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Alexandra Taraboletti, PhD

Postdoctoral Fellow, Georgetown University, Washington, DC, USA

Alexandra Taraboletti

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Biography

Dr Alexandra Taraboletti is a postdoctoral fellow training in the Tumor Biology program funded through a T32 Ruth L. Kirschstein National Research Service Award at Georgetown University Lombardi Comprehensive Cancer Center. Dr Taraboletti received her PhD from the University of Akron in Chemistry where she used mass spectrometry-based metabolomics to investigate models neurodegeneration in multiple sclerosis and myelin biology. In Dr Fornace’s lab, Alexandra applies metabolomics to assess biomarkers in easily accessible fluids following radiation injury. Specifically, she has worked on a new technology development for metabolomic-based biodosimetry, and is exploring protection methods to counter the negative impact of radiation in the brain. She uses a cell culture model of the myelin producing oligodendrocyte cell to examine in vitro effects of radiation. She is currently testing the use of dimethyl fumarate as a neuroprotectant against radiation injury. At Georgetown, Dr Taraboletti is a Co-Chair for the Georgetown Postdoc Association—an establishment that acts to foster camaraderie and organize career development activities specifically geared towards postdocs. Her work with the Georgetown Postdoc Association has led to the developing stages of a new workshop series, which she hopes will foster teaching/higher education skills in interested graduate students and postdoctoral fellows. Dr Taraboletti, also has a passion for visual science communication, and applies her skills at the graphical editor and illustrator for The POSTDOCket (the newsletter of the National Postdoctoral Association). She is an avid artist, and is always looking to grow in the bourgeoning field of #sciart.

Learn about Alexandra’s research

Title: Understanding and ameliorating radiation-induced damage to oligodendrocytes

Learning objectives

  • Understanding the current state of brain radiation treatment and patient outcomes and quality of life that can follow therapy
  • Detailing basic glial cell biology, and how to use the MO3.13 cell line to model immature and mature differentiated oligodendrocyte cells
  • Understanding mass-spectrometry based metabolomics and its use in monitoring markers of radiation injury and oligodendrocyte metabolism

Radiation therapy is a critical tool for the treatment of brain tumors, however, exposure to high doses of ionizing radiation (IR) causes numerous central nervous system side-effects, including declines in cognitive function, memory, and attention. Brain injury from IR is characterized by numerus inflammatory effects, including white matter damage from the loss of myelin-producing oligodendrocyte cells. While neuro-oncology outcomes are often concerned with survival, strategies to understand and ameliorate radiation-induced damage after IR treatment are needed to preserve and improve patient quality of life. Our lab is interested in studying the differential effects of radiation on oligodendrocyte cells, as they comprise the majority of white matter in the brain, and methods to halt radiation-induced damage. We have established a mass spectrometry-based metabolomics method to study radiation-injury in cells, tissue, and biofluids, and are applying this technique to study radiation effects on the MO3.13 oligodendrocyte cell line.

We are currently investigating the ability of dimethyl fumarate (DMF), an established neuroprotective agent, to amend damage and demyelination to oligodendrocyte cells versus glioma cells, after X-irradiation. Using metabolomics, we noted that oligodendrocyte cells upregulated tricarboxylic acid (TCA) cycle intermediates in response to DMF treatment, with sustained levels after radiation. In addition, measured levels of glutathione were elevated, and markers for generalized oxidative stress were comparably lower with DMF pretreatment. Ultimately, this information could be used to prevent radiation-induced demyelination, promoting patient quality of life.

Watch the webinar

(00:00)

Slide 1

Moderator:  Hello everyone and welcome to today's live webinar, Understanding and Ameliorating Radiation-Induced Damage to Oligodendrocytes, presented by Dr. Alexandra Taraboletti.  I'm Christy Jewell of LabRoots and I'll be your moderator for today's event. Today's educational web seminar is presented by LabRoots and brought to you by Thermo Fisher Scientific.  To learn more about our sponsor, please visit www.thermofisher.com/cellcultureheroes.  (00:34)

Now let's get started.  Today’s webinar is interactive.  We encourage you to participate by submitting as many questions as you want, at any time you want during the presentation.  To do so, simply type them into the Ask a Question box and click Send.  We will answer as many questions as we have time for at the end of the presentation. If you have trouble seeing or hearing the presentation, just use the Ask a Question box to let us know you’re having a problem. (1:04)  If you have any trouble seeing or hearing the presentation, just let us know you’re having a problem by using the Ask a Question box.

This presentation is educational and thus offers continuing education credits.  Click on the Continuing Education Credits tab and follow the process to obtain your credits.  I now present today’s speaker, Dr. Alexandra Taraboletti, Post-Doctoral Fellow, Georgetown University.  For a complete biography on our speaker, please visit the biography tab at the top of your screen. (1:38)  Dr. Taraboletti, you may now begin your presentation.

(1:50)

Dr. Alexandra Taraboletti:  Thanks, Christy, for that great introduction, and thank you so much for hosting me as the Gibco Cell Culture Hero.  Today I hope to host a really accessible talk that covers the basics of white matter damage from radiation therapy, with the title Understanding and Ameliorating Radiation-Induced Damage to Oligodendrocytes, and also go into how I use cell culture and metabolomics to better understand this issue. (2:16)  But before I dive into this topic, let me first tell you a little bit about myself.  My bachelor’s is actually in chemistry, I kind of journeyed to far away.

(2:31)

Slide 2

I got my Bachelors at University of Central Florida and I started in all things in material science, where I did mechanochemistry, doping materials with catholuminescent properties.  Tangentially, I also worked with crickets of all things, so if anyone ever needs to sex a cricket, I’m that person.  I used these to look at temperature changes in respect to global warming.  So again a very diverse background. (3:00)  But the chemistry and the biology led me to go into biochemistry.

My Ph. D. is in metabolomics and neurobiology, technically my degree is in chemistry from the University of Akron.  But I looked at global metabolomics and I investigated glial metabolism, which I’ll kind of get into the glial part a little more here within. (3:25)  But generally I looked at neurodegenerative diseases, treatments of neurodegenerative diseases and analyzing the metabolic products of some of these oligodendrocyte cells.

(3:42)

Slide 3

Now I have a post-doctoral fellowship at Georgetown University, and I study radiation metabolomics. One of my projects in particular is actually distinguishing metabolic biomarkers for radiation injury.  This was looking at a preprocessing approach for ionizing radiation injury. (4:10)  So we were able to pick out biomarkers that allowed us to indicate injury and made a type of extraction that’s useful for a radiological event to pick out injury markers. But today I am going to talk about the marriage of all of my fields of interest and knowledge, those again being the central nervous system damage, radiation injury and then using metabolomics as well, to kind of talk about all of these.  

(4:44)

Slide 4

From this overview, you should find the intersection of all three of those knowledge branches, as we discuss, again, understanding and ameliorating the radiation damage that occurs in brain oligodendrocytes following cancer radiation therapy, using the technique of metabolomics. (5:09)  Some of the learning objectives for this talk that I hope you will gain is just a general understanding of the current state of brain radiation treatment, patient outcomes and their quality of life that follows therapy.

(5:23)

Slide 5

I think this is important to always bring it back when you think about patients.  I also want to detail basic glial cell biology and how I use the MO3.13 cell line to model the immature and mature differentiated oligodendrocytes.  They’re kind of the bread and butter of my work.  And also understanding mass-spectrometry based metabolomics and how we can use it to monitor markers of radiation injury and oligodendrocyte metabolism in cell culture. (5:56)  So first I’m going to start with going into understanding the brain radiation treatment.

(6:08)

Slide 6

So to talk about radiation damage, I first have to talk about cancer.

(6:17)

Slide 7

Cancers of the CNS include cancers that affect the brain, the spinal cord and also the cranial nerves.  Today I’m going to focus mostly on talking about the brain.  Primary central nervous system cancers can derive from different cell lineages.  These include cancers like gliomas, astrocytic tumors and oligodendroglial tumors and also medullablastomas. (6:46)  Tumors can also start in another part of the body and metastasize to the brain. These tend to be the most difficult to manage cancers, and the most dangerous.  They are also more common than the primary tumors that I previously mentioned.

(7:04)

Slide 8

Treatment options for any type of CNS, whether it be a cancer that has metastasized or a primary tumor, includes things like surgery, that’s when possible to resect it from the brain.  You can do a complete or a partial resection of the tumor.  Chemotherapy as well, so chemotherapeutic drugs, but these aren’t used very often because it’s difficult for many to cross the blood-brain barrier and get into the brain. (7:38)  Then lastly is radiation, which is most commonly used.  You can penetrate the brain from the exterior, you don’t have to worry about the blood-brain barrier, and there’s no complexities of surgery, as well.  Radiation treatment consists of the conventional therapy, which uses an external beam.

(8:07)

Slide 9

It can be a beam of x-rays, gamma rays or protons, and this depends on the type of cancer, it also actually depends on the facility, and in some cases it depends on what country you’re in.  Some countries only will allow certain types of ionizing radiation.  This includes things like even carbon, which I believe is not currently being utilized in the United States right now.  But general you use CT scanning, to kind of get an idea of where to target the tumor. (8:37)  Then the ionizing beams are aimed at the tumor, to kill the cancer cells and to shrink the tumor.  This therapy is usually given over a period of several weeks, so it’s fractionated, meaning multiple doses over time.  The actual area that receives radiation treatment may be large or small, meaning a small area of the brain or a large area of the brain, and that, again, depends on the type of cancer. (9:10)  Radiation treatment is usually limited to what we call partial brain irradiation, when possible.

(9:22)

Slide 10

But whole brain radiation is also common when presented especially with metastatic cancer in the brain.  So again, metastatic cancer, which has moved from another place usually tends to form multiple areas of tumors, instead of one singular tumor.  Those are harder to singularly target, so a whole brain radiation whole beam, is used preferentially in this case.  Whole brain treatment has widespread affects, as you might guess, instead of targeting one single area, you’re just irradiating the whole brain. (9:55)  So it causes more severe damage and it also particularly impacts the stem cell population in the brain, which is a real danger.

(10:08)

Slide 11

So some of the injuries that have been documented and occur in stages that are from radiation therapy start with acute injury, and these can be things as simple as headache and drowsiness.  Then kind of early delayed effects, where demyelination has been documented, is the attention deficit and short-term memory loss.  Then late documented injuries include vascular abnormalities, again demyelination in the brain, loss of the brain cells or gliosis, and the most dangerous, cognitive impairment. (10:50)  So just general cognitive impairment seen over a long time.  This leads to poor quality of life in patients.

(11:05)

Slide 12

So currently there is a limited focus on kind of ameliorating or stopping the damage caused by radiation therapy, which again leads to this poor patient quality of life, when you have severe cognitive effects.  These are just words I’m saying, but as a reminder, I talk to patients that are involved with therapy.  I’ve spoken with advocates and those who have survived and a lot of the focus of course is put on survival in cancer. (11:34)  But not a lot of these patients now if they’re surviving, they suffer cognitive impairment, which again hasn’t really been focused on.  So that’s my goal of this study.  Some direct examples of these injuries include the radiation induced injury to children.

(12:00)

Slide 13

So a large study was done by King et al, that looked at children who received radiation therapy for cancer treatment when they were young, of course, and into adulthood. These children unfortunately had declining IQ’s if they received radiation treatment, versus if they did not receive this whole brain radiation treatment.

(12:27)

Slide 14

Studies have also shown this cognitive impairment after whole brain radiation in adults, as well.  So that's Greene-Schloesser et al. Also denotes that increasing after months of whole brain irradiation, you see almost a 85 percent of the population having cognitive impairment.  Again, this is after whole brain irradiation.

(13:06)

Slide15

One of the markers of  this cognitive impairment that was previously mentioned and is tied to the memory problems, and again cognitive impairment, has to do with demyelination or loss of white matter.  White matter is just a type of matter within the brain.  You have grey matter regions and white matter regions.

(13:31)

Slide 16

This demyelination is marked in animal models and also in humans, as well.  Here you can see, from one of my studies, this is decreased staining in the corpus callosum, the stains stain for white matter.  This is seven days after 7 gray of x-irradiation.  The corpus callosum, in particular which I’m staining, is an area that’s rich white matter area.

(14:09)

Slide 17

You can see, again denoted in a mouse model, markers of white matter cells that are lost from one day to 15 months after radiation.  You see both two markers, that’s 04 and MBP marker, which drastically decrease after radiation.  To confirm in humans, as well, tissue from humans.

(14:39)

Slide 18

These again are markers of white matter cells.  So again you see this demyelination that’s persistent in this marker, seems to go along with the cognitive decline.  So just giving the importance that the demyelination in white matter loss is connected to the cognitive decline that you see in patients.

(15:05)

Slide 19

To get a grasp on radiation-induced damage in the brain, we focus on this white matter damage, and again, its connection to cognitive impairment.  So we model the radiosensitive cells that actually make up the white matter.  This is where our lab focuses on, are the cells of the white matter and their connection to this cognitive impairment.

(15:33)

Slide 20

Which brings us into kind of our second learning objective, which is talking about glial biology, these cells of the brain, and how we use the MO3.13 cell lines to model immature and mature differentiated oligodendrocytes.  So the brain is not just made up of neuron cells, though they’re very important.  They’re important for signal transduction and memory.

(16:03)

Slide 21

There are other glial cells, we call them glial cells, that make up a bulk of brain tissue and these include things like astrocytes.  These are important for structure and metabolic support of neurons, and also the glial cell, microglia.  These are actually the specialized immune cells of the brain.  Lastly and my favorite, of course, is the oligodendrocytes.  (16:33)  These cells protect the neuronal axon and they help facilitate the signal that’s transferred through neurons.  So they act kind of like the insulation on a wire and they also serve, they have a metabolic purpose, so they metabolically serve the neurons, as well.  So a lot of important roles.

(16:58)

Slide 22

These cells, they wrap their cell membrane around neurons, around the axon, you can kind of see in this picture here, this spiral wrapping, to form what we call myelin.  So just the wrapping of the membrane around the neuron is called myelin.  And white matter regions in the brain, which we’re interested in, are highly concentrated with myelinated neurons.  So more generally saying these areas are areas with more oligodendric type cells, which is why we’re focused on the oligodendrocyte cells, they kind of are the cell of the white matter. (17:34)  So the death of oligodendrocyte cells is what causes demyelination, so what we were talking about before in the demyelination patterns.  It causes that white matter loss in radiation injury and is connected to the cognitive impairment that we’re worried about in patients.

(18:00)

Slide 23

So our lab is interested in examining these oligodendrocyte cells.

(18:08)

Slide 24

Oligodendrocytes as I noted, are also a metabolic support for neurons.

(18:11)

Slide 25

They support a lot of the metabolic demands for neurons through production of things like pyruvate, with N-Acetyl aspartate or NAA, and other small molecule metabolites.  The myelin sheath itself, which again is just the membrane of the oligodendrocyte which wraps around the neuron, is formed highly of lipids. And it also has very high energy costs, to make all of those lipids and wrap them around the neuron. (18:41)  So these cells themselves are very vulnerable to energy depletion.

(18:54)

Slide 26

Oligodendrocyte cells also start at a stem progenitor state, which is called the oligodendrocyte progenitor cell or you’ll see OPC, and they mature to the final version, which is the myelinating version, the version that actually wraps itself around the neuron.  The OPCs or stem cells are more vulnerable to oxidative stress and damage from radiation, but the mature cells are harder to regenerate once lost. (19:27)  So kind of maintaining the balance of these populations is key to the healthy white matter, and also understanding how both of these populations act in radiation is also important.

(19:44)

Slide 27

So to do this, we turn to a cell line and the advantages of an oligodendrocyte cell line, as opposed to using a primary culture, again it’s a simple system. You have reproducibility when you have a cell line, and very controlled conditions.  And in particular for when we’re doing mass-spec analysis, you can isolate individual metabolic features, which again is very key.  We’re very focused on how specifically the oligodendrocyte cell is reacting to radiation, and not necessarily astrocytes or microglia or neurons. (20:20)  Though those are all important and the components of each of those and their metabolic interests are also important, we want to first start with isolating what is making the oligodendrocyte cell die.  Looking at just the singular cell also has a less complex matrix, which is good for mass-spectrometry, and when using a cell line you have the ability to collect high cell count, which as opposed to primary cells, as they’re a bit harder to culture and to get going. (20:54)  For mass-spectrometry you need a lot of cells, and so the cell line approach in this case is very useful.

(21:07)

Slide 28

So our lab uses and has developed the model of the MO3.13 cell line, which we use in DMEM with F12.  It is a human oligodendrocyte cell line and here’s what it looks like.

(21:27)

Slide 29

This cell line, one of the nice things about it is that you’re able to look at both immature state, so more like the OPC state and a mature state of the cell.  So it has this tunable advantage, and you differentiate it simply by using PMA added to the media.

(21:51)

Slide 30

And here you can see the immature versus the differentiated state.  They both produce markers of the oligodendrocyte, but you can you can see that their membranes kind of drastically change. You also see, if you look to the right, they’re expressing this myelin basic protein, which is MPB, more highly expressed in the differentiated state, and that’s associated with the myelin forming sheath.

(22:28)

Slide 31

For the first time we’ve also used this cell line to model radiation injury.  So here you can see the cell line in a Sham state.  This is receiving no radiation, versus when undergoing 4 gray of x-ray.  This is about seven days post x-radiation.  Their proliferative capacity decreases with increasing radiation, and you can see that these cells, they apoptose generally, they hold position and they kind of flatten out. (23:04)  This is comparable to what primary cells do, as well, so this model, for the first time we’ve shown comparably modeled radiation injury in a cell line, just like how it is modeled in primary cells.

(23:27)

Slide 32

So to recap, we’ve talked about radiation therapy and how it can cause long term damage to white matter and cognitive decline.  We looked at how damaged white matter is composed of radiation-sensitive oligodendrocyte cells, and how the MO3.13 cell line can be used to model the radiation damage to oligodendrocyte cells.  So with use of this MO3.13 model in hand, we can begin to explore the impact of radiation on oligodendrocyte function and its metabolism. (24:01)  Remember, these are metabolically active cells, so understanding their metabolism is very important.  Also to model modes of protection, this is a great model to test out therapeutics.  It’s again very important to understand how the oligodendrocyte bioenergetics regulates normal brain function and how this dysfunction contributes to the N's demyelination.

(24:32)

Slide 33

That brings us to our last talking point, which is understanding mass-spectrometry based metabolomics and its use in this, its use in monitoring the markers of radiation injury and oligodendrocyte metabolism.

(24:51)

Slide 34

On the cellular level, radiation causes this oxidative damage that starts an inflammatory cascade in cells, and that leads to cell death, which then causes the demyelination or white matter loss, and potential long term cognitive impairment.  Again, oligodendrocytes, they’re very susceptible to metabolic or oxidative changes and this includes radiation. (25:23)  It seems to be why they are the most sensitive of the glial cells in the brain.

(25:32)

Slide 35

So to monitor the oscillating metabolism of the oligodendrocyte cell in response to damage, we use this technique, metabolomics,  Metabolomics is a technique that can monitor all small molecule metabolites, which are just small molecules.  So think of things like glucose or tryptophan or serotonin, all these neurotransmitters, any small molecule. (26:00)  The difference of metabolomics and the importance of it is that the information is phenotypic, and this is as opposed to things like genomics, which is measuring at a genome level, or transcriptomics or proteomics.  This is phenotypic information that also gives you kind of an instantaneous snapshot of the cell metabolism, which makes it very useful for temporal studies, as well.

(26:30)

Slide 36

So mass-spectrometry in metabolomics is utilized via two main approaches, which are targeted versus untargeted approaches.  And again, it’s useful for monitoring the injury that occurs in these oligodendrocyte cells.  So in an untargeted technique, you take your tissue or cell, in this case, and then you’re going to detect all of the metabolites that are there. (27:01)  Then we do what’s called statistical reduction and identification, we identify all of these peaks we get into actual compounds.  So we identify them as things like serotonin or tryptophan, etc.  Then we have to do a validation step, as well, just to validate, we check it against a standard.

Then a targeted approach, instead of collecting everything, you’re going to calibrate the machine to the metabolites of interest. (27:34)  So if you’re only interested in neurotransmitters or you’re only interested in things in the tryptophan pathway, then you would do a targeted approach.  Similarly, now the machine’s just calibrated to the metabolites of interest, you detect those of interest and you can quantify how much of that metabolite is present. (27:56)  So metabolomics  kind of has endless applications. In this case we usually are doing untargeted techniques to start with, just to look at what’s happening in the cell.  Then from there we move to the targeted approach, in which now we have an idea of what we’re interested in, and so we target and quantify those small molecules in the cell.

(28:23)

Slide 37

So in some of our work, we’ve utilized metabolomics to analyze the capacity of drugs to ameliorate the inflammatory damage to oligodendrocytes.  Again, it’s important that we’re using this cell line paired with metabolomics, to be able to monitor these drugs’ capacity to stop inflammatory cascade. (28:53) Generally, the idea of these projects is to induce oxidative stress and inflammation.  This can be from radiation, of course, but we can also monitor other ways of oxidative stress.  As an example, one of the drugs that we’re looking at is this dimethyl fumarate, or DMF, compared to vehicle by doing a comparative analysis.  And we use mass-spectrometry again, it’s mass-spectrometry based metabolomics. (29:24)  We’re able to analyze all of the inflammatory molecules and all the changes in small molecules that occur, whether we give a protective drug versus vehicle.

(29:43)

Slide 38

So in this example that I’ve shown, pretreatment with DMS actually altered the TCA cycle, which is an important metabolic cycle in the oligodendrocyte and it increased, you can see malate and fumarate and succinate.  This is specifically with pretreatment of the drug dimethyl fumarate, (inaudible @ 30:05) the up to 72 hours of pretreatment gave me the most increase in this TCA cycle metabolite.

(30:18)

Slide 39

This is important, because as I keep stating, oligodendrocytes have high metabolic demands and these TCA intermediates are crucial to its function.

(30:30)

Slide 40

You can see kind of the positive impact from the drug using the metabolomics on the cell line.

(30:39)

Slide 41

You also see the protection through up-regulation of glutathione.  Again, if you just pre-treat with the drug, we’ve shown that there’s increased reduced glutathione levels, an antioxidant molecule. So it’s helping to decrease the oxidation that could occur from radiation and prepare the cell for oxidative damage. (31:06) And we showed that when you pre-treat the cells with DMS, as our metabolomics led us to believe, this increase in antioxidants also led to protection of cells when they’re treated for up to 72 hours prior with the drug, from oxidative damage.  In this case shown here published is when they were treated with hydrogen peroxide, which is just another inflammation inducing treatment. (31:40)  We then moved of course to look at radiation, now that we kind of had an idea that using metabolomics in the cell line, we could see protection.

(31:49)

Slide 42

Over time, there was an initial boost, again in fumarate, succinate, and malate, from the drug dimethyl fumarate, when we used pre-treatment, but it decreased generally after radiation energy.  However, you can see that the levels of the antioxidant glutathione, the reduced glutathione, actually stays constant throughout up to 72 hours after radiation. (32:19)  This again shows the power of (metabolomics @ 32:23) [recording skip] to monitor the temporal changes in small molecules and to oligodendrocyte metabolism in this response to radiation injury.

(32:40)

Slide 43

And complimented by our metabolic data, you see that pre-treatment of this DMS actually decreases cellular apoptosis following the x-radiation at 4 gray, so decreased cell death or a loss of oligodendrocytes.  So definitely kind of modeled the candidate drug, which is DMS, that can ameliorate the damage of these white matter oligodendrocytes and we did this using mass-spectrometry based metabolomics, paired with our MO3.13 cell line.

(33:24)

Slide 44

In summation, radiation therapy can cause long term damage to white matter and cognitive decline and damaged white matter is composed of these radiation sensitive oligodendrocyte cells.  The MO3.13 cell line, which I’ve showed you, can be used as a good model for radiation damage to oligodendrocyte cells, and is well paired with mass-spectrometry based metabolomics to help model products of radiation injury in oligodendrocytes. (33:54)  I’ve also shown you a case study in which we showed that dimethyl fumarate can offer protection to oligodendrocytes through metabolic alteration.

(34:10)

Slide 45

So I’m so thankful to be, again, chosen as a Gibco Cell Culture Hero and I’m thankful to everyone that I’ve worked with at Georgetown University; in my tumor biology program at the University of Akron, where I started my Ph. D., and for all the funding that has supported me, as well.  And with that, thanks for listening and I want to open it up to any questions.  Thanks.

(34:50)

Moderator:  Thank you Dr. Taraboletti, for your informative presentation.  We will now start the live Q&A portion of our webinar.  Now, if you have a question you’d like to ask, please do so now.  Just click on the Ask a Question box located on the far left of your screen and we will answer as many of your questions as we have time for.  Okay, let’s get started, we have some great questions coming in.  Dr. Taraboletti, what exactly is the mechanism of action for DMF?

(35:16)

Dr. Alexandra Taraboletti:  Oh, great question, thanks Christy.  So DMF, let me start, actually already a known drug that’s being used.  It’s a neuroprotectant.  I’m actually using it as a repurposing drug.  So it’s FDA approved in use of multiple sclerosis.  It’s mechanism of action is actually to target Nrf2, which then leads to the up-regulation of glutathione,  which is one of the reasons we’re looking at glutathione production.

(35:49)

Moderator:  Thank you.  Now our next question.  How could these therapeutics be used for patients receiving radiation therapy?

(35:57)

Dr. Alexandra Taraboletti:  Okay, so as an actual application for the therapeutics, because most of our work was looking at a protective effect that occurred when treated prior to radiation, this would be a type of drug that would be given to patients who then are going to be going in for radiation treatment and therapy prior, about 72 hours prior to the actual radiation that they would receive. (36:28)  Then hopefully give a protective effect.

(36:38)

Moderator:  Now, are there any other fields in this research that have application to outside of cancer therapy?

(36:46)

Dr. Alexandra Taraboletti:  Yeah, so any occupational field in which people are receiving radiation and potentially could receive radiation to the brain.  It’s a great area to study for protection in those events.  One thing that I’m passionate about, because I’m actually a big space nerd, my parents worked at NASA when I was young, so one area that’s being explored also for these type of protectants is for astronauts that are going to go to Mars, (37:17) that are going to need protection from radiation, as well.  

(37:25)

Moderator:  We’ve got some great questions coming in, let’s go with this next one.  Why would you use targeted versus untargeted mass-spectrometry in your study?

(37:33)

Dr. Alexandra Taraboletti:  So I kind of touched on that, as far as what untargeted and targeted is.  Untargeted being you’re looking at exploration of all metabolites, but it’s semi-quantitative, it’s not truly quantitative in that respect.  So if you want to actually quantitate the metabolites that you’re looking at, or how much (inaudible @ 38:06) how much are there, you’d want to use a targeted experiment.  (38:09)  It’s also just more sensitive.  So once you know what you’re looking for, in this case I understood that TPA molecules are changing in my MO3.13 cells, using an untargeted first metabolomics experiment, and then I shifted over to targeted, now that I know what I’m looking for and I can actually see how much is there.

(38:34)

Moderator:  Thank you, Dr. Taraboletti.  Now what metabolites do you see in the inflammatory cascade?

(38:42)

Dr. Alexandra Taraboletti:  In the inflammatory cascade, so in this particular cell line—and again, this is because it’s an Nrf2 target—I’m looking at glutathione production.  But much of the work in my lab has been directed at this inflammatory cascade that happens with radiation, in general, but not necessarily in relation to using DMF as a protectant. (39:11)  So in  that case, you would see some of the more standard inflammatory molecules in the work they’re doing.  I suggest that you check the Fornace Lab and see what they’re working on.

(39:25)

Moderator:  Thanks, Dr. Taraboletti.  Now we’re almost out of time, we’ll have to wrap with this question.  Would DMF be protective with grade 3 to 4 brain tumors at 60 gray IR?

(39:40)

Dr. Alexandra Taraboletti:  That’s a good question, this is far in the future as far as patients that we’d be looking at.  I have looked at this in respect to glioblastoma, which is a very aggressive brain tumor.  There have been actually some patient studies that have looked at DMF not as a neuroprotectant but actually as an anti-cancer agent, I should say. (40:10)  In that case, it didn’t work as an anti-cancer agent, but they did see some protection.  I don’t know about at the 60 gray, on my end in my research, just because we haven’t tested at that amount.  Ideally right now we’re mimicking in mice, which isn’t an exact model, the amount of radiation they receive for a glioblastoma treatment. (40:41)  But I haven’t tested it at that high of radiation.

(40:46)

Moderator:  Thank you, Dr. Taraboletti.  Do you have any final comments for our audience?

(40:52)

Dr. Alexandra Taraboletti:  I just want to give a thanks again.  Thanks to you, Christy, thanks to Gibco, in general for letting me give this talk.  It’s been a really great experience.  Of course, if anyone has further questions, you want to reach out to me, feel free to look at my website at ATaraboletti.com, or you can find me all over Twitter, as well, in the science trigger bursts.  So thanks!

(41:21)

Moderator:  Thank you again.  We’d like to thank our audience for joining us today and for their interesting questions.  Again, just a reminder that the questions we did not have time for today and those submitted during the on-demand period will be addressed by our speaker via the contact information you provided at the time of registration.  Thank you again to Dr. Alexandra Taraboletti for her time today and her important research.

We’d like to thank LabRoots and also our sponsor, Thermo Fisher Scientific, for underwriting today’s educational webcast. (41:50) This webcast can be viewed on demand in LabRoots will alert you via email when it’s time for replay.  We encourage you to share that email with your colleagues who may have missed today’s live event.  That’s all for now, and we hope to see you again soon.  Have a great day!

End Presentation (42:04)

Get to know Alexandra

Why did you choose cancer research?
Like many of my peers I am innately curious—I like to tinker, and create. As a research scientist I enjoy being able to chase down life’s many, many, questions.

What motivates you to succeed in your field?
I grew up in Cape Canaveral, FL and both of my parents worked at the Kennedy Space Center for the shuttle program. I quickly became obsessed with the space program and our goals for space travel. My current work within the field of radiation injury has potential benefits for the upcoming Mars mission. The prospect of being able to help further our capabilities for space travel is what motivates me.

What are your top 3 favorite things to do outside of the lab?

  • Rock climb
  • Paint/illustrate
  • Board games/table top games

What role have the mentors you’ve had in your passion for basic research?
I have had really inspirational teachers and mentors in my life that directly inspired me to become a scientist, but also fueled my passion of science communication and outreach.

Is outreach/STEM important to you? Why?
I have always found outreach to be very important. Volunteering with young scientists is an important cause I believe we as a community must assist. My principal aim is to get children interested in these fields so they will consider STEM careers. Most students have little knowledge of the kinds of jobs and work scientists do and it is our duty to inform them.

Favorite phrase?
“The fragrance stays in the hand that gives the rose” —Hada Bejar

Why did you become a scientist?
I can’t ever remember not being a scientist!

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