Yang Jonathan Lee, Yamada-Hunter Sean A, Labanieh Louai, Sotillo Elena, Cheah Joleen S, Roberts David S, Mackall Crystal L, Ting Alice Y, Bertozzi Carolyn R
Department of Chemistry and Sarafan ChEM-H, Stanford University, Stanford, CA, USA.
Center for Cancer Cell Therapy, Stanford Cancer Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
bioRxiv. 2023 Nov 15:2023.11.14.567117. doi: 10.1101/2023.11.14.567117.
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.
溶酶体靶向嵌合体（LYTACs）是一种有前景的治疗方式，可促使细胞外蛋白质降解。然而，早期版本的LYTAC包含无法进行基因编码的合成糖肽。在此，我们展示了一种完全可基因编码的LYTAC（GELYTAC）的设计方案，使我们的工具能够与整合到治疗性细胞中兼容，以便在患病部位进行靶向递送。为实现这一目标，我们用蛋白质胰岛素样生长因子2（IGF2）取代了LYTAC的糖肽部分。在用野生型IGF2展示出初步疗效后，我们通过定向进化提高了GELYTAC的效力。随后，我们证明我们工程改造的GELYTAC构建体不仅能从人胚肾293T细胞分泌，还能从人原代T细胞分泌，从而促使各种靶标被受体细胞摄取。经工程改造分泌GELYTAC的免疫细胞因此代表了一种有前景的空间选择性靶向蛋白质降解途径。
