Recombinant adeno-associated virus virions for the treatment of lysosomal disorders

Abstract

AAV expression vectors and recombinant virions produced using these vectors, which include genes coding for enzymes defective or missing in lysosomal storage disorders, are described. These recombinant AAV virions are useful in the treatment of a variety of lysosomal storage disorders and the methods described herein provide for long-term, sustained expression of the defective or missing enzyme.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation of U.S. patent application Ser. No. 10 / 421,175, filed Apr. 22, 2003, which is a continuation of U.S. patent application Ser. No. 09 / 715,858, filed Nov. 17, 2000, now U.S. Pat. No. 6,582,692, from which applications priority is claimed under 35 U.S.C. § 120, and which claim the benefit of provisional patent applications No. 60 / 166,097, filed Nov. 17, 1999, and No. 60 / 215,430, filed Jun. 30, 2000, from which applications priority is claimed under 35 U.S.C. §119(e)(1), all of which applications are incorporated herein by reference in their entireties.[0002] This invention was made with support under NIH Grant 5 R01 DK54285-04 from the National Institutes of Health, U.S. Department of Health and Human Services. Accordingly, the United States Government may have certain rights in this invention.FIELD OF THE INVENTION [0003] The present invention relates to recombinant adeno-associated virus (AAV) expression...

AI Technical Summary

Benefits of technology

[0029] In particular, results presented here show that subjects treated with AAV virions encoding β-glucuronidase (GUS), acquire the ability to produce GUS for extended time periods. Furthermore, treatment has a beneficial effect on the subject, as it reduces or eliminates GAG storage in tissues. Morever, intrathecal injection of AAV virions into the cerebrospinal fluid to both neonatal and adult animals, results in therapeutic levels of GUS in the brain and the elimination of storage granules in brain tissue.

Problems solved by technology

Typically, the undegraded molecules accumulate in the lysosomes to form storage vesicles, which eventually distort cellular structure and compromise function.

The result is a chronic and progressive condition that causes a variety of physiological problems often leading to organ failure and premature death.

Without GUS activity, GAGs cannot be completely catabolized and therefore accumulate in the lysosomes to form storage granules or vacuoles in almost all tissues.

This in turn results in animals that have distorted facial features, defects in skeletal development, dwarfism, reduced learning capacity, and early death at approximately 5 months of age (Birkenmeier et al.

All of these treatments provided measurable improvement, but none was entirely satisfactory.

Major problems that need to be overcome are the transient nature of enzyme replacement and adenoviral vector therapies, the invasiveness and potential complications of transplantation therapies, and the difficulty in restoring therapeutic GUS activity throughout the organism.

This results in hepatosplenomegaly, bone pain and fractures, mild anemia and leukopenia, and bleeding due to displacement of platelet precursors in the bone marrow.

Pulmonary infiltration by engorged macrophages can cause respiratory failure.

Bone pain gradually decreases, but X-ray abnormalities persist.

Enzyme therapy, however, does not improve CNS symptoms and can cost as much as $200,000 per year per patient.

It is therefore cost-prohibitive for some patients.

Some patients are therefore denied treatment despite likely benefit.

Additionally, treatment schedules minimize the effectiveness of enzyme treatment.

GC has a half life of approximately 10 minutes and isolated enzyme administration creates a flood of GC during treatment sessions and a lack of GC between treatments.

The short half life of GC also limits distribution.

This regimen adds further disruption and inconvenience to patients.

As macrophages filled with sphingomyelin infiltrate the lungs, bronchitis and pneumonia occur, with respiratory difficulties increasing throughout infancy.

There is currently no treatment for Neimann-Pick Disease.

There is no current treatment for the glycoprotein-related Lysosomal Storage Diseases.

Acid Lipase Deficiency results in massive accumulation of cholesterol esters and triglycerides in most tissues of the body.

In none of these cases, however, were experiments designed to show that the AAV-mediated gene therapy actually affected the long-term course of a disease.

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