Developing antisense oligonucleotides for a TECPR2 mutation-induced, ultra-rare neurological disorder using patient-derived cellular models

Williams, Luis A.; Gerber, David J.; Elder, Amy; Tseng, Wei Chou; Baru, Valeriya; Delaney-Busch, Nathaniel; Ambrosi, Christina; Mahimkar, Gauri; Joshi, Vaibhav; Shah, Himali; Harikrishnan, Karthiayani; Upadhyay, Hansini; Rajendran, Sakthi H.; Dhandapani, Aishwarya; Meier, Joshua; Ryan, Steven J.; Lewarch, Caitlin; Black, Lauren; Douville, Julie; Cinquino, Stefania; Legakis, Helen; Nalbach, Karsten; Behrends, Christian; Sato, Ai; Galluzzi, Lorenzo; Yu, Timothy W.; Brown, Duncan; Agrawal, Sudhir; Margulies, David; Kopin, Alan und Dempsey, Graham T. (2022): Developing antisense oligonucleotides for a TECPR2 mutation-induced, ultra-rare neurological disorder using patient-derived cellular models. In: Molecular Therapy - Nucleic Acids, Bd. 29: S. 189-203

Abstract

Mutations in the TECPR2 gene are the cause of an ultra-rare neurological disorder characterized by intellectual disability, impaired speech, motor delay, and hypotonia evolving to spasticity, central sleep apnea, and premature death (SPG49 or HSAN9;OMIM: 615031). Little is known about the biological function of TECPR2, and there are currently no available disease-modifying therapies for this disease. Here we describe implementation of an antisense oligonucleotide (ASO) exon-skipping strategy targeting TECPR2 c.1319delT (p.Leu440Argfs*19), a pathogenic variant that results in a premature stop codon within TECPR2 exon 8. We used patient-derived fibroblasts and induced pluripotent stem cell (iPSC)-derived neurons homozygous for the p.Leu440Argfs*19 mutation to model the disease in vitro. Both patient-derived fibroblasts and neurons showed lack of TECPR2 protein expression. We designed and screened ASOs targeting sequences across the TECPR2 exon 8 region to identify molecules that induce exon 8 skipping and thereby remove the premature stop signal. TECPR2 exon 8 skipping restored in-frame expression of a TECPR2 protein variant (TECPR2 Delta Ex8) containing 1,300 of 1,411 amino acids. Optimization of ASO sequences generated a lead candidate (ASO-005-02) with similar to 27 nM potency in patientderived fibroblasts. To examine potential functional rescue induced by ASO-005-02, we used iPSC-derived neurons to analyze the neuronal localization of TECPR2 Delta Ex8 and showed that this form of TECPR2 retains the distinct, punctate neuronal expression pattern of full-length TECPR2. Finally, ASO-005-02 had an acceptable tolerability profile in vivo following a single 20-mg intrathecal dose in cynomolgus monkeys, showing some transient non-adverse behavioral effects with no correlating histopathology. Broad distribution of ASO-005-02 and induction of TECPR2 exon 8 skipping was detected in multiple central nervous system (CNS) tissues, supporting the potential utility of this therapeutic strategy for a subset of patients suffering from this rare disease.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Medizin > Adolf-Butenandt-Institut ; Medizin > Munich Cluster for Systems Neurology (SyNergy)
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• ISSN: 2162-2531
• Sprache: Englisch
• DFG: Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - 390857198