In some embodiments, the concentration of GAGs in the urine (e.g. urinary GAG levels) in the subject is reduced, stabilized or eliminated following administration of the composition and/or treatment according to the methods provided herein. Intravenous (IV) ERT with recombinant iduronate 2-sulfatase (IDS) protein (idursulfase; Elaprase.RTM., Shire) has been US FDA approved since 2006 for administration once every week in a dose of 0.5 mg per kg of body weight and has been shown to improve walking capacity in MPS II subjects 5 years and older. Of course this sort of brazen degeneracy is to be expected from a woman who makes her living by blasphemously whoring her body while pretending to be «tough» in the WWE. In some embodiments, the three polynucleotides are delivered to the subject with MPS II who is lacking an IDS gene such that a functional IDS protein is expressed in the subject. In further embodiments, the polynucleotides encoding the zinc finger nuclease may comprise SB-71557 (SB-A6P-ZL2, SEQ ID NO:30) or SB-71728 (SB-A6P-ZR2, Nude cam Sites SEQ ID NO:31).
In some aspects, the methods and compositions of the invention include the use of sequences encoding exogenous peptide sequences fused to eukaryotic transgene sequences. The combination of the three AAV2/6 components, including the IDS donor AAV, Left ZFN AAV and Right ZFN AAV, is collectively a composition of the invention. 32. The method of claim 1, wherein the composition comprises an article of manufacture comprising a formulation that includes three pharmaceutical compositions comprising the first, second and third AAV vectors. 36. The method of claim 35, wherein the method comprises (i) calculating the three product component volumes by multiplying the cohort dose by the patient weight before treatment and then dividing by the VG concentration as follows: (a) obtaining the cohort and patient weight at baseline from the study coordinator (b) obtaining the VG concentrations from the Clinical Certificates of Analysis; (ii) calculating the total volume by adding together the three product component volumes and the NS/PBS volume; (iii) calculating the volume of HSA intravenous solution required to achieve a final concentration of 0.25% HSA, and (iv) calculating the adjusted NS/PBS volume
> In some embodiments, the hIDS transgene (e.g. SEQ ID NO:15) useful in the compositions and methods described herein is delivered (e.g. to the hepatocyte) via AAV2/6 delivery, and an hIDS delivery vector further comprises homology arms (e.g. SEQ ID NO:13 and SEQ ID NO:16) flanking the hIDS transgene for example, with specificity for the regions flanking the ZFN cut site in the albumin locus. Compositions and methods for treating a subject with MPS II are effective to provide hIDS which is active (functional) and able to degrade mucopolysaccharides glycosaminoglycans or GAGs in vivo in the subject such that the concentration of GAGs in the urine (e.g. urinary GAG level) is reduced, stabilized or eliminated following treatment and/or provide a measurable increase in the amount of active IDS in the plasma. 3. The method of claim 1, wherein IDS levels in the plasma and/or leukocytes are stabilized and/or increased, optionally wherein IDS levels stay the same or is below the level of detectio
p> 11. The method of claim 1, further comprising measuring IDS activity and/or level in the plasma, liver, CSF or in leukocytes in the subject before and after treatment, wherein additional therapeutic procedures are delayed, reduced or eliminated if IDS activity is increased after treatment. 12. The method of claim 1, further comprising measuring total GAG levels, GAG comprising dermatan sulfate (DS GAG) levels, and/or GAG comprising heparan sulfate (HS GAG) levels (in the urine of the subject before and after treatment, wherein additional therapeutic procedures are delayed, reduced or eliminated if GAG, DS GAG and/or HS GAG levels are reduced after treatment. 15. The method of claim 1, further comprising measuring joint range of motion (JROM) before and after treatment, wherein additional therapeutic procedures are delayed, reduced or eliminated if JROM is increased after treatment. 18. The method of claim 1, wherein disability progression, organomegaly, hyperactivity, aggressiveness, neurologic deterioration, joint stiffness, skeletal deformities, heart valve thickening, hearing loss, corneal clouding and vision impairment, hernias, and/or upper respiratory infections are suppressed, reduced, delayed or eliminated in the subject after treatment. 5. The method of claim 1, wherein the additional treatment procedures that are reduced, delayed, and/or eliminated comprise enzyme replacement therapy (ERT); bone marrow transplant; and/or one or more supportive surgical procedures for orthopedic, cardiac and/or upper airway obstruction, wherein cardiac and/or upper air obstruction includes adenotonsillectomy, hernia repair, ventriculoperitoneal shunt, cardiac valve replacement, carpal tunnel release, and/orspinal decompressio