My interest in cancer research stems from my personal experience as a parent of a child with cancer. My oldest daughter was diagnosed with anaplastic Wilm's tumor when she was just 4 years old. As we progressed through her treatments, it became apparent that her prognosis was poor, and her options were limited. It became my personal mission to attempt to provide more-promising treatment options, not only for my daughter, but also for any child suffering with a rare cancer. Children with rare cancers often face a bleak prognosis with limited treatment options due to the fact that very little research has been done with their diseases. Due to their small population numbers, there is little investment in children with cancer, especially rare cancers. I am dedicated to advocating for children with rare cancers and finding ways to further research for this population of cancer patients in the hopes of providing more-effective and less-toxic therapies for children suffering from cancer. I joined the cc-TDI team in the summer of 2016, and focus on childhood kidney cancers.
Wilms tumor is a childhood kidney cancer which affects approximately 500 children per year in the U.S. Most of these will be cured with chemotherapy, but the children with anaplastic Wilms tumors do not respond well at all to current protocols. We believe that a new type of treatment(s) geared towards anaplastic disease is warranted in the hopes of improving their current survival rates. To this end, we have recently collected and validated several WT primary tumor cell cultures, cell lines and mouse explants. We have outsourced these for genetic sequencing (both RNA and DNA), and will analyze the results for unique biomarkers and mutations. We have run several of these cell cultures through drug screens and collected drug hit data. We will apply all of this collected data to our recently published probabilistic Boolean modeling method and develop a hypothesis as to an ideal drug combination which might be more effective against anaplastic Wilms tumor. We will then test this hypothesis in-vitro (in test tubes), ex-ovo (a quail egg CAM assay) and in-vivo (in mice). We hope that this method of discovering novel compounds might provide more effective treatment options for children with anaplastic disease.
Why is this important?
This project takes an in-depth look at anaplasia in Wilms tumor. While Wilms tumor in general is considered a very "cureable" cancer, the children with the anaplastic subtype do not respond well to current therapies. In order to cure the most common kidney cancer in children once and for all, we must focus on the high-risk subtypes, including those with "un-favorable" (anaplastic) histology. With this study we hope to learn more about anaplasia in cancer and why it results in tumors that are trickier to treat. We hope that this particular research project will ultimately result in more-promising and more-effective treatments for children with the anaplastic variant of Wilms tumor, while also generally advancing the knowledge of anaplastic cancers. This study uses a unique approach to develop an innovative mathematical model of an ideal drug combination which might be more effective against anaplastic disease.
Who will benefit?
Children with Wilms tumor, children with anaplastic Wilms tumor, and cancer patients with anaplastic diease.
BudgetDonations to this crowdfunding project will be used to purchase reagents for testing purposes, ship Wilms tumor research resources (cell cultures) to the lab, and continue genetically sequencing (both whole genome and transcriptome sequencing) the samples we have. This project will also provide a small stipend which will be used as "bridge-funding" and will allow me to continue working in the lab until we receive sustained funding from traditional granting sources.
Project Update - 9/15/16
I recently had the opportunity to complete a summer internship at The Children’s Cancer Therapy Development Institute in Portland, OR. My mentors were Dr. Charles Keller and Dr. Noah Berlow. Our project, “The functional genomics of Wilms’ Tumor,” explored more-promising treatments for children with Anaplastic Wilms’, a rare subtype of the disease. Wilms’ tumor (or nephroblastoma) is the most common kidney cancer found in children, and most respond favorably to traditional cancer treatments (surgery, chemotherapy, and radiation). However, those with the anaplastic version of the disease tend to respond poorly, with overall survival rates of 33% or less. Our study seeks to discover novel and more-promising treatments for children with Anaplastic Wilms’ tumor.
Exposure to the field of pediatric oncology: My summer internship at cc-TDI was amazing! Due to my family’s personal experience with childhood cancer (my oldest daughter is a Wilms’ tumor survivor), I have been interested in pediatric oncology for some time now. This summer’s internship served to strengthen my resolve to become directly involved in the childhood cancer research process. Prior to this summer’s internship, I had no experience in a laboratory setting. I am grateful for the community support and Dr. Keller for allowing me the opportunity to work directly in a lab focused on childhood cancers.
Development of practical research skills and experience with a real-time investigation: Over the course of the internship, I learned the basics of cell culture technique. I learned how to bring cell lines out of a frozen state, how to grow them into cell cultures, and how to maintain them. I also learned how to prepare them for genetic sequencing and STR analysis, how to add them to a drug screen, and how to cryopreserve them for future use. I was responsible for preparing a drug screen with 60 different compounds and testing them against cell cultures. This involved learning molarity calculations, pipetting techniques in a sterile hood, working with robotic drug screens, curve-fitting data, and careful attention to detail. Over the course of the summer, I was able to observe a mouse necropsy, western blot analysis, flow-cytometry assay, and the development of a quail egg CAM assay. Within my internship, I had the opportunity to attend tours of the OHSU medical school and pathology department, as well as the PhD program, and I also participated in cc-TDI’s Pediatric Cancer Nanocourse.
Research accomplishments: During the 6 week internship, I achieved several goals of the Anaplastic Wilms’ tumor project. We were successful in obtaining a variety of Wilms’ tumor cell lines and mouse model explants, and we worked with 4 primary cell cultures. I also facilitated several tissue donations. We were able to culture many of the cell lines, although the lines often grew slowly. I helped design and print a drug screen with 60 compounds, and tested 2 Anaplastic Wilms’ tumor cell lines on the screen.As part of the project, we prepared cell cultures for STR validation analysis and for whole exome and whole transcriptome sequencing. I also performed extensive literature research in order to gather published data on existing cell lines and known mutations found within subpopulations of Wilms’ tumor.
Future plans: My immediate plans are to continue the Anaplastic Wilms’ tumor project at cc-TDI. We will use the preliminary data generated to apply for grants and further the study. I would like to continue a career path towards cancer research with a focus on pediatric oncology, especially the children who suffer from cancers with little hope of a cure currently. It is often challenging to find ways to study these rare cancers, yet these are the children who deserve the most attention. Despite the funding difficulties inherent in studying childhood cancers, I would like to continue in this field with what I consider to be very impactful work. The connections made at OHSU during this internship period have inspired me to pursue a Master’s degree in Cancer Biology.
Scientific Methods and Results: Towards our goal of discovering more-promising treatment options for children with Anaplastic Wilms’ tumor, we first obtained and cultured several Wilms’ tumor cell lines. We then developed a drug screen of 60 anti-cancer compounds and tested these against two Anaplastic Wilms’ tumor cell lines, WiT49 and 17.94. WiT49 was generously provided as a gift from Dr. Herman Yeger at Sick Kids Hospital in Toronto, CAN. The 17.94 cell line was obtained from the German DSMZ tissue bank. Cells were maintained in DMEM and supplemented with FBS and penicillin/streptomycin in a humidified incubator with 5% CO2 at 37°C. Sixty anti-cancer reagents were purchased from Selleck Chem, Sigma Aldrich, Tocris, Active Biochem, AdooQ, Apexbio, Chemietek, EMD Millipore, LC Labs, and Xcessbio. Reagents were diluted with NBA to 20 uM and then put into wells in 1.0, 0.1, 0.01, and 0.001 dilution ratios. These were then printed onto 384-well plates with an epMotion5075 liquid handling robot, and the 2 cell lines were applied to the reagents. Cells were exposed for 72 hours and then checked for metabolic activity via a CellTiter-Glo luminescent cell viability assay. Fluorescence was measured with a BioTek Synergy HT luminometer and the resulting data was compiled in a heat map chart. Within the drug screen, activity (cancer cell growth inhibition) was noted with several compounds. The overall slow growth of most of our Wilms’ tumor cell lines was a challenge to our project.
Future Study: At the time of this report, several human Wilms’ tumor cell lines remain in culture (COG-W-408, COG-W-477, imWilms1, imWilms6, imWilms10, 4 primary cell cultures (PCB 007, PCB 053, PCB 064, and PCB 098), and mouse explants KT-13, KT-18). There are also 2 cell lines (CCG99-11 and WitP3) which are being grown up in separate labs, and will be added to our study when they are ready. These cultures will be tested against the developed drug screen and will also be genetically sequenced. We plan to integrate the chemical (drug) sensitivities data with the DNA mutations/amplifications and RNA expression data to conduct a probabilistic Boolean modeling approach developed at cc-TDI (published in BMC Bioinformatics, 2013. 14:239). Our hope is to develop a novel hypothesis which might predict drug combinations that would be effective for anaplastic Wilms’ tumor patients. I will be actively seeking grant funding for these studies that I hope to pursue full time, as soon as funding allows.