A functional RNA-origami as direct thrombin inhibitor with fast-acting and specific single-molecule reversal agents in vivo model

TY - JOUR

T1 - A functional RNA-origami as direct thrombin inhibitor with fast-acting and specific single-molecule reversal agents in vivo model

AU - Krissanaprasit, Abhichart

AU - Mihalko, Emily

AU - Meinhold, Katherine

AU - Simpson, Aryssa

AU - Sollinger, Jennifer

AU - Pandit, Sanika

AU - Dupont, Daniel M.

AU - Kjems, Jørgen

AU - Brown, Ashley C.

AU - LaBean, Thomas H.

N1 - Publisher Copyright: © 2024 The American Society of Gene and Cell Therapy

PY - 2024/7/3

Y1 - 2024/7/3

N2 - Injectable anticoagulants are widely used in medical procedures to prevent unwanted blood clotting. However, many lack safe, effective reversal agents. Here, we present new data on a previously described RNA origami-based, direct thrombin inhibitor (HEX01). We describe a new, fast-acting, specific, single-molecule reversal agent (antidote) and present in vivo data for the first time, including efficacy, reversibility, preliminary safety, and initial biodistribution studies. HEX01 contains multiple thrombin-binding aptamers appended on an RNA origami. It exhibits excellent anticoagulation activity in vitro and in vivo. The new single-molecule, DNA antidote (HEX02) reverses anticoagulation activity of HEX01 in human plasma within 30 s in vitro and functions effectively in a murine liver laceration model. Biodistribution studies of HEX01 in whole mice using ex vivo imaging show accumulation mainly in the liver over 24 h and with 10-fold lower concentrations in the kidneys. Additionally, we show that the HEX01/HEX02 system is non-cytotoxic to epithelial cell lines and non-hemolytic in vitro. Furthermore, we found no serum cytokine response to HEX01/HEX02 in a murine model. HEX01 and HEX02 represent a safe and effective coagulation control system with a fast-acting, specific reversal agent showing promise for potential drug development.

AB - Injectable anticoagulants are widely used in medical procedures to prevent unwanted blood clotting. However, many lack safe, effective reversal agents. Here, we present new data on a previously described RNA origami-based, direct thrombin inhibitor (HEX01). We describe a new, fast-acting, specific, single-molecule reversal agent (antidote) and present in vivo data for the first time, including efficacy, reversibility, preliminary safety, and initial biodistribution studies. HEX01 contains multiple thrombin-binding aptamers appended on an RNA origami. It exhibits excellent anticoagulation activity in vitro and in vivo. The new single-molecule, DNA antidote (HEX02) reverses anticoagulation activity of HEX01 in human plasma within 30 s in vitro and functions effectively in a murine liver laceration model. Biodistribution studies of HEX01 in whole mice using ex vivo imaging show accumulation mainly in the liver over 24 h and with 10-fold lower concentrations in the kidneys. Additionally, we show that the HEX01/HEX02 system is non-cytotoxic to epithelial cell lines and non-hemolytic in vitro. Furthermore, we found no serum cytokine response to HEX01/HEX02 in a murine model. HEX01 and HEX02 represent a safe and effective coagulation control system with a fast-acting, specific reversal agent showing promise for potential drug development.

KW - anticoagulant

KW - aptamer

KW - direct thrombin inhibitor

KW - in vivo

KW - murine model

KW - nanomedicine

KW - nucleic acid nanostructure

KW - reversal agent

KW - RNA nanotechnology

KW - RNA origami

UR - http://www.scopus.com/inward/record.url?scp=85193290426&partnerID=8YFLogxK

U2 - 10.1016/j.ymthe.2024.05.002

DO - 10.1016/j.ymthe.2024.05.002

M3 - Journal article

C2 - 38720458

AN - SCOPUS:85193290426

SN - 1525-0016

VL - 32

SP - 2286

EP - 2298

JO - Molecular Therapy

JF - Molecular Therapy

IS - 7

ER -