives poor outcomes for patients across many cancer types.

The manuscript, Frequent extrachromosomal oncogene amplification drives aggressive tumors, was co-authored by Boundless Bio scientists Nam-phuong Nguyen, Ph.D., and Kristen Turner, Ph.D., and scientific founders Paul Mischel, M.D., Distinguished Professor at the University of California San Diego (UC San Diego) School of Medicine and a member of the Ludwig Institute for Cancer Research; Vineet Bafna, Ph.D., Professor of Computer Science & Engineering, UC San Diego; Howard Chang, M.D., Ph.D., Virginia and D.K. Ludwig Professor of Cancer Genomics and Genetics, Stanford University; and Roel Verhaak, Ph.D., Professor and Associate Director of Computational Biology, The Jackson Laboratory.

The researchers used intensive computational analysis of whole-genome sequencing data from more than 3200 tumor samples in The Cancer Genome Atlas (TCGA) and the Pan-Cancer Analysis of Whole Genomes (PCAWG), totaling over 400 TB of raw sequencing data, to observe the impact of ecDNA amplification on patient outcomes. The researchers observed that ecDNA amplification occurs in many types of cancers, but not in normal tissue or in whole blood, and that the most common recurrent oncogene amplifications frequently arise on ecDNA. Notably, ecDNA-based circular amplicons were found in 25 of 29 cancer types analyzed, and at high frequency in many cancers that are considered to be amongst the most aggressive histological types, such as glioblastoma, sarcoma, and esophageal carcinoma. In addition, patients whose cancers carried ecDNA had significantly shorter survival, even when controlled for tissue type, than patients whose cancers were not driven by ecDNA-based oncogene amplification.

The findings demonstrate that ecDNA play a critical role in cancer, providing a mechanism for achieving and maintaining high copy number oncogene amplification and genetic heterogeneity while driving enhanced chromatin accessibility and elevating oncogene transcription. ecDNA amplifications are associated with aggressive cancer behavior, potentially by providing tumors with additional routes to circumvent current treatments and other evolutionary bottlenecks. The shorter overall survival, even when stratified by tumor type, raises the possibility that cancer patients whose tumors are driven by ecDNA may not be as responsive to current therapies and may be in need of new forms of treatment.

This important study builds on our rapidly expanding knowledge about ecDNA, showing, for the first time, that ecDNA amplifications are present in a broad range of cancer tumor types, said Jason Christiansen, Ph.D., Chief Technology Officer of Boundless Bio. These results point to the urgent need for therapies that can target ecDNA and interfere with their ability to drive aggressive cancer growth, resistance, and recurrence.

By detecting and characterizing the role that ecDNA play in driving hard-to-treat cancers, we are drawing a more accurate map of the cancer genome, said Dr. Mischel. It is our goal to take these findings and apply them to the development of powerful anti-cancer therapies for individuals with ecDNA-driven cancers.

About ecDNA

Extrachromosomal DNA, or ecDNA, are distinct circular units of DNA containing functional genes that are located outside cells chromosomes and can make many copies of themselves. ecDNA rapidly replicate within cancer cells, causing high numbers of oncogene copies, a trait that can be passed to daughter cells in asymmetric ways during cell division. Cancer cells have the ability to upregulate or downregulate oncogenes located on ecDNA to ensure survival under selective pressures, including chemotherapy, targeted therapy, immunotherapy, or radiation, making ecDNA one of cancer cells primary mechanisms of recurrence and treatment resistance. ecDNA are rarely seen in healthy cells but are found in many solid tumor cancers. They are a key driver of the most aggressive and difficult-to-treat cancers, specifically those characterized by high copy number amplification of oncogenes.

About Boundless Bio

Boundless Bio is a next-generation precision oncology company interrogating a novel area of cancer biology, extrachromosomal DNA (ecDNA), to deliver transformative therapies to patients with previously intractable cancers.

For more information, visit http://www.boundlessbio.com.

Follow us on LinkedIn and Twitter.

About Boundless Bios Spyglass Platform

Boundless Bios Spyglass platform is a comprehensive suite of proprietary ecDNA-driven and pair-matched tumor models along with proprietary imaging and molecular analytical tools that enables Boundlesss researchers to interrogate ecDNA biology to identify a pipeline of novel oncotargets essential to the function of cancer cells that are enabled by ecDNA. The Spyglass platform facilitates Boundless innovation in the development of precision therapeutics specifically targeting ecDNA-driven tumors, thereby enabling selective treatments for patients whose tumor genetic profiles make them most likely to benefit from our novel therapeutic candidates.

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Boundless Bio Announces Publication in Nature Genetics Detailing the Association Between Extrachromosomal DNA-Based Oncogene Amplification and Poor...

Cell Biology

Following the data – Washington University in St. Louis Newsroom

Kiani Arkus Gardner, AB 07 (biology), was pretty confident she had the experiment right.

An ambitious Washington University sophomore in the lab of biologist Joseph Jez, she was sure she was asking the right questions, formulating the right hypotheses, following the right procedures.

But the answer wasnt coming up the way she had hoped.

We were one or two experiments away from wrapping it up, and having a result to publish, recalls Gardner, a native Hawaiian and self-professed science fair nerd, who matriculated at WashU in the fall of 2003 to study biology and see where the science would take her academia, research, ultimately perhaps college administration.

And when the eager first-year student couldnt find a lab those first two semesters, she landed a summer internship at the Donald Danforth Plant Science Center and found herself in the lab of Jez, who is now the Spencer T. Olin Professor in Biology and chair of the Department of Biology in Arts & Sciences. She made an immediate impression, acing every task and earning the right to work alongside Jez and his graduate assistants.

She was utterly fearless and quite comfortable in the lab, recalls Jez, who at the time was in his second year as a lab head at the Danforth Center. We threw everything at her, and she responded beautifully.

Yet with her goal publishing her first paper before age 20 in sight, Gardner became frustrated when those last experiments didnt turn out the way she had envisioned. I was pretty upset, she says, almost in meltdown mode. Jez, ever the teacher, patiently listened to the young scientist and told her that the experiments were fine; it was her ideas that were getting in the way.

He said, Look. The data is what the data is, Gardner says. He told me not to be so in love with your ideas that youre willing to overlook the data, because thats where youre going to go wrong.

For Gardner, it was a light-bulb moment. I said, OK, well keep working the experiment, and well get to it, she says.

And get to it she did. Gardner studied the data, re-hypothesized and rethought the experiments, and ultimately published that paper, Mechanistic Analysis of Wheat Chlorophyllase Reveals a Connection to the Carboxyesterase Enzyme Family, as a sophomore. It was presented at a conference, won an award and appeared in the Notables section of the campus newspaper, The Record, a rare occasion for an undergraduate. And it was the first of a long line of published papers on the vitae for the now 34-year-old Gardner.

Gardner would work three years in Jezs lab and have eight published papers by the time she graduated in 2007. I had a bit of street cred, she says, entering Duke with more publications than most have when they leave the doctorate program.

I was brash, and I was bold. In fact, my program director told one P.I., If youre going to take her in the lab, you have to be sure you can handle her, she recalls. It made for some tough going early on, but she eventually ended up in the lab of Dukes Harold Erickson and earned her doctorate in cell biology.

All important lessons for a young scientist: the starts and the stops, two steps forward and one step back, stuck with her as she learned to trust the data at every turn. Jezs words, The data is what the data is, would become her catchphrase.

The words also would be the thread that would take her from WashU to Duke, to marriage and a family, to a career as a community college professor first in North Carolina, to ultimately the southern coast of Alabama in the Mobile metropolitan area.

Thats where this year, Gardner a Hawaiian, a wife, a mom, a scientist would throw her hat in the ring as a Democratic candidate for Congress in Alabamas 1st congressional district, a place that hasnt sent a Democrat to Congress since 1963. An unlikely candidate in an unprecedented time.

A force of nature is what she is, says Jez, who has kept in touch with Gardner through the years as both colleague and friend, and who remembers the day she told him she was a candidate. She was on the phone from a park, and you could hear her kids playing in the background, he says. Shes like, Yeah, Im running for Congress.

No rules

My life is a series of being in the right place at the right time, says Gardner via a Zoom call from her kitchen in her Spanish Fort, Ala., home with her two young sons, Ethan, 5, and Nolan, 3, playing nearby.

Its late June, about 3 weeks before a runoff election in which the voters of south Alabama will choose between two Democratic candidates for the November ballot. Its been almost four months since the initial primary in early March necessitated a runoff that can finally take place. Why? Because its a campaign season cloaked in COVID-19, with social distancing campaigns behind face masks and indoors with YouTube and Facebook Live providing both the messages and the medium.

Its good and its bad, she says, shrugging off the unprecedented challenge. As far as the pandemic goes, its a whole new world, but thats true for all the candidates. No one has the leg up here. I think Im a bit more tech savvy, though, and we already had a decent digital infrastructure set up.

When the campaign began in the summer of 2019, Gardner was a young mom using her PhD to teach biology at a community college, because thats where she felt she could be the most useful. Even when she was living in North Carolina, she gravitated toward teaching at that level. I really like the community college, the teaching philosophies and its role in society, she says.

But when the family moved to rural Alabama for her husband Matts job, she found a vastly different world, and she got involved in local politics because she wanted to improve the community in which she and Matt were raising their boys.

I had already begun to get involved, she says, insofar as How do I make sure the community in which I live is one that my children can enter into, and no one has to move across the world to get a good job?

When members of the local Democratic party witnessed her intellect and enthusiasm for public service, they came calling for a PhD with no political experience to run for Congress.

When members of the local Democratic party witnessed her intellect and enthusiasm for public service, they came calling for a PhD with no political experience to run for Congress. She first thought it was the craziest thing she had ever heard, but then she finally said yes with the support of her family. If youre going to do it, do it right and do it well, her husband advised her. Quit your job and make this what you do.

In July 2019, she made it official, and Gardner says the campaign was going pretty much as expected, with lots of hand-shaking and church picnics. But then you-know-what happened. The scientist simply took campaign matters in her own hands and pivoted. There are no rules in place for a pandemic campaign, she says. Since I had a following, and a platform and a message, the campaign pivoted from asking for votes to providing leadership.

The result was a Facebook Live video series called #QuarantinewithKiani, in which Gardner used a whiteboard to explain everything from what COVID-19 was, to how to safely re-enter the public sphere, to the importance of face masks. The campaign started food drives, distributed hand sanitizer and passed out masks. She offered step-by-step tutorials on absentee voting and helped explain personal finance in describing how best to use the stimulus checks.

Were making America to stop writing off south Alabama, and thats huge.

And she picked up endorsements. A few weeks prior to the July 14 runoff, Massachusetts Sen. Elizabeth Warren endorsed her candidacy, a rare occurrence of a national candidate paying attention to Democratic politics in Alabamas first district. Were making America to stop writing off south Alabama, and thats huge, Gardner says. Im very proud of that.

Jez, her old professor, wasnt surprised. Kianis one of these individuals who can do anything, he says.

It wasnt just her work in the lab while a WashU student that had an impact on Gardner. When Hurricane Katrina hit New Orleans in late August 2005, Gardner had just begun her junior year. She remembers the university taking in students from Tulane University, and the food drives and social awareness the hurricane generated on the Danforth Campus.

The next spring, she took a seminar-style class through American culture studies called Hurricane Katrina: A Case Study in Disaster Relief, which was team-taught and looked at the disaster from all sides. The course also included a trip to New Orleans for spring break that year.

That was really powerful, she says. It was really the first time I thought that my academic life could be about service, which paved the way for where I am now.

The following summer, I packed up my car, left my dog with a friend, and went back to New Orleans by myself to work.

She always had a good sense of helping other people and finding ways to give back, Jez says. She was constantly looking for ways to connect with people.

Gardner is in her kitchen a week after the runoff, in which she lost by a 57-43 percent margin to James Averhart, who will face Republican Jerry Carl in November.

Thats politics. Thats runoffs, and why theyre so terrible for so many reasons. I am proud of the race I ran and the campaign infrastructure I put in place.

Thats politics, she says. Thats runoffs, and why theyre so terrible for so many reasons. I am proud of the race I ran and the campaign infrastructure I put in place.

Its hard not to think what might have been. Gardner hints that shes not finished with politics yet, that shell remain involved through November and try to help as many candidates as she can while being a mom and helping her boys navigate through the pandemic. And shes still using her voice to educate voters, disseminating public health information on COVID-19 and encouraging blood donations.

Asked if, like that experiment in the Jez lab so many years ago, perhaps this election setback could eventually produce something bigger and better for her?

She laughs. Yes and no, she says. The better analogy is one of those lab experiments in which you go where the data takes you.

The primary election was an experiment, we got that data, moved forward. July 14 was an experiment. Got that data, moved forward. Now its time to chart a specific aim. Between the virus and the runoff, for a lot of people it exposed the weaknesses of our electoral system that day-to-day voters arent aware of.

Shes proud of the fact, for example, that her campaign pushed hard for no-excuse absentee voting, and it will be allowed in Alabama this November. When we did that work, a lot of people saw how difficult absentee balloting is, Gardner says, and how the lack of it disenfranchises communities of color, low-income communities and, in a major way, disabled communities.

Alexandria Ocasio-Cortez is so inspiring, but not everyone can go from bartender to Congress. We need narratives that are not about superstars, but just super interesting women

Whats more, she hopes her story might be inspiring to other women who are thinking about entering the political arena.

I think we are inundated with stories of these amazing political successes, she says. Alexandria Ocasio-Cortez is so inspiring, but not everyone can go from bartender to Congress. So many women who aspire to national office dont see a path for themselves, so there has to be a story thats told that says, Heres what it looks like to get involved.

It pushes back against the burdensome narrative that women can have it all. You dont have to have it all, but you can do it all because it all matters: City council. School board. Moms Demand Action. The Wall of Moms. We need narratives that are not about superstars, but just super interesting women.

And whatever comes next, Gardner will continue to define herself by being Hawaiian, and a wife, and a mom, as a scientist because the data is what the data is.

Its this really powerful idea that we live in the world we live in, Gardner says, and it doesnt necessarily matter what your idea of perfect is, or what you wish had happened.

You have to deal with what you have right now.

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Following the data - Washington University in St. Louis Newsroom

Cell Biology

Rockville cell and gene company may have found treatment for HIV – BethesdaMagazine.com

Jeff Galvin believes his Rockville cell and gene technology company has found a breakthrough treatment for people living with HIV

By David Goldstein

| Published: 2020-08-17 08:31

When Jeff Galvin was 13 years old, he came across a lone computer in the basement of Muzzey Junior High School in Lexington, Massachusetts. It was actually a teletype machine attached to a minicomputer. This was, after all, the 1970s.

He doesnt remember why it was there, but he got permission to use it and taught himself how to program. My first love affair, Galvin recalls. My head exploded with the possibility that you had this thing that never got tired. You just fed it electricity.

At 15, he was taking classes at Massachusetts Institute of Technology (MIT) in nearby Cambridge and teaching computer science to high school and college students on weekends. This may be starting to sound like Good Will Hunting, the 1997 film in which actor Matt Damon plays an MIT janitor who secretly solves complex equations on a classroom blackboard, but theres a difference. He was a math genius, Galvin says. I was a highly passionate, excited kid who had just seen the most amazing toy in history.

In his 20s, Galvin was an early recruit to Silicon Valley in those heady nascent days of the 1980s when it was fast becoming the high-tech Xanadu. His time in California included six years at Apple, but it was a visit to the National Institutes of Health in 2007 that took his career in a much different direction.

Now 61, hes the founder and CEO of a Rockville cell and gene technology company, and he believes his company stands on the cusp of a medical breakthrough. Galvins team of molecular biologists at American Gene Technologies (AGT) thinks it has developed a gene therapy procedure that can cure HIV, the virus that leads to AIDS. According to the U.S. Department of Health and Human Services, 1.1 million people are currently living with HIV in this country; the World Health Organization estimates that 75 million people across the globe have been infected with the virus since the AIDS epidemic began in 1981. About 32 million have died.

Galvin says he made HIV his target because a cure for the virus has remained elusive. People with HIV must take a daily regimen of medications that control infections and suppress the virus, but dont eliminate it. Over time, these drugs can cause everything from nausea and fatigue to more serious conditions affecting the kidneys, heart and central nervous system, according to NIH.

He believes the gene therapy procedure AGT has developed, in which genes are transferred to modify cells or tissue through the use of viral vectors, could actually be a platform to cure any number of diseases.

What cant we cure with this? Galvin says during a call to the head of a financial securities firm one afternoon in February. Hes seated at a large oval table in the AGT conference room with the phone on speaker.

Galvin spends much of his time wooing potential investors. What he brings to the task is his salesmans personality: a bit over the top, a healthy dollop of bravado, and an inclination to push the envelope. And his companys HIV treatment is getting a serious lookthe U.S. Food and Drug Administration is determining whether the procedure is safe enough for human testing.

Galvin says the gene and cell therapy industry has been exploding in recent years, causing a slowdown in the regulatory process for those kinds of treatments. But AGT is hopeful that the FDA will approve the human clinical trials by this fall. I cant imagine we cant cure almost everything in the world, he says on the call. Were going to send chemotherapy and radiation the way of bloodletting and leeches.

Galvin is a ball of energy and nonstop talker. Politics. Facebooks troubled relationship with privacy. Hell opine as long as someone will listen and he doesnt have a pressing appointment. He is 6 feet tall, has a genial smile and an eagerness to engage. And he burns with the passion of the committed.

Were down here in Rockville, Maryland. Lots of good opportunities down here, he says during the pitch, which goes on for more than an hour. Right now [we have] the beginnings of what is turning into a revolution in pharmaceuticalswere talking about the creation of probably another $3 to $5 trillion industry over the next 10 to 15 years. I think this is bigger than the dot-com boom. I think it doesnt have quite the same bubble. This isnt the kind of thing that will all go off the cliff simultaneously like dot-com because theres real science behind it. And if it works out, it can be monetized.

Galvin is a cheerleader for Montgomery Countys growth as a hub for health technology. AGT is just off Interstate 270 at Exit 6B in the life sciences corridor, where it percolates among a cluster of tech, biotech and pharmaceutical companies, as well as educational centers. Branches of the University of Maryland and Montgomery College help incubate new startups. Thats the atmosphere that got AGT off the ground in 2008.

In addition to HIV, AGT is developing therapies for several types of cancerous tumors that affect the breasts, lungs and prostate. The company hopes that its work on a treatment for liver cancer will be approved for human clinical trials in 2022. The lab is also working on a gene therapy for phenylketonuria, known as PKU, a rare inherited metabolic disorder.

Theres this great economic engine which is evolving that will make Maryland the next Silicon Valley, Galvin tells the head of the financial securities firm. I call it DNA Valley.

Galvin, it should be noted, is not a scientist. Hes an economist by training, a 1981 graduate of Harvard University. Hes a computer prodigy by pedigreehis mother was one of the rare female computer software experts in the 1960s, and his father is an MIT-trained electrical engineer who did national security work.

Galvin has this thing about disruptive technologies, he says, systems that upend the old way of doing something and change the culture in a significant way. Like how Apple co-founder Steve Jobs simplified and popularized the computer mouse. Or how GPS changed the way we get from here to there. Thats how he sees AGT and gene therapy. Ive been through a lot of technologies: computers, software, the internet, apps, IT, Galvin says.

So I understand how these technologies grow. Gene and cell therapies are bigger than any of those, and its much more emotional because its your health.

He has a fluency in arcane subjects that arent connected to his own skills. This is Galvins explanation for how HIV infects a cell through a protein known as CCR5, located on the surface of white blood cells, and why AGT believes it has developed a defense: One element of our cell product is the removal of CCR5 from the surface of CD4+ T cells. Howeverwe have also added siRNAs against conserved regions of the vif and tat HIV genes for additional protection against R5 viruses as well as extending protection to CXCR4 versions of HIV.

Heres how he puts it in plain English: We have removed thedoor handle (CCR5) that HIV uses to get into cells. Most forms of HIV use that common surface protein in cells to infect the cell, but some forms do not need that handle. AGT has added specially designed genes to our HIV treatment that are capable of producing substances inside the cell that protect against several known mutations of HIV that do not require that handle. AGT is the first company to provide this type of broad protection to the various known versions of HIV.

HIV is an insidious virus that infects a patients T cells, a type of white blood cell that helps the body fight off infection. In Montgomery County, 3,489 people were infected with HIV between 2009 and 2018, with 123 new cases in 2018, according to the Maryland Department of Health. AGTs gene therapy approach modifies the HIV-specific T cells so they can resist infections and do their job of protecting the body from pathogens that cause disease.

AGT does this by using viral vectors; the viruses are cracked open to remove the bad genes and replaced with newly modified genes that will improve the cell. Instead of a virus with the intention of infecting you and going to the next person, its been tamed to do only one part of that process, says C. David Pauza, a molecular biologist and longtime researcher in gene cloning and HIV who serves as AGTs chief science officer. We put things in it we want it to deliver and it makes one infection and stopsand doesnt go any farther. According to Galvin, once HIV T cells are able to carry out their protective work as intended, HIV patients would eventually become permanently immune to the virus and hopefully have no need to continue taking antiretroviral drugs.

AGTs concept is not new, according to Carl Dieffenbach, director of the Division of AIDS at NIHs National Institute of Allergy and Infectious Diseases. What has improved are the vectors, he says. Its reasonable to continue to watch this. Whats unknown, Dieffenbach says, is the human reaction. Once the therapy is tried on people, what you dont know is how this will actually behave.

Tami Howie, an attorney who represents tech and biotech companies, first met Galvin in 2017, when she was CEO of the Maryland Technology Council. He is one of the rare people who epitomizes the convergence in tech and biotech, she says. Hes totally cutting edge.

But messing around with the human genome can be fraught with risk. We know there are viruses out there that get into your body and activate genes that do all kinds of good things and bad things, Pauza says. [One] good thing is a very old virus thats in most of us that controls the efficiency of pregnancy in women. But then you can get other viruses that go in and they stimulate things to be made inappropriately and trigger horrible results.

To Galvin, no risk, no reward. When he made public late last year that AGT was developing a cure for HIV, he was criticized by some in the HIV/AIDS community for peddling false hope. But Galvin has no regrets and is sympathetic to their concerns. Its reasonable to experience a gut reaction to the word cure when humanity has been struggling against HIV for decades, he says. I dont fault anyone for working to protect their community. We are always clear that we will only know for sure once we prove it in a human trial, he says. He calls AGTs work the future of medicine.

Its going to be typical that many of the diseases that strike you are going to be cured by gene and cell therapy, he tells the prospective investor. And we plan to lead that revolution becausewere going to prove were the most efficient competitor in it by curing HIV this year.

If hes right, that would be seismic.

Enormous, Pauza says.

The oldest of three children, Galvin didnt get a lot of attention when he was young so he tended to get into a lot of trouble, he says. I figured out how things worked early, so it was hard to lock me in the house.

As a child, hed come home with a bloody hand, having found a razor blade, or wander into a snowstorm looking for twigs for the fireplace. His father, Aaron, says Galvin was a handful because his mind was so active. We were on a first-name basis with all the emergency wards, he says with a chuckle.

Galvin showed his entrepreneurial spirit early on, his father says. When he was 6 and the familys house in Lexington was under construction, the boy collected dirt from the excavation site and sifted it through a window screen and into plastic bags. Then he pulled his little red wagon up and down the street, selling topsoil for 50 cents a bag.

Harvard didnt offer a degree in computer science when he went there, so Galvin majored in economics and took all the computer classes he could. After graduation, he left for California to take a job with Hewlett-Packard, but he didnt like the corporate culture and went to work for Apple. Apple was more like me, Galvin says. It was in love with what computers could do.

He stayed on the West Coast for parts of the next three decades. Silicon Valley was on fire when I was there, says Galvin, whose sister, Laurie, and brother, Mark, also pursued careers in the tech world. He describes the atmosphere there as a group of people with the right ideas andpushing things at light speed. There was such a clarity of vision and purpose. There was no limit on what we could do there.

It was a fast life, though. As a marketing manager and later director of international marketing for Claris, an Apple spinoff, Galvin was always on the move: Europe, the Middle East, Indonesia, New Zealand. While in Paris on a trip to show clients how to service Apple hardware, he fell asleep at the wheel while driving through a tunnel at 70 mph after having been up for three days. When he brushed a curb, he awoke and was able to right the car. But it was a chilling experience.

How lucky can you get? Galvin says.

In 2001, after nearly two decades of 16-hour workdays, he decided he was done. At 42, hed made investments in startups, real estate, software and internet companies. I looked at my bank account and realized I didnt have to work anymore, Galvin says.

He bought a house on Maui and traveled between there and his home in San Carlos in Silicon Valley. He started dating, thinking he should settle down before he got too old. He and his wife, Cherry, married in 2004.

The couple enjoyed a carefree life in the tropics. But after five years, Galvin was bored out of my mind, so they moved back to California. He decided to tiptoe back into the game by looking for a project where he could become an angel investor, someone who puts money behind a startup often in exchange for ownership equity. The word got out, and Galvin received a proposal from a postdoctoral researcher at a lab at NIH headed by Dr. Roscoe Brady, a renowned biochemist and pioneer in the treatment of enzyme deficiencies. Galvin visited the lab in 2007 and met Brady, who explained the science behind viral vectors. It was Galvins eureka moment.

When I learned that there was a mechanism to update the DNA in a human cell, my head practically exploded, Galvin says. He reasoned that if you look at a cell as the human bodys computer, and the DNA contained in the cell as the operating system, you could employ viral vectors to convert viruses into updates for the human computer and thereby correct defects.

DNA is the instruction set for the cell, Galvin says. Your genes are just instructions to make enzymes and proteins that then react in the cell. Basically, your cell is an organic computer. You change the software, you change the cell.

Brady was retiring and NIH was closing his lab. Galvin says NIH gave him the intellectual property, free of charge, on the condition that he continue the research. I felt like he was close to making major breakthroughs, Galvin says. So he hired two of Bradys research assistants and signed Brady on as a scientific adviser. Thats how AGT was born.

In the early days, Galvin continued to live on the West Coast and funded the work out of his own pocket. His mother, Frayda, was suffering from Stage 4 pancreatic cancer. She died in 2009 after battling the disease for 15 months. Knowing what she went through bolstered Galvins conviction that AGTs work could help people. In 2010, he realized he needed to be in Rockville full time. Over the next few years, he received $1 million in grants from NIH, which was interested in innovative approaches to gene and cell therapies. Galvin says obtaining funding that way was easier than trying to lure investors, and he kicked in an additional $2 million.

AGT began as a small but determined undertaking. An early supporter was Dr. Robert Redfield, a virologist who now heads the Centers for Disease Control and Prevention and became visible during the White House coronavirus briefings in the spring. He served as an adviser to AGT beginning in 2011 and subsequently chaired the companys clinical advisory board for its HIV gene therapy program until 2018. By that time, AGT had been granted patents for its HIV therapy and had enlisted NIH as a research partner in its search for a cure. Fast forward to today, Galvin says, we have curative therapies for HIV.

Galvins day usually begins in my sweats, he says, in the kitchen of his Rockville condo where he has a three-screen computer. He calls it command central. He can work uninterrupted for about three hours, and hes usually at AGT by noon. He presides over a three-story warren of offices where employees handle regulatory affairs, marketing, finance and other administrative business.

Its in the labs, spread over 11,500 square feet on the third floor, where the microbiologists and other scientists do the research and delicate work of separating genes, the link between one generation and the next. Behind glass walls and clad in blue protective gowns and gloves, they employ an array of biological tools as they work with T cells, viral vectors and other microscopic particles. Their findings spill out of white data machines spaced at regular intervals along the work counters.

Talking up AGT requires Galvin to travel a lot. Before the coronavirus pandemic, he was often on trains to Manhattan for meetings and dinners with prospective investors, and flying around the country for conferences and other events. But the world has new rules these days. Video conference calls and Zoom meetings have become the new way of doing business.

To relax, Galvin plays Xbox or watches The Simpsons. He enjoys how the shows writers parody American culture.

I can tell these people feel the same way about the world as I do, he says. He also reads or catches up on the news. On his nightstand in February was Sapiens: A Brief History of Humankind, a New York Times bestseller.

His wife, Cherry, prefers warm weather, so she spends her winters at the couples home in Silicon Valley. They got married late in life, Galvin says, and dont have children. Cherrys sister, a single mother, lived with the couple and in 2004 sent for her daughter, who was 6 years old and staying with her grandmother in Wuhan, China. Now 21, Galvins niece, Jesse, recently graduated from the University of Washington, where she majored in molecular and cellular biology. Interning at AGT during her high school summers might have had something to do with that. I love my niece like a daughter, Galvin says.

Galvin says hes having the most fun hes ever had. Still, securing funding for new therapies and medicines can be a struggle if youre not a pharmaceutical giant. Investors generally want a quick return, and human trials for a new drug or therapy can be costly.

To be a successful entrepreneur and forging new territory, you get a lot of arrows in the back. A lot of people say no way, says Drew Palin, a physician and chief innovation officer of Intellivisit, a Madison, Wisconsin, online medical diagnosis company. Hes also one of Galvins investors. He has had to raise lots of nickels and dimes that allowed him to be a little more patient and persistent. To do that, you have to have a lot [of] drive, a lot of personal passion and a way to survive.

When Galvin isnt pitching AGTs upside to venture capitalists, blue-chip finance houses like JPMorgan Chase, big financial institutions like Citibank, and angel investors like Palin, hes championing the promise of genetic engineering.

Im basically an evangelical person that is spending his entire day either connecting people with our mission and trying to engage them [in] some way to support it or propel it, or inspiring people to achieve greatness within the mission, Galvin says.

Should AGTs experimental HIV treatment work and eventually be approved for commercial sales, Galvin says it would likely be licensed to a large pharmaceutical company with global reach to ensure the products wide availability.

Galvin doesnt have any idea how much the treatment could cost, but predicted it would be less expensive than what insurance companies now pay to cover the costs of daily antiretroviral treatments, medicine for the side effects and appointments with doctors. Hes not chasing the dollar. Hes chasing every life he can save, Tami Howie says. He figures every minute hes not working, people are dying.

Galvins fundraising has pushed investments to $40 million, and his staff has grown to 30. His commitment to AGT, however, put a strain on both his marriage and retirement nest egg. Both have survived and recovered, he says, but it took sustained engagement with high risk and giant potential downfall for me.

Now he weighs the possibility that it all could pay off. If we get a handful of cured HIV patientsI think the [National] Mall is going to look like the days of the AIDS quilt because these people have been suffering for so long, Galvin says, and I think the emotional response is going to be quite profound.

David Goldstein is a former political and investigative reporter in Washington, D.C., for McClatchy Newspapers and The Kansas City Star.

Continued here:
Rockville cell and gene company may have found treatment for HIV - BethesdaMagazine.com

Cell Biology

Scientists hope to find clues about how life emerged by tinkering with its oldest components – News-Medical.net

Reviewed by Emily Henderson, B.Sc.Aug 15 2020

"I'm fascinated with life, and that's why I want to break it."

This is how Betl Kaar, an assistant professor at the University of Arizona with appointments in the Department of Molecular and Cellular Biology, Department of Astronomy and the Lunar and Planetary Laboratory, describes her research. What may sound callous is a legitimate scientific approach in astrobiology. Known as ancestral sequencing, the idea is to "resurrect" genetic sequences from the dawn of life, put them to work in the cellular pathways of modern microbes - think Jurassic Park but with extinct genes in place of dinosaurs, and study how the organism copes.

In a recent paper published in the Proceedings of the National Academy of Sciences, Kaar's research team reports an unexpected discovery: Evolution, it seems, is not very good at multitasking.

Kaar uses ancestral sequencing to find out what makes life tick and how organisms are shaped by evolutionary selection pressure. The insights gained may, in turn, offer clues as to what it takes for organic precursor molecules to give rise to life - be it on Earth or faraway worlds. In her lab, Kaar specializes in designing molecules that act like tiny invisible wrenches, wreaking havoc with the delicate cellular machinery that allows organisms to eat, move and multiply - in short, to live.

Kaar has focused her attention on the translation machinery, a labyrinthine molecular clockwork that translates the information encoded in the bacteria's DNA into proteins. All organisms - from microbes to algae to trees to humans - possess this piece of machinery in their cells.

We approximate everything about the past based on what we have today. All life needs a coding system - something that takes information and turns it into molecules that can perform tasks - and the translational machinery does just that. It creates life's alphabet. That's why we think of it as a fossil that has remained largely unchanged, at least at its core. If we ever find life elsewhere, you bet that the first thing we'll look at is its information processing systems, and the translational machinery is just that."

Betl Kaar, Assistant Professor, University of Arizona

So critical is the translational machinery to life on Earth that even over the course of more than 3.5 billion years of evolution, its parts have undergone little substantial change. Scientists have referred to it as "an evolutionary accident frozen in time."

"I guess I tend to mess with things I'm not supposed to," Kaar said. "Locked in time? Let's unlock it. Breaking it would lead the cell to destruction? Let's break it."

The researchers took six different strains of Escherichia coli bacteria and genetically engineered the cells with mutated components of their translational machinery. They targeted the step that feeds the unit with genetic information by swapping the shuttle protein with evolutionary cousins taken from other microbes, including a reconstructed ancestor from about 700 million years ago.

"We get into the heart of the heart of what we think is one of the earliest machineries of life," Kaar said. "We purposely break it a little, and a lot, to see how the cells deal with this problem. In doing this, we think we create an urgent problem for the cell, and it will fix that."

Next, the team mimicked evolution by having the manipulated bacterial strains compete with each other - like a microbial version of "The Hunger Games." A thousand generations later, some strains fared better than others, as was expected. But when Kaar's team analyzed exactly how the bacteria responded to perturbations in their translational components, they discovered something unexpected: Initially, natural selection improved the compromised translational machinery, but its focus shifted away to other cellular modules before the machinery's performance was fully restored.

To find out why, Kaar enlisted Sandeep Venkataram, a population genetics expert at the University of California, San Diego.

Venkataram likens the process to a game of whack-a-mole, with each mole representing a cellular module. Whenever a module experiences a mutation, it pops up. The hammer smashing it back down is the action of natural selection. Mutations are randomly spread across all modules, so that all moles pop up randomly.

"We expected that the hammer of natural selection also comes down randomly, but that is not what we found," he said. "Rather, it does not act randomly but has a strong bias, favoring those mutations that provide the largest fitness advantage while it smashes down other less beneficial mutations, even though they also provide a benefit to the organism."

In other words, evolution is not a multitasker when it comes to fixing problems.

"It seems that evolution is myopic," Venkataram said. "It focuses on the most immediate problem, puts a Band-Aid on and then it moves on to the next problem, without thoroughly finishing the problem it was working on before."

"It turns out the cells do fix their problems but not in the way we might fix them," Kaar added. "In a way, it's a bit like organizing a delivery truck as it drives down a bumpy road. You can stack and organize only so many boxes at a time before they inevitably get jumbled around. You never really get the chance to make any large, orderly arrangement."

Why natural selection acts in this way remains to be studied, but what the research showed is that, overall, the process results in what the authors call "evolutionary stalling" - while evolution is busy fixing one problem, it does at the expense of all other issues that need fixing. They conclude that at least in rapidly evolving populations, such as bacteria, adaptation in some modules would stall despite the availability of beneficial mutations. This results in a situation in which organisms can never reach a fully optimized state.

"The system has to be capable of being less than optimal so that evolution has something to act on in the face of disturbance - in other words, there needs to be room for improvement," Kaar said.

Kaar believes this feature of evolution may be a signature of any self-organizing system, and she suspects that this principle has counterparts at all levels of biological hierarchy, going back to life's beginnings, possibly even to prebiotic times when life had not yet materialized.

With continued funding from the John Templeton Foundation and NASA, the research group is now working on using ancestral sequencing to go back even further in time, Kaar said.

"We want to strip things down even more and create systems that start out as what we would consider pre-life and then transition into what we consider life."

Source:

Journal reference:

Venkataram, S., et al. (2020) Evolutionary stalling and a limit on the power of natural selection to improve a cellular module. PNAS. doi.org/10.1073/pnas.1921881117.

Original post:
Scientists hope to find clues about how life emerged by tinkering with its oldest components - News-Medical.net

Cell Biology

High Point University Students, Faculty and Staff Recognized for Research and Innovation – Yes! Weekly

HIGH POINT, N.C., Aug. 14, 2020 Members of the High Point University community frequently conduct, publish and share research and creative works in a variety of ways. Below is a recap of recent research initiatives.

HPU Student, Alumna and Faculty Research Featured in National Scientific Journal

Casey Garr, HPU alumna; Candyce Sturgeon, HPU rising senior; Dr. Veronica Segarra, HPU assistant professor of biology; and Noah Franks, student at Penn Griffin School of the Arts in High Point, North Carolina; recently conducted research that was published in Autophagy, a national scientific journal.

Dr. Veronica Segarra, assistant professor of biology, recently co-authored research that was published in Autophagy, a national scientific journal.

The study, titled, Autophagy as an on-ramp to scientific discovery, examines HPUs Cell Art Collaborative program to gain understanding around how the recruitment of highly creative students into STEM fields through connections to art can be a first step in defining a specialized career path that leads to a valuable and unique contribution to science.

In addition to providing experiential learning opportunities for students at HPU to conduct hands-on research and co-author peer-reviewed articles, the Cell Art Collaborative program encourages students in the local community to explore careers that incorporate both science and art, says Segarra. This initiative continues to facilitate conversations around STEAM-based learning environments for educators to take advantage of a wider range of student talents and interests, preparing them to go forth into society as the creative thinkers and problem solvers the world needs.

HPU Students Research Featured in CBE: Life Sciences Education Journal

Clara Primus, a rising junior majoring in biology and Bonner Leader at HPU, recently collaborated with prominent scientists to conduct research that was published in CBE: Life Sciences Education, a quarterly journal published by the American Society for Cell Biology.

Clara Primus, a rising junior majoring in biology and Bonner Leader at HPU, recently collaborated with prominent scientists at the Mayo Clinic, University of California Davis and Northwestern to conduct research that was published in CBE: Life Sciences Education, a quarterly journal published by the American Society for Cell Biology. The article, titled, Scientific Societies Fostering Inclusive Scientific Environments through Travel Awards: Current Practices and Recommendations, examines how scientific societies can contribute to a diverse and inclusive workforce.

The research compares and contrasts the broad approaches that scientific societies within the National Science Foundation-funded Alliance to Catalyze Change for Equity in STEM Success (ACCESS) use to implement and assess their travel award programs for underrepresented minority (URM) trainees. Findings will improve collaboration and better position scientific societies to begin addressing some of these questions and learning from each other.

The recommendations included in this research shed light on how even scientific societies can be allies in furthering inclusion efforts, said Primus. Ive spent nearly two years studying equity and diversity, and I hope that I can take the knowledge Ive learned from all of my research to educate my peers at HPU.

HPU Exercise Science Professor Publishes Statement for the American Heart Association

Dr. Colin Carriker, assistant professor of exercise science in HPUs Congdon School of Health Sciences, recently co-authored a scientific statement for the American Heart Association (AHA).

Dr. Colin Carriker, assistant professor of exercise science in HPUs Congdon School of Health Sciences, recently co-authored an American Heart Association (AHA) scientific statement on medicinal and recreational cannabis use published in Circulation.

The statement critically reviews the use of medicinal and recreational cannabis from a clinical but also a policy and public health perspective by evaluating its safety and efficacy profile, particularly in relation to cardiovascular health. The purpose of this scientific statement was to explore the evidence and science pertaining to medical marijuana, recreational cannabis and cardiovascular health to provide physicians and health care providers with the information available to date. While cannabis may have some therapeutic benefits, these do not appear to be cardiovascular in nature. Health care providers would benefit from increased knowledge, education and training pertaining to various cannabis products and health implications, including recognition that cannabis use may, in fact, exacerbate cardiovascular events or other health problems. In this regard, the negative health implications of cannabis should be formally and consistently emphasized in policy, while aligning with the American Heart Associations commitment to minimizing the smoking and vaping of any products and banning cannabis use for youth.

It was an honor to work alongside such a high-quality team of researchers, says Carriker. I want to especially thank our committee chairs, Dr. Robert L. Page II and Dr. Larry A. Allen, as their extraordinary leadership and organization were integral components in the completion and publication of this AHA scientific statement. We publish these statements to counterbalance and debunk misinformation because the public requires high-quality information about cannabis from reputable organizations such as the American Heart Association.

Carriker is the advocacy ambassador for the American Heart Associations Council on Lifestyle and Cardiometabolic Health and served as a member of the writing committee tasked with writing this AHA Scientific Statement initiated by the AHAs Council on Clinical Cardiology.

At High Point University, every student receives an extraordinary education in an inspiring environment with caring people. HPU, located in the Piedmont Triad region of North Carolina, is a liberal arts institution with 5,400 undergraduate and graduate students. It is ranked No. 1 by U.S. News and World Report for Best Regional Colleges in the South, No. 1 for Most Innovative Regional Colleges in the South and No. 1 for Best Undergraduate Teaching in the South. The Princeton Review named HPU in the 2020 edition of The Best 385 Colleges and on the Best Southeastern Colleges 2020 Best Colleges: Region by Region list. HPU was recognized as a Great School for Business Majors and a Great School for Communication Majors. HPU was also recognized for Most Beautiful Campus (No. 18), Best College Dorms (No. 5) and Best Campus Food (No. 20). For nine years in a row, HPU has been named a College of Distinction with special recognition for business and education programs and career development, and The National Council on Teacher Quality ranks HPUs elementary education program as one of the best in the nation. The university has 60 undergraduate majors, 63 undergraduate minors and 14 graduate degree programs. It is a member of the NCAA, Division I and the Big South Conference. Visit High Point University on the web at highpoint.edu.

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Cell Biology

Super Resolution – posted by Biophotonics.World at Biophotonics.World – Biophotonics.World

Image source: Leibniz IPHT

By: Sven Dring

When trying to visualize finest details in cells, standard light microscopes reach their limits. As a doctoral student, Rainer Heintzmann discovered a method that can break this barrier. Today, he has improved the technique of super-resolution microscopy to such an extent that it is useful for applications in biology and medicine

In order to observe living cells at work, researchers have to overcome a physical law. One of the fastest techniques to overcome the resolution limit of classical light microscopy is high-resolution structured illumination microscopy.

It makes details in cells visible that are about one hundred nanometers in size, one hundred millionth of a millimeter. However, translating the recorded data back into images used to take a lot of time. Rainer Heintzmann, together with a team of researchers from Bielefeld University, has developed atechnique that allowsthe image data to be reconstructed directly.This allows researchers to watch biologicalprocesses in the cellvirtually live. "It enables completely newimaging workflows that no other high-resolution microscopy method currently allows in this way," says Rainer Heintzmann.

The graphics helps computer gamers to have a great gaming experience. Researchers use it to observe the smallest cell components in action - in real time and at a very high frame rate. "The image data can be reconstructed about twenty times faster than it would take on a PC," explains Rainer Heintzmann, who already laid the foundations for the structured illumination method in high-resolution microscopy as a doctoral student in 1998. In cooperation with the Bielefeld research team led by Thomas Huser, he further developed the technique of Super-Resolved Structured Illumination Microscopy (SR-SIM).

In the fluorescence microscopic SR-SIM method, objects are irradiated with laser light using a special pattern. It excites special fluorescent molecules in the sample so that they emit light at a different wavelength. The microscopic image then shows this emitted light. It is first recorded in several individual images and then reconstructed as a high-resolution image on a computer. "The second step in particular has taken a lot of time so far," says Andreas Markwirth from Bielefeld University, first author of the study, which the research team published in the renowned journal "Nature Communications".

For the new microscope, the research team used parallelcomputer processes on modern graphics and was thus able to significantly accelerate image reconstruction. A minimum delay of 250 milliseconds is hardly noticeable to the hu- man eye. The raw data can also be generated faster with the newly researched microscope.

Structures that are invisible to conventional microscopes

"This makes it possible to measure samples quickly and to immediately adjust test conditions during an experiment instead of having to evaluate them afterwards," says Rainer Heintzmann, describing the practical benefits of the new technology. It is only through the rapid reconstruction of images that "this type of microscopy becomes really useful for applications in biology or medicine," says Thomas Huser. "Because the problem so far is: microscopes that offer sufficiently high resolution cannot display information at the appropriate speed".

For their study, the scientists tested the method on biological cells and recorded the movements of mitochondria, the energy centers of the cells that are about one micrometre in size. "We were able to generate about 60 frames per second that's a higher frame rate than in motion pictures. There are less than 250 milliseconds between measurement and image, so the technology allows real-time recordings," says Andreas Markwirth.

Until now, super-resolution methods have often been combined with conventional methods: A conventional fast microscope is used to find structures first. These structures can then be examined in detail with a super-resolution micro- scope. "However, some structures are so small that they cannot even be found with conventional microscopes, for example special pores in liver cells. Our method provides both high resolution and speed this enables biologists to investigate such structures," said Thomas Huser. Another application for the new microscope is the investigation of virus particles on their way through the cell. "This enables us to understand exactly what happens during infection processes."

Publication: Andreas Markwirth, Rainer Heintzmann et al., Video-rate multi-color structured illumination microscopy with simultaneous real-time reconstruction, Nature Communications 10 (2019), https://doi.org/10.1038/s41467-019-12165-x

Source: Leibniz IPHT

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Super Resolution - posted by Biophotonics.World at Biophotonics.World - Biophotonics.World