PFA ependymomas are pediatric brain tumors with very poor prognosis. They represent about 10% of all pediatric tumors of the central nervous system (CNS), and up to 30% of CNS tumors in children under 3. (1) 9.4% of PFA tumors aberrantly express EZHIP (aka CXorf67), a protein that inhibits Polycomb repressive complex 2 (PRC2). PRC2 has an important function in silencing genes through trimethylating lysine 27 of histone H3, H3K27me3. (2, 3) PFA tumor cells show a reduction of this important epigenetic mark. The decrease in H3K27me3 results in deregulated gene expression and leads to cancer initiation and progression. (3) EZHIP deletion using CRISPR knockout leads to the restoration of H3K27me3. (2) These finding points at a causal relationship between EZHIP’s expression and the lack of H3K27me3, driving a subset of PFA ependymomas.
The goal of my project is to remove EZHIP from cancer cells to restore normal levels of H3K27me3. I want to use proteolysis-targeting chimeras (PROTAC) to degrade EZHIP to stop its interaction with the PRC2 complex. Below is a schematic to show the way a PROTAC would send EZHIP to the proteasome for degradation. (4)
PROTACs may work only if there is at least one domain in EZHIP that is folded into a structural module where the PROTAC can bind. To determine if that is the case, I will express and purify EZHIP, and test whether it is folded using different assays, such as differential scanning fluorimetry (5).
References:
- Pajtler,K.W., Witt,H., Sill,M., Jones,D.T.W., and Hovestadt,V. (2015) Molecular Classification of Ependymal Tumorsacross All CNS Compartments,Histopathological Grades, and Age Groups. Cancer cell, 27, 728–743
- Pajtler,K.W., Wen,J., Sill,M., Lin,T., and Orisme,W. (2018) Molecular heterogeneity and CXorf67 alterations in posterior fossa group A (PFA) ependymomas. Acta Neuropathologica, 136, 211–226.
- Hubner,J., Müller,T., Papageorgiou,D.N., Mauermann,M,. and Krijgsveld,J. (2019) EZHIP/CXorf67 mimics K27M mutated oncohistones and functions as an intrinsic inhibitor of PRC2 function in aggressive posterior fossa ependymoma. Neurooncology, 21, 878–889.
- Schapira,M., Calabrese,M., Bullock,A.N., Crews,C.M. (2019) Targeted protein degradation: expanding the toolbox. Nature Reviews Drug Discovery.
- Differential Scanning Fluorimetry (DSF). Center For Macromolecular Interactions, Harvard Medical School. ( https://cmi.hms.harvard.edu/differential-scanning-fluorimetry
Hi Setayesh, My 4 year old daughter has a PFA anaplastic ependymoma with the CXorf67 identified in her tumour. Can you please provide me with more information about when you will be conducting this study and who will qualify? We are based in Sydney, Australia. Thank you.
Hi Rachelle, Thanks for your interest in our research. I am very sorry to hear that your family is going through this situation. I am not a medical doctor, and my research is preclinical. Therefore I cannot advise you as a patient or include you in my study. After a quick search on the web, I found this website, which you might find helpful: https://www.curebraincancer.org.au/ and further, they have a page that covers childhood brain cancers: https://www.curebraincancer.org.au/page/12/childhood-brain-tumours.
Thanks again for your comment. I hope you find the help and support you need, Rachelle.
Rachelle, I just saw your comment. As it happens, I am part of a SU2C team working on brain tumours and one of our collaborators at SickKids in Toronto, Dr. Vijay Ramaswamy has started a clinical trial in children with Ependymoma using 5-azacytidine (a DNA methylation agent). You can find details here:
https://clinicaltrials.gov/ct2/show/NCT03206021
Vijay told me that there is a site in Melbourne, Australia that has joined the trial and has recently started recruitment. I suggest you get in touch with them. Here is the site and contact info:
Royal Children’s Hospital Melbourne Recruiting
Melbourne, Victoria, Australia
Contact: Jordan R Hansford, MBBS
Principal Investigator: Jordan R Hansford, MBBS
Feel free to email me if you need more details.
You can reach me at takis.prinos@utoronto.ca
Rachelle, one of our collaborators (part of our SU2C project on brain tumours) has started a clinical trial in children with Ependymoma using 5-azacytidine (a DNA methylation agent). You can find details here: https://clinicaltrials.gov/ct2/show/NCT03206021 I think a site in Melbourne, Australia has recently starting recruitment. I suggest you get in touch with them. I can also send you the contact info of the investigator in charge.
Hi Setayesh,
Do you have any results of your work yet?