Patent Application
20220175850
The field of the invention is nutritional supplements, particularly application of nutritional supplements to immunotherapies used in the treatment of cancer.
The background description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.
Immunotherapeutic approaches to treating cancer exploit elements of the patient's own immune system to treat the disease. Such immunotherapies can involve the administration of antibodies, cytokines, and/or cells retrieved from the patient and treated prior to re-administration.
Antibodies utilized in immunotherapy are typically directed to cell surface markers expressed by tumor cells, in order to activate the body's complement system or otherwise identify the cells to the immune system. Alternatively, such antibodies can be directed to cell surface receptors and interfere with down regulation of T-cell activity by cancer cells. Antibodies used for this purpose include Alemtuzumab (a monoclonal antibody directed to CD52 and which activates complement), Atezomlizumab (a monoclonal antibody directed to PD-L1 and which interferes with T-cell deactivation), and Ilipimumab (a monoclonal antibody directed to CTLA4, shifting the T-cell balance towards cytotoxicity). Unfortunately use of these therapeutic antibodies is associated with unwanted side effects, including precipitation of autoimmune disease, increased rate of infections, and neurological disorders.
Immunotherapy utilizing cytokines is directed to provoking an immune response to the tumor cells, which can themselves produce cytokines that reduce immune response. Cytokines used for immunotherapy include IFNα (used in treatment of hairy-cell leukemia, AIDS-related Kaposi's sarcoma, follicular lymphoma, chronic myeloid leukemia, and melanoma), IFNβ, and interleukin 2 (used in treatment of malignant melanoma and renal cell carcinoma). While these cytokines are known to have a variety of effects on the immune system the exact mechanism by which they attack cancer is not clear. Unfortunately, administration of these cytokines is associated with flu-like symptoms.
Immunotherapies utilizing cells involve removal of cells from the patient, activation and expansion of the cells in culture, and return of the activated cells to the patient. For example, Provenge is used to treat prostate cancer, and involves the removal of antigen presenting dendritic cells from the blood by leukapheresis, incubating them with a fusion protein made from elements of GM-CSF and a prostatic acid phosphatase, and reinfusing. The resulting improved presentation of cancer-specific antigens to the immune system is intended to improve the immune response. In another approach, CAR-T immunotherapy removes T cells and genetically modifies them to express a chimeric receptor that specifically recognizes target cancer cells. These modified T cells are returned to the patient, where it is hoped that they selectively target the cancer cells. Unfortunately, such approaches are expensive and time consuming, can cause flu-like symptoms, and have produced mixed results.
Thus, there is still a need for a simple and well tolerated immunotherapeutic approach to treating cancer.
The inventive subject matter provides compositions and methods in which a nutritional supplement is provided that modulates the expression of immune checkpoint proteins, immunotherapy targets, proteins associated with angiogenesis, and/or proteins associated with metastasis in cancer cells, which can be resistant to chemotherapy. Such cancer cells can be present in tissue culture or as a tumor. Such a nutritional supplement can be used in combination with chemotherapeutic drugs and/or radiotherapy.
One embodiment of the inventive concept is a method for providing immunotherapy to an individual with cancer (which can be drug resistant) by administering a nutritional supplement that includes selenium and fish oil (e.g. as shown in Table 1) in an amount sufficient to modify expression of a biological marker associated with an anti-neoplastic immune function. The nutritional supplement is provided in the absence of chemotherapy and/or in the absence of radiotherapy. In some embodiments the nutritional supplement is provided in combination with radiotherapy, thereby providing a synergistic effect in modifying expression of the biological marker. Suitable biological markers include AXL, HSP90, p-mTOR, PDL-1, EGFR, HDAC1, p-H2X, p-Akt, pSmad, mTOR, p-PTEN, p-STAT3, CXCR4, and STAT3. In some embodiments expression is increased for PD-1, CTLA4, FOXP3, CD8, PTEN, and/or p-P53. In some embodiments the ratio of expression of CD4 to expression of CD8 is reduced. In some embodiments the percentage of CD3+ T cells, CD3+CD4+ T cells, or CD4+CD8+ T cells in the individual's spleen is increased. In some embodiments of the inventive concept the immunotherapy provided by the nutritional supplement reduces expression of a biological marker associated with a stem cell characteristic or metastatic potential in tumor cells, such as CD24, CD29, CD31, VEGF, and MMP-9.
Another embodiment of the inventive concept is a method of activating immune cells to enhance anti-tumor activity by isolating an immune cell from an individual to be treated for cancer, contacting the isolated immune cell with an activating formulation comprising selenium and fish oil in an amount effective to modulate expression of a protein associated with immune cell activation to generate an activated immune cell, and returning the activated immune cell to the individual. A suitable formulation and/or elements thereof is shown in Table 1. In some embodiments of the inventive concept the activated immune cell is clonally expanding to generate a population of activated immune cells that is returned to the individual. In some embodiments the immune cell is clonally expanded prior to contacting with the activating formulation. In some embodiments the immune cell and/or the activated immune cell is genetically modified prior to returning to the individual. In some embodiments the individual is irradiated prior to isolating the immune cell.
Another embodiment of the inventive concept is a method of modulating expression of an immune checkpoint or immunotherapy-associated protein in a cell, by administering a supplement comprising selenium and fish oil (such as the supplement shown in Table 1). The supplement can be administered to provide a concentration of at least 200 ng/mL of selenium. The supplement can be administered to provide a concentration of at least 75 μM fish oil. Suitable fish oils include a fish oil with DHA and EPA in an about 2:3 weight ratio. The cell can be a cancer cell, which can have stem cell characteristics and/or be resistant to a chemotherapeutic drug. In some embodiments the method includes administering a chemotherapeutic drug and/or administering radiotherapy to the cell. In such embodiments the supplement can be administered prior to initiation of radiotherapy.
The modulation can be a reduction in expression, for example when the immune checkpoint or immunotherapy-associated protein is PD-L1, p-HSP27, vimentin, p-mTOR, p-p38, β-catenin, ABCG2, CD133, N-cadherin, p-MET, COX-2, GRP78, CD24, CD29, EGFR, HDAC1, p-H2X, p-Akt, MMP-9, CTLA4, CD28, CD86, C31, and/or STAT3. The modulation can be an increase in expression, and wherein the immune checkpoint or immunotherapy-associated protein is selected from the group consisting of PD-1, p-AMPKα, E-cadherin, CHOP, FOXP3, Nkp46, CD8, IL2, and/or PTEN.
Another embodiment of the inventive concept is a method for reducing circulating tumor cells in an animal with cancer by administering a nutritional supplement comprising selenium and fish oil (such as the supplement shown in Table 1) to the animal and administering chemotherapy to the animal. The supplement can be administered to provide a concentration of at least 200 ng/mL of selenium. The supplement can be administered to provide a concentration of at least 75 μM fish oil, which preferably has a DHA to EPA weight ratio of about 2:3.
Another embodiment of the inventive concept is the use of a supplement that includes selenium and fish oil to modulate expression of an immune checkpoint or immunotherapy-associated protein in a cell. The supplement can provide a concentration of at least 200 ng/mL of selenium to the cell following administration. The supplement can provide a concentration of at least 75 μM fish oil to the cell following administration. The fish oil preferably includes DHA and EPA in an about 2:3 weight ratio. The cell can be a cancer cell, such as a cancer cell that has stem cell characteristics and/or a cancer cell that is resistant to a chemotherapeutic drug. In some embodiments the supplement is used in combination with a chemotherapeutic drug and/or in combination with radiotherapy. In such an embodiment the supplement can used prior to initiation of radiotherapy.
The modulation can be a reduction in expression, such as when the immune checkpoint or immunotherapy-associated protein is PD-L1, p-HSP27, vimentin, p-mTOR, p-p38, □-catenin, ABCG2, CD133, N-cadherin, p-MET, COX-2, GRP78, CD24, CD29, EGFR, HDAC1, p-H2X, p-Akt, MMP-9, CTLA4, CD28, CD86, C31, and/or STAT3. The modulation can be an increase in expression, such as when the immune checkpoint or immunotherapy-associated protein is PD-1, p-AMPKα, E-cadherin, CHOP, FOXP3, Nkp46, CD8, and/or PTEN.
Another embodiment of the inventive concept is the use of a nutritional supplement that includes selenium and fish oil in combination with chemotherapy for reducing circulating tumor cells in an animal with cancer. The nutritional supplement provides a concentration of at least 200 ng/mL of selenium following administration. The nutritional supplement can provide a concentration of at least 75 μM fish oil following administration. In preferred embodiments the fish oil includes DHA and EPA in an about 2:3 weight ratio.
Various objects, features, aspects and advantages of the inventive subject matter will become more apparent from the following detailed description of preferred embodiments, along with the accompanying drawing figures in which like numerals represent like components.
The following description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.
The inventive subject matter provides compositions and methods in which a nutritional supplement (such as a supplement that includes chromium and certain plant-derived materials (NutraWell) and/or a supplement that includes a selenium yeast peptide complex and fish oil) is used to provide an immunotherapeutic effect, which can be demonstrated by modulation of cell-surface markers targeted by conventional immunotherapeutic drugs. In some embodiments such nutritional supplements are provided in combination with other antineoplastic therapies, such as radiation and/or chemotherapeutic agents. In some embodiments such combination therapy can surprisingly provides a significant synergistic effect in regard to modulation of cell surface markers that serve as immunotherapy targets.
One should appreciate that the disclosed techniques provide many advantageous technical effects including providing novel and well tolerated approach to modulating expression of cell surface markers associated with immunotherapy, but also in enhancing or complementing the effectiveness of current antineoplastic protocols.
The following discussion provides many example embodiments of the inventive subject matter. Although each embodiment represents a single combination of inventive elements, the inventive subject matter is considered to include all possible combinations of the disclosed elements. Thus if one embodiment comprises elements A, B, and C, and a second embodiment comprises elements B and D, then the inventive subject matter is also considered to include other remaining combinations of A, B, C, or D, even if not explicitly disclosed.
In some embodiments, the numbers expressing quantities of ingredients, properties such as concentration, reaction conditions, and so forth, used to describe and claim certain embodiments of the invention are to be understood as being modified in some instances by the term “about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable. The numerical values presented in some embodiments of the invention may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.
While the some findings described below are directed to the use of fish oil and a selenium source, the Applicant notes that the nutritional supplement formulation provided in Table 1 incorporates fish oil and selenium yeast components (such as peptides and/or amino acids prepared from selenium yeast). As such effects found in fish oil and selenium yeast studies can be extended to the use of this nutritional supplement or to formulations that include fish oil and selenium yeast in combination with one or more of the remaining components of the formulation shown in Table 1. The supplement formulation shown in Table 1 is a minimal dose formulation that has been found to have a high level of acceptance and to have unanticipated beneficial effect in regulating the expression of cell surface markers associated with antineoplastic immunotherapies. As shown below, such a nutritional supplement can also complement and/or enhance the effects of conventional antineoplastic therapies when used in combination. The formulation for a typical nutritional supplement of the inventive concept is shown below in Table 1.
Lact. Acidophilus (app. 10 billion total)
Bifido Bifidium (app. 10 billion total)
Lac. Bulgaricus (app. 10 billion total)
Bifido Longum (app. 10 billion total)
Strep. Thermophilus (app. 10 billion total)
The composition shown in Table 1 includes components that have various physiological and biochemical effects, including anti-inflammatory activity, lowering of blood glucose levels, lowering of cholesterol, and anti-tumor activity. Other components provide supplementation of necessary vitamins, minerals, and amino acids at elevated levels. Other components (e.g. enzymes, lecithin) serve to aid in digestion and absorption of components of the composition when consumed. The combination of these complementary activities provides a synergistic effect that exceeds the simple additive effect of individual components. It should be appreciated that the composition shown in Table 1 also includes certain flavorants (e.g. brown sugar, honey, vanilla flavor and masking agent) that serve to improve palatability and acceptance. Certain components (e.g. honey, brown sugar, milk, rice protein, casein) can provide both flavor and caloric energy. The Inventor has found that the combination of flavorants described above is effective in providing compliance with consumption of the nutritional supplement in effective amounts. In some embodiments, such flavorants can be excluded without negatively impacting the effectiveness of the nutritional supplement.
G to It should be appreciated that Table 1 provides a description of a formulation providing a minimum daily dosage of the cited ingredients, and that the range expressed for each of these is considered to be disclosure of intermediate ranges and/or subranges within the cited range. For example, a range of 30 μg to 4,000 μg of selenium is indicative of a minimal formulation that provides 30 μg to 4,000μ of selenium per day, and can be instructive of formulations that include various subranges that lie within this range (e.g. 500 μg to 2,000 μg, 2,000 μg to 4,000 μg. etc.). The composition shown in Table 1 can be represented by a single formulation or by two or more formulations that in sum provide the component ingredients.
It should also be appreciated that Table 1 teaches a minimal formulation (or Low dose), and that embodiments of the inventive concept can include formulations or use of formulations that include higher doses or ranges, for example a Mid dose where the composition includes 50% more of the component ingredients (i.e. 1.5× over what is shown in Table 1) or a High dose where the composition includes 100% more of the component ingredients (i.e. 2× over what is shown in Table 1). Accordingly, Mid and High dose formulations encompass formulations in which such multipliers are applied to the minimal formulation shown in Table 1. For example, a range of 30 μg to 4,000 μg of selenium in the minimal formulation can be modified by a 1.5× multiplier to 45 μg to 6,000 μg in a Mid dose formulation and to 60 μg to 8,000 μg in a High dose formulation modified by a 2× multiplier. Intermediate ranges within these Mid and High dose formulation ranges are considered disclosed as described above. Such Mid or High dose compositions can be represented by a single formulation or by two or more formulations that in sum provide the component ingredients.
In some embodiments of the inventive concept a component or ingredient listed in Table 1 can be provided as a portion of a larger molecule, salt, or complex. For example, metals such as selenium, chromium, and/or molybdenum can be provided as selenium yeast, chromium yeast, and molybdenum yeast (respectively), or as components of such yeast formulations.
In other embodiments of the inventive concept the nutritional supplement can include fish oil and a selenium-yeast preparation, or one or more selenium-containing peptides or amino acids derived from such yeast. Such a selenium peptide can include selenocysteine and/or selenomethionine.
In preferred embodiments fish oil utilized in compositions and methods of the inventive concept has a DHA to EPA weight ratio of about 2:3. Within the context of this application the term “about” in this context is inclusive of a ±10% or a ±20% deviation from the nominal value.
It should be appreciated that some embodiments of the inventive concept are compositions and methods of using a nutritional supplement containing selenium and fish oil to treat cancer, whereas other embodiments are compositions and methods that modulate the amounts of specific biological markers in living cells, whereas still other embodiments of the inventive concept are directed to the use of such a supplement in combination with one or more conventional anti-cancer therapies, in particular immunotherapy.
Some cancer cells are resistant to commonly used chemotherapeutic drugs, such as Iressa/Erlotinib. In some individuals such resistance occurs during the course of chemotherapeutic treatment, resulting in a loss of effectiveness and tumor progression. In others the cancer is resistant to the chemotherapeutic drug prior to treatment, allowing the tumor to progress as conventional chemotherapy is implemented.
The HCC827GR cell line is a lung cancer cell line that is resistant to Iressa. As shown in
As shown in
As shown in
The Inventor has found that treatment with fish oil and selenium (or with a nutritional supplement containing fish oil and selenium) can modulate expression of heat shock proteins other than HSP90. The Inventor has found that treatment with selenium and fish oil can modulate expression of heat shock proteins, for example p-HSP27. Such heat shock proteins are thought to provide a protective effect and are frequent targets of immunotherapy. As shown in
Immunotherapy is a treatment modality that remains available when tumor cells are resistant to chemotherapeutic drugs. As shown in
As shown in the left panel of
PD-1 and PD-L1 are ‘checkpoint’ proteins that regulate T-cell response and are targeted in cancer immunotherapy. PD-L1 is commonly overexpressed in cancer cells and binds to PD-1 present on activated cytotoxic T-cells, resulting their inhibition. These inhibited T-cells are ineffective in attacking the cancer cells. Immunotherapies are directed to either preventing or blocking the interaction between PD-1 and PD-L1. For example, the immunotherapeutic drug pembrolizumab is a monoclonal antibody directed to PD-1. This prevents binding of PD-L1 and blocks the inhibition of cytotoxic T-cells. Such monoclonal antibody drugs are administered by injection or infusion and are potentially immunogenic. Surprisingly, as shown in
As shown in
Inventors have also found that fish oil and selenium in combination can modulate PD-L1 in cancer sphere cells, which have cancer stem cell characteristics.
Additional studies have shown that the in vivo modulation of PD-1 and PD-L1 expression in tumor tissue and other immune checkpoint markers useful in immunotherapy by a nutritional supplement containing selenium and fish oil is dose dependent. It should be appreciated that in some embodiments a nutritional supplement of the inventive concept can provide differential effects on the expression of such immune checkpoint markers that are dependent upon the tumor location. For example modulation of immune checkpoint proteins by a nutritional supplement of the inventive concept can differ between a primary tumor site (e.g. at the site of implantation in an animal model) and a metastatic site (e.g. lung, liver, spleen, etc. in an animal model). FIG. 9A depicts a typical study design for animal model studies of this phenomena. Three different supplement formulations were used that provided low (NFS), mid (NFM-containing 1.5 times the Se content of NFS), and high (NFH, containing twice the Se content of NFS) doses of selenium and fish oil. These supplement formulations were provided in combination with a chemotherapeutic drug (such as Taxol or Adriamycin).
Vimentin, which can be expressed both intracellularly as well as at the cell surface, is another protein targeted by immunotherapies. As shown in
Inventors believe that use of a nutritional supplement that includes selenium and fish oil can replace and/or complement cancer chemotherapy and/or conventional immunotherapy by at least reducing tumor PD-L1 expression. It should also be appreciated that such a nutritional supplement is conveniently orally administered and is well tolerated.
Immunotherapies involving the use of specific monoclonal antibodies (or derivatives thereof) are finding increasing use in cancer treatment. Many of these immunotherapies target the interaction between PD-1 and PDL-1. For example, 4H2 is a chimeric murine antibody specific for PD-1.
The percentage of CD3 positive and CD4+ T cells found in the spleens of mice implanted with tumor cells and treated as shown in
The percentage of CD3 positive and CD8 positive T cells found in the spleens of mice implanted with tumor cells and treated as shown in
The percentage of dendritic cells found in the spleens of mice implanted with tumor cells and treated as shown in
As noted above, nutritional supplements can be provided that include different amounts of selenium (Se), and can also be provided with or used in combination with a chemotherapeutic agent. A typical dosing schedule used in animal studies is shown in
Epidermal growth factor receptor (EGFR) is a driver of tumorigenesis, and is frequently inappropriately activated (for example, by amplification) in lung cancer, breast cancer, and glioblastoma cells. EGFR is also associated with the development of drug resistance, as amplification has been shown to be driven by selective pressure applied by chemotherapeutic drugs. As shown in
As noted above, phosphorylated mTOR (p-mTOR) is also associated with tumor growth and drug resistance. Treatment of tumor-bearing mice with a chemotherapeutic agent e.g. taxol or avastin) in combination with a nutritional supplement containing fish oil and different amounts of selenium was found to sharply decrease expression of p-mTOR in a selenium-dose dependent manner, as shown in
Histone acetylation mediated by histone acetyltransferase represents an epigenetic modification that impacts gene expression. Aberrant expression of histone deacetylase is linked with tumor development, with altered gene expression leading to disruption of cellular functions such as cell proliferation, cell-cycle regulation, and apoptosis. Accordingly, inhibition of histone deacetylase is being investigated as a target for cancer therapy. As shown in
Phosphorylated Akt (p-Akt) is considered a marker for poor prognosis in some cancers, including breast and gastric cancer. As shown in
The Smad family of proteins is part of the TGF-β signaling pathway, which is involved in both the development and metastasis of tumors. Elevated levels of p-Smad 2 in the nuclei of cancer cells is associated with poor prognosis. As shown in
Cancer cells can have stem cell-like characteristics, the presence of which can be indicative of the ability to metastasize and/or develop resistance to chemotherapeutic drugs. CD24 and CD29 are markers that are commonly used to determine degree of stem cell character.
CD31 is another marker that is indicative of stem cell characteristics in tumor cells, which are related to their tendency to metastasize.
CD8 positive T cells are another potential target for cancer immunotherapy. The presence of such tumor infiltrating T cells in tumors correlates with overall survival in cancer patients.
CD4 positive T cells, which are active towards pre-oncogenic senescent cells, are another target for cancer immunotherapy. Such CD4 positive T cells act in concert with CD8 positive T cells, with CD8 positive T cells becoming active as senescent cells accumulate further mutations and move from senescence to oncogenesis. As such a normal balance between CD4 positive and CD8 positive T cells is desirable. The ratio between CD4 and CD8 expression provides a measure of the relative populations of these T cells in affected tissues.
CTLA4 is another immune checkpoint protein that downregulates immune responses, and is a potential target in cancer immunotherapies. For example, ipilimumab is a therapeutic antibody directed to CTLA4 that is used in cancer immunotherapy. Unfortunately use of ipilimumab can result in severe autoimmune effects.
In some embodiments of the inventive concept a nutritional supplement containing selenium and fish oil can be used to modify or regulate PD-1 and/o r PDL-1 expression
The effects of a nutritional supplement containing selenium and fish oil on the rate of change in the ratio of PD1 content of white blood cells to PDL-1 content of tumor cells has also been studied in human clinical trials.
Human clinical trials were also performed to characterize the effect of a nutritional supplement containing selenium and fish oil on the number of circulating tumor cells (CTC), which is related to metastasis from the primary tumor site. Patients were separated into three groups that received different amounts of the nutritional supplement, and the number of circulating tumor cells, serum selenium content, serum EPA content, and serum DHA content were characterized. Results are shown in Table 2.
indicates data missing or illegible when filed
As shown in Table 2, patients receiving the highest dose of a nutritional supplement containing selenium and fish oil showed the highest rate of reduction in circulating tumor cells, and also showed the highest serum concentrations of selenium, EPA, and DHA. Accordingly, providing cancer patients with a supplement that includes selenium and fish oil can result in a reduction in circulating tumor cells and, consequently, reduced tumor metastasis.
Inventors have also found that a supplement containing selenium and fish oil can enhance NK cell activity in animal models of breast cancer.
A similar study was performed in which T-cell and immune checkpoint markers were characterized in samples taken from triple negative breast cancer tumors in an animal model of human disease. Results are shown in
CXCR4 expression in tumors is associated with metastasis to CXCL12 expressing tissues, and CXCR4 inhibiting compounds have been shown to have anti-tumor activity. As shown in
Similar studies were performed in animal models of lung cancer, where the nutritional supplement was used in combination with radiotherapy. In some experiments nutritional supplementation with a supplement containing selenium and fish oil was provided for 7 days prior to the initiation of repeated rounds of radiotherapy; in other embodiments nutritional supplementation and radiotherapy began simultaneously. A typical study is shown in
CD8 is associated with cytotoxic T-cells, with an elevated CD4/CD8 ratio being associated with increased survival. As shown in
As noted above, the CD4/CD8 ratio is considered indicative of T cell activity directed to tumors. As shown in
STAT3 is observed to be elevated in cancer, and is associated with suppression of the immune system response to tumor cells. As shown in
As noted above in studies directed to animal models of breast cancer, PTEN suppression is characteristic of tumor cells.
As noted above in studies directed to animal models of breast cancer, CTLA-4 is considered an indicator of T cell activation. Results of studies characterizing CTLA-4 expression in tumor cells obtained from primary tumor cell inoculation sites in an animal model of lung cancer (as described above) are shown in
The Applicant believes that the modulation in gene expression levels found in certain tissues following the administration of a nutritional supplement that includes selenium and fish oil (without our without cotherapy using chemotherapeutic drugs and/or irradiation) can be due to changes in expression in tumor cells, changes in expression in surrounding tissue, a result of infiltration of tumor tissue and/or surrounding tissue by cells of the immune system, and/or changes in expression in cells of the immune system. The Applicant notes that many of the changes in gene expression resulting from treatment with a nutritional supplement are consistent with changes seen in activation of cells of the immune system towards anti-tumor activity.
Another embodiment of the inventive concept are methods and compositions for reducing invasiveness and/or angiogenesis in tumors, while modulating immune checkpoint proteins of the tumor. The Inventor has identified selenium and fish oil dose-dependent modulation of proteins associated with tumor angiogenesis and invasion (in addition to immune checkpoint proteins) in an animal model of triple negative breast cancer. Results are shown in
Another embodiment of the inventive concept is a method of treating cancer by treating blood or immune cells (such as NK cells) isolated from patient blood (for example, by leukophoresis) with active components of a nutritional supplement as described above (such as selenium and/or fish oil). Such ex vivo treatment can activate blood or immune cells and/or modulate expression of markers associated with improved antineoplastic immune function. Such ex vivo stimulation can take place prior to, during, or following expansion of the isolated immune cells in tissue culture. Following stimulation the activated/modified immune cells are returned to the patient where they show enhanced antineoplastic activity relative to corresponding non-stimulated immune cells. Optionally, such stimulated cells can be cultured and their numbers expanded (e.g. by clonal expansion) prior to re-implantation. In some embodiments such isolated immune cells can be modified by other methods (e.g. by genetic manipulation) to enhance anti-tumor activity in addition to exposure to active components of a nutritional supplement containing selenium and fish oil.
In some embodiments of the inventive concept ex vivo treatment of immune cells is combined with radiotherapy. In some embodiments radiotherapy can be applied to the individual prior to collection of the immune cells, such that ex vivo stimulation using active components of the nutritional supplement is directed to irradiated cells. In other embodiments ex vivo stimulation of the immune cells takes place prior to radiotherapy, such that irradiation is applied to immune cells that have been pre-treated with active components of the nutritional supplement, or to a patient following infusion of such treated cells. In still other embodiments immune cells collected from a patient can be isolated and treated with active components of a nutritional supplement of the inventive concept while they are subjected to radiotherapy. In a preferred embodiment such radiotherapy is relatively mild compared to those intended to kill tumor cells, and is sufficient to result in a degree of activation, stimulation, and/or otherwise improved anti-neoplastic function in the immune cells without generating common side effects of radiotherapy (e.g. immune suppression, gastrointestinal symptoms, hair loss, mucus membrane damage, skin lesions, etc.).
It should be apparent to those skilled in the art that many more modifications besides those already described are possible without departing from the inventive concepts herein. The inventive subject matter, therefore, is not to be restricted except in the spirit of the appended claims. Moreover, in interpreting both the specification and the claims, all terms should be interpreted in the broadest possible manner consistent with the context. In particular, the terms “comprises” and “comprising” should be interpreted as referring to elements, components, or steps in a non-exclusive manner, indicating that the referenced elements, components, or steps may be present, or utilized, or combined with other elements, components, or steps that are not expressly referenced. Where the specification claims refer to at least one of something selected from the group consisting of A, B, C . . . and N, the text should be interpreted as requiring only one element from the group, not A plus N, or B plus N, etc.
This application claims the benefit of U.S. Provisional Patent Application No. 62/895,421 filed on Sep. 3, 2019 and U.S. Provisional Patent Application No. 62/827,429 filed on Apr. 1, 2019. These and all other referenced extrinsic materials are incorporated herein by reference in their entirety. Where a definition or use of a term in a reference that is incorporated by reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein is deemed to be controlling.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US2020/026000 | 3/31/2020 | WO | 00 |
| Number | Date | Country | |
|---|---|---|---|
| 62827429 | Apr 2019 | US | |
| 62895421 | Sep 2019 | US |
