Decoupling IFNgamma functional pleiotropy with protein engineering.

Interferon gamma (IFNgamma) is a cytokine which plays a pivotal role in antitumour host immunity. IFNgamma is a potentially useful adjuvant for cancer immunotherapy based on its ability to act directly on tumour cells as well as activate cell mediated immunity. However, due to its pleiotropy, systemic administration of IFNgamma hinders its translation to the clinic. It is hypothesized that manipulation of IFNgamma binding properties could decouple its anti-tumour effects from its toxicity. However, the dimeric nature of this cytokine has made any attempts to manipulate its receptor binding properties challenging. To address this, high affinity IFNgamma monomer variants will be generated to the IFNgammaR2 subunit aiming to improve the antitumour potency of this cytokine. I will use biophysical and structural approaches to characterize how these variants engage their receptor. I will then examine the contribution of receptor affinity to IFNgamma's biological activities using RNA sequencing and
proteomic approaches.