Directed evolution of genetically encoded LYTACs for cell- mediated delivery

• 成果类型: Article
• 署名作者: Yang, Jonathan Lee; Yamada-Hunter, Sean A.; Labanieh, Louai; Sotillo, Elena; Cheah, Joleen S.; Roberts, David S.; Mackall, Crystal L.; Bertozzi, Carolyn R.; Ting, Alice Y.
• 署名单位: Stanford University; Stanford University; Stanford University; Stanford Cancer Institute; Stanford University; Stanford University; Stanford University; Stanford University; Howard Hughes Medical Institute; Stanford University; Stanford University; Chan Zuckerberg Initiative (CZI)
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-13722
• DOI: 10.1073/pnas.2320053121
• 发表日期: 2024-03-26
• 关键词: t-cells; protein; receptor; degradation; immunogenicity; efficacy

摘要

Lysosome- targeting chimeras (LYTACs) are a promising therapeutic modality to drive the degradation of extracellular proteins. However, early versions of LYTAC contain synthetic glycopeptides that cannot be genetically encoded. Here, we present our designs for a fully genetically encodable LYTAC (GELYTAC), making our tool compatible with integration into therapeutic cells for targeted delivery at diseased sites. To achieve this, we replaced the glycopeptide portion of LYTACs with the protein insulin - like growth factor 2 (IGF2). After showing initial efficacy with wild - type IGF2, we increased the potency of GELYTAC using directed evolution. Subsequently, we demonstrated that our engineered GELYTAC construct not only secretes from HEK293T cells but also from human primary T - cells to drive the uptake of various targets into receiver cells. Immune cells engineered to secrete GELYTAC thus represent a promising avenue for spatially selective targeted protein degradation.