PI: Dr. Jan Molenaar
The iTHER program
The goal of the iTHER (individualized THERapy) program is to realize personalized treatment for children with relapsed or refractory, incurable cancer. This involves 150 patients per year. The program is a METC approved clinical registry trial which is performed in collaboration with the group of Michel Zwaan. Molecular characterization includes low coverage Whole Genome Sequencing, Whole Exome Sequencing, RNAseq, an Affymetrix 133plus2 array and methylation pattern analysis using the DKFZ INFORM pipeline. Data is stored at the core facility of the Maxima and analyzed using the R2 bioinformatic platform. We have obtained an NWO precision medicine grant to further develop the iTHER program, which involves, clinical implementation of this program in the Maxima, connecting the program to targeted therapies and expanding our profiling with in vitro compound testing on patient derived tumor organoids.
Personalized cancer models
Tumors organoids from several pediatric tumor types, can now be adequately cultured. For neuroblastoma organoids the efficacy was only 50% and growth rates were low. To optimize this, we are currently systematically testing a repository of components of growth factors, pathway activating or inactivating components and various growth matrices and co-culture methods. Phenotyping is performing using single cell RNAseq on tumor biopsies and organoids. The organoids are subsequently used for automatic testing of a large compound library using robotics in a high-throughput set-up to identify optimal combination treatment options. These combination treatment options can be tested in in vivo models (PDX/GEMMS) that will be generated through an IMI2 funded program.
Target identification and validation in neuroblastoma
A highly efficient pipeline covering the full spectrum of targeted drug development is currently functional for neuroblastoma. The process starts with the identification of targets by mining high throughput data from Affymetrix mRNA profiling, RNAseq, Whole Genome Sequencing and single cell analysis. Potential new target genes are subsequently validated in in vitro systems using a variety of molecular genetic manipulation techniques. The most promising target genes are then further validated using small molecule inhibitors in vitro and in vivo. Several new models are developed to consolidate this process. Tumor organoids are used and new in vivo xenograft and transgenic models are implemented. The current studies are in various stages of development and involve BCL2, MDM2, CDK4/6, CDKN2A, the MEK pathway, ATRX and the 8 oxo-guanine repair pathway.
From class waivers to precision medicine in paediatric oncology. Pearson ADJ, Pfister SM, Baruchel A, Bourquin JP, Casanova M, Chesler L, Doz F, Eggert A, Geoerger B, Jones DTW, Kearns PR, Molenaar JJ, Morland B, Schleiermacher G, Schulte JH, Vormoor J, Marshall LV, Zwaan CM, Vassal G; Executive and Biology Committees of the Innovative Therapies for Children with Cancer European Consortium. Lancet Oncol. 2017 (IF 33,9) Jul;18(7):e394-e404. doi: 10.1016/S1470-2045(17)30442-4. Review. Pubmed PMID: 28677575
Relapsed neuroblastomas show frequent RAS-MAPK pathway mutations. Eleveld TF, Oldridge DA, V Bernard, J Koster, LC Daage, SJ Diskin, L Schild, N Bentahar, A Bellini, M Chicard, E Lapouble, V Combaret, P Legoix-Né, J Michon, TJ Pugh, LS Hart, JA Rader, EF Attiyeh, JS Wei, S Zhang, A Naranjo, JM Gastier-Foster, MD Hogarty, MA. Smith, JM Guidry Auvil. TBK Watkins, DA Zwijnenburg, ME Ebus, P v Sluis, A Hakkert, E v Wezel, CE vd Schoot, EM Westerhout, JH Schulte, GA Tytgat, MEM Dolman, , I Janoueix-Lerosey, DS Gerhard, HN Caron, O Delattre, J Khan, R Versteeg, G Schleiermacher, Molenaar JJ (corr. author), Maris JM. Nat Genet. 2015 (IF 29,4) Pubmed PMID: 26121087
TERT rearrangements are frequent in neuroblastoma and identify aggressive tumors. Valentijn LJ, Koster J, Zwijnenburg DA, Hasselt NE, van Sluis P, Volckmann R, van Noesel M, George RE, Tytgat GAM, Molenaar JJ, Versteeg R. Nat Genet. 2015 (IF 29,4) Pubmed PMID: 26523776
Sequencing of neuroblastoma identifies chromothripsis and defects in neuritogenesis genes. Molenaar JJ, Koster J, Zwijnenburg DA, van Sluis P, Valentijn LJ, van der Ploeg I, Hamdi M, van Nes J, Westerman BA, van Arkel J, Ebus ME, Haneveld F, Lakeman A, Schild L, Molenaar P, Stroeken P, van Noesel MM, Ora I, Santo EE, Caron HN, Westerhout EM, Versteeg R. Nature. 2012 (IF:41,5) Feb 22;483(7391):589-93. Pubmed PMID: 22367537
LIN28B induces neuroblastoma and enhances MYCN levels via let-7 suppression. Molenaar JJ, Domingo-Fernández R, Ebus ME, Lindner S, Koster J, Drabek K, Mestdagh P, van Sluis P, Valentijn LJ, van Nes J, Broekmans M, Haneveld F, Volckmann R, Bray I, Heukamp L, Sprüssel A, Thor T, Kieckbusch K, Klein-Hitpass L, Fischer M, Vandesompele J, Schramm A, van Noesel MM, Varesio L, Speleman F, Eggert A, Stallings RL, Caron HN, Versteeg R, Schulte JH. Nat Genet. 2012 (IF:29,4) Nov;44(11):1199-206. Pubmed PMID: 23042116