MARCH 2016 UPDATE
Neuroblastoma Organoids from Patient-Derived Cells
By Rachele Rosati - Cure First Research Fellow
I would like to share the progress that has been made on the Neuroblastoma Project:
As I previously announced, our Director, Dr. Carla Grandori, initiated a collaboration with one of the most prominent Neuroblastoma experts in the country, Dr. John Maris from the Children’s Hospital of Philadelphia (CHOP). Dr. Maris has provided Cure First with ten Patient-Derived Tumor Xenografts (PDTX), which I have been working with for the past few months. Each sample came from a patient diagnosed with Neuroblastoma and each had distinct alterations and amplifications. At Dr. Maris’s lab, the patients’ cells were injected into immunodeficiency mice and expanded. Then each tumor was chopped out and frozen. Despite the difficulty of retrieving tumor cells from the frozen pieces of tissue where the interstitial mouse cells may have spread, I was able to isolate and expand neuroblastoma tumor cells in vitro.
I started with the following PDTXs: NB-SD, which has a MYCN amplification, TP53, ALK and FLT3/4 mutations and SKNAS, which is an NRAS and TP53 mutant. I have optimized their growth, after several different experiments, with the appropriate culturing media and the appropriate dishes. The results are different for each sample and it took almost a month to achieve a perfect growth in non-adherent tumor spheres. However, this is a great achievement as you can see from these photos.
Recently I have received other 2 fresh Neuroblastoma Xenograft samples from CHOP. I processed them and for one of these, NB-Ebc1, which has KRAS and EZH2 mutations, I obtained tumor organoids as well.
Thanks to these results I can announce that we are ready to screen NB-SD and NB-Ebc1 samples, testing a library of 320 compounds compiled by CTD2 (Cancer Target Discovery and Development) Network, in order to see which drugs have an impact on the mutations associated with neuroblastoma.
I am looking forward to starting this new challenge!
We also want to thank our biggest supporter, the Soupy for Loopy Foundation, for their generous donation. I would also like to thank all the people that support us through our Global Giving Neuroblastoma Project fundraising campaign, allowing us to cover the cost of all the laboratory materials needed to carry out the screening of the cells.
Our efforts to discover new drug targets for neuroblastoma still need your help. Thank you in advance!
Proceeds from your donation will help fund Cure First's continuing efforts to discover new drug targets for children with neuroblastoma.