Deo Prakash Pandey, Dept. of Microbiology, Rikshospitalet
Contact: deo.prakash.pandey@rr-research.no, deo.prakash@gmail.com
https://www.ous-research.no/pandey/
Synopsis
Transcription is one of the key processes through which cells control regulation of gene expression. The aberrant transcriptional programs, resulting from the dysregulation at any of the steps at which transcription is regulated, are key to development and maintenance of cancer cells and consequently cancer cells are often hypersensitive to the targeting of the transcriptional machinery. For a particular cancer, often a set of transcription factors (TFs) drive the aberrant transcriptional program and a general feature of deregulated TFs is the altered transcriptional output resulting from the tumor-specific chromatin and epigenetic landscape. The idea that the specific transcriptional requirement of a particular cancer can be exploited to develop targeting strategies against tumor cells is around for some time. For example, Tamoxifen used for the treatment of nearly 70% of breast cancer patients, is one of the most successful anti-cancer drugs of all time and it works by antagonizing the proliferative function of the transcription factors, estrogen receptors specifically in breast cells, but not in other cells.
We work on tumors of central nervous systems as a cancer model system which are some of the deadliest malignancies affecting mankind. Glioblastoma multiforme (GBM) is the most prevalent and aggressive malignant tumor of the central nervous system. Glioblastoma is rapidly fatal and with the current treatment regimes, the median overall survival is less than 15 months for GBM patients. Being diagnosed with glioblastoma is an effective death sentence and desthis has not changed for last 4-5 decades. It is widely appreciated that GBM tumor propagation and resistance to existing therapies are driven by a subset of stem-like cells, which depend on a set of core developmental TFs to maintain a specific transcriptional program and sustain proliferation. These TFs include Sox2, Olig2, Sall2 and Pou3f2. Contrary to our current understanding, our preliminary data suggests that not all GBM cells express all these four TFs equally. For example, Olig2 as well as Sall2 are not always expressed in all GBM patient samples. The aim of the proposed work is to characterize the contribution of Olig2 expression in GBM transcriptional program. The MS thesis projects offered here involve application of several multi-dimensional approaches combining cancer biology, a range of biochemical approaches and use of multi-omics technologies. The potential projects are:
- Generate a profile of core TF expression in GBM
1A. Using western blotting, generate a profile of TF pression on a panel of primary patient GBM cell lines. As a control, human fetus derived neural stem (hNS) cells will be used. In addition to Olig2, these TFs will include Sox2, Sox9, Pou3f2, Sall2, Ascl1 and FoxG1.
1B. Using tissue micro-array (TMA), generate a profile of TF expression on clinical samples in collaboration with neuro-oncology dept at Radium hospital
Outcome: Expression of the master TFs in clinics
- Characterize the contribution of Olig2 towards GBM transcriptional program
We are in the process of generating and characterizing Olig2 knock-out in GBM and hNS cell lines. Using these Olig2 KO GBM and hNS cell line, we will:
2A. We will perform RIME for Olig2, Sox2, Sox9, Brn2, Sall2, Ascl1 and FoxG1 and IgG (n=8) to identify the partners of these TFs in GBMs (and in normal cells) which are dependent on Olig2 expression. These will be validated subsequently in primary GBM cells which do not express Olig2.
Outcome: Identification of the complex binding partners of GBM master TF, dependent on Olig2 expression. Do they gain or lose their partners following loss of Olig2?
2B. GBM/hNS cells are tri-potent stem cells which can be differentiated into three cell types, neurons, astrocytes, oligodendrocytes. Using Olig2 KO/control GBM and hNS cells, we will perform RNA-seq following differentiation and investigate the contribution of Olig2 in neuronal differentiation. A direct extension of this will be to perform single-cell RNA-seq following a differentiation time-course.
Outcome: Identification of the Olig2 dependent GBM and hNS transcriptional program
Supervision
We are looking to recruit one or two master students. The students will be supervised by Deo Prakash Pandey, project leader at the department of Molecular Microbiology, Oslo University Hospital (OUS) and the internal supervisor at IBV will be Prof. Rein Aasland or Prof. Pierre Chymkowitch.
Currently, our group consists of a technician and a postdoctoral fellow and 3-4 talented master students. We have the necessary resources and setup to carry out these experiments. The progress and development is monitored through regular one-to-one, group meetings and there is an opportunity the present the work at the section level.