Cancer Therapeutic Vaccines

Stress Protein Compositions and Methods for Prevention and Treatment of Cancer and Infectious Disease

U.S. Patent Number: 6,984,384; 7,378,096 (CIP)
Issue Date: January 10, 2006; May 27, 2008 (CIP)

Summary: Pharmaceutical compositions comprising a stress protein complex and related molecules encoding or cells presenting such a complex are provided. The stress protein complex comprises an hsp110 or grp170 polypeptide complexed with an immunogenic polypeptide. The immunogenic polypeptide of the stress protein complex can be associated with a cancer or an infectious disease. The pharmaceutical compositions of the invention can be administered to a subject, thereby providing methods for inhibiting M. tuberculosis-infection, for inhibiting tumor growth, for inhibiting the development of a cancer, and for the treatment or prevention of infectious disease. The invention further provides a method for producing T cells directed against a tumor cell or a M. tuberculosis- infected cell, wherein a T cell is contacted with an APC that is modified to present an hsp110 or grp170 polypeptide and an immunogenic polypeptide associated with a tumor or with the M. tuberculosis-infected cell. Included in the invention are T cells produced by this method and a pharmaceutical composition comprising such T cells. The T cells can be contacted with a M. tuberculosis-infected cell in a method for killing a M. tuberculosis-infected cell, or with a tumor cell in a method for killing a tumor cell.

Detail: The invention provides a pharmaceutical composition comprising a stress protein complex. The stress protein complex comprises an hsp110 or grp170 polypeptide and an immunogenic polypeptide. In some embodiments, the hsp110 or grp170 polypeptide is complexed wit h the immunogenic polypeptide, for example, by non-covalent interaction or by covalent interaction, including a fusion protein. In some embodiments, the complex is derived from a tumor. In other embodiments, the complex is derived from cells infected with an infectious agent. The immunogenic polypeptide of the stress protein complex can be associated with a cancer or an infectious disease. The stress protein complex of the invention can further include additional stress polypeptides, including members of the hsp70, hsp90, grp78 and grp94 stress protein families. In one embodiment, the stress protein complex comprises hsp110 complexed with hsp70 and/or hsp25. The invention additionally provides a pharmaceutical composition comprising a first polynucleotide encoding an hsp 110 or a grp170 polypeptide and a second polynucleotide encoding an immunogenic polypeptide. In some embodiments involving first and second polynucleotides, the first polynucleotide is linked to the second polynucleotide. The pharmaceutical compositions of the invention can further comprise a physiologically acceptable carrier and/or an adjuvant. The efficacy of a pharmaceutical composition can further comprise GM-CSF-secreting cells. Alternatively, GM-CSF-secreting cells can be co-administered with a pharmaceutical composition of the invention, by administration before, during or after administration of the pharmaceutical composition. The use of GM-CSF-secreting cells enhances the efficacy of the pharmaceutical composition. In some embodiments, the complex is purified from a tumor or from cells infected with an infectious agent. In such embodiments, the stress polypeptide, as purified, is complexed with one or more immunogenic polypeptides. The binding of the stress polypeptide to the immunogenic polypeptide can be altered and/or enhanced by stress, such as by exposure to heat, anoxic and/or ischemic conditions, or proteotoxic stress. In particular, a stress protein complex of the invention can comprise a stress polypeptide complexed with an immunogenic polypeptide, wherein the complex has been heated. Such heating, particularly wherein the stress polypeptide comprises a heat-inducible stress protein, can increase the efficacy of the stress protein complex as a vaccine. Examples of heat-inducible stress proteins include, but are not limited to, hsp70 and hsp 110. In some embodiments, the immunogenic polypeptide is known. The immunogenic polypeptide is a known molecule, the immunogenic polypeptide can be provided in admixture with the stress polypeptide, or as a complex with the stress polypeptide. The hsp 110 or grp170 polypeptide can be complexed with the immunogenic polypeptide by non-covalent binding. Alternatively, the complex can comprise a fusion protein, wherein the stress polypeptide is linked to the immunogenic polypeptide. Examples of immunogenic polypeptides include, but are not limited to, antigens associated with cancer or infectious disease, such as the breast cancer antigen her2/neu or the Mycobacterium tuberculosis antigens Mtb8.4 and Mtb39. Where the immunogenic polypeptide is unknown, it can be obtained incidentally to the purification of the stress polypeptide from tissue of a subject having cancer or an infectious disease. Also provided is a pharmaceutical composition comprising an antigen-presenting cell (APC) modified to present an hsp 110 or grp170 polypeptide and an immunogenic polypeptide. Alternatively, the APC can be modified to present an immunogenic polypeptide obtained by purification of hsp 110 or grp170 from disease cells, including cancer cells and cells infected with an infectious agent. Preferably, the APC is a dendritic cell or a macrophage. The APC can be modified by various means including, but not limited to, peptide loading and transfection with a polynucleotide encoding an immunogenic polypeptide. The pharmaceutical compositions of the invention can be administered to a subject, thereby providing methods for inhibiting M. tuberculosis-infection, for inhibiting tumor growth, for inhibiting the development of a cancer, and for the treatment or prevention of cancer or infectious disease. The invention further provides a method for producing T cells directed against a tumor cell. The method comprises contacting a T cell with an antigen presenting cell (APC), wherein the APC is modified to present an hsp 110 or grp170 polypeptide and an immunogenic polypeptide associated with the tumor cell. Such T cells can be used in a method for killing a tumor cell, wherein the tumor cell is contacted with the T cell. Likewise, the invention provides a method for producing T cells directed against a M. tuberculosis-infected cell, wherein a T cell is contacted with an APC that is modified to present an hsp110 or grp170 polypeptide and an immunogenic polypeptide associated with the M. tuberculosis-infected cell. Included in the invention are T cells produced by this method and a pharmaceutical composition comprising such T cells. The T cells can be contacted with a M. tuberculosis-infected cell in a method for killing a M. tuberculosis-infected cell The T cells can be CD4+ or CD8+. The invention also provides a method for removing tumor cells from a biological sample. The method comprises contacting a biological sample with a T cell of the invention. In a preferred embodiment, the biological sample is blood or a fraction thereof. Also provided is a method for inhibiting tumor growth in a subject. The method comprises incubating CD4+ and/or CD8+ T cells isolated from the subject with an antigen presenting cell (APC), wherein the APC is modified to present an hsp 110 or grp 170 polypeptide and an immunogenic polypeptide associated with the tumor cell such that T cells proliferate. The method further comprises administering to the subject an effective amount of the proliferated T cells, and thereby inhibiting tumor growth in the subject. In an alternative embodiment, the method for inhibiting tumor growth in a subject comprises incubating CD4+ and/or CD8+ T cells isolated from the subject with an antigen presenting cell (APC), wherein the APC is modified to present an hsp110 or grp170 polypeptide and an immunogenic polypeptide associated with the tumor cell such that T cells proliferate, cloning at least one proliferated cell, and administering to the patient an effective amount of the cloned T cells, thereby inhibiting tumor growth in the subject.

Use of Recombinant Heat Shock Protein Complexed to Kidney Cancer Antigen

U.S. Patent: Pending
Application Number: 11/894,970

Summary: A heat shock protein in combination with carbonic anhydrase IX and a method for improving immune response to carbonic anhydrase IX in a mammal by complexing it with a heat shock protein prior to administration to the mammal.

Detail: Carbonic anhydrase IX (CA9) has been identified recently as a potential target for immunotherapy. CA9 is present in 95-100% of clear renal carcinoma cells (RCC) and it is not present in normal, nonmucosal tissue. We have now found that HSP proteins, normally found inside a cell, are powerful immune activators when found outside a cell that are capable of stimulating an immune response against proteins complexed to the HSP.

Survivin Peptides as Cancer Vaccines

Patent(s): Pending
U.S. Patent Application Number: 12/176,052
International Pending Applications: PCT July 18, 2008

Summary: Provided are compositions and methods for treating survivin expressing cancers. The compositions contain peptide survivin peptide mimics with improved MHC-I binding characteristics. The method involves administering a survivin peptide mimic with 4 improved MHC-I binding characteristics to an individual to effect inhibition of the growth of survivin expressing cancer cells in the individual.

Detail: The present invention provides compositions and methods for treating cancers that express survivin. The compositions comprise survivin peptide mimics that contain a cysteine to methionine alteration at amino acid position 57 of the wild type survivin protein sequence. The peptides are 9-23 amino acids in length and have the sequence of SEQ ID NO:4 (ENEPDLAQMFFCFKELEGWEPDD) or a fragment thereof, wherein the fragment comprises at least 9 contiguous amino acids of SEQ ID NO:4, and wherein the fragment also comprises the sequence of SEQ ID NO:5 (QMFFCF). The peptides are capable of stimulating an improved human cell mediated immune response against survivin expressing human cancer cells, as compared to the cell mediated immune response elicited by peptides having the wild type survivin sequence. The peptides are also demonstrated to confer extended survival in a mouse model of glioma. The method of the invention comprises administering to an individual diagnosed with or suspected of having a survivin expressing cancer a composition comprising a peptide of the invention such that growth of the cancer is inhibited. Also provided is a substantially purified population of mammalian dendritic cells that are loaded with a peptide of the invention.

Method for Inhibiting Scavenger Receptor-A and Increasing Immune Response to Antigens

Patent(s): Pending
U.S. Patent Application Number: 12/104,105
International Pending Applications: PCT April 16, 2008

Summary: Provided is a method for enhancing an immune response to a desired antigen in an individual. The method is performed by administering to the individual an agent capable of inhibiting class A macrophage scavenger receptor (SR-A) and optionally administering the desired antigen. Also provided is a method for enhancing an immune response to an antigen by administering to an individual a composition containing antigen presenting cells that are characterized by specifically inhibited SR-A. Substantially purified populations of mammalian dendritic cells characterized by specifically inhibited SR-A are also provided.

Detail: The present invention provides a method for enhancing an immune response to a desired antigen in an individual. The method comprises administering to the individual a desired antigen and an agent capable of inhibiting class A macrophage scavenger receptor (SR-A). By administering the agent and the antigen to the individual, the immune response to the antigen in the individual is enhanced. In another embodiment, a method is provided for enhancing an immune response to a tumor in an individual. The method comprising administering to the individual, in an amount effective to enhance an immune response to the tumor, an agent capable of inhibiting class A macrophage scavenger receptor (SR-A), wherein the growth of the tumor is inhibited subsequent to administering the agent. The method may further comprise administering to the individual an antigen that is expressed by the tumor. The agent may be any composition of matter that can specfically inhibit SR-A. Examples of such agents include but are not limited to polynucleotides that interfere with transcription and/or translation of SR-A mRNA. The agent may also be an antibody that binds to and antagonizes SR-A. The agent may also be any of various known sulfonamidobenzanilide compounds that can be used as SR-A antagonists. Also provided is a method for enhancing an immune response to a desired antigen comprising administering to an individual a composition comprising dendritic cells, wherein the dendritic cells are characterized by specifically inhibited SR-A. The method may further comprise exposing the dendritic cells to the desired antigen in vitro prior to administration to the individual. The invention also provides a composition comprising a substantially purified population of mammalian dendritic cells, wherein the dendritic cells are characterized by specifically inhibited SR-A activity.

Compositions and Methods for Using CA9 Protein to Stimulate an Immune Response

Patent(s): Pending
U.S. Patent Application Number: 12/263,756
International Pending Applications: PCT November 3, 2008

Summary: Provided are methods for stimulating an immune response to an antigen by administering a composition to an individual in an amount effective to stimulate an immune response to the antigen. The stimulated immune response to the antigen is greater than the immune response stimulated by the antigen in the absence of CA9 protein. The compositions provided contain a complex that includes an antigen and an isolated CA9 protein.

Detail: The present invention is based upon the surprising discovery of a heat shock protein (HSP) like chaperoning function for CA9. In particular, it is disclosed herein that CA9, as well as soluble form of CA9 that is shed from the surface of certain cells and is approximately 4 kDa smaller than full-length CA9, can inhibit heat-induced protein aggregation and can facilitate protein folding. However, unlike HSPs, we show that CA9 can form immunogenic complexes with antigens at room temperature (e.g., approximately 20.degree. C.) and at human body temperature (e.g., 37.degree. C.). CA9 is also shown herein to be internalized by antigen presenting cells and to be processed primarily through the proteosomal pathway, which is believed to be important for activation of cell-mediated immune responses. Further, we demonstrate in a murine melanoma model of cancer that the method of the invention is effective for stimulating an antigen-specific antitumor response that is greater than the response generated by the antigen in the absence of CA9 protein. Accordingly, the invention provides a method for stimulating an immune response to an antigen comprising administering a composition of the invention to an individual in an amount effective to stimulate an immune response to the antigen, wherein the immune response to the antigen is greater than the immune response stimulated by the antigen in the absence of CA9 protein. The compositions of the invention comprise an antigen and an isolated CA9 protein. The isolated CA9 protein and the antigen are provided in a complex, which complex may be a non-covalent association between isolated CA9 and the antigen, or may be a chemical conjugation of isolated CA9 protein to the antigen, or may be a CA9/antigen fusion protein. The immune response stimulated by administration of the composition may be prophylactic or therapeutic, and may comprise a humoral or cell mediated immune response, or combinations thereof. The invention is useful for stimulating an immune response against a wide variety of peptide and protein antigens, but heat shock proteins are not included within the scope of antigens used in the compositions and methods of the present invention. In one embodiment, the antigen is a tumor antigen.