Fc-GDF15 glyco-engineering and receptor binding affinity optimization for body weight regulation

作者：Ella Fung, Liya Kang, Diana Sapashnik, Susan Benard, Annette Sievers, Yan Liu, Guoying Yan, Jing Zhou, Linette Rodriguez, Weijun Ma, Wayne R. Stochaj, Edward R. LaVallie, Liliana Wróblewska, Kerry Kelleher, Amy Tam, Olivier Bézy, Danna M. Breen, J Chabot, Tao He, Laura Lin, Zhidan Wu, Lidia Mosyak · 发表于：Scientific Reports · 年份：2021 · DOI：10.1038/s41598-021-87959-5 · 被引用次数：24 · 研究领域：GDF15 and Related Biomarkers、Macrophage Migration Inhibitory Factor、Nutrition and Health in Aging

GDF15 is a distant TGF-β family member that induces anorexia and weight loss. Due to its function, GDF15 has attracted attention as a potential therapeutic for the treatment of obesity and its associated metabolic diseases. However, the pharmacokinetic and physicochemical properties of GDF15 present several challenges for its development as a therapeutic, including a short half-life, high aggregation propensity, and protease susceptibility in serum. Here, we report the design, characterization and optimization of GDF15 in an Fc-fusion protein format with improved therapeutic properties. Using a structure-based engineering approach, we combined knob-into-hole Fc technology and N-linked glycosylation site mutagenesis for half-life extension, improved solubility and protease resistance. In addition, we identified a set of mutations at the receptor binding site of GDF15 that show increased GFRAL binding affinity and led to significant half-life extension. We also identified a single point mutation that increases p-ERK signaling activity and results in improved weight loss efficacy in vivo. Taken together, our findings allowed us to develop GDF15 in a new therapeutic format that demonstrates better efficacy and potential for improved manufacturability.