Directed Evolution of Genetically Encoded LYTACs for Cell-Mediated Delivery

Overview

Journal bioRxiv

Date 2023 Nov 28

PMID 38014030

Authors

Jonathan Lee Yang

Sean A Yamada-Hunter

Louai Labanieh

Elena Sotillo

Joleen S Cheah

David S Roberts

Crystal L Mackall

Alice Y Ting

Carolyn R Bertozzi

Affiliations

Soon will be listed here.

Abstract

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.

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