Involucrin-driven retroviral expression cassettes encoding human immunodeficiency virus envelope glycoproteins

Inventors

Gauduin, Marie-ClaireBlancou, Philippe

Assignees

Texas Biomedical Research Institute

Publication Number

US-9730996-B2

Publication Date

2017-08-15

Expiration Date

2033-10-24

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Abstract

The present invention provides for novel compositions and methods for delivering genes of interest to stem cells using vectors that contain differentiation-specific transcriptional regulatory elements. For example, stem cells in the internal epithelia could be transfected with a vaccine construct, which has an epithelial cell differentiation-specific promoter driving the expression of viral envelope proteins. When the promoter used is specific for terminally differentiated epithelial cells, then the viral envelope proteins will be expressed only in the upper part of the epithelia and therefore, stimulate the immune response. The infected epithelial stem cells in the basal layer will continue to produce new antigen-expressing cells, without being eliminated by the immune response. This invention will be useful in the development of vaccines against viral agents that target the internal mucosa like HIV.

Core Innovation

The invention provides novel nucleic acid compositions and methods for delivering genes of interest to stem cells using vectors containing differentiation-specific transcriptional regulatory elements. Specifically, the vectors utilize promoters such as the involucrin promoter, which is specific for terminally differentiated epithelial cells, to drive the expression of viral envelope proteins (for example, from HIV) only in particular layers of the epithelia. This targeted expression stimulates an immune response while preserving the infected epithelial stem cells in the basal layer, allowing continued generation of antigen-expressing cells.

The patent addresses the challenge of delivering antigens for prolonged periods to induce lasting and effective immunity, particularly in the context of HIV infection that typically initiates at mucosal surfaces. Existing vaccine strategies fail to maintain continuous antigen release and thus provide only transient protection. The invention overcomes this by integrating antigen-expressing genes under the control of differentiation-specific promoters into epithelial stem cells, resulting in the sustained presence of antigen-producing cells in the upper epithelial layers, which helps maintain an active immune response.

One embodiment features a retrovirus-based expression cassette lacking functional vif gene sequences and containing two short terminal repeats (lacking U3 regions), an involucrin promoter, and a gene encoding an HIV envelope protein. The promoter ensures that HIV antigens are produced exclusively in differentiated mucosal epithelial cells (such as in vaginal, rectal, oral, or nasal cavities), thus achieving localized and durable immune stimulation without depletion of the stem cell pool.

Claims Coverage

There are three identified independent claims, each outlining key inventive features regarding nucleic acid compositions with differentiation-specific promoters and retroviral expression cassettes for targeted envelope protein expression.

Expression cassette with involucrin promoter and HIV envelope gene lacking functional vif

A nucleic acid composition comprising a retrovirus-based expression cassette that includes: - An involucrin promoter. - A gene encoding an envelope protein of a human immunodeficiency virus. - Two short terminal repeat sequences that are retroviral long terminal repeat sequences lacking U3 regions. - No functional vif gene sequence. The involucrin promoter controls expression of the gene encoding the HIV envelope protein specifically in a differentiated mucosal epithelial layer in a subject.

Expression cassette with involucrin promoter targeting vaginal epithelium and lacking vif

A nucleic acid composition comprising a retrovirus-based expression cassette that includes: - An involucrin promoter. - A gene encoding an envelope protein of a human immunodeficiency virus. - Two short terminal repeat sequences that are retroviral long terminal repeat sequences lacking U3 regions. - No vif gene sequence. The involucrin promoter controls expression of the gene encoding the HIV envelope protein in a vaginal epithelial layer in a subject.

Expression cassette with selected differentiation-specific promoter for targeted HIV envelope expression lacking functional vif

A nucleic acid composition comprising a retrovirus-based expression cassette that includes: - A promoter selected from the group consisting of involucrin promoter, matrix metalloproteinase-9 promoter, keratin-10 promoter, and loricrin promoter. - A gene encoding an envelope protein of a human immunodeficiency virus. - Two short terminal repeat sequences that are retroviral long terminal repeat sequences lacking U3 regions. - No functional vif gene sequence. The selected promoter controls expression of the gene encoding the HIV envelope protein in a differentiated epithelial layer in a subject.

The inventive features focus on retrovirus-based expression cassettes with differentiation-specific promoters (including involucrin, MMP-9, keratin-10, loricrin), directing HIV envelope gene expression in differentiated epithelial tissues and lacking functional vif gene sequences. These features support localized, durable antigen expression for immunogenic applications.

Stated Advantages

Provides prolonged stimulation of the immune system with viral antigens, creating a strong barrier to viral replication.

Targets the immune response specifically at the site of primary viral replication, particularly at mucosal entry points of viruses like HIV.

Facilitates continuous antigen production by differentiated epithelial cells without elimination of the underlying epithelial stem cells, enabling sustained immunity.

Supports the development of single-dose, life-long lasting vaccines against viral agents targeting internal mucosa.

Documented Applications

Development of vaccines against viral agents, especially HIV, that target internal mucosal surfaces such as vaginal, rectal, oral, or nasal cavities.

Immunogenic compositions for eliciting immune responses in a subject, particularly for mucosal immunity.

Methods for delivering nucleic acid compositions to epithelial stem cells for sustained antigen expression.

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