Kristine M. Wadosky, PhD
Research Affiliate, Postdoctoral, Roswell Park Cancer Institute, Buffalo, NY, USA
Kristine is an expert in prostate cancer and recently published three comprehensive review articles on treatment of advanced prostate cancer and therapy resistance. She began her postdoctoral training at the Roswell Park Cancer Institute in January 2015 to study protein degradation of androgen receptor (AR), one of the factors that drives treatment-resistant prostate cancer. In September 2016, Kristine continued her training in a different laboratory at Roswell Park Cancer Institute to study how loss of the tumor suppressor retinoblastoma 1 (Rb1) promotes prostate cancer progression, continuing a project that the laboratory recently published in Science.
Kristine completed both a BS in Molecular Genetics and a BA in English with a concentration in Theater at the University of Rochester in 2009, graduating Magna Cum Laude and Phi Beta Kappa. Kristine immediately began PhD graduate training in Pathology at the University of North Carolina at Chapel Hill, defending her dissertation in 2014. Her dissertation focused on protein degradation pathways in cardiac and skeletal muscle diseases and was funded by a predoctoral grant awarded by the American Heart Association. Kristine received predoctoral trainee travel grants from both the American Society for Investigative Pathology and American Physiological Society to present her dissertation work at national meetings. Recently, her study of thyroid hormone signaling in cardiac growth was awarded a Journal Award by the Society for Endocrinology for being one of the five best papers published by its journals in 2016.
Being a first-generation student, Kristine is dedicated to community outreach. Especially in teaching young students with similar backgrounds about opportunities in science, technology, engineering, and math (STEM). In March 2017, Kristine was awarded a mentorship award by the Western New York STEM Hub, a non-profit organization dedicated to K-12 STEM education.
Learn about Kristine’s research
Title: Understanding cell reprogramming in treatment-resistant prostate cancer using organoids
- Understand the benefits of 3D organoid culture on modeling cancer
- Learn about prostate cancer cell reprogramming and how the understanding of this can lead to improved therapies
Recent studies show that cancer cells can resist treatment by changing into a different cell type. Many treatments for specific cancers, such as breast, prostate, or lung, target vital pathways active in healthy tissue. The reliance of cancer cells on these pathways suggest that they retain properties of healthy cells. A prominent example of targeted treatment is androgen deprivation therapy for advanced prostate cancer. This therapy limits the production and effectiveness of androgen hormones because prostate cancer cells depend on androgen hormones, just like their healthy counterparts. Prostate cancers that become resistant to multiple rounds of therapy often no longer express the target of therapy. These resistant or ‘reprogrammed’ tumor cells are more likely to express different cell lineage markers. These markers are expressed by neuroendocrine cells, a rare cell type in healthy and untreated cancerous prostate tissue. Once prostate cancer cells are reprogrammed, current therapies are ineffective and patients quickly succumb to their disease. Our laboratory studies reprogramming in prostate cancer cells with the aim of developing new drugs to treat these resistant patients. We use murine models and 3D organoid culture of murine and human tumors to understand how prostate cancer cells acquire the ability to reprogram and become resistant. Organoid culture is a valuable tool in our research because it allows the formation of structures that include multiple cell types. In the future, we will use organoids of aggressive prostate cancer in screens of drug candidates and assess drug effectiveness in weeks, rather than the months or years required for classic in vivo studies.
Watch the webinar
Presenter: Kristine Wadosky- Research Affiliate, Postdoctoral Roswell Park Cancer Institute Buffalo, NY
00:00 – Slide 1
Moderator: Hello and welcome. We're glad you joined us for this live webinar, Understanding Cell Reprogramming in Treatment-Resistant Prostate Cancer Using Organoids. I'm Susie Valdez of Labroots, and I'll be moderating this session. Today's education web seminar is presented by Labroots, the leading scientific social networking website and provider of virtual events and webinars advancing scientific collaboration and learning. So let's get started. (00:41)
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Without further ado I'd like to now introduce today's speaker, Dr. Kristine Wadosky. Dr. Wadosky completed both a BS in molecular genetics and a BA in English with a concentration in theatre at the University of Rochester in 2009, graduating magna cum laude and Phi Beta Kappa, she immediately began her doctoral graduate training in pathology at the University of North Carolina at Chapel Hill defending her dissertation in 2014. (02:05) Her dissertation focused on protein degradation pathways in cardiac and skeletal muscle diseases and was funded by a pre-doctoral grant awarded by the American Heart Association. Dr. Wadosky received her pre-doctoral training travel grants from both the American Society for investigate pathology and American Physiological Society to present her dissertation work at national meetings. (02:33) Recently her study of thyroid hormone signaling and cardiac growth was awarded a journal award by the Society of Endocrinology for being one of the five best papers published by its journal in 2016. Dr. Wadosky began her post-doctoral training at Roswell Park Cancer Institute in January 2015 to study protein degradation of androgen receptor, one of the factors that drives treatment resistant prostate cancer. She recently published three comprehensive review articles on treatment of advanced prostate cancer and therapy resistance. (03:11) In September 2016 Dr. Wadosky continued her training in a different laboratory at Roswell Park Cancer Institute to study how the loss of tumor suppressor retinoblastoma, RB1 promotes prostate cancer progression continuing a project that the laboratory recent published in science. Being a first generation student Dr. Wadosky is dedicated to community outreach especially in teaching young students with similar backgrounds about opportunities in STEM, also known as science technology engineering and math. (03:11) In March 2017 she was awarded a mentorship award by the Western New York STEM Hub, a non-profit organization dedicated to K12 STEM education.
I'd like to welcome Dr. Wadosky to begin her presentation. Welcome. (04:02)
Thank you for such a nice and kind introduction. My name is Kristine Wadosky and as was mentioned in my introduction I'm a post-doctoral research fellow at Roswell Park Cancer Institute Buffalo, New York. I work under my mentor, Dr. David Goodrich who is also at Rosswell Park Cancer Institute as well as our collaborator who also mentors me from Dana Farber Cancer Institute in Boston.
04:30 – Slide 2
Other than skin cancer, prostate cancer is the most highly diagnosed cancer in males representing 19% of all new cancer diagnoses estimated for 2017 by the American Cancer Society. In males prostate cancer is the third highest cause of cancer related death at 8% of all estimated cancer related deaths estimated for 2017.
05:02 – Slide 3
Some positive news about prostate cancer, however, is that 91% of all new prostate cancer diagnoses are patients that have low risk disease. So these patients have a disease that will not cause death within the first five years after diagnoses in that they have nearly 100% five-year survival rate and live for many years after their diagnosis. (05:28) However, nine out of 100 of these patients who are diagnosed with prostate cancer are initially diagnosed with advanced prostate cancer that has spread elsewhere into the body. And for those patients they only have a 29% five-year survival rate which means that only 29% of those patients will be still alive after five years after their first diagnoses of prostate cancer or 71% of those patients will have died as a result of their cancer after five years. While we study this type of prostate cancer it is currently not curable.
06:13 – Slide 4
However it is treatable and the difference is is that treatable advanced prostate cancer is cancer that responds to therapy in that the disease burden goes down after therapy. And to explain this I have this graph here. As you can see there's disease burden over time and for patients with advanced prostate cancer, they initially have a first treatment right here at which point in time almost all patients respond to this therapy for a short time but only at about two years. (06:52) And almost all patients will eventually relapse as you can see here. Disease burden will go up yet again in almost all advanced prostate cancer patients that have been treated. At that point in time there is a second type of therapy that we treat at here. And that also decreases disease burden; however, disease burden stays low for a less amount of time after the second treatment, at which point in time disease burden goes back up yet again. And at this point in time there are no targeted therapies for these patients. (07:28) Unfortunately these patients can only be treated with cytotoxic chemotherapy. And our laboratory specifically wants to find a treatable, a pathway to target in these patients that is targeted for these particular resistance prostate cancers. As you see there is multiple rounds of resistance here.
07:54 – Slide 5
But to begin to talk about our findings in our laboratory, I must first talk about what a normal prostate looks like underneath the microscope. So here is a section of a prostate. You can see that there are these round structures here. These are actually glands. So the prostate is normally functioning as a means to make seminal fluid in the male. You can see a close up on the right here. (08:28) And these glands actually produce the proteins that go into the seminal fluid. They're circular mostly in shape and you can see with all these different dots here around the glands that these are actually cells that produce the proteins that go into the empty space in the gland. So this is what a normal prostate will look like.
08:52 – Slide 6
However, in the low-grade cancer that I discuss, this is actually a curable cancer where 91% of the patients as I said before, can be cured of this cancer. This is an example of a low-grade cancer. You can see that there are still glands forming here. You can see that there are some that are circular, but then there are some that are oblong and some that have fused together. (09:20) You can see that there isn't much empty space anymore. There's much less empty space and a lot of the different cells, they are actually invading into the empty space here. And this is an example of low-grade cancer that can be treated and can be cured.
09:38 – Slide 7
As an extreme example as you can see, and very much appreciated I hope, in advanced cancer you can see that there seems no longer to be any circular glands anymore as this slide right here is actually, very, very blue which basically means that there are many, many more cells and the cells, those are packed up right next to each other. There doesn't seem to be a lot of empty space other than here where perhaps blood vessels are and there's basically no normal structure left. And this is the type of cancer that's very likely to become very advanced and resistant to the treatment that I discussed before.
10:21 – Slide 8
So what we wanted to study was how tumor suppressors are related to the resistance and the type in prostate cancer. So a tumor suppressor is a gene that protects the cell form cancer. And many different studies have shown that there are several tumor suppressors in advanced prostate cancer patients that are lost, which means that when they're lost, the cell is going to become cancerous or has the potential to become cancerous. (10:51) And so what we studied in our lab is to make a mouse model that makes prostate cancer. We have a way in this mouse to delete different genes specifically in the prostate.
11:07 – Slide 9
So we chose three tumor suppressor genes and for these tumor suppressor genes, we deleted the first tumor suppressor gene and we found that the prostate cancer that developed in these mice was curable just like the low risk prostate cancer you see in patients. However, when we deleted the second tumor suppressor gene we saw that the cancer would spread throughout the body, but it also, it responded to therapy initially but would come back like I discussed in the human prostate cancer patients. (11:38) And when all three tumor suppressor genes were deleted, we saw that these tumors are very, very aggressive and were completely resistant to therapy. So this was recently published in the Journal of Science and we've been studying these tumors specifically because this is one of the first models to be like human prostate cancer. So we can take these tumors and study how they've developed to find new ways to treat prostate cancer for humans. And one way that we've done to study these prostate cancer tumors...
12:16 – Slide 10
...is that we've looked at the cells very, very closely underneath a microscope. And so one way that we do this is that we zoom in on the cells and then we also are seeing these cells with different types of stains for proteins that will tell us what type of cells are most likely to be responsive to therapy. (12:39) So here you can see in the top in the brown is the positive cells and blue here would be negative cells. You can see for the first row many of the cells are very positive here. You see many brown cells. In the second row you also see positive cells. The staining is a little different. It's more aligned but that's the way that this is supposed to look so these two are positive. This final one is completely negative. And this is for the curable prostate cancer. So we were able to—by this data we were able to discern that these cells are responsive to therapy. And that's what we saw when we treated these mice with the treatment.
But when we knocked out the second tumor suppressor in addition to the first one, we saw a very different type of pattern. So for here you can see that for the first ones there are some cells that are positive like when only one tumor suppressor is deleted. However, these other cells here, you can see are becoming bluer. They're less brown. In the second row you can see very different staining here. (13:57) There's a lot of blue cells right here, but there's also very positive clusters of cells that stained. And finally for the third one you can see that many of these cells are going from completely blue to a little bit brown which means that that is going up, but there's also some blue cells right there, too.
So from this data we can see that there are some cells that are responsive, some cells that are non-responsive and that when you delete both of these tumor suppressors there's a combination of responsive and non-responsive cells. (14:34)
So finally when we looked at the tumors from the most aggressive tumors in this model where all three tumor suppressors we deleted, we see a very, very different scope here, very different. So we see here that almost all of these cells are completely blue and completely negative for this first row here. (14:57) For the second row there seems to be a similar patchy pattern; however, those cells within the patch seem to be a little less brown. And finally you can see that for the final one you can see there's much more brown and darker brown here on the edges of these cells. And so these data show us that most of the cells except for a few we see here are non-responsive in this tumor, which is also very similar to what we saw when we treated the tumors. They did not respond. (15:35) So this is one way that we studied these tumors to try to understand the best ways to treat these cancers and to understand the important pathways that are being activated that we can eventually target them.
So another way that we've also study these tumors that I don't have time to talk about right now is to do a gene expression analysis.
16:03 – Slide 11
And one thing that we found in the aggressive tumors with two or three tumor suppressors deleted is that these cells tended to express genes that are supposed to be expressed in stem cells. So stem cells as you may know are cells that throughout development it's a cell that can become many different cell types throughout development. (16:27) And so eventually these normal cells can also become cancerous. And as I discussed with you before, in prostate cancer we'll treat these cancer cells several times. And our findings in this data and in our paper recently published was that when there's multiple treatments or when we have multiple tumor suppressors lost, there seems to be some kind of switch that happens or reprogramming as we're calling it where these cancer cells start expressing stem cell genes. We do not understand how this is happening as of yet right now, but one thing we do know is that it's associated with treatment that completely failed in these cancers. (17:16) So there's something about becoming more stem like or more like a stem cell that lets these cancer cells evade therapy.
So another way that we are using the method in the laboratory in order to do experiments more quickly is one way that we study these types of cells and interactions is by cell culture. And cell culture is a very old method. This method has been around for decades and mostly prior to the last five years or so we would take cancer cells and grow them flat on the bottom of a dish. So that's called 2D cell culture.
18:02 – Slide 12
And unfortunately scientists have found over time that when we grow cells flat on a dish in 2D they don't really act the same way as when you grow them in 3D since they originated from a 3D person and we live in a 3D world. So there's a new method of cell culture called 3D cell culture and specifically we're using in the lab prostate organoids. (18:29) What this method is is that we can take a tumor from a patient, either from the prostate or anywhere where the tumors have spread throughout the patient. And we can take a sample just like a biopsy and we can break up the cells and instead of putting it flat on the bottom of the dish like we did before we combine these cells with a gel like, a Jell-O like substance in liquid form and then drop these gel substances on the bottom of the dish. (19:10) And this actually allows the cells to form structures that they would normally form in a body or in an organ. And these are called prostate organoids or organoids which means many little organs. And so we can grow these tiny little organs in the dish from a living patient and we can eventually hopefully be able to test different drugs that will be important for that patient's survival. (19:38) And hopefully that will lead to personalized medicine down the road. But one way that we're using this method is to try to understand what's causing these tumor suppressors—the loss of these tumor suppressors to make this prostate cancer so aggressive and so unable to be treated.
19:59 – Slide 13
This is just an example of an organoid that I've been growing in the lab. This is from a mouse tumor and you can see here that there's webbing here. This is actually an organoid that was live, so we can't see the cells but it's growing here both in all directions it's growing. And we're using this method because it takes about three weeks to grow some organoids in the lab. (20:28) However, when we use a mouse model that can take years to actually get to a point where there's a tumor that we can test drugs for. And so eventually we can use this method to test many different types of drugs much faster so they can get the patient.
20:48 – Slide 14
And finally I'd just like to discuss we're currently ongoing doing experiments with the organoids. I just wanted to show you a little bit from a beautiful paper in cancer cell, Blattner, et al. that was published earlier this year. And this is the type of experiments that we are doing. So you can see here that these are organoids that they grew and you can see that they form gland like structures like I showed you earlier in the presentation. (21:14) There's gland like structures here with cells on the outside and pink protein on the inside. That pink is actually the protein and cells are purple so that stain's the DNA. And you can also use this evasively cut the organoids. Imagine the organoids are cut directly in half you can see you can stain different stains like I showed you previously when we stain the tumors. And you can also do a very viewtiful staining with some fluorescent markers or you can stain many different things at one time. (21:49) And so we are pursuing many different avenues using the organoids that eventually we will publish. But I just wanted to show you an example of this..
So finally as research is always ongoing I wanted to show you also that if you directly examine the organoid structure compared to the structure of the glands within the tumor you can see they're very similar, so this model is a very good model to model tumors.
22:25 – Slide 15
And finally I'd like to just discuss a little bit about the Journey to a Cure in prostate cancer. This image is actually—the details are not important but the important thing I want to show here is the length of data and the length of time that so many scientists have been contributing to the Journey to a Cure for prostate cancer. (22:45) The data that we have been all building on, actually the first found in 1941 where we found—I did not find that—where we found that androgen hormones are testosterone which is the male hormone causes prostate cancer to grow. And everything as you can see all these years, discoveries are in red and drugs are in—excuse me—discoveries are in black and drugs are in red. (23:15) You can see that every year it's building upon the previous scientist's work. And I just feel as a young scientist I very much appreciate the amount of work that has gone before me and the work that I am making so that new young scientists can build upon that in order to help people with cancer.
23:39 – Slide 16
And with that I'd like to acknowledge my mentor, Dr. David Goodrich at Roswell Park Cancer Institute and the lab members who did most of this work here and led to the publish of this paper that we just published in science talking about these tumor suppressors Sheng Yu Ku and Yanquin Wang, my co-mentor and the co-senior author of this paper, Dr. Leigh Ellis at Dana-Farber Cancer Institute, our collaborators at Memorial Sloan Kettering Cancer Center, Dr. Charles Sawyers and Ping Mu. We also studied one specific gene, that's a stem cell gene. They published this also with our paper earlier this year. And of course our funding from the National Cancer Institute, funding both my mentors as well as the cancer center support grant to Roswell Park Cancer Institute.
24:41 – Slide 17
And with that I'd like to take any questions or comments. Thank you.
End Presentation: 24:52
Get to know Kristine
What motivates you to succeed in your field?
Helping patients and training young scientists
On your days off, what do you do?
Volunteer at science fairs, spend time with family, cook, play video games
Which scientist current or past would you most like to meet and why?
Rosalind Franklin, because I would want to be the first one to tell her how much her work impacted molecular biology
What is outreach/STEM to you?
Science outreach has been a regular part of my scientific life ever since the first year of graduate school
Be good to one another
Why did you become a scientist?
When I was 12 years old, I learned about DNA for the first time in science class. Ever since then, I knew I wanted to do the job that meant I got to work with DNA. Being a first generation student, I didn’t know what a scientist really did at the time. I attended an all-girls science camp in middle school at Clarkston University call Horizons and learned about science and being a scientist. That camp had a great impact on me—the camp is still running, so this year I will attend and talk to the girls as an alumna.
What’s the best way to start the day?
Who inspires you to be better?
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