Porter Lab

From the Bench to Helping the Planet: Going Green for a Better Future

fidalgo — Wed, 04 Mar 2026 19:59:55 +0000

Our names are Jeffery Martin and Vanessa Riolo, and we are the Porter lab’s Green Lab Ambassadors! Together with Dr. Fidalgo, our Green Lab representative, and Dr. Porter’s full support, we are very excited to share more about sustainability initiatives and the various steps our lab is actively taking to reduce our environmental impact and sustainability. Hopefully, we inspire others to do the same!

Porter Lab has taken the initiative to join the Green Labs Program at the University of Windsor. The Green Labs campus initiative is a certification program that aims to fuel student, staff, and faculty participation to operate with more sustainable, efficient, and responsible laboratory practices. Green Labs and the Chemical Control Centre (CCC) empower participants with tools, knowledge, and peer learning to encourage sustainable behaviours while maintaining high-quality teaching and research spaces. Key areas of this program focus on water conservation, energy conservation, waste management, green chemistry, procurement, engagement, education and awareness.

Did you know that research laboratories generate an estimated 2-3% of global plastic waste every year (ACS Publications)? Or that a single ultra-low temperature lab freezer can use as much as 14,000 kWh of energy per year, the same amount of energy as an average family home (ThermoFisher)? Statistics like these were quite alarming to discover, and that is why Porter Lab leaped to participate in the Green Labs initiative happening within the University of Windsor.

As participants in this program, we are provided with a scorecard that has actionable items which address each of the key areas listed above, and points are awarded based on completion of these items. The points are summed to then assign a ranking, ranging from bronze, silver, gold, or the top level, platinum.

As a crucial method of reducing plastic waste in our everyday work, we give a second life to single-use plastic items, which would traditionally be quickly discarded. One of the best examples of how we do this can be seen through our reuse of pipette tip boxes. Whenever a tip box runs out, we simply fill it with more tips, bake it at 140 degrees Celsius to sterilize them, and then they’re ready to be reused!

The pipette tips themselves unfortunately come in plastic bags, but luckily, we have a solution for that too! See below a list of all the ways we reuse our plastic bags and other plastic items.

When running our Western blot experiments to analyze protein expression, we reuse these bags to incubate our membranes with antibodies.
We save plastic bags to help seal and dispose of virus and cytotoxic waste.
We reuse plastic bottles (Ex. media bottles) by washing them and using them for future experiments or by using them to help dispose of virus and cytotoxic waste.
We clean and sanitize all our non-biohazardous plastic 15 mL and 50 mL conical tubes for reuse.

Further, see below a list of the ways we reuse other everyday items in our lab, like gloves and lab coats, and how we also commit to sustainable practices through using advanced technology.

Rather than using a new pair of gloves each day or each experiment, we prioritize decontamination and reuse of gloves
We reuse disposable lab coats and only replace them when the lab coat is damaged or contaminated
Using online repositories, including OneNote and Teams, we store our updated protocols and inventory for our students’ use and reference, rather than maintaining physical notebooks
At the end of each day, all equipment not in use is switched off (including biosafety cabinets, water baths, centrifuges, etc.)
As one of several labs within the CORE Research facility, the lights in our office areas go off when nobody is around.
In the CORE Research facility, we also have shared equipment rooms where expensive lab equipment can be used by many different labs. This helps to reduce the amount of equipment being produced to run each lab

Further, we truly value the efforts of each member of our Porter lab team, especially our undergraduate student volunteers who prioritize these initiatives every day, making our efforts possible. Our volunteers spend most of their time cleaning tubes and plastic bottles to allow them to be reused, baking and sterilizing tip boxes, and autoclaving biohazardous waste to ensure its proper and safe disposal. Without the help of our volunteers, our sustainability initiatives would not be possible! And last but not least, our RAs: Dr. Fifield is responsible for the fridges and freezers organization, and Dr. Lubanska takes care of the undergraduate students’ training and our lab biosafety.

We aim to nurture a lab culture that values sustainability alongside driving research advancements. One of the ways we foster this attitude is through our annual “Elf Day.” Every December, the entire lab spends one day cleaning our lab space by disposing of old items, washing items that are deemed reusable, and recycling any waste that we may find. Of course, to add a layer of holiday cheer, we do all this dressed as Santa’s elves! Additionally, when planning this event and others, we follow the UWindsor Sustainability Event Guide to ensure we host a “Green Event,” through offering sustainable food items (such as plant-based food options at our gatherings), completing zero-waste activities, and applying the 4 Rs (Reuse, Reduce, Repair, Recycle) of sustainability.

We are very pleased that, because of our sustainability initiatives listed above, we have been elevated to the Green Lab Platinum Level; however, we are not stopping there! One of the next prioritized efforts we are taking in conjunction with the goals of MyGreenLab is our participation in the ULT Freezer Challenger, to aim to reduce the energy usage of our low-temperature freezer through removing ice buildup, items that block airflow, clearing out expired or identifiable items, and more!

Going forward, we aim to continue spreading the word of this incredible and necessary program, and we encourage other labs at the University of Windsor (and nationwide) to participate! If you or someone you know works within a research lab, we encourage you to bring these program details forward to your lab supervisor and ask about signing up for this initiative. Every small task makes a big difference, and only together can we help reduce the environmental impact of research to help build a sustainable future for all.

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Research, Collaboration, and the Path Toward Better Cancer Care

Lisa Porter — Fri, 27 Feb 2026 07:40:38 +0000

On average, one in three people is diagnosed with cancer in their lifetime. Almost everyone has felt its impact, whether personally or through family and friends. That shared reality gives cancer research a weight that is impossible to ignore. Over the years, I have seen how deeply the disease affects people, and those experiences quietly shape how I approach my work. Each experiment, each discussion with students or collaborators, is guided by an awareness of what is ultimately at stake.

From Basic Biology to Cancer Research

Early in my scientific career, my attention was drawn to the basic mechanics of cell growth and division. I was fascinated by the precision required to maintain tissue health and function. As my work progressed, a clear connection emerged between these fundamental processes and the development of cancer.

The same systems that allow the body to repair itself can, conversely, drive disease when regulation breaks down. That understanding redirected my research toward focusing on where and how that balance is lost.

Helping build the WE-SPARK Health Institute has been one of the most meaningful parts of this mission. What began as a local effort has grown into a collaborative health research network connecting universities, hospitals, clinicians, and community partners across the Windsor-Essex region. Watching ideas move from discussion to discovery and then toward patient impact has reinforced the power of working together.

The Line Between Repair and Disease

One of the successes from my lab is understanding the roles of a protein called Spy1, which is elevated in several different aggressive cancers, including breast, liver, prostate, and brain cancers such as glioblastoma, and has the potential to be a novel target for more personalized, directed therapy.

As a recent example of this work, we wanted to understand the role of Spy1 in healthy brain tissue, long before a tumour forms. To do this, we focused on adult neural stem cells, which support learning, memory, and brain repair. We found that when Spy1 levels were increased in these cells, they persisted longer and divided more often. At first glance, that might sound beneficial. The results told a different story. The excess activity disrupted normal brain cell balance, interfered with learning, and created conditions that favoured abnormal growth.

These are the kinds of early changes that, over time, can increase vulnerability to cancer. This work revealed just how narrow the line can be between normal repair and disease. Even small shifts in cellular control can have lasting consequences, especially in tissues as complex as the brain.

The Importance of Working Together

Breakthrough research doesn’t happen alone. I have seen firsthand how progress depends on collaboration. My involvement with national funding agencies, such as the Canadian Institutes of Health Research, has given me a stronger appreciation of how national research priorities are set and how that support shapes what is possible in both the lab and the clinic. Serving on the board of Research Canada has also emphasized the importance of strong advocacy to sustain cancer and health research across the country. Working with charitable organizations and patient-working groups has reinforced that the most meaningful research is that which is shaped by the very people it aims to serve, ensuring our scientific efforts translate into better lives for patients and their families.

These experiences continue to shape how I think about progress. Real momentum occurs when scientists, clinicians, policymakers, patients, and communities come together, share ideas, and move forward with a common goal.

Shaping the Future of Cancer Care

Progress toward better cancer treatments is driven by patience, persistence, and a willingness to learn from unexpected results. Some discoveries answer long-standing questions, while others open new ones. Both move the field forward.

Research lays the foundation, collaboration gives it strength, and together they shape the path toward better cancer care. When discovery, clinical insight, and shared objectives come together, they create an environment where ideas move more freely, questions deepen, and understanding continues to evolve.

A Step Forward in Glioblastoma Research: What We’ve Learned About Spy1 in Stem Cells

Lisa Porter — Thu, 26 Feb 2026 07:33:20 +0000

In my lab at the University of Windsor, my team has been investigating the role of the Spy1 (Speedy) protein, a cell-cycle regulator, in aggressive brain cancers like glioblastoma, one of the most aggressive and difficult-to-treat brain cancers. Our recent work has provided new insights into how Spy1 affects neural stem cells, which are critical for brain repair and regeneration, and how this relates to cancer progression.

Spy1’s Impact on Neural Stem Cells

Neural stem cells are essential for memory, learning, and the brain’s ability to heal itself. As we age, the number of active stem cells in the brain naturally decreases, which impairs its regenerative capabilities. Spy1 has been shown to play a key role in neurogenesis by activating these stem cells, potentially aiding in brain repair.

However, we discovered that too much Spy1 causes problems. In our studies, we genetically engineered mice to overexpress Spy1 specifically in their neural stem cells. While this activation increased the number of stem cells, it also caused them to divide uncontrollably, which made them more susceptible to becoming cancerous. This was a critical finding, as it sheds light on how glioblastoma cells might develop from stem cells that have gone awry due to Spy1 overexpression.

Learning Deficits and Aging

Our research also revealed some unexpected consequences. Initially, we thought increasing neural stem cell populations might improve learning and memory. Despite the increased number of stem cells, the mice showed learning deficits. This shows that simply increasing stem cells doesn’t always improve brain function. In fact, it may even disrupt other important brain cells, leading to cognitive issues.

In addition to its role in cancer, Spy1 also appears to influence brain aging. We found that increasing Spy1 levels in mice helped maintain neural stem cell populations longer than in normal aging, which could have implications for treating neurodegenerative diseases like Alzheimer’s. However, this too must be carefully controlled, as promoting stem cell expansion without understanding the full implications can have unintended side effects.

A Path to Better Treatments

Our findings suggest that targeting Spy1 could offer a new approach to treating brain cancers such as glioblastoma. By carefully controlling Spy1 levels, we might be able to enhance brain repair without triggering excessive cell division that can lead to cancer. The challenge now is to figure out how to selectively target Spy1’s activity in a way that benefits brain health without promoting tumour growth.

A Collaborative Effort

This research has been a collective effort, and I’m grateful for the talented team that continues to work alongside me. In addition to my work at the University of Windsor, I’ve had the privilege of contributing to cancer and health research in different roles throughout my career, including formerly serving as the founding Executive Director of WE-SPARK Health Institute, a collaborative network of researchers, clinicians, and community partners across the Windsor-Essex region.

My work has also been supported by national research organizations, including the Canadian Institutes of Health Research (CIHR), and I serve on the board of Research Canada, where I advocate for advancing cancer and health research across the country.

Looking Ahead

The findings from our research are just the beginning. By continuing to study the balance between Spy1, neural stem cells, and cancer progression, we hope to develop targeted treatments for glioblastoma and find ways to improve brain health as we age. These efforts represent an exciting step toward a better understanding of cancer and more effective treatments for complex brain diseases.

Groundbreaking Prostate Cancer Research at the University of Windsor

fidalgo — Mon, 22 Sep 2025 14:07:34 +0000

My name is Jeffery Martin, and I am a second-year Master’s student working on the prostate cancer project in Dr. Porter’s lab in the Biomedical Science Department at the University of Windsor. In honour of Prostate Cancer Awareness Month, I would like to share some of the exciting prostate cancer research happening at the University of Windsor! Prostate cancer is the second most common cancer in men worldwide, affecting 76 men daily in Canada (Canadian Cancer Society). Disease management and overall patient outcomes have improved as treatment options continue to evolve, and while effective in many cases, these treatments can sometimes pressure prostate cancer cells to change into a more aggressive type of cancer, known as Neuroendocrine Prostate Cancer (NEPC). NEPC can metastasize to other parts of the body, making the current treatments available for prostate cancer much more challenging. Our research focuses on studying the progression of early-stage prostate cancer adenocarcinoma to advanced NEPC and understanding the mechanism of the cell cycle’s regulation in this process.

We believe that through better understanding prostate cancer progression and the cell cycle’s role in this process, we will become more informed on potential treatment options. As a result of our findings, we are now testing selected drugs on prostate cancer cells to try to develop novel therapeutics that can treat and prevent the progression of prostate cancer.

Additionally, we are also looking for novel biomarkers that can detect prostate cancer progression, and we are using non-invasive liquid biopsies to do so! Biomarkers are measurable indicators, like genes or proteins found within the body, that reveal a biological process or disease within the body. Common biomarkers for prostate cancer include PSA (prostate-specific antigen), PSMA (prostate-specific membrane antigen), and AR (androgen receptor). Although these markers are very effective at detecting early stages of cancer, unfortunately, as the disease progresses, these markers are often lost. This leads to the need for the development of new biomarkers to track disease progression.

Further, we hope to find biomarkers that can be detected in early stages of prostate cancer and indicate whether or not disease progression is expected. This would help to guide treatment plans and create a more personalized medicine approach going forward. As mentioned, we plan to use non-invasive liquid biopsies instead of traditional biopsies to detect these biomarkers. Although they are effective, traditional biopsies are invasive, painful for patients, and can lead to potential risks down the road. With liquid biopsies, we can detect these biomarkers in a less invasive way by using a blood, urine, or saliva sample. Through a collaboration with Dr. Kanjeekal at Windsor Regional Hospital, we will be searching for these new biomarkers with liquid biopsies collected from prostate cancer patients.

The research we conduct is mainly done in vitro, which means it’s done in a controlled environment, like in a test tube, instead of a living organism. Specifically, we work in a cell culture room for a lot of our experiments. At the moment, I’m working with 4 different prostate cancer cell lines, which are all derived from prostate cancer patients. We use these cell lines to mimic the progression of disease, and also for drug testing to see how the cells respond to treatment.

We also do some in vivo experiments, which means using living organisms. We specifically use zebrafish as a model system for testing the drugs! Using Zebrafish is becoming increasingly common around the world for drug testing and there are many benefits of this model system. First, Zebrafish are genetically similar to humans, and they are a cost-effective model that requires easy maintenance. Next, they develop quite rapidly, which is great for experimentation, and they have a transparent embryo, allowing for easy visualization. Finally, and most importantly, Zebrafish lack a functional immune system for the first 10 days of their life! This allows us to transplant human cells directly into zebrafish and then observe how these prostate cancer cells behave in a living system. From there, we can test various drugs in the Zebrafish with the prostate cancer cells and see how the cells respond to treatments in this environment. This will also help the results from our studies translate more easily and rapidly to use in humans.

This fundamental work has been supported by grants from Cancer Research Society (CRS), Ride for Dad and WE-SPARK Health Institute. Through this funding, we have made some excellent progress so far, and I look forward to the progress that we will continue to make in the upcoming years! Additionally, through submitting my project proposal to various organizations, I have been lucky enough to receive funding from NSERC (Natural Sciences and Engineering Research Council of Canada), the Windsor Prostate Cancer Scholarship, the Dr. Michael L. Petras Memorial Scholarship, and the Doctor Family Health Research Student Support Award. Each of these awards has helped fund my education and allowed me to spend more time working in the lab. Further, another Master’s student working on this project, Christian Kassa, was recently awarded a Canadian Cancer Society Research Training Award. This is a very prestigious award with only 6 recipients across all of Canada. Christian just started his graduate studies this year and will be helping advance this project over the next 2 years!

A highlight of my project has been the various connections I have made with the community and other researchers at various conferences and local events. At the WE-SPARK Health Research Conference, I had the privilege of being a LEARN program host, which stands for Lived Experience Accelerating Research Knowledge. As a LEARN host, I was paired up with two local prostate cancer survivors who were able to share their lived experiences with me and give insight into the patient perspective. Oftentimes, when working in the lab, I forget about the direct impact my work can have, and these connections remind me of the importance of research. Further, the two patient participants that I was paired with are both members of Windsor’s local Prostate Cancer Support Group, and through making a connection with them, I was invited to speak to their group this upcoming November! Presenting our important work at various conferences has attracted the interest of other researchers and physicians for possible collaborations.

It’s important to recognize everyone involved in this project as none of this work would be possible without the help of our multi-disciplinary team. This project is supervised by Principal Investigator, Dr. Lisa Porter, and Research Associates, Dr. Elizabeth Fidalgo and Dr. Bre-Anne Fifield. Our clinical collaborator is Dr. Sindu Kanjeekal, the Chief of the Department of Oncology at the Windsor Regional Cancer Centre. In-lab experiments are carried out by myself, Jeffery Martin, fellow graduate student, Christian Kassa, and undergraduate student, Kaitlin Ferraro. Previous lab work was completed by Dr. Martin Bakht while a PhD student in our lab, and undergraduate thesis student, Maria Badalova. Knowledge translation work is completed by our Clinical RA, MSc Samavia Ahmad, and Masters of Translational Health Science student, Hannah Ferasol. Finally, none of this project’s advancements would be possible without the help of everyone else in Porter lab who help to provide meaningful feedback and insights that help drive this research further.

I hope you enjoyed learning a little bit more about the prostate cancer research taking place at the University of Windsor. Feel free to watch the video here if you want to see more of the work we do. If you have any questions, please leave a comment below, and I will get back to you! And if you enjoy reading it, please share it below! Jeffery

UNDERGRADS 2025

fidalgo — Wed, 14 May 2025 14:53:46 +0000

Christopher Jaworski

Glioblastoma (GBM) stands as one of the most lethal and aggressive brain cancers, often defying current treatment strategies. The outlook for patients remains grim, with average survival hovering around just 14 months after diagnosis. A major reason for this is the tumor’s ability to resist therapy and recur, often driven by a complex network of cellular interactions within the tumor microenvironment. My research focuses on understanding the relationship between glioblastoma and a group of cells known as cancer-associated fibroblasts (CAFs). CAFs play a critical role in shaping the tumor environment by promoting tumor growth, supporting resistance to therapies, and enhancing invasive behavior. A protein of particular interest in our studies is YKL-40—a glycoprotein that is highly expressed in the mesenchymal subtype of GBM. This subtype is known for its aggressive nature and poor response to conventional treatments. By investigating how CAFs and tumor-initiating cells communicate through YKL-40, we aim to identify vulnerabilities that could lead to more effective treatment strategies. We’re also working with brain tumor organoids to model these interactions more realistically, and using techniques like shRNA knockdowns to probe the role of YKL-40 more precisely.

My journey in research began in my second year of university when I joined the Porter Lab. I entered as a volunteer, initially gaining exposure to the fundamentals of cancer biology. Through the Peer Mentor Network, I was mentored by upper-year students who helped me develop a foundational understanding of lab techniques and experimental design. As I grew more confident and capable, I began contributing more directly to ongoing experiments and even helped identify gaps in existing research that informed new directions for study. By the time I reached my upper years, I had the privilege of having my own project, from hypothesis formation to troubleshooting complex protocols. This experience has sharpened my analytical thinking and strengthened my collaborative skills, especially through interdisciplinary discussions with researchers across different fields. Being part of such a dynamic lab has deepened my appreciation for the intricacies of cancer research and sparked a commitment to lifelong learning and scientific inquiry. I’m incredibly thankful to Dr. Porter and the entire lab team for their mentorship, support, and trust in my development. This journey has not only shaped my academic path but also inspired me to continue contributing to the broader fight against cancer.

Glioblastoma (GBM) is the most aggressive type of brain tumour, characterized by its highly infiltrative and heterogeneous nature, which poses significant challenges to standard therapies. The presence of therapy-resistant Glioma Stem Cells (GSCs) contributes to GBM heterogeneity. Cancer cells (including GBM) are characterized by uncontrolled cell proliferation, which is linked to cell cycle dysregulation. SPY1 (SPDYA/RingoA) is an atypical cell cycle regulator that overrides cell cycle checkpoints, aiding in uncontrolled cell proliferation and survival through unique activation of CDKs. In GBM, elevated levels of SPY1 regulate CDK2 activity and drive clonal expansion of CD133+ GSCs. SPY1-CDK2 can also activate RNA-binding protein, Musashi-1 (MSI1), which plays a critical role in GSC maintenance through post-transcriptional regulation of NUMB and Notch pathway. MSI1 supports GSC populations to drive tumor

initiation and resistance to differentiation. This study aims to understand the role of MSI1 in maintaining GSC properties and its potential correlation with specific subgroups, and how MSI1 influences GSC self-renewal, proliferation, and response to therapies, with the goal of identifying novel therapeutic strategies to overcome treatment resistance in GBM. To demonstrate this, established GBM cell lines were infected with short hairpin (sh) MSI1 lentivirus, which resulted in reduced proliferation and self-renewal. The Zebrafish PDX platform was used to further validate the effects, confirming the impact of the MSI1 knockdown

Being part of the Porter Lab family was one of the most amazing and rewarding experiences of my undergraduate studies. It offered far more than experimental training – it provided a supportive, intellectually stimulating environment where I learned to think critically, troubleshoot experiments, and develop my intrapersonal skills, which will help me in the real world. From the beginning, I was welcomed into a community that values both scientific excellence and personal growth. Whether it was executing the experiment, analyzing data, or collaborating with your lab peers, every task contributed to my development as a researcher. The most important thing I’ve learned during the whole experience is the value of perseverance. You are bound to experimental errors that may alter your results, but you have to see these setbacks as opportunities to rethink and improve, not as failures.

Glioblastoma (GBM) is the most lethal and aggressive primary brain tumour affecting the central nervous system. Despite standard treatments, prognosis remains poor due to several barriers that limit successful treatment such as the presence of the blood-brain barrier, tumour heterogeneity, and glioma stem cells (GSCs). GSCs are known to increase aggressiveness and hinder therapy response. A notable GSC marker is the CD44 receptor, which is activated by its primary ligand, hyaluronic acid (HA), to promote cancer progression. High CD44 expression in GBM is correlated to increased aggressiveness, stemness, proliferation and, ultimately, poorer prognosis. Nanoparticle therapies are an emerging field of cancer research that allow for selective targeting of GSC populations. Our lab has shown that HA-Conjugated Nanoparticles (HA-CPNs) can selectively target CD44+ cells, eliciting anti-tumour effects both in vitro and in vivo. Within my project, I utilize HA-CPNs to target GSC populations within a biologically relevant model known as glioblastoma organoids. Organoids mimic real 3D tumour tissues making them a suitable model to study the effects of HA-CPNs on GSC regulation.

My journey within Porter Lab, starting in my second year of undergraduate studies, has provided me with some of the most rewarding experiences of my entire undergraduate career. I have been extremely fortunate to work alongside a team of talented students and RAs that I view as incredible role models and mentors. My time in lab has expanded my ability to problem solve, think critically, and above all has made me a more resilient person, appreciative of the intricacies of scientific discovery. I am extremely grateful for all of the opportunities, support, and guidance I have received from Dr. Porter, Dr. Lubanska, and the whole Porter Lab team this past year!

My project focuses on determining the potential of the CDK1/2 inhibitor Dinaciclib as a therapeutic direction for prostate cancer (PC). Adenocarcinoma prostate cancer (AdPC) represents 95% of all PC cases and has several treatment options available, such as surgery, chemotherapy, and hormone therapy, like androgen deprivation therapy (ADT). ADT works by blocking the production of androgens in the body, such as testosterone, to reduce the proliferation of prostate cancer cells. However, some patients do become resistant to ADT, leading to treatment-resistant populations of PC to emerge, called castration-resistant prostate cancer (CRPC). CRPC has a median-survival rate of 1-2 years and is much more aggressive than AdPC. A common treatment method used for CRPC is AR inhibitors, which block the activity of the androgen receptor (AR) outright to slow the proliferation of the prostate cancer cells. However, patients can also become resistant to this form of therapy, and differentiate into a treatment-resistant form called neuroendocrine prostate cancer (NEPC). NEPC is the most aggressive and deadly subtype of prostate cancer, and has a 7-month median survival rate. Furthermore, the downregulation of prostate-specific membrane antigen (PSMA), an important detection target, and AR in the transdifferentiation to NEPC further contribute to the poor prognosis of this type of cancer. Our lab proposes targeting the cell cycle to slow the proliferation of prostate cancer, using a CDK1/2 inhibitor named Dinaciclib. My project uses three prostate cancer cell lines to test the efficacy of Dinaciclib in vitro in stopping the proliferation of prostate cancer cells: LNCaP-FBS, an AdPC cell line, LNCaP-CSS, a CRPC cell line, and NCI-H660, an NEPC cell line. My project also uses zebrafish as an in vivo model to test Dinaciclib’s effect on tumour burden, where zebrafish have been injected with prostate cancer cell lines and treated with Dinaciclib. Zebrafish are a viable model for prostate cancer because they develop rapidly, have transparent embryos, which makes them easy to see under a microscope, and also lack an immune system within their first 10 days of life, decreasing the chance of rejection if they were to be given human cells.

My time in Porter Lab has been one of the most important and fulfilling experiences of my life because it has allowed me to grow not only as a scientist but also as a learner. I would like to express my sincere gratitude to my supervisors Dr. Lisa Porter and Dr. Elizabeth Fidalgo da Silva, as well as PhD and master’s students for their mentorship, encouragement, and guidance in developing my research ability. I wish the best of luck to the next group of thesis undergraduates!

Undergrads 2024

fidalgo — Tue, 21 May 2024 16:25:44 +0000

Glioblastoma (GBM) is the most frequent and aggressive primary malignant brain tumour. Despite rigorous therapeutic efforts, the prognosis of GBM remains extremely low, with a median survival of approximately 14 months after diagnosis. Our lab studies a cyclin-like protein called Spy1 which has been found to be elevated in GBM. Spy1 overrides cell-cycle checkpoints and avoids conditions that would normally cause cell cycle arrest, leading to increased and uncontrolled cell proliferation. Our research has demonstrated Spy1’s involvement in regulating senescence, a prolonged state of cell cycle arrest, specifically within the context of GBM. We have shown that targeting Spy1 leads to an increase in the number of senescent cells, and importantly, these senescent populations can be effectively eliminated using senolytics, drugs that can target and eliminate senescent cells.

I joined the Porter Lab at the beginning of my first year of university as an outstanding scholar. I started as a lab volunteer and a junior student in the Peer Mentor Network (PMN) where I learned the basics of cancer research from more experienced students. Over time, I took on a more active role, identifying knowledge gaps, assisting with experiments, and collaborating with peers to address current research challenges. Working on my own project in my upper years allowed me to develop my critical thinking and problem-solving skills, and taught me the benefits of working in a collaborative environment. This experience has ignited a passion within me for research and lifelong learning, and I am truly grateful to have had the opportunity to pursue research under the guidance of Dr. Porter and the research associates.

My project looked at the functional consequences of Tuberin mutations, more specifically the A614D and C696Y missense mutations. Mutations in this protein result in a disease called Tuberous Sclerosis Complex (TSC). TSC causes benign tumour formation in various organs of the body including the brain, heart, eyes, and lungs among others. These point mutations result in Tuberin losing its regulatory function over protein synthesis and mitotic onset. I investigated the punctate perinuclear localization pattern that is seen with these point mutations by looking at the lysosome as a potential cause for this. We hypothesize that Tuberin mislocalization to the lysosome results in a loss of Tuberin’s regulatory role over mitotic onset. I also aimed to develop cells that express these Tuberin mutations at physiological levels so we can better understand the interactions of these Tuberin mutants at endogenous levels.

I joined Porter lab in my 4^th year of undergrad to do a thesis and my experience in the lab was great, being able to work in the lab allowed me to gain valuable skills in not only critical thinking and problem solving but also oral and written communication, every member of the lab was very kind and generous with their guidance. The skills that I developed in the lab will be extremely beneficial in the future and I want to thank everyone in the lab for their continued support throughout the year.

My project aimed to illuminate the role of the protein Spy1 in genomic instability and drug resistance in Glioblastoma. Investigating how Spy1 deregulates DNA damage checkpoints and increases treatment resistance can help us validate Spy1 as a potential therapeutic target and improve future treatment for Glioblastoma.

Being in Porter Lab was an incredible experience. I learned a lot about dedication, resilience and overcoming failure, though I must also credit my success to the help of my supervisor Dorota, my mentor Hema, and the rest of the lab for their assistance and kindness. In the future, I plan to continue in a career in the health sciences.

The cell cycle is a highly regulated process consisting of cyclins, CDKs, and checkpoints that control cellular progression. p53, a tumor suppressor protein is activated in the presence of DNA damage, halting the cell cycle, thereby allowing DNA repair to occur prior to cell division. Tuberin (TSC2), another tumor suppressor protein, regulates the G2/M transition and negatively regulates mammalian target of rapamycin complex 1 (mTORC1) by binding to the GTPase activating protein (GAP) domain, effectively suppressing protein synthesis and cell growth. Previously, our lab has shown that Tuberin regulates mitotic onset by retaining Cyclin B1 (the G2/M cyclin) in the cytoplasm depending on nutrient availability. Due to this unique ability, and Tuberin’s ability to sense nutrient status in the cell, we aim to investigate whether Tuberin could be playing a role in DNA damage response by delaying the onset of mitosis and allowing for DNA repair mechanisms to occur. My project focused on creating Tuberin-null U2OS cell lines using CRISPR-Cas9 editing technology. Tuberin-ΔGAP lines were also constructed to clarify whether Tuberin’s role at arrest is independent of mTORC1 activity. To induce DNA damage, etoposide, a topoisomerase II drug was used, and cells were analyzed by flow cytometry using DAPI staining to determine cell cycle profiles. This project will provide further insight into the role of Tuberin at mitotic onset and elucidate the mechanisms of DNA damage induced carcinogenesis, TSC, and other cancers.

“My time in the Porter Lab has truly been one of the most pivotal experiences of undergraduate years. I joined the lab in my second year where my first exposure to the lab’s research was through the Peer Mentor Network. I was blown away by the amazing projects being investigated in the lab and honestly, found the work to be quite daunting. I then began my own project in third year, actively conducting experiments and engaging in research. Despite the challenges and hours of troubleshooting, I can confidently say that this experience has been instrumental in shaping my academic and personal growth. Not only have I gained a multitude of technical skills, but I have also honed my critical thinking, problem solving, and resilience. This journey has been truly gratifying, from the realization that my hands had become steady while pipetting, to finally imaging a successful western blot. Most importantly, the level of mentorship I have received throughout my thesis is unparalleled. I would be lost without the support, advice, and patience of the many grad students in our lab. Their guidance made such a supportive learning environment and helped me gain confidence in my own abilities as a researcher. I am also sincerely grateful to Dr. Lisa Porter and Dr. Elizabeth Fidalgo da Silva for all the opportunities for growth, and I wish all the best to the next group of thesis undergrads!”

My project focuses on tool development and neuroendocrine prostate cancer. A standard treatment protocol for castrate-resistant prostate cancer involves the use of androgen receptor inhibition therapies. While effective, in some cases, androgen deprivation through treatment can pressure prostate cancer cells to differentiate into neuroendocrine prostate cancer cells (NEPC). Current treatment options for NEPC remain limited, and understanding the molecular mechanisms guiding this differentiation will be essential in developing future therapies. My research questions focuses on whether we can stop the cells from transforming into such an aggressive form of NEPC cells by using Cyclin-dependent kinase (CDK) inhibitors. I use LNCaP cell lines and the zebrafish animal model to test the hypothesis that elevated levels of Cyclin A and/or Cyclin B1 support the evolution to NEPC and that select targeting of their kinases, Cdk1 and/or Cdk2, at the G2M checkpoint, could represent an effective therapy for preventing progression and treating NEPC. Furthermore, I constructed fluorescent splitFAST vectors that allow real-time monitoring of the interaction between Cyclin B1 and Tuberin. Our lab has observed that Tuberin expression plays a role in embryonic neurodifferentiation and could be a potential player within NEPC differentiation. The fluorescent vectors allow us to visualize the effects of the CDK inhibitor. While research is constantly testing the unknown, there is strong rationale and data to support that this is a promising direction that could dramatically impact overall outcomes on a global scale.

I joined Porter Lab and my third year and it has been one of the most fulfilling experiences in my university career so far. I’m incredibly thankful to Dr. Porter and Ras, especially Dr. Elizabeth Fidalgo de Silva for supporting me and helping me in my journey as a scientist. I’m incredibly grateful for the Porter Lab community for supporting and being a great team. My time in Porter Lab has taught me invaluable skills that I am looking forward to carry on with me into my next steps for my science career.

Undergrads – 2023

fidalgo — Thu, 04 May 2023 18:47:44 +0000

Amy Llancari

My project focuses on differentiating the progression of Non-alcoholic Fatty Liver Disease (NALFD) into Hepatocellular Carcinoma (HCC) between male and female mice. HCC is the sixth most prevalent cancer in the world, yet it ranks as the third leading type of cancer deaths with an accelerated increase in incidence rates. NAFLD is a major risk factor for HCC, and is emerging as the most common chronic liver condition in the Western world. Prevalence of both HCC and NAFLD is significantly higher in men than women worldwide, suggesting sex-specific differences may play a role in disease susceptibility. My project will focus on investigating these sex-specific morphological and cell expression pattern differences. This will provide insight on potential protective mechanisms found in female mice inhibiting liver injury, in addition to potential diagnostic markers and molecular treatment targets during NAFLD progression.

I have been part of the Porter since my first year of undergrad. My time in the Porter Lab has been the highlight of my university career. I cannot put into words how thankful I am for the unwavering support and guidance I’ve received throughout my project. Dr. Porter and the RA’s have been exceptional role models, their dedication to the scientific community is truly inspiring, and their passion for research is contagious. Being a part of this incredible team has been an honor, and has provided me with so many opportunities both in research and community involvement. My time in the lab has taught me valuable research skills, as well as transferable skills that I will carry with me throughout my career. As I pursue my Masters in the Porter Lab, I am excited to continue to learn and grow as a researcher.

Alan Cieslukowski

Efficient targeting of multiple components of a tumour might be a successful strategy in aggressive types of cancer such as glioblastoma (GBM), which remains the most common and malignant primary brain tumour with an extremely poor patient survival of less than 15 months. The significant therapeutic challenge posed by GBM stems from its genetic and phenotypic heterogeneity fueled by characteristics such as aggressive and treatment-resistant populations of Tumour Initiating Cells (TICs) and high levels of angiogenesis contributing to tumour evolution. TICs, which are responsible for GBM patient relapse, thrive in niches close to blood vessels where they interact with endothelial cells (ECs), exit the cell cycle, and evade therapies. Targeted antiangiogenic drugs, preventing GBM cells from recruiting new blood vessels are only temporarily effective in 50% of patients, resulting in acquired secondary resistance by the tumour. This project explores the TIC-EC interplay and its role in propagating tumour aggressiveness and therapy resistance. This project investigates the impact of ECs on the aggressive characteristics of individual, specific populations of TICs using GBM patient-derived systems, including 3D organoid models and zebrafish patient-derived xenografts (PDXs). Considering that NOTCH1 signaling regulates both TIC and EC populations in GBM, this project dissects the role of NOTCH pathway in TIC-EC interaction. Elucidating the details of cell cycle signaling and specific populations of aggressive TICs with dependence on the EC component will contribute to the identification of new and better therapeutic targets and personalized approaches to treatment of patients with GBM in the future.

Depen Sharma

Tuberous Sclerosis Complex (TSC) is a genetic disorder affecting 1 in 6,000 births, leading to benign tumour formation in various body parts such as the brain, kidney, skin, and lungs. This condition arises from mutations in the TSC1 or TSC2 genes and presents a wide range of neurological symptoms, making diagnosis challenging. Tuberin, a protein encoded by the TSC2 gene, is critical in regulating cell growth, proliferation, and the cell cycle. A key element in TSC is the interaction between Tuberin and Hamartin, encoded by the TSC1 gene. Hamartin ensures Tuberin stability by inhibiting its interaction with the E3-ligase HERC1, which would otherwise lead to Tuberin degradation. Tuberin also regulates the G1/S and G2/M checkpoints in the cell cycle, which are crucial for controlling cell growth and division.

Our research explores the potential role of Cyclin B1, a cell cycle protein, in stabilizing Tuberin levels without Hamartin. Cyclin B1 is essential for mitotic entry, and its function has yet to be entirely understood. It has five serine residues that can be mutated to create a phospho-null Cyclin B1 construct (Cyclin B1 5A), which we utilize in our investigation. Previous work has shown that Tuberin can bind Cyclin B1 during the G2 phase of the cell cycle, but more is needed to know about their interactions when Hamartin is present. Cyclin B1 may be essential in Tuberin stabilization during the G2/M transition. Understanding the relationship between Cyclin B1 and Tuberin in TSC could provide valuable insights into the disorder’s molecular mechanisms and potentially lead to novel therapeutic approaches. By elucidating the role of Cyclin B1 in Tuberin stabilization, we hope to contribute to the ongoing efforts in understanding and treating Tuberous Sclerosis Complex.

Emmanuel Boujeke

My research focuses on Glioblastoma (GBM). GBM is the most frequent and aggressive primary brain tumour in adults. GBM patients have an extremely poor prognosis, with survival averaging only 14 months post-diagnosis. Our lab has been investigating the role of a novel cell cycle regulator (Spy1) in the development and progression of many cancers, including GBM. The focus of my project is to investigate the interplay between Spy1 and some of the known drivers of GBM. Using a transgenic mouse model created by the lab, I study the impact of Spy1-mediated effects on gene expression, self-renewal potential, stemness, and oncogenic transformation in the face of aberration in selected oncogenes and tumour suppressors known to drive glioma progression in neural stem cells.

I joined the Porter Lab at the beginning of my second year of University as an outstanding scholar. Having no prior research experience, I felt intimidated and unsure, worried that I would not be good enough for the lab. Fast forward to two years later, I can now say that joining the Porter Lab was one of the best decisions I made in university. Being surrounded by intelligent, motivated, and driven people inspired me to be persistent and hardworking both inside and outside of the lab. Through my thesis, I learned to think critically and creatively, mentor others, and overcome obstacles. The RA’s and Dr. Porter were exceptional role models and provided unconditional support despite my frequent mistakes. Through this experience, I learned what it means to do research and found my place in it.

Lia Oschanney

Liver cancer is one of the leading causes of cancer death globally, of which 90% of cases are Hepatocellular Carcinoma (HCC). One risk factor of HCC is Non-Alcoholic Fatty Liver Disease (NAFLD), often triggered by sedentary lifestyle, poor diet, and obesity. Most cases of HCC are cirrhotic and can start from excess fat accumulation in NAFLD. Chronic fat can trigger inflammatory responses that damage cells, leading to the liver initiating a proliferative response to recover. However, over time the liver shifts to a fibrotic state to maintain structural integrity with low levels of proliferation, characteristic of Non-Alcoholic Steatohepatitis (NASH). As fibrosis becomes extensive, the liver becomes cirrhotic, and the environment increases HCC risk; however, 20% of HCC cases are non-cirrhotic arising directly from NAFLD. The atypical cyclin-like protein Spy1 is capable of binding to CDKs, increasing cell division and overriding checkpoints. Spy1 has been shown to have a role in cancers, including HCC. Previous research has shown that mice with an overexpression of Spy1 in the liver develop NAFLD and HCC with decreased fibrosis, suggesting that elevation of Spy1 drives a proliferative response in the presence of cell damage, thus increasing HCC risk. To better understand the mechanisms that govern this response and the role of Spy1, this project examines NAFLD development in a transgenic mouse model driving Spy1 overexpression in the liver. This work may lead to the use of Spy1 as a potential drug target in the treatment of HCC.

In my first year, I knew I had a strong interest in research, but I was unsure how to take the first steps towards getting involved. Then, I discovered Dr. Porter’s website and was immediately captivated by the amazing work being conducted in the lab. However, just as I was eager to get started, the pandemic hit, and I was unable to set foot in the lab for nearly two years. Despite this obstacle, the lab kept me engaged through peer mentoring, which continued to fuel my curiosity and teach me the fundamental principles underlying research. When I finally had the opportunity to work in the lab as a third-year student, I had limited practical experience due to the majority of my undergraduate studies occurring online. However, under the guidance of the Porter lab team, I learned a multitude of techniques and quickly progressed from a hesitant and uncertain beginner to a confident and independent researcher capable of working on my own project. This experience has not only prepared me for future research endeavors but has also equipped me with invaluable skills that will undoubtedly serve me well in my future career in medicine. Completing my undergraduate thesis was an incredibly rewarding experience, and it would not have been possible without the support of my peers and the guidance of Dr. Bre-Anne Fifield and Dr. Lisa Porter. I am grateful for the opportunity to have worked in such an inspiring environment and to have learned from some of the brightest minds in the field.

Vanessa Vuong

The cell cycle contains checkpoints that delay mitotic progression and allow for DNA repair before cell division. Our cells receive thousands of DNA lesions a day and this damage can be caused by radiation, drugs, and other processes, leading to cellular mutations and carcinogenesis. There are two types of damage, single-stranded DNA breaks or the more lethal double-stranded DNA breaks (DSB). Upon receiving DSBs, the DNA damage response (DDR) pathway activates p53, which is a tumour suppressor protein that induces apoptosis or cell cycle arrest for cellular repair. Tuberin (encoded by the TSC2 gene) is another tumour suppressor protein that regulates the G2/M transition in the cell cycle and negatively regulates protein synthesis and cell growth. Mutations in tuberin can lead to the multisystem autosomal dominant disease known as tuberous sclerosis (TSC). Porter lab has previously shown that regulates mitotic onset through cellular localization of the G2/M Cyclin, Cyclin B1. My project focuses on the Tuberin/Cyclin B1 complex and its role in G2/M arrest for DNA damage repair. In this study, tuberin knockout NIH3T3 (mouse) cells were created using CRISPR editing technology. The cells will then be treated with etoposide, which is a chemotherapy agent that forms a complex with DNA and topoisomerase II to prevent DNA religation and this induces DNA damage. Wildtype NIH3T3 cells are compared to the knockout line and their cell cycle profile and DNA damage will be visualized using flow cytometry. Results have shown that without the prescence of tuberin, cells will return to G1 much faster since tuberin is not present to bind to cyclin B1 to prolong G2/M arrest. This project will provide greater insight into DNA damage induced carcinogenesis, TSC, and other proliferative diseases.

Joining Porter lab as a member of the peer mentor network and eveutally moving up to complete my undergraduate thesis was truly the best experience! I got to meet many wonderful people who help guide me throughout my struggles with getting data, especially those in the tuberin group who I had to see everyday. The problem-solving, critical thinking, and hands-on techniques I learned during my thesis are skills that I will certainly use in my future career. Best of luck to the incoming thesis students and new undergrads!

Undergrads 2022 – part 3

fidalgo — Tue, 24 May 2022 20:36:48 +0000

Determining the Dependency of Spy1 based on Rb Status in Triple Negative Breast Cancer

Breast cancer is the second most common cancer worldwide and the most common cancer among women. Triple Negative Breast Cancer (TNBC) is a particularly aggressive form of breast cancer with many subtypes based on gene expression profiles. There are currently no targeted treatments for TNBC due to its molecular characteristics, urging the discovery of new therapeutic targets. Potential therapeutic avenues are the cell cycle and its mediators which play an important role in cancer formation and progression. Spy1, a cyclin-like protein, promotes cell proliferation through the G1/S and G2/M checkpoints. Spy1 promotes proliferation even in the presence of DNA damage, overriding checkpoints and increasing cancer susceptibility. While Spy1 has been found to be elevated in breast cancer, its unique binding structure makes for an ideal candidate for cell cycle inhibition therapy. The retinoblastoma tumor suppressor protein (Rb) is known to regulate the DNA damage response system and is key in regulating the cell cycle. However, studies have shown that Rb is often mutated in TNBC inducing deregulated cell cycle progression potentially leading to tumor development. For some breast cancer subtypes the presence or absence of Rb (Rb status) can dictate response to treatment by cell cycle inhibitor drugs. Using in vitro TNBC models (MDA-MB-231 & Bt549 cell lines), this study aims to determine if Spy1 can override checkpoints independently of Rb status, and if elevated levels of Spy1 alter this response. These results could provide further guidance in developing cell cycle inhibition targeted therapies and potentially better TNBC patient outcomes.

“Before transferring to the University of Windsor, I wasn’t sure of how to get involved in research. Being in second year at the time, I was intimidated by research and unsure about my capabilities as a scientist. When I enrolled in CURES (cancer undergraduate research experience students), a course with Dr. Lisa Porter, I learned about the diversity of local cancer research taking place in Windsor-Essex. This course fuelled my motivation to learn, and genuine passion to contribute in this field. I found my confidence as a researcher when I joined Porter Lab in my fourth year, where I was thrilled to be given the opportunity to complete my undergraduate thesis. Up until my last year of University, I never knew myself to be as confident and disciplined as I am now. Being in a research lab has accelerated my learning, competency, and knowledge translation in such a short time. The atmosphere of Porter Lab is extremely motivating, and the people are supportive and patient. Porter Lab offers each member numerous opportunities to move forward science within the community and has helped me adapt to becoming a leader in science- a field I never felt I fit in before. During my time in lab, I have learned a multitude about the nature of science and what is involved in basic science research, including all the time invested in planning experiments and writing grants. Completing my undergraduate thesis was extremely rewarding and I am excited to continue my research in a master’s degree with Porter Lab, working alongside all the Porter Lab rats!”

The Impact of Cyclin B1 on Tuberin Stabilization

Tuberous Sclerosis Complex (TSC) is an autosomal dominant disorder caused by mutations in either TSC1 or TSC2, genes that encode the proteins Hamartin and Tuberin respectively. Hamartomas (benign tumours), skin lesions, neurological symptoms, renal dysfunctions, and retinal malformations are often present in TSC patients with varying severity. Tuberin and Hamartin regulate protein synthesis through mTORC1 inhibition. Tuberin can also delay mitotic onset at the G2/M cell cycle transition by binding to Cyclin B1. Hamartin has been shown to stabilize Tuberin by inhibiting its ubiquitination by HERC1 and subsequent degradation. Preliminary data from our lab suggests that Cyclin B1 may also contribute to the stabilization of Tuberin levels during the G2/M transition. Phosphorylation status of the cytoplasmic retention sequence (CRS) of Cyclin B1 plays an important role in the formation of the Tuberin/Cyclin B1 complex. The unphosphorylated CRS form of Cyclin B1 (Cyclin B1 5xA) binds stronger to Tuberin compared to the phosphorylated form (Cyclin B1 5xE). My thesis investigates the role of Cyclin B1 in Tuberin stabilization. HEK293-TSC1 null cell lines (IC2) were transfected with varying concentrations of Cyclin B1 5xA DNA and Tuberin protein levels were quantified by Western blot techniques. After blocking HEK293 cells in G2/M using thymidine and collecting them at different timepoints, endogenous Tuberin levels at different stages of the cell cycle were also quantified with Western blot techniques. Understanding the role of Cyclin B1 in Tuberin stabilization will shed light on cell proliferation and growth mechanisms that underlie tumorigenic disorders.

Characterization of Multiple Myeloma Cells Treated with CDK Inhibitors

Multiple myeloma (MM) is an aggressive hematopoietic cancer that is caused by the abnormal growth of plasma cells in the bone marrow. MM has a poor prognosis and most patients will eventually relapse with a more aggressive and untreatable form of the disease.
Cyclin Dependent Kinase Inhibitors (CKis) are an emerging targeted therapeutic option for Relapsed MM and have been investigated in many clinical trials. Flavopiridol is a first-generation non-specific CKI that inhibits CDK1, CDK2, CDK4, and CDK7. Flavopiridol has had high pre-therapeutic efficacy, but limited clinical success as a monotherapy. Dinaciclib is a second-generation CKI that is more selective and inhibits CDK1, CDK2, CDK5, and CDK9. Dinaciclib has also had low efficacy in clinical trials. In this study, we compared MM cells (U266B1) treated with Dinaciclib to Flavopiridol in-vitro by examining cell proliferation, apoptosis, and cell cycle profiles. Our data shows that there is no significant difference in cell proliferation or apoptosis of U266B1 cells treated with Dinaciclib and Flavopiridol after 24 hours, 48 hours, and 72 hours. Cells treated with Flavopiridol were more frequently arrested at the G2M checkpoint compared to Dinaciclib after 72 hours. Better characterization of MM cells treated with different types of CKIs will help to better understand their lack of clinical success as a monotherapy.

Undergrads 2022 – part 2

fidalgo — Tue, 17 May 2022 23:16:14 +0000

Applying CRISPR/Cas9 and Fluorescent Tools to Dissect the Role of Tuberin in Cell Cycle Regulation

How cells regulate their growth and division involves a tightly controlled integration of many mechanisms. In cells, Tuberin (gene – TSC2) is a protein in the Tuberous Sclerosis Complex (TSC) which modulates cellular growth, size, and proliferation. Mutations in the proteins forming the TSC can cause Tuberous Sclerosis Complex, an autosomal dominant disorder characterized by multisystem pathologies, and is often associated with benign hamartomas in the brain, kidney, lungs, and skin. The focus of my research is to clarify the role of Tuberin in the regulation of cell size and proliferation at the G2/M cell cycle checkpoint. During late G2, Tuberin retains Cyclin B1 (gene – CCNB1), a mitotic cyclin, in the cytoplasm thereby prolonging mitotic onset. We constructed six TSC2 mutants that harbour clinically relevant mutations which are known to destabilize the TSC. Interestingly, these mutations fall within the Tuberin Cyclin B1 binding domain. By over-expressing these Tuberin mutants in Tuberin null cells, an increased mitotic index is observed indicating a dysregulation of the G2/M transition. The resultant phenotypes are analyzed by flow cytometry, co-immunoprecipitation, and immunofluorescence. To aid in the temporal study of the cell cycle, we aim to validate successful CRISPR/Cas9-mediated knock-in of a near infrared (iRFP720) tag within the TSC2 gene of HEK293 cells, creating an endogenously expressed fluorescent Tuberin-RFP fusion protein. This new cell line will be a powerful tool to dissect the roles of Tuberin in regulating cellular growth and division and can provide a deep understanding of proliferative diseases like TSC and cancers.

“When I first applied to be part of the Porter Lab back in the Spring of 2019, I knew that I was joining a well-recognized and large research lab on campus which has continuously been on the forefront of scientific discovery. However, I did not foresee the new family I would make during my time here. Over the past few years, I have grown from being a student volunteer that started off by filling ethanol bottles and pipette tip boxes to working on a project that uses CRISPR technology to create a modified HEK-293 cell line! My progress as a young scientist has been a direct result of the mentorship and teaching opportunities in the Porter lab ecosystem. It took no time for me to get my hands dirty with experiments at the start, and while the COVID-19 pandemic had put a slight pause on my progress, the lab still provided a means of learning new skills and leadership opportunities through the Peer Mentor Network! I have made many friendships during my time here, and while some of these friends are also starting new chapters in their careers and lives, our relationship will prove to withstand the test of time. As I look forward to a life in medicine, these past few years have created a special place in my heart for biomedical research and it will certainly be a pillar of my future career. I am beyond grateful for the constant support that Dr. Lisa Porter and Dr. Elizabeth Fidalgo da Silva have given me this past year while also pushing me to do expand my scholarly boundaries. While embarking on my thesis this year, I have learned so many skills from troubleshooting experiments (shoutout to Adam Pillon for literally always being there to help) to communicate my work to both a general and scientific audience. As this chapter in my life nears an end, I will forever cherish the friendships and skillsets I have developed and will always have a special place in my heart for my fellow Porter lab rats. #porterlab #lifechanging #mentorship #futureinscience”

Spy1 Exacerbates the Long-Term Effects of Parity on the Mammary Gland

Age is a significant variable in cancer development, and approximately 1/3 of breast cancer cases occur in patients older than 70. Another emerging risk factor is parity, or childbearing, which may be linked to cellular changes that affect a woman’s risk of developing breast cancer over the course of her lifetime. These changes are thought to result from the mammary gland not reverting to normal

after lactation and involution – a developmental remodeling process where the milk secreting cells are cleared and replaced with adipocytes after weaning. Aberrant expression of the cyclin-like protein Spy1 has been shown to stimulate precocious development, resulting in disrupted morphology and oncogenesis within the mammary gland. Preliminary data suggests that the mammary glands of mice overexpressing Spy1 do not fully regress following lactation and involution, which may predispose them to breast cancer. We hypothesize that the overexpression of Spy1 exacerbates the long-term effects of parity on mammary gland morphology. To investigate this, we performed hematoxylin and eosin (H&E) staining as well as immunohistochemistry (IHC) on paraffin embedded sections, and whole mount staining of MMTV-Spy1 mice, a transgenic mouse model that overexpresses Spy1 within the mammary gland. We then compared the mammary gland morphology of parous MMTV-Spy1 mice to nulliparous MMTV-Spy1 mice, parous control FVB

mice, and nulliparous control FVB mice. This research begins to improve our understanding of Spy1’s role in regulating proliferation and apoptosis, contributes to our overall knowledge of breast cancer dynamics, and further solidifies Spy1 as an important target for treatment.

“When I first joined Porter lab, I was a volunteer with very little experience under the unprecedented circumstances of a pandemic. The amount of growth I have experienced as a scientist since then is astonishing, and I could not have imagined the full extent of it before I started as a thesis student. The environment and learning opportunities at Porter lab have been exceptional thanks to everyone’s commitment to our lab’s values, and I have met people and learned skills that will stay with me for the rest of my life. It still seems surreal to me to have been lucky enough to apply concepts, techniques, and an understanding of the scientific method from undergraduate courses to a real project with Implications on the field of mammary development and breast cancer research under the guidance of my mentors. I’m excited to pass the torch onto the next group of undergrads at the end of this journey, and excited to keep up with the lab’s future accomplishments.”

Undergrads 2022 – part 1

fidalgo — Mon, 09 May 2022 23:16:25 +0000

Control of Stemness in Glioma via Nanoparticle-Mediated Regulation of CD44 Receptor

Glioblastoma, also known as glioma or GBM, is the most common and aggressive malignant primary brain tumor which continues to puzzle researchers as decades of efforts to find an effective therapy has proven to fail. Barriers to success of the available treatment include impermeability of the blood brain barrier (BBB) to drugs, tumor heterogeneity, and existence of aggressive and resistant Tumour Initiating Cells (TICs). Collaborative studies in our lab show that conjugated polymer nanoparticles (CPNs) can serve as a potential therapeutic approach to overcome these barriers. CPNs are labeled with hyaluronic acid (HA-CPNs) which is a ligand for one of the most prominent receptors found on TICs, CD44. Our data to date has demonstrated that HA-CPNs cause selective decrease in proliferation, migration potential, and expression of markers of aggressiveness and self renewal in CD44+ glioma cells. This study investigates the mechanism by which HA-CPNs exert their effects and how the uptake of the nanoparticles is regulated. We study the status of CD44 receptor on the protein and mRNA levels as well as how manipulation of its levels affects glioma aggressiveness. We explore inhibition of Clathrin Mediated Endocytosis as responsible for the uptake of HA-CPNs and use it to validate selected effects of HA-CPNs. This work will reveal molecular mechanism behind HA-CPN anti-glioma effects which will further contribute to more in- depth evaluation of this nanoparticle system as a potential therapy for patients with GBM.

“Since joining Porter Lab in my second year of undergrad, I would have never thought this experience would have been so rewarding. Before my time in the lab, I had little knowledge of what the research side of academia was like. Through the COVID-19 pandemic, I found myself thinking largely about my future and through working in the lab via virtual platforms and maintaining my interest in research, I began my thesis in my forth year in person. Although this experience was such a learning curve, I have never been so appreciative and lucky to have this opportunity. Learning and gaining valuable skills in the lab such as trouble-shooting, team work, and resiliency, I can honestly say research has become such a passion of mine.

In my time in Porter Lab, I have created so many friendships with my fellow lab mates that I know will last forever. If you asked me in my first year what I thought I would accomplish during my undergrad, I would have never thought I could say that on a micro level, I am contributing to the advancement of cancer research in my community. I am excited to pursue my masters degree in biological sciences with Porter lab and eager to see all the change each and every member contributes to cancer research.”

Novel Cell Cycle Therapeutic Strategy Against Type II Medulloblastoma

Medulloblastoma (MB) is one of the most common malignant brain tumours, occurring mainly in children between the ages of 1 to 9 years. The standard of care which is currently being applied for MB includes combination of radiation, surgery, and chemotherapy. This treatment is aggressive and is only effective for a small percentage of patients due to the presence of aggressive and therapy resistant populations of Tumour Initiating Cells (TICs). A recent approach in clinical trials is the use of cell cycle regulator proteins, CKIs, which are synthetic cyclin-dependent kinase inhibitors. Our lab focuses on a specific cell cycle regulatory protein, Speedy (Spy1) which promotes proliferation and overrides cell cycle checkpoints established by natural CKIs. Spy1 plays a role in the maintenance and expansion of stemlike populations of TICs in different types of brain tumour. It is our hypothesis that Spy1 can drive tumour initiating cells in medulloblastoma in face and despite ongoing therapy. In this study we investigate and reveal the role of Spy1 in MB in response to known therapeutic agents and Spy1 mediated effects on the dangerous TIC populations in the treatment setting. In addition, Sonic hedgehog (Shh) MB, a type 2 medulloblastoma, is studied and manipulated to assess potential effects of Shh pathway in the regulation of Spy1. This project is the first one to demonstrate that standard of care treatment of MB leads to increased levels of Spy1 and selection of resistant TIC populations. We show that a key molecular driver in MB plays a role in the regulation of Spy1 levels which is an important finding in further studies seeking for novel effective therapeutic strategies against MB. Overall, my project will contribute to better understanding of the role of Spy1 in MB and its application as a potential therapeutical target in the future.

“Joining the Porter Lab at the end of my first year was the best decision I made in my University experience; it was life changing. I met a lot of amazing, professional and talented people that I am proud to call them my family now. I gained a lot of unique skills like patience, problem solving and critical thinking in addition to a lot of new techniques. I learned to never give up and most importantly passion in cancer research as without hope there is no meaningful research. My work in the lab was affected by Covid-19 but with the help of Dr. Porter, the RAs, the grad students and the mentor network I was able to overcome many challenges. We are a true family that stands which each other. Joining the Porter Lab helped me better understand the importance of cancer research and value the importance of hard work. My experience in the lab helped realize that I want to pursue a career that involves research in the future. I learned something new every minute I spent in the lab and this experience increases my desire in continuing to learn more and more every day. I am very grateful for my experience in the Porter Lab and I am proud to call myself a Porter Lab rat.”

The Role of Tuberin in DNA Damage Repair During Cell Proliferation

The cell cycle contains DNA damage checkpoints that delay mitotic progression and allow for DNA repair before cell division. DNA damage can be caused by radiation, drugs, and other processes which lead to cellular mutations and carcinogenesis. The tumour suppressor protein p53 is activated in the presence of DNA damage. It induces apoptosis or cell cycle arrest which allows cells to repair themselves. Tuberin (TSC2), another tumour suppressor protein, regulates the G2/M transition in the cell cycle and negatively regulates protein synthesis and cell growth. Mutations in tuberin can lead to the multisystem autosomal dominant disease known as tuberous sclerosis (TSC). Previously, our lab has shown that Tuberin regulates mitotic onset through cellular localization of the G2/M Cyclin, Cyclin B1. My project focuses on the Tuberin/Cyclin B1 complex in relation to G2/M arrest and DNA damage repair. In this study, we will overexpress Tuberin-WT and Tuberin clinical mutants in NIH-3T3 (mouse) and U2OS (human) p53 wild type cells. Etoposide, a topoisomerase II drug, will be used to induce DNA damage. Cells will then be analyzed by flow cytometry, TUNEL assay, and western blot to assess their cell cycle profile, apoptotic levels, and protein expression. Using CRISPR-Cas9 technology, a NIH-3T3 null TSC2 cell line will be created to clarify the role of Tuberin during DNA repair. Preliminary results have determined that 4μM of etoposide treatment at 8 hours arrests 50% of NIH-3T3 cells at G2/M. This project will provide greater insight into DNA damage induced carcinogenesis, TSC, and other proliferative diseases.

“Joining the Porter lab in my first year has been one of the most rewarding experiences of my undergraduate career. Not only was I able to learn about advanced scientific techniques in class, but I was able to put them in action throughout my own research in lab. The problem-solving, critical thinking, and hands-on techniques I learned during my thesis are skills that I will carry forever. I was also lucky enough to make life-long friendships during my research journey. There were some challenging moments, but I could always rely on my lab team members and mentors to guide me. The last three years have been an amazing experience, and I wish the best for the next team of brilliant undergrads.”

The Essentiality of Spy1 in Cooperation with Hepatocellular Carcinoma Drivers to Promote Tumour Formation

Liver cancer is the fourth leading cause of cancer-related deaths worldwide, standing at an estimation of 800,000 deaths annually. Among the various subtypes, hepatocellular carcinoma (HCC) is the most common primary liver malignancy. HCC is known to develop through a series of genetic and epigenetic alterations of proto-oncogenes and tumour suppressor genes in the liver environment. These changes ultimately lead to the malignant transformation of hepatocytes, the primary cells of the liver. Various HCC drivers are known to cause disruption of cellular pathways and promote tumour formation. Importantly, several cell cycle mediators cause dysregulation, thereby stimulating tumour formation and progression. The cyclin-like protein Spy1, promotes cell cycle progression and overrides apoptosis. Recent reports have detected increased levels of Spy1 in human HCC, which directly correlates to severity of the disease and poor prognosis. We hypothesize that Spy1 plays a critical role along with hepatocellular carcinoma drivers to advance tumour development.  We will test the essentiality of Spy1 on HCC development by first investigating potential gRNAs to use for Spy1 knockout in vivo in the liver, for hydrodynamic tail vein injections in wildtype mice. Simultaneously, in vitro testing of HCC cells (HepG2) will study the importance of Spy1 in HCC cell characteristics in combination with HCC drivers such as c-Myc, p53, and 𝛽-catenin. This project will assist in understanding the essentiality of Spy1 in HCC, which may reveal insight into the molecular mechanism of the tumour suppressors and proto-oncogenes connected with this subset of liver cancer. 

“Applying to Porter lab during the start of my second year was genuinely a spontaneous decision but it turned out to the best decision I’ve made in my life. Before joining the lab, I had no idea what I wanted to do after graduating but after stepping foot into Porter Lab, I was able to unlock my passion for research. My time in the lab learning new skills, meeting peers from various walks of life and finally executing my undergraduate thesis made me realize the potential for myself to pursue a career in research. I have learned an incredible amount of information during the years and the passion to learn continues to grow. I learned to appreciate research as not just a career but as a lifelong passion which at times is underapprecaited. It may be time consuming, require numerous amounts of trials and errors and force you to step out of your comfort zone but the journey is worth it. I was privileged to work alongside such an amazing team of research assistants and colleagues that supported me at every point of my journey. Seeing how much investment is made into cancer research, and how much of an impact every single attribute plays into the final goal, makes me realize that I chose the correct path and I can’t wait to delve more into it. The Porter lab has wonderful incoming students that will continue to make the lab’s name shine, can’t wait to see what more they can accomplish. I will always cheer on for the lab and its members!!. “