Every night I read to my daughters at bedtime, then head back to the lab for a second shift. One night, my youngest daughter, then 4 and a half years old, asked, “Daddy, you leave for your research every night...but have you ever saved anyone’s life from the research you do in your lab, ever?” My answer was, unfortunately, “No. not from my research.” I could have said that nobody really ever has - the path from basic science to clinical applicability seemed too complicated to explain. But she had a valid question, and it got me thinking: wouldn’t children with cancer ask the same question, too?
Fast forward to Spring 2014, at the annual Children’s Oncology Group meeting. The Chair announced dramatic cuts in the National Institute of Health’s budget for the COG... suggesting that not every childhood cancer could have an open clinical trial: only the ones with preclinical justification. The standard approach (dropping in adult cancer drugs into trials for children) simply hadn’t worked. At the same meeting, the NCI announced that as a result of the sequester, the budget of the Pediatric Preclinical Testing Program was cut earliest, and deepest, of any NCI program (by 40 percent)!
Among colleagues who were leaders in pediatric oncology at academic centers, the feeling was that preclinical testing of basic science findings, to move exciting discoveries to clinical trials, was too tedious and narrow-scoped for university laboratories and government programs. As a result, the best and brightest scientific discoveries for childhood cancers never actually make it into the clinic. Rare cancers are the hardest hit, with survival rates remaining stagnant for decades. Knowledge that could save kids’ lives simply languishes in this black hole: the preclinical gap.
By chance, my reading material for the airplane ride home was A Life Decoded, the book by J. Craig Venter. In this story of the first group to sequence the human genome, Dr. Venter achieved remarkable speed and cost efficiency by “going outside the box” of academia. Curious, I drove straight from the airport to a biotech incubator. Renting a 250 sq ft lab space for per year: a mere $10,500. The “what if’s” began: what if we could change the existing paradigm of research grants leading to publications (leading to more grants and papers, but never tangible results)?
What if we could bridge the preclinical gap as a mission...with scientists partnering with families to achieve the cures they so desperately desired? What if science driving drugs into the clinic existed as a singular mission? My research team and I simply wanted to know how a non-profit biotech could answer my daughter’s question. The result is the Children’s Cancer Therapy Development Institute. We value our pharmaceutical partners, give parents a seat at the table, and really listen to the clinical trialists. All in the name of converting scientific discovery into clinical trials for children with rare and underserved cancers.
WOW - our thanks to the anonymous donor who is offering a 1:1 match up to $250,000 for this project!
A question families often ask is, how does lab research on embryonal rhabdomyosarcoma apply to me or my child? Our lab doesn't offer medical advice, but the question is still valid. This project's goal is to determine the major types of embryonal rhabdomyosarcoma - and determine the best treatments for each "archetype". In a children's block letter toy set, There are 26 letters. If we divide those into 5 major groups (A-E, F-J, K-O, P-T and U-Z), we can look for treatments with available and emerging new drugs that treat each group in a more "personalized" way.
Our approach is as follows:
We will use publicly available RNA (and DNA) sequencing data to divide embryonal rhabdomyosarcoma into 5 groups. We will use Pax7:Foxo1 rhabdomyosarcoma as a ERMS-similar outgroup. We’ll know that these groups cover the full breadth of embryonal rhabdomyosarcoma (eRMS) because these biopsy samples represent all children across North America who were enrolled in Children’s Oncology Group (COG) clinical trials.
What’s next? We need experimental model systems to test drugs in. One kind of a model system is a cell line. The RD cell line is very popular for studying eRMS, but it was made in 1968. That’s a long time ago! We need a broader range of cell lines to match up to the COG biopsy samples. We’ve cataloged and have in our lab a great number of other eRMS cell lines (for details, see https://www.ncbi.nlm.nih.gov/pubmed/23665679).
We also develop new primary tumor cell cultures from families willing to share tissue from surgery (and sometimes from autopsies). Since these tumor cell cultures are made in recent months or years, they are perhaps especially reflective of contemporary eRMS patients.
Can we do more? Absolutely. We want to be able to go beyond cell lines in petri dishes for testing drugs: we aim to test drugs against the different types of eRMS using mice. We can implant mice with tumors from children, adolescents and young adults. Once tumors have grown, we can treat these mice with the drugs (or drug combinations) that showed the most promise in petri dish (cell line/cell culture) studies. Many models exist that we have already generated with our partners at The Jackson Laboratory. Our collaborators at Champions Oncology have also created a number of these models. We continue to make new models as families donate tissue to this effort.
THE RESULT: Generating data as efficiently and accurately as possible, we plan to have drug – genetic matches for each type of embryonal rhabdomyosarcoma … and to publish this data, but also to share it in real time with pediatric oncologists of eRMS patients who may have scientific interest.
Why is this important?
This project is important because many children, teens and young adults with embryonal rhabdomyosarcoma need new options. When metastatic, the five-year survival rate is only 43%. We want eRMS to be universally survivable --- with the least side effects and highest quality of life after treatment.
As a grandparent recently put it,
"Childhood cancer is vastly underfunded (for a variety of reasons), even though it is a particularly tragic variant of cancer. To watch a two-year-old undergo a drastic surgery and chemotherapy is difficult. But what is really heartbreaking is when the best and most aggressive treatments are unable to save a child's life. Each child saved may have 70 years added to their lifespan, including many of the best years. Community support is urgently needed to work towards breakthroughs."
As a mother conveys,
"My son was diagnosed with an aggressive and deadly form of cancer called rhabdomyosarcoma when he was just 19 months old. When he was first diagnosed, and nearly every week since then, I have taken to reading the medical literature about rhabdomyosarcoma, looking for new treatments that could possibly help improve the odds of my son beating this cancer. Sadly, very little research is being done on rhabdomyosarcoma. First off all, there is not enough government funding of cancer research in general (and my understanding is the funding situation is getting worse and worse). Second, very little--we're talking around 4%--of funding for cancer research in the US goes to pediatric cancer. Of that limited pediatric cancer research funding, only a tiny amount goes to rhabdomyosarcoma because it is considered a "rare" cancer. (However, there are 400-500 kids diagnosed with rhabdomyosarcoma each year... they do matter.... and research on "rare" cancers can help our understanding of other cancers, too.) So, the treatments that are currently used for rhabdo are very old treatments, with high toxicity, and they only cure rhabdomyosarcoma about 60% of the time. This is terrifying and unacceptable."
Who will benefit?
Children, teens and young adults with embryonal rhabdomyosarcoma will benefit from new drug treatments, matched to a child’s tumor’s genetics, being introduced into clinical trials. Our longstanding work on eRMS also tells us that eRMS is related to certain other soft tissue sarcomas (https://linkinghub.elsevier.com/retrieve/pii/S1535-6108(10)00532-5), making it possible that both children & adults affected by eRMS and undifferentiated sarcomas will benefit from this two-year project.
BudgetThis project in its basic form runs two years at $125,000 per year ($250,000 total). The expenditure category breakdown is given below. It is supervised by Dr. Keller, and led first-hand by a biomedical engineer. The project has both bioinformatics/computational biology and wetlab experiment studies. If this crowdfunding project reaches its goal, our anonymous donor will match each dollar 1:1, doubling the funds available allowing us to hire a biologist to work alongside our biomedical engineer ... doubling the speed and throughput towards our mission find the best treatment option for each group of children with embryonal rhabdomyosarcoma.
Our engineer Noah and bioinformatics intern Chandler are busy building archetype groups across child, teen and young adult embryonal rhabdomyosarcoma. Above is a pictograph representation of their preliminary results.
Pictured is team leader, Noah, and bioinformatics interns Nick (front) and Chandler (back) working on the integration of ERMS cell lines into the Archetypes above. more soon!
Work over the past few weeks has been on "data aggregation". Specifically, we're trying to fill in the gap in the last archetype/tumor family tree diagram by adding in typical ERMS cell lines. First we get permission for the data, then we download it, then we run it through an analysis pipeline for each cell line, then we re-analyze all sequencing data (from biopsies, cell lines, GEM mouse models and PDX mouse models). We are constantly being vigilante to use the right "gene list" so that mouse and human ERMS can be compared directly. We're just about there - stay tuned!
For the family tree studies, there are a number of algorithms that can be used, each with slightly different (but important) subgrouping effects that we observe. We are currently using DNA mutations for known outgroups to pick the best dendogram (subgrouping) algorithm. Another next step, by our new engineer Cora, is to integrate Pax7:Foxo1 rhabdomyosarcomas into this study as an ERMS-related outgroup. This is an important "control group" to getting the family trees right.
from Cora (engineer project lead):
Over the past week we have worked to finalize the "database overview", "get analyzed data", and "data aggregation" portions of the project. We are ensuring that users will have the ability to download analyzed data for each sample and obtain a statistical summary of the data without any problems. Next, we will be focusing on integrating Pax7:Foxo1 rhabdomyosarcoma into the study.
Pax7:Foxo1 rhabdomyosarcomas are now integrated into the archetype! (they will serve as an 'outgroup'). As mentioned previously, there are many types of subgrouping algorithms. We are currently deciding the best method of clustering with the Pax7:Foxo1 rhabdomyosarcomas now integrated into the archetype. Once that is completed we can begin to utilize the dendogram for drug discovery.
This week has been dedicated to figuring the best subgrouping algorithm. We have added a table matching specific mutations to each sample as well as the diagnosis of each sample. By using the table, we can decipher which method of clustering groups together specific mutations of interest (Pax7:Foxo1 being one of them). We still have some work to do, but we are well on our way to finalizing the dendogram!
We have also been working on some preliminary experiments. We will be using cell lines derived from eRMS tumors to look at alterations in the cells' phenotype once applied with a small molecule. In a process known as high content analysis (HCA) staining we can label proteins with fluorescent tags and detect changes at the subcellular level using imaging. This will be an especially useful tool for drug discovery in the later stages of this project.
This week has been focused on making small changes to the dendrogram so that we can ultimately decide the best subgrouping algorithm. Here's a closer look at the program we've been working with (above). We also have been continuing our preliminary experiments on the staining protocol. We hope to identify a variety of cellular alterations, such as proliferation, apoptosis, and myo-differentiation. More to come soon!
Dr. Keller and I were lucky enough to attend a wonderful event called Golf Fights Cancer (GFC). This organization gets golfers together and raises money for cancer, and cc-TDI was their chosen mission to support for this event! Moreover, this event was in support of this very project. I was also able to see my initial inspiration for joining cc-TDI's mission- my cousin, Kiley Sullivan, who was diagnosed with stage 4 alveolar rhabdomyosarcoma in 2014. Pictured is Brian, co-founder of GFC, the Sullivan family, Dr. Keller, and myself at GFC. It was amazing and heart-warming to see so much support from so many people at this event.
In terms of the dendrogram, we are close to completing it. We are still working on choosing the best subgrouping algorithm and ensuring that we have the right set of genes that fully recapitulates mouse and human eRMS tumors.