Exploring healthy glial cell and

neuron interactions with glioma and

associated implications of
Kynurenine pathway
 ABSTRACT

• GBM and astroglioma are common, largely incurable

and malignant CNS tumours originating from the glial
cells. Generally, all gliomas are classified between
grades 1 – 4 depending on the levels of invasiveness
and malignancy.
• While the effects of neurons and microglia on the
tumour microenvironment and regulation of glioma
growth is being extensively characterised, several
aspects of neuronal and glial structure and function
impairment in glioma microenvironment remains to be
characterised.
• This project aims to study morphological and
functional changes in healthy microglia, astrocytes and
cortical neurons in exposure to tumour
microenvironment using C6 cell line as well as evaluate
the implication of kynurenine pathway in glioma.

Source: MSD Science

AIMS AND OBJECTIVES

1. To evaluate how C6 CM affect neurons and healthy glial cells in

terms of
• morphology
• function
• viability

2. To evaluate the implications of KP in glioma

• Evaluate potential reversal of abnormalities upon IDO-1 inhibition
 BACKGROUND
• Gliomas exist in a highly selective and poorly accessible
microenvironment and exhibit tumour heterogeneity and
intercellular communication contributing to their poor
prognosis.
• Chemokine mediated communication is implicated in
tumour metastasis, angiogenesis and immune tolerance.
• The typical approaches towards glioblastoma treatment
either target the tumour or the TME.
• KP is the catabolic pathway of L-Trp that leads to the
production of neurotoxic (3-hydroxykynurenine, quinolinic
acid) and neuroprotective metabolites.
• C6 cell line is commonly adapted in glioma research due to
its invasiveness, angiogenesis capacity and ability to mimic
human GBM.
 BACKGROUND
• Astrocytes: The same mechanisms
that have been implicated in
astrogliosis have been known to
promote tumour proliferation
under the influence of tumour
cells. (O’Brien et al., 2013)
Heiland et al., 2019
• Microglia are commonly found in
association with glioma cells,
although co-proliferation of microglia
with the tumour cells remains
uncertain. Co-culture of microglial
cells and glioma cells have shown that
microglial cells show fluctuations in
their phagocytic properties upon
exposure to glioma (Voisin et al.,
2010)
• Neurons: Hyperexcitability of
neurons is considered a driver of
tumour proliferation. Increased
glutamate secretion, formation of
neuron-glia synapses and NLGN-3
conc are known to aid tumour
proliferation.
Yan et al., 2011
WORK
PLAN
 Expected outcomes
Primary outcomes
• Changes in astrocyte morphology: swollen cell bodies, unipolar or bipolar
processes. (Difference btw RA and TAA?)
• Microglia: More retracted and ovoid. Increased F-Actin density on staining.
Proliferation and activity changes? (Voisin et al., 2010).
• Neurons: reduced complexity, less branched processes, reduced synaptogenesis.
• Reduction in abnormalities with 1-MT treatment

Secondary outcomes
• Reduced Glu conc in TME? (Guilemmin et al., 2011)
• Other possible catalysts of KP (Du et al, 2020)
RESULTS - NEURONS
RESULTS - ASTROGLIA
CURRENT SCHEDULE STATUS
 REFERENCES
• O’Brien, E. R., Howarth, C., & Sibson, N. R. (2013). The role of astrocytes in CNS tumors: pre-clinical
models and novel imaging approaches. Frontiers in Cellular Neuroscience, 7.
• Voisin, P., Bouchaud, V., Merle, M., Diolez, P., Duffy, L., Flint, K., … Bouzier-Sore, A.-K.
(2010). Microglia in Close Vicinity of Glioma Cells: Correlation Between Phenotype and Metabolic
Alterations. Frontiers in Neuroenergetics, 2. doi:10.3389/fnene.2010.00131 
• Henrik Heiland, D., Ravi, V.M., Behringer, S.P. et al. Tumor-associated reactive astrocytes aid the
evolution of immunosuppressive environment in glioblastoma. Nat Commun 10, 2541 (2019).
https://doi.org/10.1038/s41467-019-10493-6
• Yan, T., Skaftnesmo, K. O., Leiss, L., Sleire, L., Wang, J., Li, X., & Enger, P. Ø. (2011). Neuronal markers
are expressed in human gliomas and NSE knockdown sensitizes glioblastoma cells to radiotherapy
and temozolomide. BMC cancer, 11, 524. https://doi.org/10.1186/1471-2407-11-524
• Du, L., Xing, Z., Tao, B. et al. Both IDO1 and TDO contribute to the malignancy of gliomas via the
Kyn–AhR–AQP4 signaling pathway. Sig Transduct Target Ther 5, 10 (2020).
https://doi.org/10.1038/s41392-019-0103-4