Molecular Biology

Supervisory Team:

Dr Abhishek Gupta

School of Pharmacy, Faculty of Science and Engineering (FSE), University of Wolverhampton, Wulfruna Street, Wolverhampton, UK.

Research Institute of Healthcare Science (RIHS), Faculty of Science and Engineering, University of Wolverhampton, Wulfruna Street, Wolverhampton, UK.

Dr Aikaterini Karakoula

School of Pharmacy, Faculty of Science and Engineering (FSE), University of Wolverhampton, Wulfruna Street, Wolverhampton, UK.

Research Institute of Healthcare Science (RIHS), Faculty of Science and Engineering, University of Wolverhampton, Wulfruna Street, Wolverhampton, UK.

About the Project:

Glioblastoma remains as the most common and aggressive malignant brain tumour, standing with a poor prognosis and treatment prospective. Despite the aggressive standard care, such as surgical resection and chemoradiation, median survival rates are low; about 15 months from the diagnosis and a 5-year survival rate of only 5%. Treatment resistance arises from a wide variety of mechanisms, including the blood–brain barrier (BBB), inter- and intra-tumoral heterogeneity, and a profoundly immunosuppressive tumour microenvironment.

Curcumin (CUR), chemically known as diferuloylmethane, obtained from Curcuma longa plant, is a polyphenolic compound that is well known for its pharmacological benefits like, antitumor, anti-inflammatory, and antioxidant. The anticancer properties of CUR are attributed to its unique abilities of inducing apoptosis and inhibiting proliferation and invasion of tumours by suppressing cellular signalling pathways in various cancers, including malignant gliomas. Furthermore, several studies have demonstrated that CUR is able to cross the BBB as well as to interact synergistically with commonly used chemotherapeutics making it an attractive therapeutic agent for malignant brain tumours. However, the biomedical application of CUR is greatly hindered by its hydrophobicity and low bioavailability. The poor bioavailability of CUR might be associated with its poor absorption, quick metabolism and rapid systemic elimination. Several research attempts have been made to produce structural modifications in CUR molecule to enhance its solubility and selective toxicity towards cancer cells. Moreover, different delivery systems have been produced to improve its physiochemical properties.

Microencapsulation of medicinal substances in a suitable carrier is a common practice in pharmaceutics for drug delivery. Cyclodextrins (CDs) are naturally occurring cyclic oligosaccharide obtained from starch by enzymatic cyclisation that are used as pharmaceutical adjuvants. Our group has reported the production of water-soluble CUR inclusion complex with hydroxypropyl-β cyclodextrins (HPβCD) by solvent evaporation method. CUR:HPβCD has demonstrated reduced viability (in vitro) against the range of tested cancer cell lines demonstrating that CUR maintains its anticancer properties in the inclusion complex.

Techniques associated with this project:

The current project will involve the production of a CUR:HPβCD complex and evaluating its anticancer properties against malignant glioma cells using a wide range of cellular and molecular biology/genetics techniques.

Key references:

A. Gupta, et al., 2019. Production and characterisation of bacterial cellulose hydrogels loaded with curcumin encapsulated in cyclodextrins as wound dressings. European Polymer Journal 118: 437-450

Giordano A. and Tommonaro G, 2019. Curcumin and Cancer. Nutrients, 11:2376.

Mansouri, K. et al., 2020. Clinical effects of curcumin in enhancing cancer therapy: A systematic review. BMC Cancer, 20: 791.

Perry M. et al., 2010. Curcumin inhibits tumor growth and angiogenesis in glioblastoma xenografts. Mol Nutr Food Res, 54: 1192–1201.

Purkayastha S. et al., 2009. Curcumin blocks brain tumor formation. Brain Res, 1266: 130–138.

Ramachandran C. et al., 2012. Potentiation of etoposide and temozolomide cytotoxicity by curcumin and turmeric force™ in brain tumor cell lines. J Complement Integr Med, 9: article 20.

Weissenberger J. et al., 2010. Dietary curcumin attenuates glioma growth in a syngeneic mouse model by inhibition of the JAK1,2/STAT3 signaling pathway. Clin Cancer Res, 16: 5781–5795.

Zanotto-Filho A. et al., 2013. Curcumin-loaded lipid-core nanocapsules as a strategy to improve pharmacological efficacy of curcumin in glioma treatment. Eur J Pharm Biopharm, 83: 156–167.

Zanotto-Filho A. et al., 2015. Autophagy inhibition improves the efficacy of curcumin/temozolomide combination therapy in glioblastomas. Cancer Lett, 358: 220–231.

Zhuang W. et al., 2012. Curcumin promotes differentiation of glioma-initiating cells by inducing autophagy. Cancer Sci, 103: 684–690.

For more information: For an informal discussion please contact via direct email to Dr Abhishek Gupta (a.gupta@wlv.ac.uk) and/or Dr Aikaterini Karakoula (a.karakoula@wlv.ac.uk).

Supervisory Team

Dr Omar Hafid, School of Life Sciences and Research Institute of Healthcare Science (RIHS), Faculty of Science and Engineering, University of Wolverhampton, Wulfruna Street, Wolverhampton, UK.

Dr Aikaterini Karakoula, School of Pharmacy and Research Institute of Healthcare Science (RIHS), Faculty of Science and Engineering, University of Wolverhampton, Wulfruna Street, Wolverhampton, UK.

About the Project:

Glioblastoma remains as the most common and aggressive malignant brain tumour, standing with a poor prognosis and treatment prospective. Despite the aggressive standard care, such as surgical resection and chemoradiation, median survival rates are low. Treatment resistance arises from a wide variety of mechanisms, including the blood–brain barrier (BBB), inter- and intra-tumoral heterogeneity, and a profoundly immunosuppressive tumour microenvironment.

Metabolic adaptation processes are believed to provide cancer cells, including glioblastoma, with proliferation and survival benefits over normal cells. However, these processes can also make cancer cells selectively dependent or addicted to certain nutrients and metabolic pathways. The overlapping metabolic reprogramming of cancer and immune cells is a putative determinant of the antitumor immune response in cancer. Increased evidence suggests that cancer metabolism not only plays a crucial role in cancer signalling for sustaining tumorigenesis and survival, but also has wider implications in the regulation of antitumor immune response through both the release of metabolites and affecting the expression of immune molecules.

Recent work in our lab has confirmed that overexpression of a key glycolytic enzyme hexokinase 2 (HK2) is associated with a shorter overall survival in glioblastoma patients. HK2 is an important facilitator of aerobic glycolysis in GBM, enabling survival and proliferation of the tumour microenvironment. A significant decrease in cell proliferation was established through CRISPR-mediated knockout of the HK2 gene in our patient-derived glioblastoma cell cultures, suggesting the potential of HK2 as a therapeutic target in glioblastoma patients. Results from CRISPR-treated glioblastoma cultures using RNA-sequencing analysis revealed significant deregulated immune-related signalling pathways including Toll-like receptors

(TLR1, TLR2, TLR4, TLR5, TRL6), immune-checkpoint inhibitors (HAVCR2 and PDCD1LG2), JAK-STAT signalling pathway (STA1, 2, 4, 5A, 6), cytokine-mediated signalling and response to inflammation (IL6R, IL6ST). Activation of TLRs indicates an inflammatory profile within the tumour microenvironment. This can lead to the recruitment of inflammatory immune cells and the production of proinflammatory cytokines, chemokines and growth factors which can promote tumour development and progression. Reduction in glioblastoma low-grade inflammation may render tumour cells susceptible to cancer drugs and anti-tumour immune responses.

More recently, attention has been given to the impact of alterations in glycolytic pathways on not only proliferating tumour cells but also the tumour microenvironment and resulting changes in immune cell metabolism and function. Metabolic reprogramming within tumour cells diminishes the function of effector immune cells through depletion of essential metabolites and promotes enrichment of suppressive immune populations.

In this 3-year research project, we plan to further explore the effects of GBM aerobic glycolysis and immunosuppression through HK2 to identify potential therapeutic targets and pathway interactions within the heterogeneous tumour microenvironment. We also plan to assess the inflammatory profile within the glioblastoma microenvironment by investigating the production of proinflammatory as well as Th2 type cytokines and chemokines that signal via the JAK-STAT pathway. This project will provide extensive training in a wide range of cell and molecular biology, and analytical techniques, including but not limited to RT2 Profiler PCR Arrays, flow cytometry, confocal, immunofluorescence and Luminex technology.

Key references:

Bi J., Bi F., Pan X., Yang Q. (2021) Establishment of a novel glycolysis-related prognostic gene signature for ovarian cancer and its relationships with immune infiltration of the tumor microenvironment. J. Transl. Med., 19:382.

Coussens L.M., Werb Z. (2022). Inflammation and cancer. Nature, 420(6917):860-867.

Choudhary N., Osorio R.C., Oh J.Y., Aghi M.K. (2023). Metabolic Barriers to Glioblastoma Immunotherapy. Cancers 15(5):1519.

DePeaux K., Delgoffe G.M. (2021). Metabolic barriers to cancer immunotherapy. Nat. Rev. Immunol. 21:785–797.

Jarmuzek P. et al. (2023) Cytokine Profile in Development of Glioblastoma in Relation to Healthy Individuals. Int J Mol Sci. 24(22):16206.

Muir A., Vander Heiden M.G. (2018). The nutrient environment affects therapy. Science, 360:962–963.

Ni Y., Low J.T., Silke J., O'Reilly L.A. (2022). Digesting the Role of JAK-STAT and Cytokine Signaling in Oral and Gastric Cancers. Front Immunol.,13:835997.

For more information: For an informal discussion please contact via direct email to Dr Aikaterini Karakoula (A.Karakoula@wlv.ac.uk) and/or Dr Hafid Omar (H.Omar6@wlv.ac.uk)