During our lab meeting today one of our graduate students, Isabelle Hinch, presented a great seminar about Mental Health in Academia.

Academic life, for both faculty and students, has always been stressful, but the pandemic has made it worse. In her presentation, Isabelle share resources that can help to deal with the insecurities and stress in a University community.

Check here to watch her presentation.

Producer and Director – Our fabulous graduate student Isabelle Hinch.

Just in case you missed, below are the previous videos:

Part 1 – Lisa and Adam

Part 2 – Nick and Sami

Part 3 – Jackie and Jill

Part 4 – Isabelle

Part 5 – Emily

Part 6 – Samaneh

Just in case you missed, below are the previous videos:

Part 1 – Lisa and Adam

Part 2 – Nick and Sami

Part 3 – Jackie and Jill

Part 4 – Isabelle

Part 5 – Emily

Check out:

Part 1 Part 2 Part 3 Part 4 videos!

Identifying Molecular Markers of Progression to Muscle Invasive Bladder Cancer

An estimated 9,000 Canadians are diagnosed with bladder cancer each year making it the 5th most common cancer in Canada, the 12th most common among women and the 4th among men. Most patients are initially diagnosed with non-muscle invasive bladder cancer (NMIBC), which in some cases can progress to muscle-invasive bladder cancer (MIBC). MIBC is associated with a significantly poorer prognosis than NMIBC, and it is unclear why some progress to MIBC while others do not. Thus, a better understanding of the molecular progression of bladder cancer is needed. DDR2 is a tyrosine kinase that functions as a cell surface receptor for collagen and regulates cell differentiation, cell migration, invasion, and proliferation. It is known that an increase of DDR2 is ninety percent specific for muscle invasion in bladder cancer and that DDR2 is overexpressed in MIBC compared to NMIBC. Our results showed that MIBC is associated with an increase/gain of DDR2. With that being said, this project sought to demonstrate that DDR2 can be used as a prognostic indicator for the progression to MIBC and can be used as a therapeutic target preventing progression to MIBC.

Treatment Timing of Triple-Negative Breast Cancer

Breast cancer is the second highest cause of death from cancer in Canadian women, and triple-negative breast cancer (TNBC) accounts for 10-15% of these cases. Since TNBC lacks three main receptors (estrogen receptor, progesterone receptor, and HER2) that are normally used to target breast cancer, generalized chemotherapy treatments must be used instead. Chemotherapy drugs currently used to treat TNBC include adriamycin (A), cyclophosphamide (C), paclitaxel (T), and carboplatin (Ca). In current treatments, patients receive AC for 4 cycles biweekly followed by T (or T+Ca) for 4 cycles biweekly. Each drug targets various aspects of the cancer cells and causes the cells to arrest in various phases of the cell cycle. This study looked into how the timing and pattern of the administration of the drugs influence the overall results in treating TNBC. To test this, MDA-MB-231 TNBC cells were treated in vitro with one or more of the drugs (AC, T, or T+Ca) at various time-points and the proliferation rate and cell cycle profile of the cells were retrieved using Flow cytometry and Trypan Blue exclusion assay. The results indicated that the pattern of chemotherapy drug treatments that leads to the highest response of TNBC cells was when cells were first treated with T followed by T+Ca. This denotes how the pattern of drugs influences the effectiveness of treatments, and that the order found contains the highest efficiency. The information found may help improve the 5-year survival rate of patients with TNBC and save many lives in the future.

Fatima has completed two theses back to back.

CD44-Selective Conjugated Polymer Nanoparticles: New Theranostics for Glioblastoma Multiforme (2020)

 The most aggressive type of brain tumour is Glioblastoma multiforme (GBM). GBM encompasses 17% of all brain tumours, and the median survival for patients does not exceed 15 months. The diffuse nature of the disease and the high level of heterogeneity leads to ineffectiveness of surgical resection and diminished success of established therapies. A novel therapeutic option includes utilizing nanoparticles. My project specifically focuses on the molecular properties of Conjugated Polymer Nanoparticles (CPNs), which are Diketopyrrolopyrrole-based (DPP) CPNs, that are fluorescently labeled with a Hyaluronic Acid (HA) coating. I will first study the CPN concentration-dependent effects on glioma cells in vitro by studying the cell cycle phase accessibility, cell proliferation, death, and metabolic activity. I will investigate the selectivity of targeting CD44, a HA receptor, positive cells by CPNs, comparing CD44+ and CD44- cell populations, separated using Fluorescence-Activated Cell Sorting (FACS), or by using CD44 knockdown. Finally, the bioavailability of CPNs will be assessed using an in vivo Zebrafish system. I will first evaluate any potential toxicity of CPNs on the Zebrafish, and then I will determine the optimal concentration and timing of CPN treatments. I will then investigate the effects of CPNs on xenografted U251 tumours in vivo. This project will enhance our understanding of the CPN-mediated effects not only on the GBM cells but also in an in vivo setting allowing for further development of this therapeutic approach to potentially improve clinical outcomes in patients with GBM.

The Role of the Microenvironmental Landscape in Glioblastoma Multiforme Progression and Therapy Resistance (2019)

Glioblastoma multiforme (GBM) is a type of brain tumour that is categorized as having the highest degree of aggressiveness, accounting for 60% of adult brain tumours with poor prognosis. Despite extensive chemo- and radiotherapy treatments, patients’ relapse. Therefore, a better understanding of GBM biology is crucial to the advent of effective therapeutic interventions. The tumour niche, also known as the cancer stroma, is composed of the extracellular matrix and several types of recruited cells including fibroblasts. Fibroblasts secrete diverse molecules that were found, in other types of cancer, to contribute to the maintenance of the malignant characteristics of the tumour mass. Therefore, we hypothesize that fibroblast activation plays a crucial role in the aggressiveness and progression of GBM. We will first study the characteristics and content of the fibroblast populations in sections obtained from GL261 glioma cell line-derived brain tumours, in comparison to normal brain tissue. We will employ commercially available mouse embryonic fibroblasts to establish co-cultures with GL261 cells in-vitro. Both monolayer and 3D culture models will be utilized to study the activation and the role of the fibroblast component in the control of GBM progression and therapy resistance. In summary, my project will not only contribute to a better understanding of the mechanisms regulating the GBM microenvironment, but it will also identify potential novel treatment approaches.

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

“Being part of the Porter Lab has meant so much to me and has been the highlight of my undergraduate career. I remember receiving Elizabeth’s email that I had been accepted into the lab after completing an interview during Winter 2017. I was simply over the moon! I started my journey in the lab by helping out with lab maintenance and interacting with several students at different stages in their careers. This gave me insight into what really goes on behind the scenes in a research setting where everyone is a team player and serves an integral role in the overall success of the lab – whether this is through baking filter tips for an RNA extraction or by offering feedback when your IHC doesn’t work. With each semester, I began to appreciate the intricacies of research even more. I greatly admired the determination, grit, and hard work that my fellow lab mates displayed. All these factors and qualities that I gained from my lab team were traits that I knew would help me in my thesis and future career. When I started my thesis, it was definitely a steep learning curve, but I was eager to continue learning and knew that I had my lab team to support me every step of the way. The lab’s continuous support led me to complete two theses featuring projects that I am very passionate about, and which taught me so much about the vast and extensive field of GBM research. I knew how debilitating this disease could be on a personal level as I grew up in an underserved community and witnessed this disease affecting several of my family members. The perseverance of my lab team and knowing that there are so many individuals involved in cancer research is very reassuring and gives me so much hope for those who may be affected by this disease.“

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. “

Just in case you missed:

part 1, part 2, and part 3 videos!

Hope you enjoyed it and stay safe!

Jackie Fong shows us how to create an office space at home and Jillian Brown is sharing recipes for making wonderful cocktails. I guess it is a great combo ‘science and drinks’ for this difficult time! 🙂

Just in case you missed the previous videos, part 1 is here and part 2 is here.