Research Projects

Glioblastoma Multiforme (GBM) is a central nervous system (CNS) cancer arising from glial cells. 54% of the global CNS tumour prevalence is from Asia, with India as the second largest contributor(https://gco.iarc.fr/). GBMs are associated with high mortality even after surgery, chemo and radiotherapy [1], [2]. GBMs are heavily infiltrated (~30%) by myeloid-derived cells such as Microglia and macrophages [3, 4]. Increasing evidence suggests that microglia and macrophages interact with tumour cells, promoting their growth, migration, invasiveness and angiogenesis [5]. Extracellular traps (ETs) are web-like structures released from immune cells composed of nuclear or mitochondrial DNA[6-8]. (ETs) were first reported in neutrophils in 2004. Neutrophil extracellular traps (NETs) are central to infection and immunity. ETs are decorated with multiple antimicrobial proteins like myeloperoxidase and neutrophil elastase [9-11] and are capable of trapping and killing bacteria [6], fungi [12], parasites [13] and viruses [14] It is now known that innate immune cells such as monocytes [15], macrophages [16], eosinophils [17], basophils [18, 19] and mast cells [20] can also form ETs. NETs play a role in the pathogenesis of many diseases, including cancers such as breast and lung cancers [21]. In 2021, our laboratory was the first to report microglia extracellular traps (miETs) induced by dopamine [22]. Our published research showed an increased presence of miETs in human GBM tissue [22]. Even though ETs have been reported to play a role in cancers, there is no report of a link between dopamine-induced miETs in glioma pathophysiology. Interestingly, GBM cells express dopamine receptor 2 (DRD2) with elevated expression in the glioma-initiating cell population [23]. DRD2 signalling activates the hypoxia response and alters metabolism[23]. This raises interesting possibilities for the role of dopamine and miETs in GBM pathophysiology. This proposal aims to elucidate the role of dopamine-induced miETs in GBM pathophysiology. There are three main objectives of the proposal: 1) Understanding the molecular composition of dopamine-induced miETs. PCR and omics approaches will be utilised to assess the nucleic acid (DNA, RNA, mitochondrial DNA or a mixture) and protein composition of Dopamine-induced miETs. 2) Functional assessment of dopamine-induced miETs in GBM cell proliferation, migration and invasiveness. Microscopy, wound healing and transwell migration assays will be utilised. 3) Understanding the role of dopamine-induced miETs in inflammation and cell death in GBM. ETs are known to cause activation of NLRP3 inflammasome in lupus [24]. Nlrp3 deficient microglia and pharmacological inhibitors will be utilised to understand NLRP3 signalling-mediated cell death and cytokine release in GBM. The discovery of dopamine-induced miETs and their preliminary characterization has already been achieved by our lab (TRL1: Basic principles observed). Aim1 will result in a validated list of potential targets and pathways modulated by miETs in GBM microenvironment (TRL2: Technology Concept and/or Application Formulated). Aim2 will provide proof that modulation of miETs or their pathways can alter GBM cell behaviour, establishing biological relevance and therapeutic potential (TRL3: Analytical and Experimental Proof-of-Concept). Finally, Aim3 will validate in vitro functional assay platforms demonstrating controlled, reproducible modulation of GBM pathology via miET-targeting strategies (TRL 4: Technology Validated in Lab Environment). Future partnerships with clinical partners and biotech firms will aid transition and progression of our research outcomes from TRL 5-7 by proof-of-concept trials or therapeutic screening programs. The outcomes of the proposal will lay the foundation for linking dopamine signalling to immune modulation in GBM for future drug development aimed at modulating neuroinflammation and improving GBM therapy.