Three of these students are sharing the highlight of their work in the Porter Lab.

Isabelle joined Porter Lab in 2017 as a volunteer student and in September she will start a Master’s thesis project in our lab.

The Role of Spy1 During Mammary Gland Involution

One in eight women will be diagnosed with breast cancer in her lifetime. From puberty to menopause, factors attributed to breast cancer fluctuate with the natural development of the breasts. A period of increased breast cancer risk, metastatic potential, and higher mortality rates occur following childbirth. The propensity of postpartum breast cancer (PPBC) is in part due to the process of involution: when the mammary gland reverts to non-lactating tissue through intense remodeling, balancing high rates of cell death and regeneration. The increased amount of cellular activity occurring in the tissue during mammary involution creates the perfect microenvironment for tumorigenesis. Our lab studies the Spy1 protein, which plays a role in promoting cell growth and stemness as well as override cell death. Importantly Spy1 is elevated in a host of human breast cancers and correlates with more aggressive forms of cancer. Interestingly, levels of Spy1 are also elevated during involution. We hypothesized that Spy1 protects the cell population necessary for normal mammary gland reconstitution post involution. To address this, an in vitro involution model was deployed with the mouse epithelial cell line (HC11) over a time course of delivery and withdrawal of hormonal cues to study post-differentiation (or mock ‘involution’) cell proliferation and death. This was paired with in vivo tissue collection of the mouse model overexpressing Spy1 in the mammary gland (MMTV-Spy1). At peak lactation, pups were removed to trigger involution and mammary glands were collected during an involution time course for analysis. In vitro results suggest the intrinsic ability of Spy1 as capable of maintaining high proliferative abilities and stemness post-differentiation; whereas in vivo data indicates decreased proliferation and apoptosis in involution while maintaining higher epithelial content. This research begins to articulate the role of Spy1 during normal mammary involution in maintaining the survival of epithelial cell populations, and how overexpression could potentially play a role in the predisposition of the breast to oncogenesis.

Maheen joined Porter Lab in 2017 as a volunteer student and in September she will start a Genetic Counselling Graduate Program at Wayne State University School of Medicine.

Exploring the Characteristics of Spy1 Overexpression in Glioblastoma Multiforme

Glioblastoma Multiforme (GBM) is an aggressive type of brain cancer that develops from glial cells. It is highly malignant and common, accounting for more than 60% of all brain tumors in adults with an average prognosis of only fourteen months. A cell cycle regulator protein, Spy1, has been found to be highly expressed in GBM. Spy1 is an unusual cell cycle protein because it has been observed to overpass the normal cell-cycle checkpoints and elevated levels of Spy1 have been implicated in poor prognosis in GBM patients. The aim of this project is to characterize the role of Spy1 protein in GBM. The mouse glioma GL261 cell line which was employed in all in vitro and in vivo experiments performed in this study contains several molecular alterations characteristic of GBM. We infected GL261 cells using lentivirus to overexpress Spy1 protein. Using neurosphere formation assays we found that GL261 cells with increased Spy1 expression demonstrated elevated self-renewal capacity as compared to control. Through stereotactic injection technique, we generated GL261 Spy1 and control tumours in GL261 line syngeneic mice to better understand the role of Spy1 protein in vivo. Using immunohistochemistry, we demonstrated increased angiogenesis in Spy1 tumours compared to controls. We found that increased Spy1 expression both in vitro and in vivo correlated with some severe pathological features seen in GBM patients; hence, implicating the role of Spy1 in GBM progression and aggressiveness. In summary, the results of this project contribute to a better understanding of Spy1 functionality in GBM and provide insight for the future identification of potential therapeutic strategies.

Nick joined Porter Lab in 2017 as a volunteer student and in September he will start a Master’s thesis project in our lab.

A Potential Driving Force of the Breast Cancer Stem Cell Population in Triple Negative Breast Cancer

My project focused on Triple Negative Breast Cancer (TNBC), a rare subtype that occurs in 10-15% of breast cancer diagnoses. Of importance, this subtype tends to be more aggressive and have a poorer prognosis for patients. One reason for this is a greater proportion of cells known as breast cancer stem cells (BCSC) which are capable of not only self-renewing but becoming the various cell types that make up the bulk tumour mass. This work sought to investigate the role of manipulated levels of Spy1, an atypical cell cycle mediator on the BCSC population. The results indicated that Spy1 was capable of expanding this population in vitro as well as in an in vivo mouse model, leading to a greater disease state. This work serves as a potential platform for utilizing this protein as a potential targeted therapy in order to increase the prognosis of patients diagnosed with TNBC.

Below, Nick is sharing with us a little bit of his experience in the Porter Lab.

“Undertaking the journey of an undergraduate thesis has been one of the most rewarding experiences I have been a part of. Not only did this opportunity allow me to find my passion for research but it allowed me to develop a variety of skills. It allowed me to learn how to manage my time effectively, develop my role in a team environment, and understand the importance of patience. Research can be a very trying endeavour with failures more prevalent than successes. I have repeated the same experiment numerous times due to the finicky nature of science. However, these obstacles allowed me to appreciate the instances when I succeeded even more so. This is something I have applied to my outlook on everyday life. Lastly, my experience as a thesis student has allowed me to see the difference I am making in the cancer research realm. Even though the contribution I am making may represent an infinitely small piece of the puzzle that we are trying to solve, I find the fact that I am able to contribute the coolest aspect of this opportunity. “

Click here to check out part 1.

The first video is from Nick, a student in the Breast research group that will start a Master thesis in September to continue his exciting research focusing on breast cancer.

The second one if from Sami, a student in the Brain research group that will start his undergrad thesis in September focusing on brain cancer.

I hope you enjoyed these videos, and stay tuned for more next week.

The videos below show 4 of our summer students talking about their summer projects. These videos were produced by our awesome research assistant Samira Bashiri and posted on Instagram! Yes, our lab is on Instagram @porter_lab_uwindsor! Please, visit, follow and like our photos and videos, it’s another way to be in touch with our lab and research.

Hope you enjoyed the visit!

The Porter Lab was a success at the WCRG2018 this weekend. Our lab marked presence with one of nine Rapid Fire Session presentations and had a total of 10 posters presentations from a total of 58 conference posters. One of our students received a Student Research Excellence Award for his poster presentation and a Porter Lab alumni also received a Student Research Excellence Award for her poster presentation.

Thanks for visiting and please leave comments below.

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One more time Dr. Porter made us proud!

Here is the video of her brilliant talk.

Please leave your comment below.

Elizabeth Fidalgo, Ph.D

The Porter lab has always enjoyed great diversity in the mix of personalities and talents in the lab. At a fundamental level our four Research Associates (RA) are the force that keeps our lab running smoothly and each RA brings invaluable expertise that benefits our entire team. Here are some highlights for each of them this year (there are too many to cover them all):

As the head of our Tuberin group Elizabeth has continued our NSERC funded work focusing on the importance of the novel interaction between Tuberin and Cyclin B1. Elizabeth’s discovery that this interaction plays a role in regulating mitosis has begun to get the attention of the research community and was the focus of an Oncology Letters paper in 2017 published by the Wang Lei group. It is Elizabeth’s patience, care and guidance that is allowing us to dissect this complicated interaction – the data that will be revealed in her upcoming manuscript provides very clear confirmation that this is a novel checkpoint regulating how a cell controls size according to available nutrients. In addition to her work on our NSERC program, Elizabeth has collaborated with the Gauld group on a Seeds4Hope funded project to model the structure of Tuberin when bound to either Cyclin B1 or Hamartin. She has also started a collaboration with the Swan lab to study how Tuberin and Cyclin B1 interact in a Drosophila model system (a new grant submitted to Seeds4Hope). Elizabeth has continued to train students both in our lab and surrounding labs on microscopy, and she runs and maintains the flow cytometry facility. Of course she is also the Yoda for all complex cloning projects in our lab, for which we are so grateful! I’m always constantly amazed by the scope of talent that Elizabeth has – now taking over the Porter lab website and kicking off this awesome blog!

The brain group was excited to have Dorota back from maternity leave this fall (welcome baby Anthony!). While juggling 2 kids at home Dorota’s review article summarizing the implications of CDK inhibitors in brain tumour treatment came out in Drugs in R&D Jun; 17(2):255. Dorota also came back with a paper written (of course she did!) and a new patent application in hand – both of which are in progress now. As part of our CCSRI funded work Dorota has developed a platform for studying individual patient brain tumours using her own advances on organoid modeling. She has also established a collaboration with Dr. Tirupati Bolisetti in Engineering to use mathematical modeling to predict how individual patient tumours respond to drug treatment (Seeds4Hope and CCSRI LOI submitted). She has advanced a collaboration with Dr. JR Ewing in Physics at Henry Ford to study how mechanical forces impact brain tumour properties – this work received a grant from Henry Ford in the summer and will be submitted as an NIH RO1 grant in the spring.

Bre-anne and Rosa co-lead our CIHR funded work and the breast group. Bre-anne also juggles our constantly expanding mouse colony – managing over 23 mouse lines! 2017 has been a productive year for BreAnne – she published an EMBO paper in collaboration with the Rubin lab (UCSC) solving the structure of Spy1 when bound to Cdk2, which incidentally is one of my favorite papers ever! She has also submitted 2 very important papers characterizing the phenotypes of one of our Spy1 transgenic mouse models – these are both in review at Oncogene. One of the surprising results from this work is that elevated levels of Spy1 induces liver tumorigenesis in male mice, this work is the subject of our upcoming CRS application. Breanne is part of a Seeds4Hope funded project led by Dr. Sindu Kanjeekal from the Windsor Regional Hospital to help advance personalized medicine here locally. Breanne is a leader on a full US patent secured this year (with Dorota and Ingrid) for a new mouse model of brain cancer.

Rosa has been instrumental in setting up our new zebrafish drug screening platform (although we learned that writing ‘drug screening’ on a door sign is not a wise move! … door is finally fixed). We obtained funding from Caesars Windsor to expand this platform as part of a core facilities network led by the Windsor Cancer Research Group (WCRG) termed NUCLEUS. Rosa used this model in her Oncotarget paper that came out in April this year, which showed that elevated levels of Spy1 contribute to Tamoxifen resistance in ER+ breast cancer patients (Oncotarget 8(14): 23337). The first Spy1 clinical trial in collaboration with Dr. Caroline Hamm from Windsor Regional Hospital is now complete … we are excited for this data to be published in 2018! Rosa has mastered creating our own tissue microarray panels – and we now have a large cohort of local Triple Negative Breast Cancer patients tissues in these panels with clinical information. Rosa has some exciting data from these patients that will come out in 2018!

Our graduate students have been working hard and making very important progress on their projects. Janice is pulling her data together for publications and thesis write up while on maternity leave with her latest addition baby Joseph. Ingrid published a paper in Genomics (in press) in collaboration with the Reuda lab. This work used computational biology to isolate potential biomarkers that indicate progression of prostate cancer. Ingrid also published a book chapter (in Methods in Molecular Biology) to teach about using flow cytometry to study the cell cycle in brain cancer stem cell populations. Ingrid has two more publications that she aims to get out on her brain cancer work early in 2018 as she is targeting a late spring/early summer graduate date (sniff sniff). She presented her unpublished work at the London Oncology Day – winning the top poster award! Frank has pulled together some fascinating data showing the role for Spy1 in aggressive gyneaecological cancers and continues to use his pathology expertise to advance many areas of our research program. Our newest PhD Martin is expanding our focus on prostate cancer and has found some very important data to support a role for Spy1 in prostate cancer progression. Martin published a paper in Current Pharm Design and has another in late stage preparation for submission in 2018.

Within our MSc cohort, Ellen is putting the finishing touches on her thesis with graduation date early in 2018 and Iulian has made headway on demonstrating a molecular role for Spy1 in one of our mouse mammary phenotypes. In 2017 we welcomed a new MSc student Adam, originally from Windsor recruited back from Western Ontario. Adam is working with Elizabeth on the Tuberin project and will dig into the collaboration with Dr. Swan.

Undergrads are always a big part of the Porter lab environment and contribute to our ideas, energy and FUN!. We said farewell to our thesis students and long time lab members John Kelly (now doing a MSc at Western), and Melanie Grondin (currently in the MD/PhD program in Ottawa) – John and Melanie we wish you both well and don’t forget to visit!! We were lucky to get back our talented artist and scientist Phil Habashy as a growing collaboration with the Zhang lab. Our new group of EIGHT thesis students is the largest group in the tenure of the Porter lab – Amy, JT, JO, Jackie, Youshaa, Gillian, Dalton and Phil – each has teamed up with a research associate or graduate student and are well on their way to answering their research questions set out in Sept. This year we had 10 outstanding scholars in the lab (Amy, JT, Jackie, Youshaa, Jake, Catalin, Isabelle, Anne, Melanie and John). Summer 2017 we had 4 awesome NSERC USRA students (John, Amy, Anne and Catalin), a very bright and keen SWORP medical student (Joshua Samsoondar) and we were very proud of Alex Rodzinka for securing a Brain Tumour Foundation Scholarship. Because of this group of undergrads we held our first ever Christmas gift exchange – yes I gave the only blooper gift (sorry Adam – hows that fart gun?) – and I particularly loved Ingrid’s gift – providing her with an RA contract and mug that will bind her as a Porter lab member forever.

In addition to our progress on papers, patents and advancing science – our lab puts a lot of effort into communicating our research in many different ways and advocating for the research community. Porter lab has been an instrumental component in the organization and delivery of the WCRG quarterly ‘think tanks’ that advance cancer research ideas and partnerships. These think tanks have advanced 24 research projects and have brought together 233 researchers from across 4 different hospitals, 7 universities/colleges and 4 industrial partnerships. I have delivered 9 talks to the public and our group has hosted countless lab tours to interested students and community members. Ingrid and Breanne kicked off a RIOT (Research Information Outreach Team) in collaboration with the Canadian Cancer Society (making Windsor one of only 4 in Ontario). Ingrid was the guest speaker for the CCS Volunteer night for both Windsor and Chatham this year. Ingrid has also helped in organizing a Windsor “Lets Talk Cancer” and has been a true leader in advocacy for research – her efforts in this area were recognized by winning the Faculty of Science Ambassador Award. Ingrid and I helped to organize a Windsor effort to meet with our local politicians and community leaders to inform about the changes to research funding and the important impacts that this would have on Canada. Rosa made us all proud by presenting her research at SoapBox Science at York and is now leading a similar event for Windsor in 2018. Our lab participated in events like the Brain Tumour Spring Sprint, CCS Relay For Life, Science Rendezvous, Katelyn Bedard Bone Marrow Bowl-a-Thon and swab events, Devonshire Mall Research Showcase – with the motivation to educate, empower and better our local cancer community. The extra mile that my group goes to support our community in this way truly makes me proud!

When I look over the scope of research ideas and activities that our group covers I’m always blown away – and admittedly a little terrified. A visiting scientist once lectured me on ‘staying focused’. I’ve thought a lot about this since then and I’ve concluded that there is a clear difference between not successfully following through on a good idea – and not doing the idea just because its out of your comfort zone. Cancer is a complex problem and it requires bold aggressive ideas to move the field forward. We aren’t going to get amazing cures by ignoring good ideas – or skating around unexpected results to play it safe. I’m thankful for my group who never raise an eyebrow when I throw out one of my “can’t we just make a liver?” comments – but rather research the idea and come back with a plan of how to do it better than I imagined.

Cheers to a great 2017 – I have the best group ever and I’m excited for where our ideas and results are going to take us in 2018!!

Lisa Porter

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Would you ever believe that a fish no bigger than your fingernail could provide information on the best form of therapy for cancer treatments? Well, it’s true! In the Porter lab, the zebrafish group is a part of the Breast Group with our focus being on using fish, specifically zebrafish, to answer questions about which form of therapy is best for individual breast cancer patients. We focus on how drugs work, the pathways inside the cells that they affect, and how the drugs get metabolized within cell culture and our zebrafish model.

Zebrafish are a great model system to use to study heart diseases, neural development and neurotoxins, Alzheimer’s, muscular dystrophy, cancer, and much more! But, do fish have any similarities to humans to study human diseases? Interestingly, yes, they do! They are vertebrates, which are animals with backbones and they share more than 75% of the genome with humans. They have organs similar to humans and have a brain and spinal cord. With an internal environment similar to humans, their responses to cancerous cells and cancer treatments are true to how humans will respond. Zebrafish embryos and larvae are translucent, so we can track the cells throughout the zebrafish daily. We can watch tumours form and regress without any invasive procedures. What’s even greater about using these fish is they have an innate immune system when they are born, but their fully developed immune system doesn’t come into play until 2 weeks after they are born. What this means is we can inject human cells into the fish and they won’t get rejected, but unlike other animal models, the cells are still exposed to some immune system regulations. With all of this, we can now approach new biological questions that I’m so excited to share with you!

Breast Cancer & Treatments

Breast cancer isn’t just one disease. It varies with each patient, which is what makes it difficult to treat at times. We can simplistically separate it into two separate groups or subtypes; hormone receptor positive and triple negative. Each group is controlled by many different factors.

Hormone Receptor Positive: Patients grouped into this subtype in the clinic means their cancer has at least one of three hormone receptors; estrogen, progesterone, and/or human epidermal growth factor 2 (Her2). A benefit of this subtype is these receptors can be targeted with specific drugs to stop the cells from dividing; however, 10-20% of the patient population can develop resistance to these targeted therapies over 10 years and relapse can still occur. A major question is how does resistance or relapse occur? While there are many possible roles, we were able to dissect one mechanism earlier this year.

In February 2017, we published a paper in Oncotarget 2017; 8:23337-52 which described how our protein of interest, Spy1, regulates the estrogen receptor and its pathway changing its response to hormone therapy. As mentioned in previous posts, Spy1 is a protein that promotes increased cell division by binding to CDKs.  I was able to show that when I expressed high amounts of Spy1 in breast cancer cells, the estrogen receptor stopped responding to the hormone therapy Tamoxifen. Using our zebrafish model, we were able to confirm this finding by injecting Spy1 elevated breast cancer cells into the zebrafish followed by treatment with Tamoxifen and watching little to no effect on the tumour. When I inhibited the Spy1-activated pathway (specifically inhibiting the kinase ERK1/2), this showed a significant response to Tamoxifen therapy. Knowing Spy1 levels in patients and targeting Spy1 activated pathways could help us understand what treatments will be more successful.

Triple Negative Breast Cancer: Approximately 10-20% of breast cancers are found to be triple negative. Patients in this subtype do not have high levels or any presence of the estrogen, progesterone, or Her2 receptors. These cancers grow independently of these hormones. Without these receptors, targeted therapies are not useful. These patients are usually treated with chemotherapy. Response to chemotherapy is initially high; however, 34% of triple negative patients have a high recurrence rate in only 2.6 years. One of the reasons for this relapse is the mixed population of cells within the tumour. Therapies normally treat the tumour as one whole product with the hopes that all the cells stop dividing and eventually die. But, the mixed population of cells include a high percentage of special cells called breast cancer stem cells. Breast cancer stem cells have been found to evade treatment, making them a dangerous population of cells. Trying to find new treatment options for triple negative patients that increases overall survival and years of disease free status is one of the most important aspects that we are trying to tease apart.

In collaboration with Windsor Regional Hospital, the Porter lab is a part of two ongoing clinical trials. Since Spy1 has been shown to be elevated in more aggressive and invasive breast cancers (BMC Cancer 2012 12:45), and since elevated levels in hormone positive breast cancers affects treatment, we are interested in determining whether high levels of Spy1 in triple negative patient samples can affect their response to chemotherapy regimens as well as new forms of treatment currently entering clinical trials. Our first clinical trial was to look at the levels of Spy1 in hormone positive and triple negative breast cancer. For this, I received patient tissue in paraffin blocks and created multiple tissue microarrays. These tissue microarrays are then stained for different proteins, which can create a signature for each patient. The goal of this trial is to retroactively look at the patient’s response and levels of Spy1 and see if there is a correlation.

For our portion in the second clinical trial, we will receive fresh patient tissue from the hospital, bring it back to the lab and isolate the cells. One of the first experiments we will perform will be injecting the cells into the zebrafish where we can test out the standard chemotherapy most triple negative patients receive in the clinic as well as some new treatments just entering clinical trials, such as synthetic CDK inhibitors.

CDK inhibitors (CKIs) are naturally found in your body and regulate cell division. They stop your cells from dividing too much or too fast. In cancer, there is a decrease in the amount or levels of these inhibitors, meaning cells have less control in stopping cell divisions from occurring. Pharmaceutical companies have found a way to combat this problem by creating synthetic CKIs and re-introducing them to the body as a form of cancer therapy. Annie, a very hardworking and dedicated 3^rd year undergrad who has received the NSERC Undergraduate Student Research Award for the last 2 years, is currently testing out a number of these CKIs using cell culture work and our zebrafish model on triple negative cell lines. She has already begun to see exciting results and has started combining CKIs with other therapies to test their efficacy. Once patient samples are received from the hospital we can then utilize the protocols she has worked so hard to perfect and we can begin seeing results within one week of testing!

Another aspect of our portion of the clinical trial is to determine what effect breast cancer stem cells may have on treatment. A masters student in the lab, Ellen, is working on multiple tools for our lab to use to determine how much of a population is made up of stem cells. Current work around the world use different kits with labelling techniques, but these have shown to be inconsistent. What Ellen is working on will allow for a more standardized and reliable way to detect stem cell populations. With this new detection method, we can sort out the stem cell population and find new ways to treat these cells to increase response rates in patients. After testing this model in cell lines, we will be able to inject the newly sorted stem cells into the zebrafish model for further treatment testing.

Did you ever wonder how cancer could start in one place, like the breast, and move to another area of the body? Well, cancer cells must go through multiple steps to migrate from one tissue to another and then invade that second tissue. In our group, we have a PhD student, Janice, working on what signals are active in all those steps that makes a cell successfully migrate and invade new tissue. She is trying to figure out whether Spy1 increases the rate of migration and the chances of survival when cells reach the new tissue. She is doing a lot of work in cell culture testing out many different pathways to see which ones are driving these processes. She utilizes the zebrafish model to test out what she sees in cell culture, showing an increase in cell migration when Spy1 levels are increased.  Janice has secured a Mitacs Accelerate Studentship through collaboration with Acenzia here in Windsor based on her work with the zebrafish model. Using the zebrafish, she is testing out the toxic effects chemotherapy drugs can have on the zebrafish and looking at how therapies can alter the immune system. Understanding the immune system regulations and changes is important for understanding the responses we see in the clinic.

Prostate Cancer: Why Movember is Important

Our breast group is always growing, with our newest PhD student Martin, coming to us in 2016 after receiving the Ontario Trillium Scholarship (OTS). Martin has just begun looking at a small subtype of triple negative breast cancer that have the androgen receptor and what role this receptor plays in other subtypes of breast cancers. The androgen receptor, like the others already mentioned, is a hormone receptor and is activated by androgens/testosterone. Martin’s interest in this specific subtype comes from his interest and background in studying Prostate Cancer. Prostate and breast cancer are not that different from each other. Hormonal regulation plays a major role in the development of both cancers and resistance to hormone therapy can happen in both. Prostate cancer is one of the most common cancers found in men; however, research, earlier detection methods, and better treatment options have significantly decreased the death rate by 3.3% per year since 2001. Martin has already completed an abundance of work studying the role Spy1 plays in prostate cancer and if there is any correlation between Spy1 and the androgen receptor. His work is focusing on the subtype of prostate cancer, called neuroendocrine transdifferentiation prostate cancer, which forms after treatment with anti-androgen deprivation therapy and loss of the androgen receptor. He is focusing on Spy1 in 3 main pathways; AKT, β-catenin, and CDK1 activation, to determine if other therapies can be used when androgen dependence is gone. He is also focusing on the role Spy1 plays in the migration and invasion of prostate cells using our zebrafish model.

In our cells and our zebrafish our main goal is to find new and better therapeutics for cancer patients. We understand that each patient is different and should be treated based on their specific cancer and genetics. Our Canadian Institute for Health Research (CIHR) funded work is trying to shed light on what role Spy1 plays in multiple pathways that could lead to patient resistance in breast cancer. We are developing new tools and using a new model system to help us get to answers faster and more reliably.

Research can be enjoyable, but it can also be tough, so we try to follow Dory’s motto while we work and we “just keep swimming!” I hope you enjoyed learning about the second half of the Breast Group! Let us know what you think in the comments below!

Rosa Ferraiuolo, PhD

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