FORCE platform overcomes barriers of oligonucleotide delivery to muscle and corrects myotonic dystrophy features in preclinical models
Timothy Weeden, Tyler Picariello, Brendan Quinn, Sean Spring, Peiyi Shen, Qifeng Qiu, Benjamin Vieira, L. Schlaefke, Ryan J. Russo, Ya-An Chang, Jin Cui, Monica Yao, Aiyun Wen, Nelson Hsia, Tama Evron, Katy N. Ovington, Pei-Ni Tsai, Nicholas C. Yoder, Bo Lan, Reshmii Venkatesan, John K. Hall, Cody A. Desjardins, Mohammed Qatanani, Scott Hilderbrand, John Najim, Zhenzhi Tang, Matthew Tanner, Romesh R. Subramanian, Charles A. Thornton, Oxana Ibraghimov‐Beskrovnaya, Stefano Zanotti
Abstract
We developed the FORCETM platform to overcome limitations of oligonucleotide delivery to muscle and enable their applicability to neuromuscular disorders. The platform consists of an antigen-binding fragment, highly specific for the human transferrin receptor 1 (TfR1), conjugated to an oligonucleotide via a cleavable valine-citrulline linker. Myotonic dystrophy type 1 (DM1) is a neuromuscular disorder caused by expanded CUG triplets in the DMPK RNA, which sequester splicing proteins in the nucleus, lead to spliceopathy, and drive disease progression. Multiple surrogate conjugates were generated to characterize the FORCE platform. DYNE-101 is the conjugate designed to target DMPK and correct spliceopathy for the treatment of DM1. HSALR and TfR1hu/mu;DMSXLTg/Tg mice were used as models of myotonic dystrophy, the latter expresses human TfR1 and a human DMPK RNA with >1,000 CUG repeats. Cynomolgus monkeys were used to determine translatability of DYNE-101 pharmacology to higher species. In HSALR mice, a surrogate FORCE conjugate achieves durable correction of spliceopathy and improves myotonia to a greater extent than unconjugated ASO. In patient-derived myoblasts, DYNE-101 reduces DMPK RNA and nuclear foci, consequently improving spliceopathy. In TfR1hu/mu;DMSXLTg/Tg mice, DYNE-101 reduces mutant DMPK RNA in muscle, thereby correcting splicing. Reduction of DMPK foci in cardiomyocyte nuclei accompanies these effects. Low monthly dosing of DYNE-101 in TfR1hu/mu;DMSXLWT/Tg mice or cynomolgus monkeys leads to a profound reduction of DMPK expression in muscle. These data validate FORCE as a drug delivery platform and support the notion that DM1 may be treatable with low and infrequent dosing of DYNE-101. Oligonucleotides are small pieces of DNA or RNA that can be used to modify expression of genes. Myotonic dystrophy type 1 (DM1) is a severe disorder caused by an abnormal gene that affects multiple organs, including muscle. We developed the FORCE platform to deliver oligonucleotides to muscle. Here we evaluate the impact of this platform on muscle cells from people living with DM1, myotonic dystrophy mouse models, and healthy non-human primates. Our results show that FORCE can deliver oligonucleotides to muscle and provide beneficial effects in animal models of DM1. In the future, FORCE could potentially be used to treat people living with DM1. Weeden et al. use a method called FORCE to deliver oligonucleotides to muscle and evaluate the platform’s utility in models of myotonic dystrophy. An antisense oligonucleotide targeting DMPK is guided to muscle tissue with an antibody fragment and beneficial results are seen in mouse models of disease and cynomolgus monkey.