Through their lens: Erica Keane does research to better understand Williams syndrome

This summer we’re sponsoring high school interns in stem cell labs throughout California. We asked those students to contribute to our Instagram photos and YouTube videos about life in the lab, and write about their experiences.

Erica Keane working in the lab of Ken Kosik at UCSB. She submitted this photo through Instagram to CIRM’s #CIRMStemCellLab collection

In my research I verified a patient specific induced pluripotent stem cell line that will be used to model a neurodevelopmental disorder. Induced pluripotent stem cells (iPSCs) were adult somatic cells but they are been genetically reprogrammed into an undifferentiated state. The cells that I am working with were skin cells from patients with Williams syndrome. My job is to make sure that these skin cells were successfully reprogrammed into an undecided state. If they are in fact pluripotent they will have the ability to turn into any type of cell: liver cells, blood cells, skin cells, etcetera. If I am able to validate this stem cell line, the cells will be turned into neurons to gain a better understanding of Williams syndrome. Scientists can also use these cells for disease modeling, drug screening, and transplantation studies. One promising method of treatment is regenerative medicine, in which scientists will use these iPSCs to grow tissues and organs that can be transplanted into patients.

Williams syndrome effects 1 in 10,000 people born in the US. People with Williams syndrome die early due to heart attacks, strokes, and other complications. Despite mental retardation, people with Williams syndrome exhibit very social personalities and highly developed verbal skills. Scientists are not sure why there is the combination of low IQs and high verbal skills but hopefully future research will shed light on this peculiarity.

To verify the stem cell line I carried out a number of tests. The Williams syndrome skin cells that had been genetically reprogrammed into an undecided state were first Karyotyped for a quality check. This was to make sure that no macroscopic damage had occurred to the chromosomes during reprogramming. Then I carried out bisulfite conversion to check the methylation status of the Oct4 promoter. This was to make sure that the reprogramming was successful on an epigenetic level. Next, we used ICC (immunocytochemistry) to check for self-renewal marker expression. This consisted of staining the cells and looking for certain embryonic stem cell markers under a fluorescent microscope. These markers are necessary to maintain the undecided state. Lastly, we made sure that the iPSCs could differentiate into the three germ layers that are found in an embryo. We did this by again using ICC but this time looking for markers in each germ layer.

Since all of our tests proved that the cells are in fact induced pluripotent stem cells, the line can now be used to model Williams syndrome. The line can also be used to model other disease but my research will first have an impact of gaining a better understanding of Williams syndrome.

Erica Keane

Erica sent us this video of her experience:

Through their lens: Dan Roman studies the role of genes in the single-cell embryo

This summer we’re sponsoring high school interns in stem cell labs throughout California. We asked those students to contribute to our Instagram photos and YouTube videos about life in the lab, and write about their experiences.

Dan Roman looking at nematode worms under the microscope. He submitted this photo through Instagram to CIRM’s #CIRMStemCellLab collection

Dan Roman worked in the lab of Joel Rothman at University of California, Santa Barbara

This summer I worked on a research project that included the identification of new cell-cycle regulators in C. elegans. I knocked-down seven candidate genes using an RNA interference experiment and observed the result of this on the single-cell embryo. More specifically, I looked at the mitotic division of this cell as the chromosomes of the sperm and oocyte met at the prenuclei meeting point, chromosomes condensed and then segregated. Timing all of these events gave us insight into the mitotic cell-cycle regulating function of these genes in somatic cells. However, six weeks was not quite enough time to be able to identify the definite role of these candidate genes in mitotic cell-cycle regulation. In fact, this was a very important lesson for me as I learned more about the research process. Instead of doing this project for only six weeks, researchers would likely carry out this experiment and run trials for several years in order to confirm the definite function of these genes!

What I enjoyed most about my project was looking at the single-cell embryos through the advanced microscope as they divided. In doing so, I was actually able to observe the first signs of life in an organism, and I never thought I would be able to see something like that. Not only did I witness this phenomenon, but I was also able to contribute to our scientific understanding of the mechanisms and genes involved in this process of cellular development. What I found most challenging was dissecting the worm in order to extract the single-cell embryo. In doing so, we used two needles and, looking through the microscope, used the needles to cut the worms in half without scratching the slides, which was difficult for me because it required a very steady hand. Given the experience I have had this summer, a career in research would be very thrilling because one would go to work each day not knowing what they are going to find or discover and how that could improve our overall scientific knowledge.

If I was an established stem cell researcher in Californian in 2040, I would probably look back at my CIRM Creativity Award Internship as the beginning of my path. Without it, perhaps I would not be there at all and would have entered a less fulfilling career. This internship has truly exposed me to the world of research and all the thrills that it has to offer.

Dan Roman

Dan sent us this video of his experience:

Stem cell Stories that caught our eye: Stem cells aiding cancer therapy, first clinical trial with reprogrammed cells and Alzheimer’s

Here are some stem cell stories that caught our eye this past week. Some are groundbreaking science, others are of personal interest to us, and still others are just fun.

Enlisting stem cells to tolerate chemotherapy. When we talk about stem cells and cancer we are usually talking about the dark side of stem cells, the cancer stem cells that many researchers think are responsible for the spread of tumors in our bodies. This report provides a nice counterpoint. Most tumors could be killed if doctors could give large enough doses of chemotherapy or radiation. Problem is, those doses too often would kill the patient. A team at the University of Michigan found a molecule that revs up the resident stem cells in the gut and greatly enhances their ability to repair damage to the lining of the gut caused by the therapy. More than half of the mice treated with the molecule survived doses of chemo that killed all the mice that did not get the stem cell-directed treatment. Here’s more about the work.

First clinical trial with reprogrammed stem cells set. A team at Japan’s Riken institute announced that they would begin this week the early steps of the first ever clinical trial using stem cells created by reprogramming adult cells. They said they would begin recruiting six patients with Age-related Macular Degeneration (AMD). They will then reprogram skin cells from those patients into iPS type stem cells. Following that will be a long period of testing those cells to make sure they don’t cause cancer. So, the first patient will not receive a transplant of retinal cells made from their own skin until next summer.

This type of personalized therapy using cells that are an immunological match for the patient has been a touch stone goal for our field. This is faster progress than I would have guessed. iPS cells were not even created from human cells until 2007. Here is a report on the project from an official Japanese news service.

CIRM funds a number of teams looking into therapies for AMD and other forms of blindness. You can read about that work here.

CIRM team launches Alzheimer’s project. Never underestimate the difficulty in moving a project from the lab bench to the clinic. There are decisions to make and roadblocks to get around along every stretch of the path to clinical trials. How do you design the pre-clinical safety tests to assure the Food and Drug Administration (FDA) that your therapy is safe. You have to make sure your therapy—cells in this case—are pure and consistent. Also you have line up all the financing you need to carry out all these steps. CIRM granted nearly $20 million to Stem Cells Inc. late last summer to conduct all the preclinical work to get FDA permission to begin a trial in Alzheimer’s disease within four years. They have worked for the last nine months or so to get all these things coordinated. So it was nice to see this press release officially announcing the commencement of our project.

The work of the team, which includes CIRM grantees at UC Irvine, is described here along with other research we fund on the devastating disease.

The following day the company issued another press release, this one providing two-year follow-up data on its trial using the same type of stem cells to treat a rare neurologic disease in children. That trial, for Pelizaeus-Merzbacher disease, sought to replace some of the myelin sheath that normally protects neurons and is lost in the disease. The new data showed that the myelin replacement they saw after one year was maintained at two years, and somewhat increased. Money News picked up the press release here.

Guilty plea in questionable stem cell therapy. A doctor who admits he has no training in processing stem cells pleaded guilty in federal court to illegally shipping stem cells across state lines. He harvested stem cells from umbilical cord blood and processed them for shipment to firms in other states that are also under investigation for offering stem cell therapies for ALS (Lou Gehrig’s disease), Parkinson’s disease and multiple sclerosis even though there is no FDA approved stem cell therapies for any of those diseases. Here is one article on the case.

CIRM has long cautioned against participating in unauthorized clinical procedures. Here is our web page offering suggestions on gaging various clinical options.

Don Gibbons

Guest blogger Alan Trounson — July’s stem cell research highlights

Alan Trounson, CIRM President

Each month CIRM President Alan Trounson gives his perspective on recently published papers he thinks will be valuable in moving the field of stem cell research forward. This month’s report, along with an archive of past reports, is available on the CIRM website.

My report this month discusses five journal articles including one that describes potentially game changing work in the field of cell reprogramming. That team succeeded in reprograming skin cells into iPS-type stem cells with only chemicals instead of genetic factors, which could be safer, easier and more cost-effective than current methods of reprogramming.

But, I want to focus this blog on two papers that both contribute to an important trend in stem cell science. They both coaxed stem cells into becoming complex structures by giving them an environment to grow in that more closely mimicked the environment of the embryo than standard lab cultures. This was one of the theme’s that ran through this year’s meeting of the International Society for Stem Cell Research in Boston in June. A colleague wrote about some related work from the meeting here.

One team from Yokohama in Japan became the first to create complex functional organ tissue from pluripotent stem cells. They turned iPS cells into the precursors of the main components of liver. But rather than growing them by themselves in the lab, they grew them along with two other types of cells that would have been their neighbors in the developing embryo. That combination of cells self-assembled in culture into small spheres that are precursors to mature liver, which the team called liver buds. When they transplanted those liver buds into the abdomen of mice they continued to mature forming functional liver tissue including the blood vessels that are essential for a liver to work. They showed that these rudimentary livers were able to produce proteins only produced in the liver and to metabolize drugs.

The second team, from London, created precursors to photoreceptors, light sensory cells in the eye, and got them to mature into cells that had the properties of functional photoreceptors. Others have succeeded in turning embryonic stem cells into early-stage photoreceptors, but until now no one has coaxed those cells to develop the layered structure of mature photoreceptors. Those cells need to include an outer segment packed with visual pigment that is needed to turn light into electrical signals that can be sent to the brain. The team got the more complete maturation by growing the cells embedded in a gel rather than growing on a flat surface. Again, this came closer to mimicking the environment the cells would have experienced in a developing embryo.

As our field moves increasingly into pre-clinical and clinical applications of the science, these sorts of more complex laboratory procedures will likely become commonplace.

My full report is available online, along with links to my reports from previous months.

Alan Trounson