Krotova, et al. (2026)
Presented at ASGCT – 2026
Abstract
In vivo CAR T-cell therapy offers a transformative approach that generates CAR T-cells directly inside the patient’s body, eliminating complex production steps and the need for lymphodepletion. For this purpose, the CAR delivery vehicle must efficiently target T cells in the presence of serum and exhibit minimal or no off-target effects.
G-Link is a trimerizing adapter protein that blocks the natural tropism of VSV-G and redirects entry to other receptors. The trimerization design allows three binding arms of the adapter to simultaneously engage each of the protomers of the mature VSV-G protein, significantly stabilizing the interaction by enhancing its avidity.
The high-avidity trimeric G-link adaptors were derived from the monomeric engagers developed at the Weizmann Institute (Israel Patent application No. 296547, 2022, inventors: M. Rubinstein, G. Schreiber, D. Gataulin). These fused cell-targeting polypeptides bind to the LDL receptor (LDLR) via its CR2 and CR3 binding domains. Our G-link adaptor protein incorporates three cysteine-rich domains (CR1, CR2, and CR3) of the LDLR, linked to a CD3-targeting domain via a trimerizing peptide (Vyriad Patent application No. US63904491, 2025, inventors: SJ. Russell, GN. Nenavath, MS.Hasan). Trimeric G-link adaptor proteins can associate with virally displayed VSV-G glycoproteins either during LV production through co-expression, or by post-mixing as purified protein with preformed LVs. Here, we evaluate the stability and robustness of the interaction between the G-link adaptors and their G protein targets in both formats.