Charge-altering releasable transporters enhance mRNA delivery in vitro and exhibit in vivo tropism

Zhijian Li; Laura Amaya; Ruoxi Pi; Sean K Wang; Alok Ranjan; Robert M Waymouth; Catherine A Blish; Howard Y Chang; Paul A Wender

doi:10.1038/s41467-023-42672-x

Charge-altering releasable transporters enhance mRNA delivery in vitro and exhibit in vivo tropism

Nat Commun. 2023 Nov 1;14(1):6983. doi: 10.1038/s41467-023-42672-x.

Authors

Zhijian Li^#¹, Laura Amaya^#^{2

3}, Ruoxi Pi⁴, Sean K Wang^{2

5}, Alok Ranjan¹, Robert M Waymouth¹, Catherine A Blish^{4

6}, Howard Y Chang^{2

7}, Paul A Wender^{8

9}

Affiliations

¹ Department of Chemistry, Stanford University, Stanford, CA, 94305, USA.
² Center for Personal Dynamic Regulomes, Stanford University, Stanford, CA, 94305, USA.
³ Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, 94305, USA.
⁴ Division of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford, CA, 94305, USA.
⁵ Department of Ophthalmology, Stanford University School of Medicine, Stanford, CA, 94305, USA.
⁶ Chan Zuckerberg Biohub, San Francisco, CA, USA.
⁷ Howard Hughes Medical Institute, Stanford University, Stanford, CA, 94305, USA.
⁸ Department of Chemistry, Stanford University, Stanford, CA, 94305, USA. wenderp@stanford.edu.
⁹ Department of Chemical and Systems Biology, Stanford University, Stanford, CA, 94305, USA. wenderp@stanford.edu.

^# Contributed equally.

Abstract

The introduction of more effective and selective mRNA delivery systems is required for the advancement of many emerging biomedical technologies including the development of prophylactic and therapeutic vaccines, immunotherapies for cancer and strategies for genome editing. While polymers and oligomers have served as promising mRNA delivery systems, their efficacy in hard-to-transfect cells such as primary T lymphocytes is often limited as is their cell and organ tropism. To address these problems, considerable attention has been placed on structural screening of various lipid and cation components of mRNA delivery systems. Here, we disclose a class of charge-altering releasable transporters (CARTs) that differ from previous CARTs based on their beta-amido carbonate backbone (bAC) and side chain spacing. These bAC-CARTs exhibit enhanced mRNA transfection in primary T lymphocytes in vitro and enhanced protein expression in vivo with highly selective spleen tropism, supporting their broader therapeutic use as effective polyanionic delivery systems.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Gene Editing*
RNA, Messenger / metabolism
T-Lymphocytes* / metabolism
Transfection
Tropism

Substances

RNA, Messenger

Abstract

Publication types

MeSH terms

Substances

Grants and funding