Nanocourse publication

Congratulations to the 2012 Participants of the OHSU Pediatric Cancer Nanocourse whose peer-reviewed commentary, "A Diffuse Intrinsic Pontine Glioma Roadmap: Guiding Research Toward a Cure" is published in the journal, Pediatric Blood & Cancer.  
  
It should be emphasized that this scholarly work is that of members of the community: parents, survivors and students.  We are grateful to have been able to work with this talented and insightful group of individuals.  

Banbury Conference on Rhabdomyosarcoma

stay tuned for additional information on an upcoming Banbury Meeting on Rhabdomyosarcoma, hosted by the Cold Spring Harbor Laboratory, May 13-16.

Tumor Cells in a Hurry: Duct Tape will do!

In alveolar rhabdomyosarcoma, 70-75% of tumors carry the Pax3:Foxo1a oncogene.  This oncogene seems to account for a large difference in the response to chemotherapy and radiation relative to rhabdomyosarcomas that do not carry this fusion gene.  

  

In a report just released in PLoS Genetics, Ken from our lab reports that Pax3:Foxo1a levels fluctuate during tumor cell replication, and that a primary function of Pax3:Foxo1a seems to be “checkpoint adaptation” – the process of giving the cell permission to continue cell division despite damage from therapy… with the hopes that this damage can be later repaired, simply tolerated – or may offer an advantage to the tumor cell as a result of newly gained mutations/properties. The paper is freely available online at:  http://www.plosgenetics.org/doi/pgen.1004107. 

Fishermen helping kids with cancer

This incredible group of commercial fishermen in British Columbia support children with cancer and childhood cancer research while having a fun time. Their website and 2013 herring sale video can be seen at www.fishermenhelpingkidswithcancer.com. 

Lemon Ball!

On Saturday Jan 10th was the Alex's Lemonade Stand "Lemon Ball". What a fabulous event, and a terrific community of supporters.  In attendance was not only Charles, but also our collaborator DR. David Langenau from MGH (fish rhabdomyosarcoma expert) and his wife, Brenna.  Dave and Brenna are pictured here.  

International group of pediatric brain cancer researchers receives $300K toward a cure

press release

Parents of children who died from rare cancer believe greater investment in quality research is critical

PORTLAND, Ore. – An international consortium of researchers focused on identifying new molecularly targeted drugs to treat the most fatal form of childhood brain tumor, diffuse intrinsic pontine glioma, or DIPG, has been awarded nearly $300,000 by The Lyla Nsouli Foundation for Children's Brain Cancer Research, based in London, England. The foundation was established in memory of 3-year-old Lyla Nsouli, who died in January 2012 after a devastating five-month battle with DIPG.

Lyla’s parents, Nadim and Simone Nsouli, are hopeful their contribution will help bring researchers closer to a cure: “Facing her sudden, brutal diagnosis without any real option for treatment or survival is not an experience any child or their family should ever have to bear. Significantly greater investment in quality research is vital to improving the prognosis for children like Lyla. We are determined that research can provide treatments and eventually a cure for this cruel childhood cancer.”

To date, no treatment does more than incrementally increase survival of children with DIPG. One day a child may have a headache or unsteadiness, but the next day a family's life and plans are tragically changed. A group of international researchers called the DIPG Preclinical Consortium hopes to change this.

“Our first phase of drug screening and tumor DNA sequencing couldn't have been possible without the support of the Lyla Nsouli Foundation, the Cure Starts Now, Accelerate Brain Cancer Cures and CureSearch Foundations. Now that we have drug leads, the hard work of validating these begins. The Lyla Nsouli Foundation has been with us every step of the way, both in terms of support and accountability — both matter,” said consortium coordinator Charles Keller, M.D., associate professor of pediatrics at Oregon Health & Science University Doernbecher Children’s Hospital and the OHSU Knight Cancer Institute.

“The members of the Children’s Oncology Group CNS committee express a deep sense of gratitude to the Lyla Nsouli Foundation for funding the DIPG Preclinical Consortium,” said Amar Gajjar, M.D., chair of the brain tumor committee for the National Cancer Institute-supported Children's Oncology Group (COG). “The grant from the foundation has sparked a global effort to find new and effective therapies using the latest technologies currently available against diffuse intrinsic pontine glioma. The rapid translation of information from research laboratories to a clinical protocol is an often sought aim for advancing cancer cure rates – the grant from the Lyla Nsouli Foundation has made this dream a reality.”

Gajjar and Maryam Fouladi, M.D., co-chair of the COG brain tumor new agents committee and leader of its Pediatric Brain Tumor Consortium, conceived the DIPG Preclinical Consortium with Keller: “Real-time science in partnership with the community for a shared goal of finding a two-drug combination to put into international clinical trials for DIPG.”

Consortium member Jacques Grill, M.D., Ph.D., Institut Gustave-Roussy, Villejuif, France, innovates by creating living cell cultures not from autopsy-derived tumor samples, but from biopsies from the brainstem. This novel approach was initially controversial but is winning acceptance, Keller noted. 

“Grill and his colleague, Dr. Darren Hargrave at Great Ormond Street Hospital, London, keep a clear line of communication so that the consortium's results are reported in real time to the European clinical trial groups to inform on that side of the Atlantic,” said Keller. “The scientific teams are diverse due to the pressing nature of the need to understand and treat DIPG.”

Each member will take a different but complementary role to ensure the results of the robotic drug screen of 17 autopsy- or biopsy-derived DIPG cultures can be validated in mouse models.

"This collaboration has been a wonderful opportunity to work together as a community to move the field closer to an effective therapy for this terrible disease. I am hopeful that, together and with the immense support from Lyla Nsouli Foundation, we will make real strides forward now," said Michelle Monje, M.D., Ph.D., Stanford University Beirne Faculty Scholar in Pediatric Neuro-Oncology, Stanford Cancer Institute, Lucile Packard Children's Hospital.

"The DIPG Preclinical Consortium offers hope where once there was very little. When my son, Andrew, was diagnosed with DIPG in October 2007, I never imagined that such a collaboration would exist a few short years later — a collaboration where exceptional science and a remarkable patient community meet in partnership to change the future for children like Andrew and Lyla," said DIPG parent Sandy Smith.  

# # #

Consortium members include: Keller, Kellie Nazemi, M.D., Nathan Selden, M.D., Ph.D., Doernbecher Children’s Hospital, Oregon Health & Science University; Monje, Stanford University; Grill, Institut Gustave-Roussy; Oren Becher, M.D., Duke University Medical Center; Cynthia Hawkins, M.D., Ph.D., University of Toronto; Xiao-Nan Li, M.D., Ph.D., Baylor College of Medicine; Esther Hulleman, VU Cancer Center Amsterdam; Eric H. Raabe, Johns Hopkins University; Katherine Warren, Paul Meltzer and Martha Quezado, NIH; and Marta Alonso, University of Navarra, Madrid, Spain.  

The DIPG Preclinical Consortium is funded by the Lyla Nsouli Foundation for Children’s Brain Cancer Research, the Cure Starts Now, Accelerate Brain Cancer Cures and CureSearch Foundation.

ABOUT OHSU DOERNBECHER CHILDREN’S HOSPITAL

OHSU Doernbecher Children's Hospital ranks among the nation’s "Best Children’s Hospitals."* It is one of only 22 National Institutes of Health-designated Child Health Research Centers in the country and ranks 39th for NIH awards to children's hospitals and their university-affiliated department of pediatrics.**

Nationally recognized physicians and nurses at OHSU Doernbecher provide a full range of pediatric care to tens of thousands of children each year from Oregon, Southwest Washington and around the nation in a family-centered environment. OHSU Doernbecher specialists also travel throughout Oregon and Southwest Washington, caring for more than 3,000 children at more than 200 outreach clinics in 15 locations. Neonatal and pediatric critical care experts provide round-the-clock consultations to community hospitals statewide through OHSU Doernbecher's state-of-the-art telemedicine network.

*U.S. News & World Report 2013-14 Best Children’s Hospitals

** National Association of Children’s Hospitals and Related Institutions (NACHRI)

ALSF REACH Grant!

We are grateful to the Alex's Lemonade Stand Foundation for funding to enable the translation of a basic science discovery to preclinical trials so that Children's Oncology Group phase I and II trials might be possible.  The discovery of EphB4 as a therapeutic target in alveolar rhabdomyosarcoma was work of former HHMI medical student fellow, Imran Aslam (now a JHMI internal medicine resident).  Working in collaboration with the Druker laboratory, Imran uncovered the EphB4-EphrinB2 as the potentially highest value target in rhabdomyosarcomas.  This ALSF REACH award allows us to investigate an anti-EphB4 antibody in embryonal rhabdomyosarcoma, as well as an EphB4-nanoparticle sump in alveolar rhabdomyosarcoma, embryonal rhabdomyosarcoma and osteosarcoma.  This work will be carried on by our star muscle- and mouse models expert, Megan.  Check back regularly for updates on Megan's progress!

project updates:

01/14/2014:  Project Funded!
  
02/06/2014:  The immunocompromised mice we will be using to test the efficacy of an anti-EphB4 antibody have arrived. These mice will be used to create pilot orthotopic xenograft models of eRMS in the coming weeks.  
[ picture:  at center is Megan. ]  
  
02/20/2014:  This week, the first set of immunocompromised mice were injected with Rh18 cells (a human derived eRMS cell line). This begins a pilot study to demonstrate the latency with which a xenograft from this cell line will successfully create tumors in the strain of mice that we will use for future Alex’s Lemonade Stand experiments. In other strains of immunocompromised mice, Rh18 cells have generated tumors in 6-8 weeks- we’ll have to wait and see if these mice follow suit!
  
02/27/2014:  Currently we are growing a new cell line to inject into the mice which are part of our pilot experiment. To be consistent between experiments, we need the same amount of cells for each mouse we use. This new cell line is a little slow growing; as it is with many other research experiments, the hardest part is waiting!
  
03/14/2014:  more watchful waiting (nothing new).
  
03/20/2014:  A second round of cells have been injected into immunocompromised mice this week to continue our pilot experiments for this Alex’s Lemonade Stand project. Our next steps will be to observe these mice over the next few weeks and take note of when the tumor cells engraft and begin to grow.
  
03/27/2014:  no new news.  still in watchful waiting mode. 
  
04/04/2014:  still awaiting tumor development before initiating drug trials. 
  
04/10/2014:  Our pilot study is progressing nicely this week, with 3 of our orthotopic xenograft mice developing tumors. We will continue to observe our other injected mice for tumor development over the next several weeks.
  
04/14/2014:  See the blog post  http://kellerlabblog.blogspot.com/2014/04/imrans-paper-published-in-pnas.html for a link to today's published PNAS paper on this topic.  The studies in Imran's paper were the basis of this "next steps" project.  
  
04/17/2014:  The growth of the current tumors is progressing as expected. Our second group of mice have not grown any tumors yet, but they are still within the expected latency time for development. We continue to monitor them daily.

04/24/2014:  New batch of EphinB2-neutralizing biological agent received from pharma partner, facilitating the next series of therapeutic studies. 
  
05/08/2014:  mouse studies about to start for EphinB2-neutralizing biological agent. 
  
05/15/2014:  This week our official investigation on the efficacy of an EphrinB2-neutralizing agent has begun. The study mice arrived on Wednesday and were injected with Rh18 cells, a well characterized human eRMS cell line. Using our pilot data, we expect tumors to form within the next 2-3 months. Additionally, our second pilot study is still ongoing; we are continually monitoring the mice for development of tumors.
  
05/21/2014:  Rh18 xenograft studies still ongoing. 
  
05/28/2014:  not unexpectedly, still waiting on tumors to develop before treatments can begin...
  
06/05/14:  No new news to report this week. We do not expect treatment on our experimental mice to begin for a few more weeks, and we are continually monitoring both our initial pilot mice and current experimental for tumor development.
  
06/12/2014:  Still patiently waiting this week for mice in both experimental groups to develop tumors.  

06/26/2014:  Tissue microarrays of EphB4 and EphrinB2 on embryonal rhabdomyosarcoma are complete (it will take 1-2 weeks to analyze these with our pathologist collaborator).  We are also still patiently monitoring our immunocompromised mice for development of tumors. We are checking them 3 times a week so as soon as tumors develop we can begin treatment. Stay tuned!

  
07/10/2014:  Exciting news from the Keller Lab for our Alex’s Lemonade Stand project this week! The immunocompromised mice have started growing small tumors for the Rh18 eRMS cell line. In the coming weeks, most will begin receiving either an EphrinB2-neutralizing agent or a control drug.  Each mouse will be monitored daily and tumors measured three times per week.  This experiment is progressing on schedule and the next few months should be full of interesting data.

  
07/18/2014:  The Alex’s Lemonade Stand experiment is progressing nicely this week. All mice are tolerating the dosage of the drug well, and are not exhibiting any side effects/toxicity.  Expression of EphB4 and EphrinB2 in a small set of human osteosarcoma samples verified (request for TMA sent to COG); early studies of neuroblastoma are promising.  
  
07/31/2014:  This week, our experimental mice continue to be monitored daily. We will be wrapping up the in vivo testing in the coming weeks, then moving on to the data analysis phase.
  
08/08/2014:  EphB4 and EphrinB2 immunohistochemistry optimized for canine osteosarcoma (for pets with spontaneous osteosarcoma a potential preclinical model to help the dogs, and plan for a human osteosarcoma trial.)  Request sent to NCI comparative oncology program for canine TMA slides.  Meanwhile, the Rh18 eRMS mouse studies with the EphrinB2-neutralizing agent are wrapping up. 

  
08/21/2014:  The in vivo portion of our Alex's Lemonade Stand project is slowly coming to a close. This week some of the raw data analysis and number crunching was started, although it will be several weeks until that is finished. Additionally, there are various supporting experiments that still need to be executed to fully determine the efficacy of the EphrinB2 blocking agent. 
  
09/11/2014:  This week our supplemental experiments continue. We are now using RT-PCR to examine the amount of metastasis in the harvested lungs of our treated and control mice. These experiments are time consuming, but will help us decide if the EphrinB2 neutralizing agent administered to the mice was effective in diminishing the metastatic capability of RH18 cells.  

10/02/2014:  Canine osteosarcoma TMA's for staining received; COG neuroblastoma TMA slides approved but MTA still to perform; COG osteosarcoma TMA proposal still in review (since July). rt-pcr strategy for above EphrinB2 blocking agent mouse studies (completed) is in revision.  Some delays occurred as Megan spent the last 3 weeks in a collaborator's lab at Cold Spring Harbor Laboratory for a new strategic collaboration.    
  
10/16/2014:  A few setbacks this week owing to the particularly specific requirements of some molecular biology techniques. We have ordered a few new reagents in an attempt to complete the RT-PCR experiments, and should be back at the bench soon.
  
10/30/2014:  Our new reagents for the revised experiments have just arrived, and soon we should have a clear picture about the efficacy of the EphrinB2 blocking agent on lung metastasis.
  
12/04/2014:  Preclinical trial of sEphB4-HSA in Rh18 orthotopic xenografts is now complete.  The next step is preclinical testing in a patient-derived xenograft (see the description of this autopsy-derived model).  

  
12/31/2014:  Project on short term hold pending transfer of grant from OHSU to our new, bold adventure at cc-TDI. 
  
01/06/2014:  New lab at cc-TDI started!
  
01/07/2014:  Collaborators Ayeza Bajwa and Atiya Mansoor have completed analysis of human eRMS tissue microarrays with unexpected results that point to key differences in aRMS and eRMS with respect to EphB4 and EphrinB2 biology. 
  
01/08/2015:  ALSF REACH grant year 2 funding received... this is the first grant funding for cc-TDI ... thank you, ALSF!
  
01/15/2015:  Strategic meeting (teleconference) held including pharma and CSU partners to consider a companion animal trial for osteosarcoma using EphB4/EphrinB2 therapeutics.  
  
01/27/2015:  Integration of animal study underway by Matthew, including ongoing tissue analysis from these studies.  
  
02/06/2015:  We have begun considering approaches to separating cell-autonomous and tumor microenvironment contributions of EphB4-EphrinB2 signaling in eRMS.  

02/13/2015:  analysis of tumor cell vs stroma cell expression of EphB4 and EphrinB2 reveals distinct differences in alveolar and embryonal rhabdomyosarcomas.

02/20/2015:   For Rh18 xenograft studies (completed), lung metastasis counts are in progress and pharmacodynamic studies are planned.
  
02/27/2015:  COG TMA requests for osteosarcoma and neuroblastoma slides are approved, but MTAs are still in progress. 
  
03/04/2015:  Rh18 xenograft pharmacodynamics studies in progress.
  
05/06/2015:  Rh18 xenograft pharmacodynamics studies done; Rh18 lung metastasis count in progress. 
  
05/20/2015:   COG osteosarcoma TMA slides arrived May 8; working to verify EphrinB2 expression in PDX models.
  
06/03/2015:  Rh18 xenograft lung metastasis count done.
  
07/29/2015:  CTEP approved neuroblastoma TMA slides protocol July 26.
  
08/07/2015:  westerns on PDXs still ongoing; neuroblastoma TMA slides received.