The United States of America, as Represented by the Secretary, Department of Health and Human Services

The United States of America as Represented by the Department of Health

Brian Safer

John A Chiorini

Robert Michael Kotin

Stephen M Wiener

Roland A Owens

Matthew D Weitzman

Robert F Hoyt, Jr

John R Deleonardis

Randall R Clevenger

Elena Urcelay

Robert J Lutz

Robert Kotin

Douglas V Christini

The present invention provides an adeno-associated virus 4 (AAV4) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAV4 vectors and particles.

AAV4 vector and uses thereof

Methods and devices are disclosed for selective delivery of therapeutic substances to specific histologic or microanatomic areas of organs. Introduction of the therapeutic substance into a hollow organ space (such as an hepatobiliary duct or the gallbladder lumen) at a controlled pressure, volume or rate allows the substance to reach a predetermined cellular layer (such as the ephithelium or sub-epithelial space). The volume or flow rate of the substance can be controlled so that the intralumenal pressure reaches a predetermined threshold level beyond which subsequent subepithelial delivery of the substance occurs. Alternatively, a lower pressure is selected that does not exceed the threshold level, so that delivery occurs substantially only to the epithelial layer. Such site specific delivery of therapeutic agents permits localized delivery of substances (for example to the interstitial tissue of an organ) in concentrations that may otherwise produce systemic toxicity. Occlusion of venous or lymphatic drainage from the organ can also help prevent systemic administration of therapeutic substances, and increase selective delivery to superficial epithelial cellular layers. Delivery of genetic vectors can also be better targeted to cells where gene expression is desired. The access device comprises a cannula with a wall piercing tracar within the lumen. Two axially spaced inflatable balloons engage the wall securing the cannula and sealing the puncture site. A catheter equipped with an occlusion balloon is guided through the cannula to the location where the therapeutic substance is to be delivered.

Method for pressure mediated selective delivery of therapeutic substances and cannula

A composition for delivering at least one DNA sequence encoding a desired protein or polypeptide (such as a therapeutic agent) to a cell. The composition comprises an adeno-associated virus rep protein (or a nucleic acid sequence encoding an adeno-associated virus rep protein) and a genetic construct including at least one DNA sequence encoding a protein or polypeptide or genetic transcript of interest and a promoter controlling the at least one DNA sequence. The genetic construct also includes a first adeno-associated virus ITR or protein or derivative thereof and a second adeno-associated virus ITR or a portion or derivative thereof. The first and second adeno-associated virus ITRs or portions or derivatives thereof flank the at least one DNA sequence encoding the protein or polypeptide or genetic transcript of interest and the promoter controlling the at least one DNA sequence encoding the protein or polypeptide or genetic transcript of interest. Such a composition provides for integration of genetic material at a specific locus in the human chromosome, while minimizing the possibility of inadvertent inactivation of host genes and minimizing the possibility of viral contamination.

Methods of using adeno-associated virus rep protein

Lipid vesicles containing adeno-associated virus rep protein for transgene integration and gene therapy

A vector system comprising a first vector, which is an adeno-associated viral vector, and which includes an adeno-associated virus 5'ITR, an adeno-associated virus 3'ITR, and at least one heterologous DNA sequence. The vector system also includes a second vector which includes an inducible origin of replication, such as an SV40 origin of replication, which is capable of being induced or activated by an agent, such as the SV40 T-antigen. The second vector also includes DNA sequences encoding the adeno-associated virus rep and cap proteins. When induced by an agent, the second vector may replicate to a high copy number, and thereby increased numbers of infectious adeno-associated viral particles may be generated.

Vector systems for the generation of adeno-associated virus particles

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Silver Spring, MD, United States of America

Brian Safer