Inducible interleukin-12
Inventors
Morgan, Richard A. • Rosenberg, Steven A. • Zhang, Ling • Restifo, Nicholas P.
Assignees
US Department of Health and Human Services
Publication Number
US-8556882-B2
Publication Date
2013-10-15
Expiration Date
2030-04-22
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Abstract
The invention provides an isolated or purified nucleic acid comprising a nucleotide sequence encoding a nuclear factor of activated T-cells (NFAT) promoter operatively associated with a nucleotide sequence encoding IL-12. The invention also provides a nucleic acid comprising a nucleotide sequence encoding a nuclear factor of activated T-cells (NFAT) promoter operatively associated with a nucleotide sequence encoding IL-12, wherein the NFAT promoter is located 3′ of the nucleotide sequence encoding IL-12. Also provided are related recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions. The invention further provides the use of the inventive nucleic acids or related materials in the treatment or prevention of cancer or an infectious disease in a mammal and in the induction of IL-12 expression in a mammal.
Core Innovation
The invention provides nucleic acids comprising a nucleotide sequence encoding a nuclear factor of activated T-cells (NFAT) promoter operatively associated with a nucleotide sequence encoding interleukin-12 (IL-12). The NFAT promoter can be located either 5′ or 3′ of the IL-12 nucleotide sequence. These nucleic acids enable inducible expression of IL-12, where IL-12 is expressed only upon specific stimulation of the host cell, such as T-cell receptor (TCR) stimulation by an antigen or non-specific stimulation by agents like PMA/Ionomycin.
The core problem addressed by the invention is the limited clinical efficacy and severe systemic toxicity of recombinant IL-12 in cancer treatment. Recombinant IL-12 has shown potent anti-cancer activity in animal models but causes severe toxicity and limited results in human clinical trials when administered systemically. Thus, there is a need for compositions and methods that effectively treat cancer or infectious disease while reducing IL-12-associated toxicity.
By linking IL-12 expression to the NFAT promoter, the invention allows controlled, localized, and inducible IL-12 expression within host cells, such as T cells, only in the presence of relevant antigen stimulation. This strategic expression aims to enhance cytolytic activity and effector cytokine production while reducing systemic exposure to IL-12, thereby mitigating toxicity. The invention also includes various recombinant expression vectors, host cells transduced with these vectors, cell populations, and related pharmaceutical compositions for use in treating or preventing cancer or infectious diseases and for inducing IL-12 expression in mammals.
Claims Coverage
The patent includes sixteen independent claims covering nucleic acids, recombinant expression vectors, host cells, and therapeutic methods involving an NFAT promoter operatively associated with IL-12 sequences.
Nucleic acid encoding NFAT promoter operatively associated with IL-12
An isolated or purified nucleic acid comprising a nucleotide sequence encoding a nuclear factor of activated T-cells (NFAT) promoter operatively associated with a nucleotide sequence encoding IL-12.
Nucleic acid with NFAT promoter located 3′ of IL-12
A nucleic acid comprising a nucleotide sequence encoding a NFAT promoter operatively associated with a nucleotide sequence encoding IL-12, wherein the NFAT promoter is located 3′ of the IL-12 nucleotide sequence.
Recombinant expression vectors comprising nucleic acids encoding NFAT promoter and IL-12
Recombinant expression vectors comprising nucleic acids encoding NFAT promoter operatively associated with IL-12, including vectors selected from SEQ ID NOs: 5-8, 11-12, and 14, and employing vectors such as retroviral, lentiviral, herpesviral, adeno-associated viral, transposon, or adenoviral vectors.
Host cells comprising recombinant expression vectors
Host cells comprising a recombinant expression vector with a nucleotide sequence encoding an NFAT promoter operatively associated with IL-12, optionally further comprising a recombinant expression vector encoding a T cell receptor (TCR) with antigenic specificity for infectious disease or cancer antigens.
Methods for treating cancer or infectious diseases
Methods for treating a mammal with cancer or infectious disease involving isolating autologous T cells expressing a specific TCR, transducing them with the recombinant expression vector encoding NFAT promoter operatively associated with IL-12, and administering the modified T cells to the mammal to achieve inducible IL-12 expression stimulated by antigen presence.
Methods for inducing IL-12 expression in mammals
Methods for inducing IL-12 expression in a mammal by isolating autologous T cells, transducing them with a recombinant expression vector encoding NFAT promoter operatively associated with IL-12 and a second vector encoding a TCR specific for an antigen, administering the transduced cells, and stimulating the TCR by antigen to induce IL-12 expression.
The claims cover isolated nucleic acids encoding NFAT promoters operatively linked to IL-12 sequences in various vector forms, host cells containing these vectors optionally with TCR expression, and therapeutic or inducible IL-12 expression methods for treating cancer and infectious diseases.
Stated Advantages
Enables controlled expression of IL-12 only upon cell stimulation, reducing systemic toxicity associated with excess IL-12.
Enhances cytolytic activity and production of effector cytokines for improved anti-cancer or anti-infectious disease responses.
Permits inducible and localized IL-12 expression at tumor sites or infection sites, potentially improving therapy efficacy while minimizing side effects.
Allows for the expansion of transduced T cells without the deleterious effects seen in constitutive IL-12 expression systems.
Documented Applications
Treatment or prevention of cancer in mammals by administering nucleic acids, recombinant vectors, or host cells encoding inducible IL-12 under NFAT promoter.
Treatment or prevention of infectious diseases in mammals using compositions encoding inducible IL-12 expression.
Induction of IL-12 expression in mammals by transducing autologous T cells with vectors encoding NFAT promoter operatively associated with IL-12 and antigen-specific TCRs followed by cell administration and antigen stimulation.
Adoptive T cell therapy for melanoma and other cancers, demonstrated by regression of B16 melanomas in mice treated with T cells transduced with NFAT-controlled IL-12 vectors.
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