AAV-mediated gene therapy for maple syrup urine disease (MSUD)
A kind of allelic, base pair technology, applied in the composition of promoting the expression of functional BCKDHA protein in the subject, the composition of promoting the expression of functional BCKDHB protein in the subject, treating the subject with MSUD It can solve the problems of imperfect efficacy, difficulty in implementation, long-term immunosuppression accompanied by surgery, etc.
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Embodiment 1
[0188] Example 1: Analysis of bckdha gene therapy
[0189] MSUD is a rare genetic disease that affects branched amino acids (BCAA; leucine, isoleucine and proline) and degradation of ketoconic derivatives. It is caused by the mutation of the double allele in one of the three genes encoding the branzone sour dehydrogenase complex subunit (BCKDHA, BCKDHB and DBT). Serious (classic) MSUD is fatal in the absence of treatment. Diet BCAA restrictions are the main methods of treatment, but it is difficult to implement, uninterexably efficacy, and cannot provide the protection of an encephalopathy crisis against paroxysmia. Liver transplant is an effective alternative to diet therapies, but accompanied by surgery and long-term immunosuppression.
[0190] The impact of MSUD in the world is about 1 case per 185,000 birth population and is screened in most states and most developed countries. Due to the common Bckdha starting variant of the 4.5% group-specific carry frequency (C.1312T> a; p...
Embodiment 2
[0192] Example 2: Gene therapy by maplexia (MSUD) caused by bckdha or bckdhb
[0193] It is an aim to develop a AAV-mediated gene replacement therapy for MSUD caused by bckdha or bckdhb double allele mutation. Firstly, the AAV vector (OPTI-BCKDHA; SEQ ID NO: 1) or BCKDHB gene (OPTI-BCKDHB; SEQ ID NO: 4), which gene encodes the BCKD complex E1- α and E1-β subunits. BCKDHA activity is not effectively recovered in the art of the functional BCKDC E1 component (Alpha2-Beta2 "and BCKDHA or BETA2") and BCKDHA or BETA2 "and BCKDHA or BCKDHB. And BCKDHB Dual Carrier (Dual-Vector) Figure 1B And Figure 2). The protein expression of these carriers was verified in the HEK 293T cell line ( Figure 3A . Based on BCKDC enzyme activity assay, it is found that these carriers are functional. Figure 3b . Active recovery is more effective when bckdha and bckdhb are circulated. Pack Opti-Bckdha, Opti-BckDHB and Dual -Opti-Bckdha / BckDHB box is packaged in AAV9, which effectively targeted liver and ske...
Embodiment approach 1
[0197] Embodiment 1. A method for promoting functional BCKDHA protein in the subject, the functional BCKDHA protein is an E1-alpha subunit of branched alpha-ketoacly (BCAA) dehydrogenase complex, said method A nucleic acid comprising an effective amount comprising a casing is administered to the subject comprising an engineered nucleic acid to express BCKDHA in the liver and / or skeletal muscle of the subject, wherein the subject comprises at least An endogenous Bckdha allele, the endogenous Bckdha allele having a functionally lost mutation associated with maplexuria (MSUD).
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