Abstract
Clinical trials for Duchenne muscular dystrophy (DMD) are assessing the therapeutic efficacy of systemically delivered adeno-associated virus (AAV) carrying a modified DMD transgene. High vector doses (>1E14 vg/kg) are needed to globally transduce skeletal muscles; however, such doses trigger immune-related adverse events. Mitigating these immune responses is crucial for widespread application of AAV-based therapies. We used single-cell RNA sequencing and T cell receptor (TCR) sequencing on peripheral blood mononuclear cells from five participants prior to, and after, dosing. One subject in the high-dose cohort experienced thrombotic microangiopathy (TMA). Few changes in cell frequencies occurred after treatment; however, differential gene expression demonstrated induction of interferon response genes in most T cell types. T cell clonotype and clumping analysis showed the expansion or appearance of groups of related TCR sequences in the post-treatment samples. Three of these expanded clumps could be assigned to prior human herpesvirus infections, two of which were present in the participant that exhibited TMA. These data provide insight on the mechanistic basis of human immune-AAV interactions and lay a foundation for improved understanding of why TMA arises in some patients and not others.
| Original language | English (US) |
|---|---|
| Article number | 101349 |
| Journal | Molecular Therapy Methods and Clinical Development |
| Volume | 32 |
| Issue number | 4 |
| DOIs | |
| State | Published - Dec 12 2024 |
Funding
The authors thank and acknowledge the use of services from the following cores: the UCLA Technology Center for Genomics & Bioinformatics (TCGB) and the UCLA Institute for Quantitative and Computational Biosciences (QC Bio). This study was supported by the Department of Defense (to M.J.S. and S.A.V.); NIH (NINDS-R01NS117912 to M.J.S.; NIAMS-R01NS120060 to S.A.V. and P50AR052646 (to E.M.M. and M.J.S.); R01HL061322 and R01NS047726 (to E.M.M.); NIAID R01AI136514, U01AI150747, and R21AI169085 (to P.G.T.); the Cystic Fibrosis Foundation (PTAC postdoc-faculty transition award to M.A.B.); and the American Lebanese Syrian Associated Charities (to P.G.T.). The authors thank and acknowledge the use of services from the following cores: the UCLA Technology Center for Genomics & Bioinformatics (TCB) and the UCLA Institute for Quantitative and Computational Biosciences (QC Bio). This study was supported by the Department of Defense (to M.J.S. and S.A.V.); NIH ( NINDS - R01NS117912 to M.J.S.; NIAMS - R01NS120060 to S.A.V. and P50AR052646 (to E.M.M. and M.J.S.); R01HL061322 and R01NS047726 (to E.M.M.); NIAID R01AI136514 , U01AI150747 , and R21AI169085 (to P.G.T.); the Cystic Fibrosis Foundation (PTAC postdoc-faculty transition award to M.A.B.); and the American Lebanese Syrian Associated Charities (to P.G.T.). The five subjects described in this manuscript participated in a phase 1b clinical trial designed to test the safety of systemically delivered AAV9-mini-dystrophin (PF-06939926 fordadistrogene movaparvovec) in DMD patients. The trial was sponsored by Pfizer. The study was conducted in compliance with ethical principles of the Declaration of Helsinki and all International Conference on Harmonisation Good Clinical Practice Guidelines. The protocol was approved by the relevant institutional review board/independent ethics committee at each study site. All participants (or parent/legal guardian) provided written informed consent.
Keywords
- AAV
- Duchenne
- T cell receptor sequencing
- adeno-associated virus
- gene therapy
- immune response
- muscular dystrophy
- neuromuscular disorders
- single-cell sequencing
ASJC Scopus subject areas
- Molecular Medicine
- Molecular Biology
- Genetics