Nutritional supplement containing fish oil and selenium

Abstract

To provide a nutritional supplement to modulate the expression of a hypoxia marker in a primary tumor or an apoptosis marker in a tumor.SOLUTION: Both a nutritional supplement containing fish oil and selenium and a cotherapy with such a supplement and radiotherapy have been found to cause a reduced incidence of metastasis from the primary implantation site relative to radiotherapy alone and also modulate the expression of genes associated with apoptosis in tumor cells. Such a supplement that is well tolerated and palatable is provided.SELECTED DRAWING: Figure 13B

Description

[Technical Field]

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 519,087, filed June 13, 2017. These and all other cited external materials are incorporated herein by reference in their entirety. In the event that the definition or use of a term in a reference incorporated by reference is inconsistent or contrary to the definition of a term provided herein, the definition of the term provided herein shall control.

[0002] (Technical field) The field of the invention relates to nutritional supplements containing fish oil and selenium. [Background technology]

[0003] The background discussion includes information that may be useful in understanding the present invention. None of the information provided herein is admitted to be prior art or related to the invention claimed herein, or that any publication specifically or implicitly mentioned is prior art.

[0004] Although radiation therapy and chemotherapy protocols utilized in the treatment of cancer can clearly benefit patients, they may be ineffective or less effective in some cancers. Furthermore, both radiation therapy and chemotherapy are associated with significant side effects, including nausea, weight loss, hair loss, damage to the gastrointestinal tract, and dermatitis.

[0005] Attempts have been made to enhance the effectiveness of radiation therapy. For example, gold nanoparticles modified to target tumor cells have been used to enhance radiation therapy (Yang et al., ACS Nano, 2014, 8(9):8992-9002). All publications herein are incorporated by reference to the same extent as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. In the event that a definition or use of a term in an incorporated reference is inconsistent with or contrary to a definition of a term provided herein, the definition of the term provided herein applies, and the definition of the term in the reference does not apply. Similarly, COX-2 inhibitors have been used to selectively sensitize tumor cells to the effects of radiation (Choy and Milas, J. Natl Cancer Inst (2003) 95(19):1140-1452). However, such approaches may have selectivity issues and may not be effective against all tumor types. To date, attempts to reduce the side effects of radiation therapy have primarily focused on dividing the total radiation dose into multiple smaller radiation doses (with time between to allow for recovery), targeting the tumor using shielding, identifying the tumor's borders, and localizing radiation therapy to that site.Unfortunately, such approaches cannot adequately treat all tumor cells.

[0006] Attempts have also been made to enhance the effectiveness of chemotherapy. Some studies suggest that taking fish oil can improve chemotherapy outcomes, while others suggest that fish oil can inhibit them (Daenen et al., JAMA Oncol (2015) 1(3):350-358). Formulation of chemotherapy agents as nanoparticles has also been attempted (Xu et al., Coll. Surf. B: Biointerfaces (2006) 48(1):50-57). However, it is unclear whether all chemotherapy agents are suitable for such reformulation. Codelivery of chemotherapy agents with siRNA designed to prevent multidrug resistance has also been investigated. However, such siRNAs are sequence-specific and may not be suitable for some tumors.

[0007] The reduction of the side effects of chemotherapy generally results in symptom reduction.For example, antiemetic drugs can be used to reduce nausea, along with improving diet and avoiding certain foods and eating small meals frequently.Unfortunately, this approach is not always effective.In some cases, chemotherapeutic agents are selected to reduce toxicity in order to reduce side effects, but these agents may also reduce the effectiveness of tumor cells. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Yang et al,ACS Nano,2014,8(9):8992-9002 [Non-patent document 2] Choy and Milas, J. Natl Cancer Inst(2003)95(19):1140-1452 [Non-patent document 3] Daenen et al, JAMA Oncol (2015)l(3):350-358 [Non-patent document 4] Xu et al,Coll.Surf.B:Biointerfaces(2006)48(l):50-57 Summary of the Invention [Problem to be solved by the invention]

[0009] Thus, there remains a need for safe and effective compositions and methods that enhance the efficacy and / or reduce the side effects of cancer radiation and / or chemotherapy. [Means for solving the problem]

[0010] (Summary of the Invention) The subject invention provides compositions and methods for synergistically enhancing and improving tumor radiotherapy through the use of nutritional supplements containing fish oil and selenium.

[0011] One embodiment of the inventive concept is a method of treating a tumor by providing a patient in need of treatment with a nutritional supplement comprising fish oil and selenium (e.g., as shown in Table 1) in amounts that provide a synergistic effect in reducing tumor volume or weight while administering a radiation therapy protocol to the patient. In some embodiments, the nutritional supplement is provided to the patient prior to the initiation of radiation therapy. Such a nutritional supplement can be formulated so that two or more components of the supplement are provided in the amounts set forth in Table 1.

[0012] Another embodiment of the inventive concept is a method of providing a patient with a nutritional supplement formulated to include fish oil and selenium (e.g., as set forth in Table 1) in amounts that reduce the side effects of a radiation therapy protocol while reducing the side effects. In some embodiments, the nutritional supplement is provided to the patient prior to the initiation of radiation therapy. Such a nutritional supplement can be formulated to provide two or more components of the supplement in amounts set forth in Table 1.

[0013] Another embodiment of the inventive concept is a method of modulating gene expression in a tumor (e.g., angiogenesis-related genes, apoptosis-related genes, etc.) by providing a nutritional supplement to a tumor or a tumor-bearing animal, wherein the nutritional supplement comprises fish oil and selenium (e.g., as set forth in Table 1), and the nutritional supplement is provided in an amount that modulates gene expression in the tumor. In some embodiments, the nutritional supplement is provided to a patient prior to the initiation of and during radiation therapy, while in other embodiments, such a supplement is provided during or prior to the start of radiation therapy administration. In a preferred embodiment, the nutritional supplement is formulated such that two or more components of the supplement are provided in the amounts set forth in Table 1.

[0014] Another embodiment of the inventive concept is a method of reducing metastasis from a tumor by providing a patient with a metastatic tumor with a nutritional supplement comprising fish oil and selenium (e.g., Table 1), wherein the nutritional supplement is provided in an amount that reduces the metastatic activity of the tumor. In some embodiments, such a nutritional supplement is provided either before or at the start of radiation therapy, and can be provided throughout the course of radiation therapy. In a preferred embodiment, the nutritional supplement is formulated so that two or more components of the supplement are provided in the amounts set forth in Table 1.

[0015] Another embodiment of the inventive concept is a method of reducing angiogenesis in a tumor, comprising providing a patient having said tumor with a nutritional supplement comprising fish oil and selenium (e.g., as set forth in Table 1) in combination with radiation therapy, wherein said nutritional supplement is provided in an amount that reduces the angiogenic activity of said tumor. In some embodiments, such a nutritional supplement is provided simultaneously with the initiation of radiation therapy and can be provided to the patient during the administration of radiation therapy. In a preferred embodiment, the nutritional supplement is formulated so that two or more components of the supplement are provided in the amounts set forth in Table 1.

[0016] Another embodiment of the inventive concept is a method of reducing the incidence of cancer stem cells in a tumor by providing a patient with a tumor with a nutritional supplement comprising fish oil and selenium (such as in Table 1), wherein the nutritional supplement is provided in an amount that reduces the incidence of stem cells in the tumor. In a preferred embodiment, the nutritional supplement is provided concurrently with the administration of radiation therapy. The nutritional supplement can be formulated so that two or more components of the supplement are provided in the amounts set forth in Table 1.

[0017] Specifically, the present invention provides the following:

[0018] 1. A method of treating a tumor, comprising administering a radiation therapy protocol to a patient in need of treatment, and providing said patient with a nutritional supplement comprising fish oil and selenium in amounts that provide a synergistic effect in reducing tumor volume or weight; Preferably, the nutritional supplement is provided to the patient prior to the initiation of radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0019] A method for reducing side effects of radiation therapy, comprising administering a radiation therapy protocol to a patient in need of treatment, and providing the patient with a nutritional supplement comprising fish oil and selenium in an amount that reduces the side effects of the radiation therapy protocol; Preferably, the nutritional supplement is provided to the patient prior to the start of the radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0020] providing a method for modulating gene expression in a tumor, the method comprising providing a nutritional supplement to a tumor or an animal having a tumor, the nutritional supplement comprising fish oil and selenium, the nutritional supplement provided in an amount that modulates gene expression in the tumor; Preferably, said nutritional supplement is provided to said patient prior to the initiation or during the administration of said radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the nutritional supplement are provided in the amounts set forth in Table 1. Preferably, the supplement comprises three or more ingredients as set forth in Table 1. Preferably, the gene encodes an angiogenic factor or encodes an apoptotic factor.

[0021] A method for reducing metastasis from a tumor, comprising providing a nutritional supplement comprising fish oil and selenium to a patient having a metastatic tumor and administering a radiation therapy protocol to the patient, wherein the nutritional supplement is provided in an amount that reduces metastatic activity of the tumor; Preferably, said nutritional supplement is provided concurrently with said radiation therapy protocol. Preferably, the nutritional supplement is formulated so that the components of the nutritional supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0022] providing a method for reducing angiogenesis in a tumor, the method comprising providing a nutritional supplement to a patient having a tumor, the nutritional supplement comprising fish oil and selenium, and administering a radiation therapy protocol to the patient, wherein the nutritional supplement is provided in an amount that reduces the angiogenic activity of the tumor; Preferably, the nutritional supplement is provided concurrently with radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the nutritional supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0023] 1. A method for reducing cancer stem cells in a tumor, comprising providing a patient having a tumor with a nutritional supplement comprising fish oil and selenium, and administering a radiation therapy protocol; wherein the nutritional supplement is provided in an amount effective to reduce the occurrence of cancer stem cells in the tumor; Preferably, said nutritional supplement is provided concurrently with said radiation therapy protocol. Preferably, the nutritional supplement is formulated so that the components of the nutritional supplement are provided in the amounts set forth in Table 1. Preferably, the nutritional supplement comprises three or more ingredients as set forth in Table 1.

[0024] A method for reducing PDL-1 expression in cancer cells is provided, comprising administering a radiation therapy protocol to said cancer cells, Preferably, the method further comprises contacting the cancer cells with a nutritional supplement comprising fish oil and selenium. Preferably, said nutritional supplement is provided prior to the commencement of said radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0025] Provided is a method for reducing PDL-1 expression in cancer cells, the method comprising contacting the cancer cells with a nutritional supplement comprising fish oil and selenium; Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the nutritional supplement comprises three or more ingredients as set forth in Table 1.

[0026] A method of enhancing anti-tumor immunotherapy in an individual is provided, comprising administering an anti-tumor immunotherapy protocol to said patient and administering a radiation therapy protocol to said patient; Preferably, the method further comprises administering to the patient a nutritional supplement comprising fish oil and selenium. Preferably, said nutritional supplement is provided prior to the initiation of said radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0027] A method of enhancing anti-tumor immunotherapy in an individual, the method comprising administering an anti-tumor immunotherapy protocol to the patient and providing the patient with a nutritional supplement comprising fish oil and selenium; Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the nutritional supplement comprises three or more ingredients as set forth in Table 1.

[0028] A method of reducing EGFR expression in cancer cells, comprising administering a radiation therapy protocol to said cancer cells, Preferably, the method further comprises contacting the cancer cells with a nutritional supplement comprising fish oil and selenium. Preferably, the nutritional supplement is provided prior to the start of the radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0029] 2. A method of reducing EGFR expression in cancer cells, comprising contacting the cancer cells with a nutritional supplement comprising fish oil and selenium; Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the nutritional supplement comprises three or more ingredients as set forth in Table 1.

[0030] A method of enhancing anti-EGFR therapy in an individual with cancer is provided, comprising administering an anti-EGFR therapy protocol to the patient and administering a radiation therapy protocol to the patient; Preferably, the method further comprises administering to the patient a nutritional supplement comprising fish oil and selenium. Preferably, said nutritional supplement is provided prior to the commencement of said radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0031] A method of enhancing anti-EGFR therapy in an individual with cancer, comprising administering an anti-EGFR therapy protocol to the patient and providing the patient with a nutritional supplement comprising fish oil and selenium, Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the nutritional supplement comprises three or more ingredients as set forth in Table 1.

[0032] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for improving radiation therapy in an individual with cancer, wherein the fish oil and selenium are provided in amounts that provide a synergistic effect in reducing tumor volume or weight when used in combination with a radiation therapy protocol; Preferably, the nutritional supplement is formulated for provision to the patient prior to the commencement of the radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0033] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for reducing side effects associated with radiation therapy in an individual undergoing radiation therapy for cancer, wherein the fish oil and selenium are provided in amounts that provide a synergistic effect in reducing the side effects of the radiation therapy protocol; Preferably, the nutritional supplement is formulated to be effective when administered to the patient prior to the initiation of radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement comprises three or more ingredients as set forth in Table 1.

[0034] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for modulating gene expression in a tumor in a patient undergoing radiation therapy, wherein the nutritional supplement is provided in an amount that modulates gene expression in the tumor; Preferably, the nutritional supplement is formulated to be provided to the patient prior to or during the initiation of radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the nutritional supplement are provided in the amounts set forth in Table 1. Preferably, the supplement comprises three or more ingredients as set forth in Table 1. Preferably, the gene encodes an angiogenic factor or encodes an apoptotic factor.

[0035] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for reducing metastasis from a tumor in a patient undergoing radiation therapy, wherein the nutritional supplement is provided in an amount that reduces metastatic activity of the tumor; Preferably, the nutritional supplement is formulated to be given concurrently with a radiation therapy protocol. Preferably, the nutritional supplement is formulated so that the components of the nutritional supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0036] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for reducing angiogenesis in a tumor in a patient undergoing radiation therapy, wherein the nutritional supplement is provided in an amount that reduces angiogenic activity in the tumor; Preferably, the nutritional supplement is provided concurrently with radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the nutritional supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0037] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for reducing cancer stem cell content in a tumor of a patient undergoing radiation therapy, wherein the nutritional supplement is provided in an amount effective to reduce the occurrence of cancer stem cells in the tumor; Preferably, providing said nutritional supplement so that it is administered concurrently with a radiation therapy protocol. Preferably, the nutritional supplement is formulated so that the components of the nutritional supplement are provided in the amounts set forth in Table 1. Preferably, the nutritional supplement comprises three or more ingredients as set forth in Table 1.

[0038] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for reducing PDL-1 expression in cancer cells undergoing radiation therapy, wherein the nutritional supplement is provided in an amount effective to reduce PDL-1 in the cancer cells; Preferably, the nutritional supplement is formulated to be administered prior to the commencement of the radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the nutritional supplement comprises three or more ingredients as set forth in Table 1. Preferably, the cancer cells are exposed to radiation therapy.

[0039] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for enhancing immunotherapy in an individual undergoing immunotherapy for cancer, wherein the nutritional supplement is provided in an amount effective to reduce PDL-1 in the cancer cells; Preferably, the nutritional supplement is provided before the start of radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0040] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for reducing EGFR expression in cancer cells, wherein the nutritional supplement is provided in an amount effective to reduce EGFR in the cancer cells; Preferably, the cancer cells are subjected to radiotherapy. Preferably, the nutritional supplement is formulated to be administered prior to the initiation of radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0041] Use of a nutritional supplement comprising fish oil and selenium for the manufacture of a medicament for enhancing anti-EGFR therapy in an individual with cancer, wherein the nutritional supplement is provided in an amount effective to reduce EGFR in cancer cells; Preferably, the individual is undergoing radiation therapy. Preferably, the nutritional supplement is formulated to be administered prior to the initiation of radiation therapy. Preferably, the nutritional supplement is formulated so that the components of the supplement are provided in the amounts set forth in Table 1. Preferably, the supplement contains three or more ingredients as set forth in Table 1.

[0042] 1. A nutritional supplement for reducing H1f-α expression at primary tumor sites, comprising fish oil and selenium yeast, providing a nutritional supplement, wherein the nutritional supplement is provided in an amount effective to reduce H1f-α expression at the primary tumor site; Preferably, it contains 150 mg to 10,000 mg of fish oil and 30 μg to 4,000 μg of selenium yeast. Preferably, the nutritional supplement is formulated for use in combination with radiation therapy. Preferably, the nutritional supplement further comprises 10,000mg to 50,000mg of maltodextrin, 5,000mg to 60,000mg of whey protein isolate, 1,000mg to 50,000mg of whey protein concentrate, 40mg to 15,000mg of fructooligosaccharides / insulin, 1,000mg to 9,000mg of granulated honey, 500mg to 15,000mg of oat fiber, 500mg to 20,000mg of natural French vanilla flavor, 500mg to 50,000mg of soy protein, 500mg to 1 ... of brown powdered sugar, 500mg-5,000mg of natural vanilla masking flavor, 200mg-10,000mg of lecithin, 50mg-5,000mg of nonfat milk, 50mg-5,000mg of rice protein powder, 50mg-2,000mg of calcium caseinate, 100mg-7,000mg of flax oil, 100mg-7,000mg of canola oil, 100mg-7,000mg of borage oil, 100mg-7,000mg of olive oil, 100mg-1,000mg of pure lemon oil, 50mg-1,000mg of pure orange oil, 0.5mg ~200mg mixed tocopherols, 200mg-1,500mg potassium phosphate, 100mg-5,000mg calcium carbonate, 150mg-2,500mg choline bitartrate, 100mg-2,000mg sodium chloride, 100mg-2,000mg tribasic calcium phosphate, 50mg-3,000mg ascorbic acid, 50mg-2,000mg potassium chloride, 50mg-500mg magnesium oxide, 30μg-3,000μg chromium yeast, 30μg-2,000μg molybdenum yeast, 10mg-5,000mg Inositol, 5mg-200mg zinc sulfate monohydrate, 5IU-2,000IU dried vitamin E acetate, 5mg-500mg niacinamide, 3mg-100mg ferric orthophosphate, 3mg-200mg calcium pantothenate, 3mg-100mg manganese sulfate monohydrate, 1mg-100mg beta-carotene, 1mg-15mg copper gluconate, 25IU-5,000IU vitamin D3, 2μg-1,000μg vitamin K2, 0.5mg-200mg pyridoxine HCl, 0.5mg-1,500mg potassium iodide, 0.5mg-1,000mg riboflavin, 0.5mg-2,500mg thiamine hydrochloride, 1μg-500μg dried vitamin K1, 500IU-100,000IU vitamin A acetate, 100μg-10,000μg folic acid, 10μg-10,000μg d-biotin, 1μg-3,000μg vitamin B12, 300mg-30,000mg L-carnitine, 500mg-60,000mg L-glutamine, 500mg-30,000mg 0mg L-arginine base, 50mg-2,000mg taurine, 50mg-2,000mg L-lysine, 10mg-1,000mg alpha lipoic acid, 15mg-1,500mg resveratrol, 10mg-5,000mg coenzyme Q10, 5mg-1,000mg glycine, 5mg-1,000mg proline, 2mg-500mg Lact. Acidophilus (total approx. 10 billion), 2mg-500mg Bifidobacterium (total approx. 10 billion), 2mg-500mg Lac. Bulgaricus (total approx. 10 billion), 2mg-500mg Bifidobacterium Longum (approximately 10 billion in total), 2mg-500mg of Strep. Thermophilus (approximately 10 billion in total), 5mg-100mg of papain, 5mg-100mg of pepsin, 5mg-100mg of lipase, 5mg-100mg of bromelain, 0.5mg-100mg of pancreatin 4X, 1mg-100mg of lactase, 3mg-100mg of betaine HCl, 2mg-500mg of pineapple juice powder, 2mg-500mg of papaya fruit powder, 30mg-3, It contains 1,000mg of quercetin, 25mg to 600mg of EGCG, 15mg to 500mg of OPC, 15mg to 5,000mg of anthocyanins, 10mg to 300mg of ellagic acid, 2mg to 90mg of astaxanthin, 20mg to 1,500mg of fucoidan, 5mg to 6,000mg of cordyceps, 15mg to 10,000mg of Ganoderma lucidum, 40mg to 15,000mg of shiitake mushroom, 30mg to 15,000mg of maitake mushroom, and 30mg to 15,000mg of trautauraceae.

[0043] 1. A nutritional supplement for increasing Bax expression in tumors, comprising fish oil and selenium yeast, providing a nutritional supplement, wherein the nutritional supplement is provided in an amount effective to increase Bax expression in tumors; Preferably, it contains 150 mg to 10,000 mg of fish oil and 30 μg to 4,000 μg of selenium yeast. Preferably, the nutritional supplement is formulated to provide a synergistic effect in combination with radiation therapy in increasing Bax expression in tumors. Preferably, the nutritional supplement further comprises 10,000mg to 50,000mg of maltodextrin, 5,000mg to 60,000mg of whey protein isolate, 1,000mg to 50,000mg of whey protein concentrate, 40mg to 15,000mg of fructooligosaccharides / insulin, 1,000mg to 9,000mg of granulated honey, 500mg to 15,000mg of oat fiber, 500mg to 20,000mg of natural French vanilla flavor, 500mg to 50,000mg of soy protein, 500mg to 1 ... of brown powdered sugar, 500mg-5,000mg of natural vanilla masking flavor, 200mg-10,000mg of lecithin, 50mg-5,000mg of nonfat milk, 50mg-5,000mg of rice protein powder, 50mg-2,000mg of calcium caseinate, 100mg-7,000mg of flax oil, 100mg-7,000mg of canola oil, 100mg-7,000mg of borage oil, 100mg-7,000mg of olive oil, 100mg-1,000mg of pure lemon oil, 50mg-1,000mg of pure orange oil, 0.5mg ~200mg mixed tocopherols, 200mg-1,500mg potassium phosphate, 100mg-5,000mg calcium carbonate, 150mg-2,500mg choline bitartrate, 100mg-2,000mg sodium chloride, 100mg-2,000mg tribasic calcium phosphate, 50mg-3,000mg ascorbic acid, 50mg-2,000mg potassium chloride, 50mg-500mg magnesium oxide, 30μg-3,000μg chromium yeast, 30μg-2,000μg molybdenum yeast, 10mg-5,000mg Inositol, 5mg-200mg zinc sulfate monohydrate, 5IU-2,000IU dried vitamin E acetate, 5mg-500mg niacinamide, 3mg-100mg ferric orthophosphate, 3mg-200mg calcium pantothenate, 3mg-100mg manganese sulfate monohydrate, 1mg-100mg beta-carotene, 1mg-15mg copper gluconate, 25IU-5,000IU vitamin D3, 2μg-1,000μg vitamin K2, 0.5mg-200mg pyridoxine HCl, 0.5mg-1,500mg potassium iodide, 0.5mg-1,000mg riboflavin, 0.5mg-2,500mg thiamine hydrochloride, 1μg-500μg dried vitamin K1, 500IU-100,000IU vitamin A acetate, 100μg-10,000μg folic acid, 10μg-10,000μg d-biotin, 1μg-3,000μg vitamin B12, 300mg-30,000mg L-carnitine, 500mg-60,000mg L-glutamine, 500mg-30,000mg 0mg L-arginine base, 50mg-2,000mg taurine, 50mg-2,000mg L-lysine, 10mg-1,000mg alpha lipoic acid, 15mg-1,500mg resveratrol, 10mg-5,000mg coenzyme Q10, 5mg-1,000mg glycine, 5mg-1,000mg proline, 2mg-500mg Lact. Acidophilus (total approx. 10 billion), 2mg-500mg Bifidobacterium (total approx. 10 billion), 2mg-500mg Lac. Bulgaricus (total approx. 10 billion), 2mg-500mg Bifidobacterium Longum (approximately 10 billion in total), 2mg-500mg of Strep. Thermophilus (approximately 10 billion in total), 5mg-100mg of papain, 5mg-100mg of pepsin, 5mg-100mg of lipase, 5mg-100mg of bromelain, 0.5mg-100mg of pancreatin 4X, 1mg-100mg of lactase, 3mg-100mg of betaine HCl, 2mg-500mg of pineapple juice powder, 2mg-500mg of papaya fruit powder, 30mg-3, It contains 1,000mg of quercetin, 25mg to 600mg of EGCG, 15mg to 500mg of OPC, 15mg to 5,000mg of anthocyanins, 10mg to 300mg of ellagic acid, 2mg to 90mg of astaxanthin, 20mg to 1,500mg of fucoidan, 5mg to 6,000mg of cordyceps, 15mg to 10,000mg of Ganoderma lucidum, 40mg to 15,000mg of shiitake mushroom, 30mg to 15,000mg of maitake mushroom, and 30mg to 15,000mg of trautauraceae.

[0044] 1. A nutritional supplement for increasing Bcl-2 expression in tumors, comprising fish oil and selenium yeast, The nutritional supplement, wherein the nutritional supplement is provided in an amount effective to increase Bcl-2 expression in tumors. Preferably, it contains 150 mg to 10,000 mg of fish oil and 30 μg to 4,000 μg of selenium yeast. Preferably, the nutritional supplement is formulated for use in combination with radiation therapy. Preferably, the nutritional supplement further comprises 10,000mg to 50,000mg of maltodextrin, 5,000mg to 60,000mg of whey protein isolate, 1,000mg to 50,000mg of whey protein concentrate, 40mg to 15,000mg of fructooligosaccharides / insulin, 1,000mg to 9,000mg of granulated honey, 500mg to 15,000mg of oat fiber, 500mg to 20,000mg of natural French vanilla flavor, 500mg to 50,000mg of soy protein, 500mg to 1 ... of brown powdered sugar, 500mg-5,000mg of natural vanilla masking flavor, 200mg-10,000mg of lecithin, 50mg-5,000mg of nonfat milk, 50mg-5,000mg of rice protein powder, 50mg-2,000mg of calcium caseinate, 100mg-7,000mg of flax oil, 100mg-7,000mg of canola oil, 100mg-7,000mg of borage oil, 100mg-7,000mg of olive oil, 100mg-1,000mg of pure lemon oil, 50mg-1,000mg of pure orange oil, 0.5mg ~200mg mixed tocopherols, 200mg-1,500mg potassium phosphate, 100mg-5,000mg calcium carbonate, 150mg-2,500mg choline bitartrate, 100mg-2,000mg sodium chloride, 100mg-2,000mg tribasic calcium phosphate, 50mg-3,000mg ascorbic acid, 50mg-2,000mg potassium chloride, 50mg-500mg magnesium oxide, 30μg-3,000μg chromium yeast, 30μg-2,000μg molybdenum yeast, 10mg-5,000mg Inositol, 5mg-200mg zinc sulfate monohydrate, 5IU-2,000IU dried vitamin E acetate, 5mg-500mg niacinamide, 3mg-100mg ferric orthophosphate, 3mg-200mg calcium pantothenate, 3mg-100mg manganese sulfate monohydrate, 1mg-100mg beta-carotene, 1mg-15mg copper gluconate, 25IU-5,000IU vitamin D3, 2μg-1,000μg vitamin K2, 0.5mg-200mg pyridoxine HCl, 0.5mg-1,500mg potassium iodide, 0.5mg-1,000mg riboflavin, 0.5mg-2,500mg thiamine hydrochloride, 1μg-500μg dried vitamin K1, 500IU-100,000IU vitamin A acetate, 100μg-10,000μg folic acid, 10μg-10,000μg d-biotin, 1μg-3,000μg vitamin B12, 300mg-30,000mg L-carnitine, 500mg-60,000mg L-glutamine, 500mg-30,000mg 0mg L-arginine base, 50mg-2,000mg taurine, 50mg-2,000mg L-lysine, 10mg-1,000mg alpha lipoic acid, 15mg-1,500mg resveratrol, 10mg-5,000mg coenzyme Q10, 5mg-1,000mg glycine, 5mg-1,000mg proline, 2mg-500mg Lact. Acidophilus (total approx. 10 billion), 2mg-500mg Bifidobacterium (total approx. 10 billion), 2mg-500mg Lac. Bulgaricus (total approx. 10 billion), 2mg-500mg Bifidobacterium Longum (approximately 10 billion in total), 2mg-500mg of Strep. Thermophilus (approximately 10 billion in total), 5mg-100mg of papain, 5mg-100mg of pepsin, 5mg-100mg of lipase, 5mg-100mg of bromelain, 0.5mg-100mg of pancreatin 4X, 1mg-100mg of lactase, 3mg-100mg of betaine HCl, 2mg-500mg of pineapple juice powder, 2mg-500mg of papaya fruit powder, 30mg-3, It contains 1,000mg of quercetin, 25mg to 600mg of EGCG, 15mg to 500mg of OPC, 15mg to 5,000mg of anthocyanins, 10mg to 300mg of ellagic acid, 2mg to 90mg of astaxanthin, 20mg to 1,500mg of fucoidan, 5mg to 6,000mg of cordyceps, 15mg to 10,000mg of Ganoderma lucidum, 40mg to 15,000mg of shiitake mushroom, 30mg to 15,000mg of maitake mushroom, and 30mg to 15,000mg of trautauraceae.

[0045] 1. A nutritional supplement for increasing the ratio of Bax expression to Bcl-2 expression in tumors, comprising fish oil and selenium yeast, providing a nutritional supplement provided in an amount effective to increase the ratio of Bax expression to Bcl-2 expression in a tumor; Preferably, it contains 150 mg to 10,000 mg of fish oil and 30 μg to 4,000 μg of selenium yeast. Preferably, the nutritional supplement is formulated to provide a synergistic effect in combination with radiation therapy in increasing the ratio of Bax to Bcl-2 expression in tumors. Preferably, the nutritional supplement further comprises 10,000mg to 50,000mg of maltodextrin, 5,000mg to 60,000mg of whey protein isolate, 1,000mg to 50,000mg of whey protein concentrate, 40mg to 15,000mg of fructooligosaccharides / insulin, 1,000mg to 9,000mg of granulated honey, 500mg to 15,000mg of oat fiber, 500mg to 20,000mg of natural French vanilla flavor, 500mg to 50,000mg of soy protein, 500mg to 1 ... of brown powdered sugar, 500mg-5,000mg of natural vanilla masking flavor, 200mg-10,000mg of lecithin, 50mg-5,000mg of nonfat milk, 50mg-5,000mg of rice protein powder, 50mg-2,000mg of calcium caseinate, 100mg-7,000mg of flax oil, 100mg-7,000mg of canola oil, 100mg-7,000mg of borage oil, 100mg-7,000mg of olive oil, 100mg-1,000mg of pure lemon oil, 50mg-1,000mg of pure orange oil, 0.5mg ~200mg mixed tocopherols, 200mg-1,500mg potassium phosphate, 100mg-5,000mg calcium carbonate, 150mg-2,500mg choline bitartrate, 100mg-2,000mg sodium chloride, 100mg-2,000mg tribasic calcium phosphate, 50mg-3,000mg ascorbic acid, 50mg-2,000mg potassium chloride, 50mg-500mg magnesium oxide, 30μg-3,000μg chromium yeast, 30μg-2,000μg molybdenum yeast, 10mg-5,000mg Inositol, 5mg-200mg zinc sulfate monohydrate, 5IU-2,000IU dried vitamin E acetate, 5mg-500mg niacinamide, 3mg-100mg ferric orthophosphate, 3mg-200mg calcium pantothenate, 3mg-100mg manganese sulfate monohydrate, 1mg-100mg beta-carotene, 1mg-15mg copper gluconate, 25IU-5,000IU vitamin D3, 2μg-1,000μg vitamin K2, 0.5mg-200mg pyridoxine HCl, 0.5mg-1,500mg potassium iodide, 0.5mg-1,000mg riboflavin, 0.5mg-2,500mg thiamine hydrochloride, 1μg-500μg dried vitamin K1, 500IU-100,000IU vitamin A acetate, 100μg-10,000μg folic acid, 10μg-10,000μg d-biotin, 1μg-3,000μg vitamin B12, 300mg-30,000mg L-carnitine, 500mg-60,000mg L-glutamine, 500mg-30,000mg 0mg L-arginine base, 50mg-2,000mg taurine, 50mg-2,000mg L-lysine, 10mg-1,000mg alpha lipoic acid, 15mg-1,500mg resveratrol, 10mg-5,000mg coenzyme Q10, 5mg-1,000mg glycine, 5mg-1,000mg proline, 2mg-500mg Lact. Acidophilus (total approx. 10 billion), 2mg-500mg Bifidobacterium (total approx. 10 billion), 2mg-500mg Lac. Bulgaricus (total approx. 10 billion), 2mg-500mg Bifidobacterium Longum (approximately 10 billion in total), 2mg-500mg of Strep. Thermophilus (approximately 10 billion in total), 5mg-100mg of papain, 5mg-100mg of pepsin, 5mg-100mg of lipase, 5mg-100mg of bromelain, 0.5mg-100mg of pancreatin 4X, 1mg-100mg of lactase, 3mg-100mg of betaine HCl, 2mg-500mg of pineapple juice powder, 2mg-500mg of papaya fruit powder, 30mg-3, It contains 1,000mg of quercetin, 25mg to 600mg of EGCG, 15mg to 500mg of OPC, 15mg to 5,000mg of anthocyanins, 10mg to 300mg of ellagic acid, 2mg to 90mg of astaxanthin, 20mg to 1,500mg of fucoidan, 5mg to 6,000mg of cordyceps, 15mg to 10,000mg of Ganoderma lucidum, 40mg to 15,000mg of shiitake mushroom, 30mg to 15,000mg of maitake mushroom, and 30mg to 15,000mg of trautauraceae.

[0046] Various objects, features, aspects and advantages of the inventive subject matter will become more apparent from the following detailed description of preferred embodiments, taken in conjunction with the accompanying drawings, in which like numerals represent like components. [Brief explanation of the drawings]

[0047] [Figure 1] Figure 1 shows the treatment protocol in which nutritional supplementation was provided beginning either 7 days before administration of radiation therapy or concurrently with tumor cell implantation. Mice were sacrificed 21 days after tumor cell implantation. [Figure 2A] Figure 2A shows the change in body weight over time for the various treatment groups, where body weight is reported after removal of the primary tumor mass. [Figure 2B] Figure 2B shows the change in body weight in the various treatment groups over the course of the study, where body weight is reported after removal of the primary tumor mass. [Figure 3A] Figures 3A-3D show the typical synergistic effect of radiation therapy and combination therapy with a fish oil and selenium supplement on tumor volume and weight. Figure 3A shows the change in tumor volume over time for the various treatment groups. [Figure 3B] Figures 3A-3D show the typical synergistic effect of radiation therapy and combination therapy with a fish oil and selenium supplement on tumor volume and weight. Figure 3B provides a detailed view of the first 11 days of treatment as shown in Figure 3A. [Figure 3C] Figures 3A-3D show the typical synergistic effect of radiation therapy and combination therapy with a supplement containing fish oil and selenium on tumor volume and weight. Figure 3C provides a histogram of tumor weight for the various treatment groups. [Figure 3D] Figures 3A-3D show the typical synergistic effects of radiation therapy and combination therapy with a supplement containing fish oil and selenium on tumor volume and weight. Figure 3D provides photographs of exemplary tumors from various treatment groups. [Figure 4A] FIG. 4A: Effects of treatment with radiation therapy, a supplement containing fish oil and selenium, and combination therapy on gastrocnemius muscle mass 21 days after tumor cell injection. [Figure 4B] Figure 4B: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on soleus muscle mass 21 days after tumor cell injection. [Figure 4C] FIG. 4C: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on lung (including metastatic tumor) weight 21 days after tumor cell injection. [Figure 4D] FIG. 4D: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on liver (containing metastatic tumors) weight 21 days after tumor cell injection. [Figure 4E] FIG. 4E: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on spleen (containing metastatic tumors) weight 21 days after tumor cell injection. [Figure 4F] FIG. 4F provides the results of a quantitative study of lung metastases in animal subjects treated with a supplement containing fish oil and selenium and / or radiation therapy. [Figure 4G] FIG. 4G shows the results of a study of Ki-67 expression at primary tumor and metastatic sites in animal subjects treated with a supplement containing fish oil and selenium and / or radiation therapy. [Figure 5A] FIG. 5A: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on platelet counts 21 days after tumor cell injection. [Figure 5B] Figure 5B: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on white blood cell counts 21 days after tumor cell injection. [Figure 5C] FIG. 5C: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on lymphocyte counts 21 days after tumor cell injection. [Figure 5D] Figure 5D: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on granulocyte counts 21 days after tumor cell injection. [Figure 5E]FIG. 5E: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on neutrophil / lymphocyte percentage 21 days after tumor cell injection. [Figure 6A] FIG. 6A: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on serum albumin 21 days after tumor cell injection. [Figure 6B] Figure 6B: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on serum creatinine 21 days after tumor cell injection. [Figure 7A] FIG. 7A: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on serum IL-6 21 days after tumor cell injection. [Figure 7B] Figure 7B: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on serum IL-Ιβ 21 days after tumor cell injection. [Figure 8A] FIG. 8A: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on VEGF gene expression in implanted tumors 21 days after tumor cell injection. [Figure 8B] FIG. 8B: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on BAX gene expression in implanted tumors 21 days after tumor cell injection. [Figure 8C] FIG. 8C: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on Bcl-2 gene expression in implanted tumors 21 days after tumor cell injection. [Figure 8D] FIG. 8D: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on caspase-3 gene expression in implanted tumors 21 days after tumor cell injection. [Figure 8E] FIG. 8E: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on BAX gene expression in the lungs 21 days after tumor cell injection. [Figure 8F]FIG. 8F: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on Bcl-2 gene expression in the lungs 21 days after tumor cell injection. [Figure 8G] FIG. 8G: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on caspase 3 gene expression in the lungs 21 days after tumor cell injection. [Figure 9A] Figure 9A shows the experimental protocol in which mice receive radiation therapy on days 8, 10, and 12, with nutritional supplementation commencing at the same time as radiation therapy begins. Mice are culled on days 14 or 24 after tumor cell implantation. [Figure 9B] FIG. 9B: Treatment groups derived from the protocol shown in FIG. 9A. [Figure 10A] FIG. 10A: Serum albumin concentrations 14 and 24 days after tumor cell implantation in mice treated as shown in FIG. 9A. [Figure 10B] FIG. 10B: Lymphocyte counts at 14 and 24 days after tumor cell implantation in mice treated as shown in FIG. 9A. [Figure 10C] FIG. 10C shows the N / L ratios at 14 and 24 days after tumor cell implantation in mice treated as shown in FIG. 9A. [Figure 11A] Figure 11A: VEGF expression within tumor masses 24 days after tumor cell implantation in mice treated as shown in Figure 9A. FITC represents VEGF-specific staining. [Figure 11B] Figure 11B: VEGF expression in the lungs (metastases) 24 days after tumor cell implantation in mice treated as shown in Figure 9A. FITC represents VEGF-specific staining. [Figure 11C] Figure 11C: EGFR expression in lungs (metastases) 24 days after tumor cell implantation in mice treated as shown in Figure 9A. FITC represents EGFR-specific staining. [Figure 11D] Figure 11D: EGFR expression within tumor masses 24 days after tumor cell implantation in mice treated as shown in Figure 9A. FITC represents EGFR-specific staining. [Figure 12]CD31 (cancer stem cell marker) expression within tumor masses and lung tissue (metastasis) 24 days after tumor cell implantation in mice treated as shown in Figure 9A. FITC represents CD31-specific staining. [Figure 13A] Figure 13A shows the presence of H1F1-α (hypoxia marker) protein within the tumor mass and in lung tissue (metastasis) 24 days after tumor cell implantation in mice treated as shown in Figure 9A. FITC represents H1F1-α-specific staining. [Figure 13B] FIG. 13B shows the results of a gene expression study of tumor samples from subjects treated with a nutritional supplement containing fish oil and selenium and / or radiation therapy. [Figure 14] Expression of apoptotic markers 24 days after tumor cell implantation in mice treated as shown in Figure 9A. [Figure 15A] FIG. 15A shows the results of a study characterizing PDL-1 gene expression at primary and metastatic tumor sites in an animal model of human disease treated with a supplement containing fish oil and selenium and / or radiation therapy. [Figure 15B] Figure 15B shows the results of a study characterizing PD-1 gene expression at primary and metastatic tumor sites in an animal model of human disease treated with supplements containing fish oil and selenium and / or radiation therapy. [Figure 16A] Figure 16A shows the experimental protocol in which mice receive radiation therapy on days 8, 10, and 12, with nutritional supplementation provided before, at the time of, or at the start of radiation therapy. Mice are sacrificed 21 days after tumor cell implantation. [Figure 16B] FIG. 16B: Treatment groups derived from the protocol shown in FIG. 16A. [Figure 17] Photomicrograph of a cross section of the intestine from a mouse treated by the protocol described in Figure 16A. [Figure 18A] FIG. 18A: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on VEGF gene expression 21 days after tumor cell injection. [Figure 18B]FIG. 18B: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on BAX gene expression 21 days after tumor cell injection. [Figure 18C] FIG. 18C: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on Bcl-2 gene expression 21 days after tumor cell injection. [Figure 18D] FIG. 18D: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on caspase-3 gene expression 21 days after tumor cell injection. [Figure 18E] FIG. 18E: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on Bcl-2 gene expression in the lungs (i.e., metastases) 21 days after tumor cell injection. [Figure 18F] FIG. 18F: Effect of treatment with radiation therapy, NutraWell supplement, and combination therapy on caspase 3 gene expression in lungs (i.e., metastases) 21 days after tumor cell injection. [Figure 19A] Figure 19A shows the experimental protocol in which mice are provided with nutritional supplements for 7 days prior to tumor cell implantation and then undergo radiation therapy on days 8, 10, and 12. Mice are culled 24 days after tumor cell implantation. [Figure 19B] Figure 19B: Treatment groups from the protocol shown in Figure 19A. [Figure 20A] FIG. 20A: Effect of a nutritional supplement containing fish oil and selenium on body weight in mice receiving repeated radiation therapy after tumor cell implantation, using the protocol shown in FIG. 19A. [Figure 20B] FIG. 20B: Effect of treatment with radiation therapy, a nutritional supplement containing fish oil and selenium, and combination therapy using the protocol shown in FIG. 19A on gastrocnemius muscle mass 21 days after tumor cell injection. [Figure 21A]Figure 21A shows the experimental protocol mice were provided with nutritional supplements for 7 days prior to tumor implantation, on the day of tumor cell implantation, or 8 days after tumor cell implantation, and then received radiation therapy on days 8, 10, and 12. Mice were culled 21 days after tumor cell implantation. [Figure 21B] FIG. 21B: Treatment groups derived from the protocol shown in FIG. 21A. [Figure 22] Effect of NutraWell supplementation on body weight of mice undergoing repeated radiation therapy after tumor cell implantation using the protocol shown in Figure 21A. [Figure 23] Time course of tumor volume upon repeated radiation therapy in combination with NutraWell supplement in mice treated as in the protocol shown in Figure 21A. [Figure 24A] FIG. 24A: Effect of repeated radiation therapy in combination with NutraWell supplements and treatment using combination therapy on serum TNF-α 21 days after tumor cell injection in mice treated using the protocol shown in FIG. 21A. [Figure 24B] FIG. 24B: Effect of repeated radiation therapy in combination with NutraWell supplements and treatment using combination therapy on serum IL-6 21 days after tumor cell injection in mice treated using the protocol shown in FIG. 21A. DETAILED DESCRIPTION OF THE INVENTION

[0048] (Detailed explanation) The following description includes information that may be useful in understanding the present invention. No admission is made that any of the information presented herein is prior art or relevant to the present invention, or that any publication specifically or implicitly cited is prior art.

[0049] The subject matter of the invention provides compositions and methods for using a nutritional supplement containing fish oil and selenium (e.g., a supplement containing fish oil, selenium derived from selenium yeast, and certain vitamins, minerals, amino acids, and sugars, e.g., "NutraWell") in combination with radiation therapy. Surprisingly, combined therapy using radiation and such a supplement provides significant synergistic effects in reducing tumor size. Additionally, side effects of radiation therapy (e.g., neutropenia, weight loss, muscle loss, damage to gastrointestinal acyl cells, etc.) are reduced and / or alleviated compared to the administration of radiation therapy without such a supplement. Surprisingly, gene expression related to angiogenesis and apoptosis was also found to be regulated in tumor cells when using a supplement containing fish oil and selenium, both with and without the administration of radiation therapy. Additionally, metastasis was found to be prevented, and the growth and proliferation of cancer stem cells was reduced.

[0050] It will be appreciated that the disclosed technology provides many beneficial technical effects, including enhancing the effectiveness of current radiation treatment protocols used in the treatment of cancer, while reducing the side effects associated with these approaches.

[0051] The following discussion provides many example embodiments of the inventive subject matter. While each embodiment represents a single combination of inventive elements, it is understood that the inventive subject matter includes all possible combinations of the disclosed elements. Thus, if one embodiment includes elements A, B, and C and a second embodiment includes elements B and D, it is understood that the inventive subject matter also includes any other remaining combinations of A, B, C, or D, even if not explicitly disclosed.

[0052] In some embodiments, numbers expressing properties such as amounts of ingredients, concentrations, and reaction conditions used to describe and claim certain embodiments of the invention should be understood to be modified in some instances by the word "about." Accordingly, in some embodiments, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending on 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.

[0053] In one embodiment of the inventive concept, the nutritional supplement composition shown in Table 1 ("Nutrawell") is provided to improve the outcome of radiation therapy.

[0054] [Table 1-1]

[0055] [Table 1-2]

[0056] [Table 1-3]

[0057] The compositions shown in Table 1 contain ingredients with various physiological and biochemical effects, including anti-inflammatory activity, blood glucose lowering, cholesterol lowering, and anti-tumor activity. Other ingredients provide high levels of necessary vitamin, mineral, and amino acid supplementation. Other ingredients (e.g., enzymes, lecithin) function to aid in the digestion and absorption of the composition components when ingested. The combination of these complementary activities provides a synergistic effect beyond the simple additive effects of the individual components. It should be understood that the compositions shown in Table 1 also contain certain flavorants (e.g., brown sugar, honey, vanilla flavor, and masking agents) that function to improve palatability and acceptability. Certain ingredients (e.g., honey, brown sugar, milk, rice protein, casein) can provide both flavor and caloric energy. The inventors have found that the combination of flavoring agents is effective in providing compliance with the intake of the nutritional supplement at an effective amount. In some embodiments, such flavoring agents can be omitted without negatively affecting the efficacy of the nutritional supplement.

[0058] The ingredients shown in Table 1 can be provided as a single formulation (e.g., as a pill, tablet, capsule, powder, liquid, suspension, etc.) or can be divided into different formulations (e.g., as a pill, tablet, capsule, powder, liquid, suspension, or combinations thereof). The amounts shown in Table 1 are exemplary and are typical daily dosages provided to an adult of normal height and otherwise normal health. These amounts can be adjusted to account for differences in weight, gender, medical condition, etc. For example, a relatively small patient weighing 40 kilograms or less may benefit from a dosage provided at or below the lower end of the provided range, while a relatively large patient weighing 100 kilograms or more may require a dosage provided at the upper end (or higher) of the indicated range. In some embodiments, such daily amounts can be distributed as multiple doses throughout the day. In some such embodiments, the composition of each such distributed dose can be the same. In other embodiments, the composition of such distributed doses can differ, provided that the sum of such doses provides the required supplementation.

[0059] In an exemplary embodiment, human tumor cells (implanted into nude mice) are treated with 1 gram / day of the nutritional supplement, radiation therapy, or 1 gram / day of the nutritional supplement and chemotherapy. The mice are weighed during treatment to characterize side effects such as nausea and anorexia. After several weeks, the mice are sacrificed and the tumors are characterized. Tumor volume is measured to determine the effect of therapy on organ and muscle volume. The degree of neutropenia is also characterized. A typical protocol is shown in Table 2.

[0060] [Table 2]

[0061] Weight / Waste A typical treatment regimen is outlined in Figure 1. Some subjects receive treatment with a supplement containing fish oil and selenium before tumor cell implantation (PTN, PTRN), and some of these (PTRN) receive radiation therapy. Other subjects begin treatment with such supplements at the time of tumor cell implantation (TN, TRN), and some of these (TRN) receive radiation therapy. The results of a weight study comparing control subjects (C) with tumor cell-implanted but otherwise untreated subjects (T) are shown in Figures 2A and 2B. As shown, mice receiving both radiation therapy and a supplement containing fish oil and selenium gained weight at a significantly higher rate than mice receiving radiation therapy alone, indicating a reduction in the usual side effects associated with this treatment modality. Treatment with a nutritional supplement containing fish oil and selenium had a particularly striking effect.

[0062] Tumor size Figures 3A-3D show typical synergistic effects on tumor volume and weight of combined radiation therapy and a fish oil and selenium supplement for similar treatment groups in a murine model of human cancer. Figure 3A shows the effect of various treatment protocols on tumor volume over the course of three weeks, while Figure 3B provides a magnified view of the effect over the first 11 days. As shown, treatment with a fish oil and selenium supplement alone provides an approximately 60% reduction in tumor volume. Treatment with radiation alone provides a similar reduction in tumor volume. Therefore, without synergy, a reduction in tumor volume to approximately 25% of that of untreated tumors would be expected. Surprisingly, what is observed is a greater than 90% reduction in tumor volume, compared to approximately 7% of the untreated tumors, demonstrating significant synergy. Figure 3C shows that a similar effect is found when tumor weight is characterized. Figure 3D shows a typical example of tumors removed from test animals after treatment, and the effect of the combined therapy using nutritional supplements containing fish oil and selenium and radiation therapy is easily apparent by visual inspection.Therefore, it is clear that the combined therapy using nutritional supplements containing fish oil and selenium and radiation therapy can have a synergistic effect on reducing tumor volume and / or mass.

[0063] Muscle wasting Side effects of radiation therapy go beyond loss of appetite and weight loss and can include damage to internal organs, loss of muscle mass, anemia, neutropenia, and decreased kidney function. To determine the protective effect of combined treatment with a supplement containing fish oil and selenium with respect to such side effects on muscle mass and organs, the body weight of treated mice was also characterized after treatment. The results are shown in Figures 4A-4E. Figures 4A and 4B show the effects of treatment with a supplement containing fish oil and selenium, radiation therapy, and combined radiation therapy with such supplements (with or without supplement pretreatment) on the weight of the gastrocnemius and soleus muscles (respectively) for the test groups described above. As shown, combined treatment with such supplements and radiation therapy provided nearly complete preservation of muscle weight.

[0064] metastasis Figures 4C, 4D, and 4E show the effects of treatment as described above on the weight of the lung containing any metastatic tumors (Figure 4C), the liver containing any metastatic tumors (Figure 4D), and the spleen containing any metastatic tumors (Figure 4E). As shown, the absence of treatment leads to an increase in weight, due at least in part to the presence of metastatic tumors. This increase is not completely attenuated by radiation therapy alone (TR), but subjects receiving treatment with a supplement containing fish oil and selenium had organ weights similar to those of controls, both with and without radiation therapy.

[0065] Similarly, metastatic sites were quantified in the lungs of an animal model of human disease treated with a nutritional supplement containing fish oil and selenium and / or radiation therapy. Typical metastatic sites are shown in the photographs on the left panel of Figure 4F and depicted in the histograms on the right panel. As shown, treatment with either such a supplement (TN) or radiation therapy (TRN) reduced the number of metastatic sites to some extent, but treatment with both the supplement and radiation therapy (TRN) resulted in a complete absence of obvious metastatic sites in most subjects.

[0066] Without wishing to be bound by theory, the inventors believe that this effect on metastasis (and, as mentioned above, on tumor size and mass) may be due to the synergistic effect on tumor cell proliferation brought about by the use of a nutritional supplement containing fish oil and selenium in combination with radiation therapy. As shown in Figure 4G, the expression of the proliferation marker Ki-67 is suppressed at the primary tumor site upon treatment with either a supplement containing fish oil and selenium (TN), radiation therapy (TR), or both modalities (TRN). Metastatic sites (right panel) showed a trend associated with treatment with either the nutritional supplement or radiation therapy alone, resulting in a moderate decrease in the expression of this proliferation marker, but surprisingly, showed a synergistic effect in reducing Ki-67 expression at the metastatic site. This means that the use of a supplement containing fish oil and selenium can reduce tumor metastasis, including metastases not prevented by radiation therapy, and can do so synergistically.

[0067] Neutropenia / anemia Anemia and neutropenia are often seen in cancer and as side effects of radiation therapy, due at least in part to suppressed bone marrow activity. The effects of combined treatment with a supplement containing fish oil and selenium on various blood cell populations are shown in Figures 5A-5E. Figure 5B shows the effects of a nutritional supplement containing fish oil and selenium, radiation therapy, and the combined treatment with such a supplement and radiation therapy on white blood cells, as shown in Figure 5B, and on lymphocytes, as shown in Figure 5C. This means that a nutritional supplement containing fish oil and selenium may be useful for ameliorating the suppression of red blood cells, granulocytes, white blood cells, and / or lymphocytes caused by the presence of tumors and the radiation therapy used to treat such tumors.

[0068] As shown in Figure 5A, platelets were found to be elevated in untreated tumor-bearing animals (T). Platelet concentrations were reduced by treatment with either a fish oil and selenium supplement or radiation therapy, but were most pronounced in animals receiving a combination therapy with such supplements and radiation therapy. This effect was particularly pronounced in subjects receiving pretreatment with a fish oil and selenium supplement, which reduced platelet concentrations to the levels seen in control animals. Granulocyte concentrations were similarly elevated in untreated tumor-bearing animals (as shown in Figure 5D) and further elevated in those animals receiving radiation therapy alone. Combination therapy using a fish oil and selenium nutritional supplement and radiation therapy was found to reduce this effect. As shown in Figure 5E, tumor-bearing animals exhibited an elevated neutrophil-to-lymphocyte ratio (NLR) compared with control animals, which was even more elevated in similar subjects receiving radiation therapy alone. Treatment with a fish oil and selenium supplement, both as a monotherapy and as part of a combination therapy with radiation therapy, was found to be effective in shifting this ratio toward more normal values. This means that a nutritional supplement containing fish oil and selenium may be useful in reducing platelet concentrations, granulocyte concentrations, and / or neutrophil / lymphocyte ratios that are elevated due to the presence of tumors and due to radiation therapy used to treat such tumors.

[0069] serum proteins The effects of combined therapy using a supplement containing fish oil and selenium on various serum biochemical markers that provide information about kidney function, liver function, and / or nutritional status are shown in Figures 6A and 6B. Figure 6A shows the effects of a nutritional supplement containing fish oil and selenium, radiation therapy, and combined therapy using such a supplement and radiation therapy on an animal tumor model. As shown, serum albumin levels (an indicator of nutritional status) decrease in untreated tumor-bearing animals, and are only slightly improved by radiation therapy alone. Treatment with nutritional supplements comprising fish oil and selenium improves serum albumin concentration, either as a monotherapy or as a combination therapy with radiotherapy, especially when such supplements are provided as pretreatment.This is improved by the combination therapy with nutritional supplements comprising fish oil and selenium.This means that treatment with nutritional supplements comprising fish oil and selenium can improve the nutritional status of tumor-bearing subjects, either as a monotherapy or as a combination therapy with radiotherapy.

[0070] Figure 6B shows the results of a similar study in which serum creatinine (a measure of kidney function) was characterized. As shown, subjects with untreated tumors exhibit elevated creatinine levels, indicating kidney damage. This is slightly improved by radiation therapy alone. However, combined therapy using a nutritional supplement containing fish oil and selenium and radiation therapy shows synergistic effects in reducing serum creatinine levels, especially when such nutritional supplements are provided as pretreatment.

[0071] cytokines Tumor growth and spread, as well as the administration of radiation therapy, are associated with inflammation. Surprisingly, the inventors found that combined treatment with a supplement containing fish oil and selenium was effective in reducing the levels of proinflammatory cytokines, suggesting that such combined treatment is effective in reducing inflammation associated with tumors and tumor radiation therapy. The effect of NutraWell supplements on serum levels of proinflammatory cytokines is shown in Figures 7A and 7B. Figure 7A shows the levels of IL-6 in control animals, untreated tumor-bearing animals, and tumor-bearing animals treated with a nutritional supplement containing fish oil and selenium, radiation therapy, or a combination of such supplements and radiation therapy. As shown, untreated tumor-bearing animals exhibit a significant increase in serum IL-6 levels, which are only slightly reduced by radiation therapy. The use of a nutritional supplement containing fish oil and selenium, either as monotherapy or in combination with radiation therapy, was found to reduce serum IL-6 levels in tumor-bearing animals. Figure 7B shows the results of a similar study of IL-Ιβ. The results for IL-Ιβ are similar to those found for IL-6. This means that a nutritional supplement containing fish oil and selenium, either as a monotherapy or in combination with radiation therapy, may be effective in reducing serum concentrations of inflammation-related cytokines in tumor-bearing subjects. The inventors believe that such a reduction is accompanied by a reduction in inflammation in such animals.

[0072] Tumor gene expression Surprisingly, the inventors have also found that treatment with a nutritional supplement comprising fish oil and selenium can modify gene expression in tumor cells in vivo and provide a synergistic effect against such changes in gene expression caused by radiation therapy. In some embodiments, the genes are cytokine-related and / or apoptosis-related. Examples of the effects of radiation therapy, treatment with a nutritional supplement comprising fish oil and selenium, and combination therapy with radiation therapy on gene expression in transplanted tumor cells in vivo are shown in Figures 8A-8G. Figure 8A shows the results of such treatments on the expression of VEGF in tumor cells. As shown, monotherapy with a nutritional supplement comprising fish oil and selenium and radiation therapy reduced VEGF expression. Combination therapy with a nutritional supplement comprising fish oil and selenium and radiation therapy dramatically reduced VEGF expression.

[0073] Figures 8B and 8D show the effects of treatment with a nutritional supplement containing fish oil and selenium, radiation therapy, and combination therapy using such a supplement and radiation therapy on tumor BAX expression and metastatic (lung) tumor BAX expression, respectively. BAX is considered a marker of apoptosis. As shown, BAX expression in untreated tumors is low and unaffected by radiation therapy. Monotherapy using a nutritional supplement containing fish oil and selenium resulted in a dramatic increase in BAX expression, and when used in combination with radiation therapy (especially when the supplement was provided as a pretreatment), it also elevated BAX expression. As shown in Figures 8C and 8E, Bcl-2 expression was found to be elevated in tumor cells and metastatic (lung) tumor cells (respectively) and reduced by either treatment with a nutritional supplement containing fish oil and selenium as monotherapy or radiation therapy. When such supplements and radiation therapy were used as a combination therapy, an even greater decrease in Bcl-2 expression was observed. Expression of caspase 3 (associated with apoptosis) was elevated in tumors treated with radiation therapy or a nutritional supplement containing fish oil and selenium alone (Figure 8D), and also elevated by combined radiation therapy with such supplements (especially when the supplements were provided as a pretreatment). As shown in Figure 8F, caspase 3 expression was reduced in metastatic (lung) tumors compared with control lung tissue. As shown, caspase 3 expression in such tumors was increased by radiation therapy or treatment with a nutritional supplement containing fish oil and selenium, as well as by combined therapy.

[0074] Repeated radiation therapy Similar studies were performed using modified treatment protocols. One modified treatment protocol utilizing multiple radiation treatments, as is typical in human radiation therapy, is shown in Figures 9A and 9B.

[0075] Figures 10A-10C show results from serum albumin and blood cell characterization following treatment with NutraWell, radiation therapy, and combined treatment with a nutritional supplement containing fish oil and selenium and radiation therapy using the protocol shown in Figure 9A. As shown in Figure 10A, serum albumin levels decline in tumor-bearing animals, particularly at later time points. This is improved by treatment with a nutritional supplement containing fish oil and selenium as monotherapy or radiation therapy, as well as by combined treatment. As shown in Figure 10B, tumor-bearing animals exhibited suppressed lymphocyte counts compared to controls, particularly at later time points. This was only slightly improved by radiation therapy alone, whereas treatment with a nutritional supplement containing fish oil and selenium (either as monotherapy or in combination with radiation therapy) was effective in increasing lymphocyte levels, particularly at later time points. Figure 10C shows results from a similar study characterizing the neutrophil-to-lymphocyte ratio (NLR). As shown, tumor-bearing animals exhibit a dramatic increase in this value at later time points. It is reduced by treatment with nutritional supplements containing fish oil and selenium, radiation therapy, and combination therapy using such supplements and radiation therapy.

[0076] Surprisingly, treatment with a nutritional supplement containing fish oil and selenium, radiation therapy, and combined treatment with such supplements and radiation therapy using the protocol shown in Figure 9A also have an effect on both tumor cell marker expression and tumor cell metastasis. In the following study, the tumor cells selected for implantation were derived from lung tumors and have a strong tendency to metastasize from the implantation site to the lung. Figures 11A-11D show the results of immunocytochemistry studies of various tissues from mice treated with the protocol shown in Figure 9A. It is noteworthy that NutraWell supplementation alone reduced or eliminated metastases.

[0077] As shown in Figure 11A, VEGF expression in the tumors of untreated mice (left panel, FITC staining) is evident. Treatment with either a nutritional supplement containing fish oil and selenium or radiation therapy alone dramatically reduced VEGF expression, as did the combination therapy. Numerical results are presented in the right panel of Figure 11A. Similar results are seen in tumors that metastasized to the lung, as shown in Figure 11B. As shown, VEGF expression was evident in untreated metastatic tumors (left panel, FITC staining) and was clearly reduced in animals treated with a nutritional supplement containing fish oil and selenium, radiation therapy, or both. Numerical results of these studies are presented in the right panel of Figure 11B.

[0078] Similar results were observed for the expression of EGFR, whose overexpression is associated with tumors. As shown in Figure 11C, elevated EGFR expression (left panel, FITC staining) was evident in untreated tumors and decreased in subjects treated with a nutritional supplement containing fish oil and selenium, radiation therapy, or a combination therapy using such a supplement and radiation therapy. Numerical results are shown in the right panel of Figure 11C. Similar results were observed for EGFR expression in tumors metastasized to the lung, as shown in Figure 11D. As shown, elevated EGFR expression (left panel, FITC staining) was evident in untreated metastatic sites and decreased in subjects treated with a nutritional supplement containing fish oil and selenium or radiation therapy as monotherapy, or a combination therapy using such a supplement and radiation therapy. This suggests that the use of a nutritional supplement containing fish oil and selenium, radiation therapy, and / or a combination of treatment with such a supplement and radiation therapy may be effective in enhancing EGFR-directed treatment protocols in individuals undergoing cancer treatment.

[0079] Surprisingly, treatment with a nutritional supplement containing fish oil and selenium, radiation therapy, and combined treatment with such supplements and radiation therapy using the protocol shown in Figure 9A also have an effect on tumor stem cells. Such stem cells are associated with the development of metastasis and resistance to various cancer therapies. In the following study, the tumor cells selected for transplantation were derived from lung tumors and have a strong tendency to metastasize from the transplant site to the lung. Figure 12 shows the results of immunocytochemistry studies of various tissues from mice treated with the protocol shown in Figure 9A. The left panel of Figure 12 shows staining results for CD31 (FITC staining), a stem cell marker, in tumor cells. The right panel shows similar results for metastatic cells in the lung. Untreated subjects show many cells with elevated CD31 expression. Surprisingly, treatment with a nutritional supplement containing fish oil and selenium in the absence of radiation therapy reduces or eliminates the development of cancer stem cells at both the tumor transplant site and the lung metastasis site.

[0080] Treatment with a nutritional supplement containing fish oil and selenium, radiation therapy, and combined NutraWell supplementation and radiation therapy using the protocol shown in Figure 9A also affect the hypoxia often found in or between tumor cells. HIF1-α is a marker associated with hypoxia. In the following study, the tumor cells selected for implantation were derived from lung tumors and have a strong tendency to metastasize from the implantation site to the lung. Figure 13A shows the results of immunocytochemistry studies of various tissues from mice treated with the protocol shown in Figure 9A. The left panel shows the results of immunocytochemistry staining (FITC staining) of HIF1-α in tumor cells, while the right panel shows the results of similar staining at lung metastatic sites. Untreated subjects show many cells with elevated levels of HIF1-α. Surprisingly, treatment with a nutritional supplement containing fish oil and selenium in the absence of radiation therapy reduces or eliminates the occurrence of this hypoxia marker at both the tumor implantation site and the lung metastatic site. Figure 13B shows typical numerical results from a similar study characterizing gene expression in tumor samples. As shown, both treatment with a supplement containing fish oil and selenium (TN) and one week of pretreatment with a nutritional supplement containing fish oil and selenium (PTN) reduce tumor expression of HIF1-α, similar to radiation therapy (TR). When radiation therapy and such nutritional supplements are used in combination (PTRN, TRN), a dramatic reduction in tumor HIF1-α expression is particularly evident in pretreated subjects (PTRN), demonstrating a synergistic effect. It is clear that the use of a nutritional supplement containing fish oil and selenium in combination with radiation therapy can reduce both HIF1-α protein content and gene expression at tumor sites, making them more susceptible to hypoxia.

[0081] Treatment with a nutritional supplement containing fish oil and selenium, radiation therapy, and combined supplementation and radiation therapy using the protocol shown in Figure 9A also affect apoptotic activity in tumor cells. Figure 14 shows the results of a qPCR study of the expression of various apoptosis markers (Bax, Bcl-2, and caspase-3) 24 days after tumor cell implantation in mice. As shown, the Bax / Bcl-2 expression ratio was low in tumor cells and only slightly improved by radiation therapy. Surprisingly, treatment with a nutritional supplement containing fish oil and selenium alone significantly increased this ratio. In addition, a significant synergistic effect was observed with the combined use of such supplements and radiation therapy. Caspase-3 expression was actually somewhat suppressed by radiation therapy alone, but dramatically increased by treatment with a nutritional supplement containing fish oil and selenium, both as monotherapy and in combination with radiation therapy.

[0082] Treatment using a nutritional supplement containing fish oil and selenium in combination with radiation therapy has also been found to regulate PDL-1 and PD-1 expression at primary and metastatic (lung) tumor sites in an animal model of human disease. As shown in Figure 15A, both primary (left panel) and metastatic (right panel) tumor cells exhibit high levels of PDL-1 expression (T). Surprisingly, these are significantly reduced by treatment with a nutritional supplement containing fish oil and selenium both pre-treatment (PTN) and at the time of transplantation (TN). Therefore, the inventors believe that such supplements can be used to enhance immunotherapeutic approaches to cancer treatment. Treatment with radiation therapy (TR) also demonstrated a reduction in PD-L1 gene expression at both primary and metastatic sites, suggesting that radiation therapy alone can be used to enhance immunotherapeutic approaches to cancer treatment. Combination therapy using such nutritional supplements and radiation therapy is effective in reducing PDL-1 expression at both primary and metastatic tumor sites, regardless of whether the supplements are provided before transplantation (PTRN) or at the time of transplantation (TRN). Surprisingly, the effect is more pronounced at the primary tumor site than at metastatic sites, suggesting that combined therapy using a nutritional supplement containing fish oil and selenium in combination with radiation therapy can make tumor cells at both primary and metastatic sites more susceptible to the patient's immune system and / or enhance the effect of immunotherapy approaches to cancer treatment.

[0083] Similar studies were performed on PD-1 expression. As shown in Figure 15B, PD-1 expression was reduced in primary tumor cells treated with radiation therapy (TR) and enhanced in primary tumor cells in animals treated with a supplement containing fish oil and selenium either before transplantation (PTN) or immediately after transplantation (TN) (left panel). Combination therapy with the nutritional supplement and radiation therapy provided differential results depending on whether the supplement was provided before transplantation (PTRN) or at the time of transplantation (TRN). As shown in the right panel of Figure 15B, tumor cells at metastatic sites showed reduced PD-1 expression compared with samples from control animals. Surprisingly, radiation therapy resulted in increased PD-1 expression at metastatic sites when treatment with a supplement containing fish oil and selenium was performed either before transplantation or at the time of transplantation (right panel). Combination therapy with radiation therapy and such a supplement resulted in higher levels of PD-1 expression than those observed with monotherapy. It appears that treatment with a nutritional supplement containing fish oil and selenium, particularly in combination with radiation therapy, can shift PD-1 expression in tumor-bearing subjects from the reduced expression levels seen, particularly at metastatic sites.

[0084] Initiation of radiation therapy supplementation before transplant, before radiation therapy, and with repeat radiation therapy Another treatment protocol combining both pretreatment with a nutritional supplement containing fish oil and selenium and multiple radiation treatments typical of clinical applications is shown in Figures 16A and 16B. In this protocol, treatment with the supplement was initiated before tumor cell implantation, at the time of tumor cell implantation, and at the start of radiation therapy.

[0085] Loss of intestinal absorption and resulting malnutrition are well-known side effects of radiation therapy, especially repeated radiation therapy. Figure 17 shows micrographs showing the effect of treatment with a nutritional supplement containing fish oil and selenium on the intestinal cellular structure during radiation therapy. The intestinal cellular structure is shown for an untreated control (upper left panel), a tumor-implanted subject treated with multiple radiation therapy sessions (right panel), a tumor-implanted subject pretreated with a nutritional supplement containing fish oil and selenium and then administered radiation therapy (lower left panel), and a tumor-implanted subject treated with a nutritional supplement containing fish oil and selenium at the start of radiation therapy (lower right panel). As shown in the figure, treatment with a nutritional supplement containing fish oil and selenium can maintain the gastrointestinal brush border during radiation therapy and significantly strengthen the gastrointestinal brush border. This indicates that treatment with such supplements, especially pretreatment, can effectively counter the side effects of radiation therapy.

[0086] Such treatment protocols have been shown to modify the expression of certain genes in tumor cells in vivo. qPCR studies of gene expression (e.g., angiogenesis-related, apoptosis-related, etc.) in tumor cells from mice treated with the protocol shown in Figure 16A are shown in Figures 18A-18F. As shown in Figure 18A, treatment with a nutritional supplement containing fish oil and selenium reduces VEGF expression in tumor cells, similar to repeated radiation therapy. Combination therapy using such supplements with radiation therapy enhances the reduction of VEGF expression, especially when the nutritional supplement containing fish oil and selenium is provided before the start of radiation therapy.

[0087] Figure 18B shows that BAX expression in tumor cells is increased by treatment with a nutritional supplement containing fish oil and selenium, while remaining relatively unaffected by repeated radiation therapy alone. Pretreatment with such a supplement in combination with repeated radiation therapy also increased BAX expression. Similar results were observed for the expression of another apoptosis-related gene (caspase 3), as shown in Figure 18D. Results of a caspase 3 expression study in metastatic (lung) tumors are shown in Figure 18F. As shown, caspase 3 expression in such metastatic tumors is elevated by treatment with a nutritional supplement containing fish oil and selenium and by repeated radiation therapy, and is similar to that in control cells when the supplement is provided as a combination therapy prior to the start of radiation therapy.

[0088] The expression of Bcl-2, another apoptosis-related gene in tumor cells, is reduced by treatment with a nutritional supplement containing fish oil and selenium and repeated radiation therapy alone (see Figure 18C). This reduction in Bcl-2 expression is even more pronounced for combination therapy using such a supplement and repeated radiation therapy, especially when the supplement is provided before the start of radiation therapy. Figure 18E shows the results of a similar study conducted on metastatic (lung) tumors. As shown, the reduction in Bcl-2 expression is synergistic for combination therapy using a nutritional supplement containing fish oil and selenium and repeated radiation therapy, especially when the supplement is provided before the start of radiation therapy.

[0089] Pretransplant supplementation and repeated radiation therapy 19A and 19B show the treatment protocol and associated test groups (respectively) in which a nutritional supplement containing fish oil and selenium was provided 7 days before tumor cell implantation, and radiation therapy was administered on days 8, 10, and 12 after implantation. Mice were sacrificed 24 days after implantation.

[0090] A well-known side effect of both cancer and repeated radiation therapy is weight loss. This may be due to the weakness associated with the disease and the side effects of radiation therapy. The effect of treatment with a nutritional supplement containing fish oil and selenium on weight loss and muscle mass loss after repeated radiation therapy is shown in Figures 20A and 20B. It should be understood that weight was characterized after removal of the tumor mass. As shown in Figure 20A, upon removal of the tumor mass, a significant decrease in remaining body weight is evident compared to the control. This is ameliorated by treatment with a nutritional supplement containing fish oil and selenium and radiation therapy as monotherapy. Surprisingly, weight gain exceeds that of the subject when such supplements and repeated radiation therapy are used in combination. Figure 20B shows the weight of the gastrocnemius muscle in the various test groups. A decrease in muscle mass is evident in untreated tumor-bearing animals. This is slightly ameliorated by repeated radiation therapy alone. Surprisingly, treatment with a nutritional supplement containing fish oil and selenium provides significant muscle mass preservation. The combination of nutritional supplements containing fish oil and selenium and repeated radiation therapy results in a synergistic improvement in muscle mass compared to subjects with untreated tumors. It is clear that pretreatment with nutritional supplements containing fish oil and selenium effectively reverses the weight loss (compared to controls) and muscle mass caused by both the presence of the tumor and repeated radiation therapy.

[0091] Pretransplant supplementation and repeated radiation therapy Another treatment protocol related to the treatment groups is shown in Figures 21A and 21B, which is similar to the protocol shown in Figures 16A and 16B. In this protocol, radiation therapy was administered 8, 10, and 12 days after tumor cell implantation. Treatment with a nutritional supplement containing fish oil and selenium was administered either 7 days before implantation, on the day of implantation, or at the start of radiation therapy. Mice were sacrificed 21 days after tumor cell implantation.

[0092] A well-known side effect of both cancer and radiation therapy is weight loss. Figure 22 shows the effect of treatment with a nutritional supplement containing fish oil and selenium on weight loss and muscle mass loss when provided before and simultaneously with the start of repeated radiation therapy. It should be understood that weight was characterized after the removal of the tumor mass. As shown, the weight gain over time of subjects with untreated tumors (after tumor resection) is dramatically reduced compared to controls. A similar reduction is evident with respect to repeated radiation therapy when used alone. Surprisingly, treatment with a nutritional supplement containing fish oil and selenium provides significantly improved weight gain over time, both as a monotherapy and when used in combination with repeated radiation therapy. This is particularly evident when such supplements are used as pretreatment.

[0093] The inventors also found that pretreatment with a nutritional supplement containing fish oil and selenium enhanced the reduction in tumor volume seen in response to repeated radiation therapy, as shown in FIG. As shown in the figure, both repeated radiation therapy and treatment with a nutritional supplement containing fish oil and selenium are moderately effective in reducing tumor volume compared to tumors in animals with untreated tumors. Surprisingly, tumor volume shows little change over time when repeated radiation therapy and a nutritional supplement containing fish oil and selenium are used as combined therapy, indicating a synergistic effect.

[0094] The presence of tumors and repeated radiation therapy can also lead to the development of inflammation, which can be characterized by the presence of proinflammatory cytokines in serum. Figures 24A and 24B show the effects of administering a nutritional supplement containing fish oil and selenium and repeated radiation therapy on the levels of proinflammatory cytokines in mice treated as in the protocol shown in Figure 21A. Figure 24A shows serum TNF-α levels. It is clear that animals bearing untreated tumors exhibit very high levels of TNF-α, which are reduced to some extent by repeated radiation therapy. Treatment with a nutritional supplement containing fish oil and selenium also resulted in a decrease in serum TNF-α, especially when such supplements were used in combination with repeated radiation therapy. Figure 24B shows the results of a similar study characterizing the serum levels of IL-6, which show similar results.

[0095] Beyond those already described, it will be apparent to those skilled in the art that many further modifications are possible without departing from the inventive concepts herein. The inventive subject matter, therefore, is not limited 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 to refer to elements, components, or steps in an inclusive manner, meaning that the referenced element, component, or step may be present, utilized, or combined with other applications, components, or steps not specifically referenced. When the specification or claims refer to at least one selected from the group consisting of A, B, C, ..., and N, the context should be interpreted as requiring only one element from the group, not A+N, or B+N, etc.

Claims

[Claim 1] 1. A nutritional supplement for reducing H1f-α expression at primary tumor sites, comprising fish oil and selenium yeast, The nutritional supplement is provided in an amount effective to reduce H1f-α expression at the primary tumor site.

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