WO2008034129A2 - Treatment of cancer or obesity with conjugated linoleic acids - Google Patents

Abstract

The present invention discloses a method of screening cancer or obesity patients to determine whether the form of cancer or obesity they have is suitable for treatment with conjugated linoleic acids as well as treating patients for cancer based on that determination. The screening identifies patients with tumor cells with a disregulation of lipid metabolism, patients who overexpress the Thyroid Hormone Responsive Spot 14 gene, and patients who overexpress the Sterol Responsive Element Binding Protein gene.

Description

TREATMENT OF CANCER OR OBESITY WITH CONJUGATED

LINOLEIC ACIDS

[0001] This application claims the benefit of U.S. Provisional Patent

Serial No. Serial No. 60/844,815, filed September 15, 2006, which is hereby incorporated by reference in its entirety.

[0002] The subject matter of this application was made with support from the United States Government under the United States Department of Agriculture, Grant 2006-35206-16643. The U.S. Government may retain certain rights in this invention.

FIELD OF THE INVENTION

[0003] The present invention is directed to treatment of cancer or obesity with conjugated linoleic acids.

BACKGROUND OF THE INVENTION

[0004] Mammalian milk fat concentration and composition are variable and responsive to nutritional factors. First described over a century ago, low- fat milk syndrome, commonly referred to as milk fat depression (MFD), is characterized by a marked decrease in milk fat yield classically observed in ruminants fed highly fermentable diets or diets that contain plant or fish oils. Recent investigations have established that the basis relates to an inhibition of mammary synthesis of milk fat by specific fatty acid intermediates produced during rumen biohydrogenation under certain dietary situations (Bauman et al, "Nutritional Regulation of Milk Fat Synthesis," Annu Rev Nutr 23:203-27 (2003)). Trans-10, cis-Yl conjugated linoleic acid (CLA) was the first of these unique biohydrogenation intermediates to be identified (Baumgard et al., "Identification of the Conjugated Linoleic Acid Isomer that Inhibits Milk Fat Synthesis," Am J Physiol Regul Integr Comp Physiol 278 :R179-84 (2000)).

[0005] During MFD, transcription of mammary genes involved in milk fat synthesis are coordinately down-regulated (Bauman et al., "Nutritional Regulation of Milk Fat Synthesis," Annu Rev Nutr 23:203-27 (2003)). Molecular mechanisms mediating this inhibition are not well understood, but a role for sterol response element-binding protein (SREBP) family of transcription factors was proposed (Baumgard et al, "Trans-10, cis-12 Conjugated Linoleic Acid Decreases Lipogenic Rates and Expression of Genes Involved in Milk Lipid Synthesis in Dairy Cows," J Dairy Sci 85:2155-63 (2002)) based on their function as global regulators of expression for many genes involved in lipid synthesis (Eberle et al., "SREBP Transcription Factors: Master Regulators of Lipid Homeostasis," Biochimie 86: 839-48 (2004)). This was supported in studies with a bovine mammary epithelial cell line where trans- 10, cis-12 CLA decreased abundance of the nuclear active SREBPl protein (Peterson et al., "The Inhibitory Effect of trans- 10, cis-12 CLA on Lipid Synthesis in Bovine Mammary Epithelial Cells Involves Reduced Proteolytic Activation of the Transcription Factor SREBP-I," J Nutr 134:2523-27 (2004)). An additional potential mechanism was disclosed when thyroid hormone responsive spot 14 (S 14) was identified as a trans- 10, cis- 12 CLA responsive candidate gene in microarray analysis of bovine mammary epithelial cell cultures. Although its exact biochemical function is not known, S 14 is a gene that encodes a nuclear protein that is closely associated with the regulation of fatty acid synthesis in lipogenic tissues (Cunningham et al., ""Spot 14" Protein: A Metabolic Integrator in Normal and Neoplastic Cells," Thyroid 8:815-25 (1998)).

[0006] The objective was to investigate the expression of SREBPl and

S 14 in the mammary tissue of lactating cows under two situations where milk fat synthesis is reduced, diet induced-MFD and administration of trans- 10, cis-12 CLA. Down-regulation of SREBPl, SREBPl regulatory proteins and SREBP- regulated enzymes during milk fat depression is shown. Moreover, the studies revealed a previously unrecognized involvement of S 14 in the regulation of mammary synthesis of milk fat and a broader role in CLA-related regulation of lipid synthesis.

[0007] The present invention is directed to overcoming these and other deficiencies in the art. SUMMARY OF THE INVENTION

[0008] The present invention relates to a method of screening cancer patients to determine whether the form of cancer they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for tumor cells with a disregulation of lipid metabolism, and identifying patients with tumor cells with a disregulation of lipid metabolism as having a form of cancer suitable for treatment with conjugated linoleic acids.

[0009] The present invention also relates to a method of treating a patient for cancer. This involves selecting a patient having tumor cells with a disregulation of lipid metabolism and administering conjugated linoleic acids to the selected patient under conditions effective to treat cancer.

[00010] The present invention also relates to a method of screening cancer patients to determine whether the form of cancer they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for overexpression of the Thyroid Hormone Responsive Spot 14 gene, and identifying patients who overexpress the Thyroid Hormone Responsive Spot 14 gene as having a form of cancer suitable for treatment with conjugated linoleic acids.

[00011] The present invention also relates to a method of treating a patient for cancer. This involves selecting a patient overexpressing the Thyroid Hormone Responsive Spot 14 gene and administering conjugated linoleic acids to the selected patient under conditions effective to treat cancer.

[00012] Another aspect of the present invention relates to a method of screening cancer patients to determine whether the form of cancer they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for overexpression of the Sterol Responsive Element Binding Protein gene, and identifying patients who overexpress the Sterol Responsive Element Binding Protein gene as having a form of cancer suitable for treatment with conjugated linoleic acids. [00013] The present invention also relates to a method of treating a patient for cancer. This involves selecting a patient overexpressing the Sterol Responsive Element Binding Protein gene and administering conjugated linoleic acids to the selected patient under conditions effective to treat cancer.

[00014] Another aspect of the present invention relates to a method of screening obesity patients to determine whether the form of obesity they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for overexpression of the Thyroid Hormone Responsive Spot 14 gene, and identifying patients who overexpress the Thyroid Hormone Responsive Spot 14 gene as having a form of obesity suitable for treatment with conjugated linoleic acids.

[00015] The present invention also relates to a method of treating a patient for obesity. This involves selecting a patient overexpressing the Thyroid Hormone Responsive Spot 14 gene and administering conjugated linoleic acids to the selected patient under conditions effective to treat obesity.

[00016] Another aspect of the present invention relates to a method of screening obesity patients to determine whether the form of obesity they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for overexpression of the Sterol Responsive Element Binding Protein gene, and identifying patients who overexpress the Sterol Responsive Element Binding Protein gene as having a form of obesity suitable for treatment with conjugated linoleic acids.

[00017] The present invention also relates to a method of treating a patient for obesity. This involves selecting a patient overexpressing the Sterol Responsive Element Binding Protein gene and administering conjugated linoleic acids to the selected patient.

[00018] Milk fat synthesis in dairy cows can be inhibited by unique fatty acid intermediates produced during rumen biohydrogenation. One of these inhibitory intermediates is trans-10, cis-\2 conjugated linoleic acid (CLA) and this milk fat depression (MFD) involves a coordinated decrease in mammary expression of lipogenic enzymes. The SREBP transcription factor system was investigated in mammary tissue of cows during MFD induced by a low forage, high oil diet (LF/HO) and trans-10, cis-ll CLA infusion. LF/HO diet and CLA treatment decreased milk fat yield by 38 and 24%, respectively. Treatments causing MFD decreased expression of SREBPl and insulin responsive gene

(INSIG) 1, consistent with decreased abundance of active SREBPl. The LF/HO diet also decreased expression of INSIG2 and SREBP cleavage activating protein (SCAP). In addition, the involvement of thyroid hormone responsive spot 14 (S 14) was identified in the regulation of mammary synthesis of milk fat. A broader role for S 14 in the trans- 10, cis-Yl CLA-mediated decrease in fat synthesis was explored by mining publicly available microarray datasets and found mouse adipose expression of S 14 was decreased in response to CLA treatment. Overall, the decreased mammary expression of SREBPl, SREBP activation protein and the coordinated reduction in SREBPl -responsive lipogenic enzymes provides strong support for a central role of SREBPl in the regulation of milk fat synthesis. In addition, the results provide evidence for an involvement of S 14 in mammary regulation of milk fat synthesis and a possible broader role for S 14 in the reported anti-obesity effects of CLA.

BRIEF DESCRIPTION OF THE DRAWINGS

[00019] Figure 1 shows the effects of trans-\0, cis-Yl conjugated linoleic acid (CLA) and a low forage, high oil diet (LF/HO) on mammary mRNA abundance for key lipogenic enzymes [fatty acid synthase (FASN), lipoprotein lipase (LPL), and stearoyl-CoA desaturase (SCD)]. Values represent means ± SE relative to control. Significant difference from control indicated for P < 0.10(+), P < 0.01(**), and P < 0.001 (***).

[00020] Figure 2 shows the effects of trans-10, cis-Yl conjugated linoleic acid (CLA) and a low forage, high oil diet (LF/HO) on mammary mRNA abundance for key transcription factors (sterol regulatory element binding protein 1 (SREBPl) and thyroid hormone responsive spot 14 (S 14)). Values represent means ± SE relative to control. Significant difference from control indicated for P < 0.01(**) and P < 0.001(***). [00021] Figure 3 shows the effects of trans-10, cis-Yl conjugated linoleic acid (CLA) and a low forage, high oil diet (LF/HO) on mammary mRNA abundance for genes associated with SREBPl processing (insulin induced gene 1 and 2 (INSIGl and INSIG2), and SREBP cleavage-activating protein (SCAP)] and transcriptional coactivators (PPAR gamma coactivator 1-apha and beta (PGCIa, and PGCIb)). Values represent means ± SE relative to control. Significant difference from control indicated for P < 0.10(+), P < 0.05 (*), P < 0.01(**), P < 0.001(***).

[00022] Figure 4 shows a tissue profile of Spot 14 (S 14) and midline 1 interacting protein 1 (MIDlIPl). Values are shown means (n 3-6 per tissue) and plotted on a log axis. With the exception of non-lactating mammary, all samples came from cows in the mid to late stages of the lactation cycle. Tissues include subcutaneous adipose tissue (AT), liver (Liv), lactating mammary gland (Lact), non-lactating mammary tissue (Nlact), uterine (Uter), lung (Lung), brain, skeletal muscle (Muse), and heart.

[00023] Figure 5 shows the effects of trans-10, cis-Yl conjugated linoleic acid (CLA) on adipose mRNA abundance for thyroid hormone responsive Spot 14 (S 14) using online microarray data from studies by House et al. (House et al., "Functional Genomic Characterization of Delipidation Elicited by trans- 10, cis- 12-conjugated Linoleic Acid (£lO,cl2-CLA) in a Polygenic Obese Line of Mice," Physiol Genomics 21 :351-61 (2005), which is hereby incorporated by reference in its entirety). Values indicated by means ± SE relative to control. Significant difference from control indicated by P < 0.05(*) and P < 0.001(***).

DETAILED DESCRIPTION OF THE INVENTION

[00024] The present invention relates to a method of screening cancer patients to determine whether the form of cancer they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for tumor cells with a disregulation of lipid metabolism, and identifying patients with tumor cells with a disregulation of lipid metabolism as having a form of cancer suitable for treatment with conjugated linoleic acids.

[00025] The term "conjugated linoleic acid" (CLA) is meant to include a family of many isomers of linoleic acid, which are found primarily in the meat and dairy products of ruminants. As implied by the name, the double bonds of CLAs are conjugated.

[00026] Conjugated linoleic acid is a trans fat. Unlike most trans fatty acids found in the human diet, CLA occurs naturally, produced by microorganisms in the fore-stomach of ruminants. Non-ruminants, such as humans, may be able to produce some isomers of conjugated linoleic acid from non-conjugated ruminant fats. One such example is vaccenic acid, which could be converted to CLA by delta-9-desaturase (Banni et al., "Vaccenic Acid Feeding Increases Tissue Levels of Conjugated Linoleic Acid and Suppresses Development of Premalignant Lesions in Rat Mammary Gland," Nutr Cancer 41(l-2):91-7 (2001), which is hereby incorporated by reference in its entirety).

[00027] The conjugated linoleic acids may be selected from the group consisting of trans-10, cis-12 conjugated linoleic acid, cis-9, trans-11 conjugated linoleic acid, and mixtures thereof.

[00028] Conjugated linoleic acid comes in many isomers (forms) with two having been more extensively studied, cis-9, trans-11 CLA and trans-10, cis-12 CLA. Of these, it is the trans-10, cis-12 isomer which primarily prevents lipogenesis (synthesis of milk fat during lactation or storage of fat in adipose tissue in non-lactating state) (Bauman et al., "Nutritional Regulation of Milk Fat Synthesis," Annu Rev Nutr 23:203-227 (2003); Brown et al., "Trans-10, Cis-12, but Not Cis-9, Trans-11, Conjugated Linoleic Acid Attenuates Lipogenesis in Primary Cultures of Stromal Vascular Cells from Human Adipose Tissue," J Nutr 131(9):2316-21 (2001); Wang et al., "Conjugated Linoleic Acid and Obesity Control: Efficacy and Mechanisms," Int J Obes Relat Metab Disord 28(8):941-55 (2004) and Faulconnier et al., "Isomers of Conjugated Linoleic Acid Decrease Plasma Lipids and Stimulate Adipose Tissue Lipogenesis Without Changing Adipose Weight in Post-prandial Adult Sedentary or Trained Wistar Rat," J Nutr Biochem 15(12):741-8 (2004), which are hereby incorporated by reference in their entirety).

[00029] Cis-9, trans- 11 conjugated linoleic acid is also described in

Peterson et al., "Analysis of Variation in Cis-9, Trans- 11 Conjugated Linoleic Acid (CLA) in Milk Fat of Dairy Cows," J Dairy Sci 85(9):2164-2172 (2002) and Perfield et al., "Trans-9, Cis-11 Conjugated Linoleic Acid Reduces Milk Fat Synthesis in Lactating Dairy Cows," J Dairy Sci 90(5) :2211-2218 (2007), which are hereby incorporated by reference in their entirety.

[00030] Trans-10, cis-12 conjugated linoleic acid is also described in

Bauman et al., "Nutritional Regulation of Milk Fat Synthesis," Annu Rev Nutr 23:203-27 (2003) and Lock et al., "Trans-10 Octadecenoic Acid Does Not Reduce Milk Fat Synthesis in Dairy Cows," J Nutr 137(l):71-76 (2007), which are hereby incorporated by reference in their entirety.

[00031] Various antioxidant and anti-tumor properties have been attributed to conjugated linoleic acid, and studies on mice and rats have shown reduction in mammary, skin, and colon tumor growth (Belury, MA., "Inhibition of Carcinogenesis by Conjugated Linoleic Acid: Potential Mechanisms of Action," J Nutr 132(10):2995-8 (2002), which is hereby incorporated by reference in its entirety).

[00032] Many studies on conjugated linoleic acid in humans show a tendency for reduced body fat (Thorn et al., "Conjugated Linoleic Acid Reduces Body Fat in Healthy Exercising Humans," J Int Med Res 29(5):392-6 (2001), Erratum in (correction of dosage error in abstract): J Int Med Res 30(2):210

(2002), which are hereby incorporated by reference in their entirety), particularly abdominal fat, changes in serum total lipids and decreased whole body glucose uptake. The maximum reduction in body fat mass was achieved with a 3.4 g daily does (Blankson et al., "Conjugated Linoleic Acid Reduces Body Fat Mass in Overweight and Obese Humans," J Nutr 130(12):2943-8 (2000), which is hereby incorporated by reference in its entirety).

[00033] Conjugated linoleic acid may be obtained from several dietary sources. Food products derived from ruminants (e.g., lamb, beef, milk, and dairy products) are the main human dietary sources. In these foods there are over 20 different isomers (forms) of CLA but cis-9, trans- 11 CLA is the major form often comprising over 90% of total CLA (Bauman et al., "Conjugated Linoleic Acid: Biosynthesis and Nutritional Significance," Advanced Dairy Chemistry, Vol. 2: Lipids, Chapter 3, pages 93-136 (2006), which is hereby incorporated by reference in its entirety). The CLA content of ruminant derived foods can also be markedly increased by certain diets (e.g., grazing or feeding plant oils or plant seeds), but in all cases the predominant CLA isomer is cis-9, trans- 11 and this CLA isomer has no effect on rates of lipid synthesis (Bauman et al., "Conjugated Linoleic Acid: Biosynthesis and Nutritional Significance," Advanced Dairy Chemistry, Vol. 2: Lipids, Chapter 3, pages 93-136 (2006), which is hereby incorporated by reference in its entirety).

[00034] A method (performed as a diagnostic assay) to identify cancer cells in a human by providing nucleic acid from a suspected tumor suppressor gene which specifically hybridizes to RNA expressed from such a gene, involves the use of Northern blot or in situ hybridization assays. These assays are used to measure the level of expression for normal cells and suspected cells from a tissue sample. Labelling of the nucleic acid sequence allows for the detection and measurement of relative expression levels. By comparing the level of expression between normal cells and suspected cells from a tissue sample, a pre-cancerous or cancerous state may be identified by the reduced expression level of the gene product.

[00035] Performance of this method may involve the use of a labeled nucleic acid probe comprising a nucleotide sequence of at least 8 nucleotides in length, more preferably at least 15 nucleotides in length, and even more preferably at least 40 nucleotides in length, up to and including all or nearly all of the coding sequence of a suspected tumor suppressor gene. A first tissue sample comprising cancerous cells is obtained and a second tissue sample comprising non-cancerous cells is also obtained. Using a Northern blot format, the nucleic acid probes are contacted under high-stringency hybridizing conditions with RNA of each of the first and second tissue samples and then the degree of hybridization between the probes and the RNA samples is measured (i.e., detecting the presence of any type of conventional label). Using an in situ hybridization format, the labeled probe is inserted into morphologically intact cells present within a tissue sample and hybridization is carried out using hybridizing conditions well known to those skilled in the art. The labeled probes will identify non-cancerous cells while cancerous cells remain unlabeled. As corroborating evidence of cancer, the aforementioned techniques are useful to provide an estimation of the number of cancerous cells present in a given tissue sample, and/or whether a given cell is likely to be cancerous.

[00036] An approach to detecting the presence of a given sequence or sequences in a polynucleotide sample involves selective amplification of the sequence(s) by polymerase chain reaction. PCR is described in U.S. Patent No. 4,683,202 to Mullis et al. and Saiki et al., "Enzymatic Amplification of Beta- globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia," Science 230:1350-1354 (1985), which are hereby incorporated by reference in their entirety. In this method, primers complementary to opposite end portions of the selected sequence(s) are used to promote, in conjunction with thermal cycling, successive rounds of primer- initiated replication. The amplified sequence(s) may be readily identified by a variety of techniques. This approach is particularly useful for detecting the presence of low-copy sequences in a polynucleotide-containing sample, e.g., for detecting pathogen sequences in a body-fluid sample.

[00037] More recently, methods of identifying known target sequences by probe ligation methods have been reported. The methods have been described in U.S. Patent No. 4,883,750 to Whiteley et al., Wu et al., "The Ligation Amplification Reaction (LAR)-- Amplification of Specific DNA Sequences Using Sequential Rounds of Template-dependent Ligation," Genomics 4:560-569 (1989), Landegren et al., "A Ligase-mediated Gene Detection Technique," Science 241 :1077-1080 (1988), and Winn-Deen et al, "Sensitive Fluorescence Method for Detecting DNA-ligation Amplification Products," Clin Chem 37:1522-1523 (1991), which are hereby incorporated by reference in their entirety).

[00038] The present invention also relates to a method of treating a patient for cancer. This involves selecting a patient having tumor cells with a disregulation of lipid metabolism and administering conjugated linoleic acids to the selected patient under conditions effective to treat cancer.

[00039] According to one embodiment, the patient has breast cancer, prostate cancer, ovarian cancer, colon cancer, endometrium cancer, lung cancer, bladder cancer, stomach cancer, osteophagus cancer, oral tongue cancer, oral cavity cancer, skin cancer, mesotheliomas, retinoblastomas, and/or nephroblastomas .

[00040] The disregulation of lipid metabolism may be in the form of: overexpression of fat synthesis enzymes, high lipogenic activity, or accumulation of lipids in tumor cells.

[00041] The present invention also relates to a method of treating a patient for cancer including selecting a patient overexpressing the Thyroid Hormone Responsive Spot 14 gene and administering conjugated linoleic acids to the selected patient under conditions effective to treat cancer.

[00042] The conjugated linoleic acids of the present invention may be orally administered, for example, with an inert diluent, or with an assimilable edible carrier, or they may be enclosed in hard or soft shell capsules, or they may be compressed into tablets, or they may be incorporated directly with the food of the diet. For oral therapeutic administration, the therapeutic may be incorporated with excipients and used in the form of tablets, capsules, elixirs, suspensions, syrups, and the like. Various other materials may be present as coatings or to modify the physical form of the dosage unit. [00043] This aspect of the present invention can be carried out by formulating and administering the conjugated linoleic acids in substantially the same manner as described above.

[00044] The present invention also relates to a method of screening cancer patients to determine whether the form of cancer they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for overexpression of the Thyroid Hormone Responsive Spot 14 gene, and identifying patients who overexpress the Thyroid Hormone Responsive Spot 14 gene as having a form of cancer suitable for treatment with conjugated linoleic acids.

[00045] The term "Thyroid Hormone Responsive Spot 14 (THRSP) gene is meant to include a transciption factor involved in the regulation of adipogenic enzymes by 3 thyroid response elements in the promoter region. Thyroid Hormone Responsive Spot 14 is described in Zhan et al, "Molecular Cloning and Expression of the Duplicated Thyroid Hormone Responsive Spot 14 (THRSP)

Genes in Ducks," Poult Sci 85(10): 1746-1754 (2006); Cunningham et al., ""Spot 14" Protein: A Metabolic Integrator in Normal and Neoplastic Cells," Thyroid 8:815-25 (1998); LaFave et al., "S14: Insights from Knockout Mice," Endocrinology 147(9):4044-4047 (2006); Kinlaw et al., "Spot 14: A Marker of Aggressive Breast Cancer and A Potential Therapeutic Target," Endocrinology 147(9):4048-4055 (2006); Harvatine et al., "SREBPl and Thyroid Hormone Responsive Spot 14 (S 14) are Involved in the Regulation of Bovine Mammary Lipid Synthesis During Diet-induced Milk Fat Depression and Treatment with CLA," JNutr 136(10):2468-2474 (2006), which are hereby incorporated by reference in their entirety.

[00046] In carrying out this aspect of the present invention the above- described screening formats can be utilized.

[00047] This aspect of the present invention can be carried out by treating a patient for cancer and administering conjugated linoleic acids as described above. [00048] Another aspect of the present invention relates to a method of screening cancer patients to determine whether the form of cancer they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for overexpression of the Sterol Responsive Element Binding Protein gene, and identifying patients who overexpress the Sterol Responsive Element Binding Protein gene as having a form of cancer suitable for treatment with conjugated linoleic acids.

[00049] The term "Sterol Responsive Element Binding Protein" (SREBP) gene is meant to include transcription factors that help activate a variety of sterol responsive genes. These include genes encoding enzymes in the cholesterol biosynthetic pathway and the saturated and unsaturated fatty acid biosynthetic pathway, as well as the low-density lipoprotein receptor (LDLR) gene (Chang TY., "SREBPs, Membrane Lipid Biosynthesis, and Fatty Acids," J Clin Invest 100 (8)1905-1906 (1997), which is hereby incorporated by reference in its entirety).

[00050] SREBPs were first discovered as specific transcription factors that bind to the 10-bp sterol regulatory element (SRE) within the promoters of LDLR and 3-hydroxy-3-methylglutaryl Co-A synthase (an enzyme in the sterol synthesis pathway). It is now known that there are at least two different genes that produce at least three different SREBP proteins: SREBP- l_a and SREBP- l_c (two isoform proteins) and SREBP-2. Although the functions of these three proteins are similar, they are not identical (Brown et al., "The SREBP pathway: Regulation of Cholesterol Metabolism by Proteolysis of a Membrane-bound Transciption Factor," Cell 89:331-340 (1997), which is hereby incorporated by reference in its entirety). The SREBP-I gene was discovered independently by its ability to stimulate fatty acid synthesis and adipocyte differentiation, in a rat preadipose cell line (Tontonoz et al., "ADDl : A Novel Helix- loop-helix Transcription Factor Associated with Adipocyte Determination and Differentiation," MoI Cell Biol 13:4753-4759 (1993), which is hereby incorporated by reference in its entirety). Subsequently, it was shown that the promoters of genes that encode enzymes in the fatty acid synthesis pathway contain the SRE element and are regulated by the SREBP proteins (Lopez et al., "Sterol Regulation of Acetyl CoA Carboxylase: A Mechanism for Co-ordinate Control of Cellular Lipid," Proc Natl Acad Sci USA 93:1049-1053 (1996), which is hereby incorporated by reference in its entirety). SREBPs are also described in Horton et al., "SREBPs: Activators of the Complete Program of Cholesterol and Fatty Acid Synthesis in the Liver," J Clin Invest 109: 1125-1131 (2002); Sekiya et al., "Polyunsaturated Fatty Acids Ameliorate Hepatic Steatosis in Obese Mice by SREBP-I Suppression," Hepatology 38:1529- 1539 (2003); Shimano et al., "Sterol Regulatory Element-binding Proteins (SREBPs): Transcriptional Regulators of Lipid Synthetic Genes," Prog Lipid Res 40:439-452 (2001) and Yahagi et al., "Absence of Sterol Regulatory Element- binding Protein- 1 (SREBP-I) Ameliorates Fatty Livers But Not Obesity or Insulin Resistance in Lep(ob)/Lep(ob) mice," J Biol Chem 277:19353-19357 (2002) which are hereby incorporated by reference in their entirety.

[00051] SREBPs are also described in Smith et al., "The Sterol Response

Element Binding Protein Regulates Cyclooxygenase-2 Gene Expression in Endothelial Cells," J Lipid Res 46(5):862-871 (2005); Chouinard et al., "Sterol Regulatory Element Binding Protein- 1 Activates the Cholesteryl Ester Transfer Protein Gene In Vivo but is Not Required for Sterol Up-regulation of Gene Expression," J Biol Chem 273(35): 22409-22414 (1998); Swinnen et al., "Androgens, Lipogenesis and Prostate Cancer," J Steroid Biochem MoI Biol 92(4):273-279 (2004); Canbay et al., "Lipid Metabolism in the Liver," Z

Gastroenterol 45(1):35-41 (2007) and Harvatine et al., "SREBPl and Thyroid Hormone Responsive Spot 14 (S 14) are Involved in the Regulation of Bovine Mammary Lipid Synthesis During Diet-induced Milk Fat Depression and Treatment with CLA," J Nutr 136(10):2468-2474 (2006), which are hereby incorporated by reference in their entirety.

[00052] In carrying out this aspect of the present invention the above- described screening formats can be utilized.

[00053] Another aspect of the present invention relates to a method of treating a patient for cancer. This involves selecting a patient overexpressing the Sterol Responsive Element Binding Protein gene and administering conjugated linoleic acids to the selected patient under conditions effective to treat cancer. [00054] This aspect of the present invention can be carried out by treating a patient for cancer and administering conjugated linoleic acids as described above.

[00055] Another aspect of the present invention relates to a method of screening obesity patients to determine whether the form of obesity they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for overexpression of the Thyroid Hormone Responsive Spot 14 gene, and identifying patients who overexpress the Thyroid Hormone Responsive Spot 14 gene as having a form of obesity suitable for treatment with conjugated linoleic acids.

[00056] In carrying out this aspect of the present invention the above- described screening formats can be utilized.

[00057] Another aspect of the present invention relates to a method of treating a patient for obesity including selecting a patient overexpressing the Thyroid Hormone Responsive Spot 14 gene and administering conjugated linoleic acids to the selected patient under conditions effective to treat obesity.

[00058] This aspect of the present invention can be carried out by administering conjugated linoleic acids as described above.

[00059] Another aspect of the present invention relates to a method of screening obesity patients to determine whether the form of obesity they have is suitable for treatment with conjugated linoleic acids. This involves providing a tissue or fluid sample from the patient, evaluating the sample for overexpression of the Sterol Responsive Element Binding Protein gene, and identifying patients who overexpress the Sterol Responsive Element Binding Protein gene as having a form of obesity suitable for treatment with conjugated linoleic acids.

[00060] In carrying out this aspect of the present invention the above- described screening formats can be utilized.

[00061] Another aspect of the present invention relates to a method of treating a patient for obesity including selecting a patient overexpressing the Sterol Responsive Element Binding Protein gene and administering conjugated linoleic acids to the selected patient.

[00062] This aspect of the present invention can be carried out by administering conjugated linoleic acids as described above.

EXAMPLES

[00063] The following examples are provided to illustrate embodiments of the present invention but are by no means intended to limit its scope.

Materials and Methods for Examples 1-4

Animals and treatments

[00064] All experimental procedures were approved by the Cornell

University Institutional Animal Care and Use Committee. Nine mid-lactation cows (193 ± 32 d postpartum; mean ± SD) from the Cornell University Teaching and Research Center were assigned randomly to replicated 3 x 3 Latin squares. Experimental periods were 14 d and specific treatments were control, short term administration of trans- 10, cis-\2 CLA, and feeding a low forage, high oil diet (LF/HO). During the control and CLA treatment periods, cows received a base diet that was formulated to meet or exceed nutrient requirements with minimal fat content (Table 1). The LF/HO treatment received a slightly lower forage diet that included 3.0% soybean oil and 1.5% fish oil. Diets were fed ad lib as a total mixed ration and daily intakes determined. Feed ingredients were sampled weekly, dried (55⁰C forced-air oven for 72 h), ground (Wiley mill with 1-mm screen; Arthur H. Thomas), and nutrient composition determined by wet chemistry procedures (AOCS Official Methods of Analysis, 17th ed. AOCS, Arlington, VA. (2000), which is hereby incorporated by reference in its entirety; Dairy One Cooperative, Ithaca, NY). TABLE 1

A. Ingredient and nutrient composition of experimental diets

l The base diet was feed during the control and CLA treatment periods. The low forage, high oil diet was feed during the LF/HO treatment period.

² Corn silage contained 31.3% DM (as fed).

³ Alfalfa silage contained 32.2% DM (as fed).

⁴ Grain mix contained 28.8% corn glutten feed, 20.5% Amino Plus, 20.0% wheat midds, 14.3% soybean hulls, 3.7% sodium bicarbonate, 3.5% blood meal, 2.7% limestone, 1.9% soybean meal, 1.75% salt, 1.0% urea, 0.9% calcium sulfate, 0.6% magnesium oxide, 0.22% selenium (0.06% selenium), 0.1% trace mineral (12.3% Ca, 12.5% sulfur, 0.32% magnesium, 19,000 PPM copper, 90,000 PPM manganese, 2500 PPM cobalt, 1900 PPM iodine, 110,000 PPM zinc and 0.395 IU/kg vit E), 0.1% vitamin ADE (3600 IU/g vit A, 7000 IU/g vit D and 70000 IU/kg vit E), and 0.01% vitamin E.

[00065] The CLA treatment involved a 3 d i.v. infusion of a CLA-

Intralipid emulsion (d 12 to 14 of experimental period). Jugular catheters were installed on d 11 and the infusion supplied 10 g/d of trans- 10, cis-Yl CLA in equal doses infused every 6 h at a rate of 1 mL emulsion/min. The trans- 10, cis-Yl CLA methyl ester stock (BASF Corporation) contained 88.3% total CLA (98% trans- 10, cis-Yl isomer), 6.8% palmitic acid, 2.7% oleic acid and 2.0% stearic acid. Previous investigations have shown that the methyl ester and free fatty acid forms of trans- 10, cis-Yl CLA are equally effective in inhibiting milk fat synthesis (de Veth et al., "Effect of CLA on Milk Fat Synthesis in Dairy Cows: Comparison of Inhibition by Methyl Esters and Free Fatty Acids, and Relationships Among Studies," Lipids 39:365-72 (2001), which is hereby incorporated by reference in its entirety). Intralipid (Baxter Healthcare

Corporation) emulsion contained 10% soybean oil and the CLA stock was added to give a final solution containing 10% CLA. Methyl esters of CLA were rapidly emulsified during homogenization and remained in solution.

Milk sampling and analysis

[00066] Cows were milked 3x/d and yields recorded. Samples were taken at each milking on the last day and analyzed for fat and true protein using a mid- infrared spectrophotometer (AOCS Official Methods of Analysis, 17th ed. AOCS, Arlington, VA. (2000); Dairy One Cooperative). Additional samples for fatty acid analysis were composited based on milk fat yield; lipid was extracted, transmethylated and methyl esters quantified by GC according to Perfield et al. (Perfield et al., "Trans-10, trans-Yl Conjugated Linoleic Acid Does Not Affect Milk Fat Yield But Reduces Delta-9 Desaturase Index in Dairy Cows," J Dairy Sci 89:2559-2566 (2006), which is hereby incorporated by reference in its entirety).

Tissue biopsy

[00067] Mammary biopsies were performed 1-3 h after milking on d 14.

Cows were given intravenous xylazine (15-25 mg) and a 15 mL lidocaine HCL sub-dermal block was administered above the incision site. A 0.5 cm incision was made in the skin at the midpoint of the rear quarter and the Magnum Biopsy Gun system (Bard Biopsy Systems) was used. Briefly, a 10 g cannula with trocar was inserted 5 cm into the mammary gland through the incision. The trocar was removed and a 12 g biopsy needle mounted in the biopsy gun was inserted through the cannula and fired. Tissue samples (30 mg tissue/biopsy) were washed with 0.9% saline solution, inspected to verify tissue homogeneity and snap frozen in liquid nitrogen. Samples were stored at -80⁰C until RNA extraction. Multiple biopsies were routinely collected by reinserting the biopsy needle through the cannula. Immediately upon removal of the biopsy needle, the trocar was replaced and a purse string suture was placed around the cannula with #1 Nylon. The suture was tied as the cannula was removed and pressure applied to reduce collection of blood under the skin. The biopsy procedure resulted in minimal bleeding and milk appeared normal in 2 to 4 milkings following the biopsy; no intra-mammary infections or loss of production were encountered.

RNA isolation and Real-Time PCR

[00068] Total RNA was isolated from ~30 mg mammary tissue from one biopsy using the RNeasy Lipid Kit (Qiagen) and DNA contamination was removed by on-column DNase treatment (RNase-Free DNase Set; Qiagen). RNA concentration and integrity were determined by an Agilent 2100 Bio Analyzer (Agilent Technologies). Total RNA was reverse transcribed using Superscript III First Strand Synthesis kit (Invitrogen) with random primers. Quantitative real- time reverse transcriptase PCR (qRT-PCR) assays were developed for genes of interest (Table 2). Briefly, primers were designed on or spanning exon boundaries when possible using PrimerExpress v2.0 (Applied Biosystems) and optimal primer pairs were selected using Primer3 (Rozen et al., "Primer3 on the WWW for General Users and for Biologist Programmers," In: Krawetz S, Misener S, editors. Bioinformatics Methods and Protocols: Methods in Molecular Biology. Totowa, NJ: Humana Press; p. 365-86 (2000), which is hereby incorporated by reference in its entirety) and PerlPrimer (Marshall OJ., "PerlPrimer: Cross- platform, Graphical Primer Design for Standard, Bisulphite and Real-time PCR," Bioinformatics 20:2471-72 (2004), which is hereby incorporated by reference in its entirety). qRT-PCR reactions included iTaq SYBR Green Supermix with ROX (Bio-Rad Laboratories) and 400 nM of gene specific forward and reverse primers (Invitrogen). cDNA (5-25 ng) was amplified using a two step program (95°C for 15 s and 60⁰C for 60 s) with an ABI PRISM 7000 Sequence Detection System (Applied Biosystems). Dissociation curves were generated at the end of amplification to verify presence of a single product. Sample message level was determined relative to a dilution curve of pooled mammary cDNA (ABI Prism. Relative Quantification of Gene Expression. 7700 Sequence Detection System User Bulletin 2. Applied Biosystems. pp. 1-36 (2001), which is hereby incorporated by reference in its entirety).

Data mining

[00069] Publicly available data from two microarray experiments were downloaded. Specifics of animals and experimental procedures were previously published (House et al., "Functional Genomic Characterization of Delipidation Elicited by trans-10, cώ-12-conjugated Linoleic Acid (?10,cl2-CLA) in a Polygenic Obese Line of Mice," Physiol Genomics 21 :351-61 (2005) and Hargrave et al., "Effect of Dietary Conjugated Linoleic Acid on Adiposity and the Adipose-transcriptome," J Dairy Sci 88(Suppl. l):280 (2005), which are hereby incorporated by reference in their entirety). Briefly, House et al. (House et al., "Functional Genomic Characterization of Delipidation Elicited by trans- 10, cis- 12-conjugated Linoleic Acid (£lO,cl2-CLA) in a Polygenic Obese Line of Mice," Physiol Genomics 21 :351-61 (2005), which is hereby incorporated by reference in its entirety) fed 9 wk old mice (NCSU M16 line) diets containing 1% trans- 10, cis-\2 CLA or control (1% linoleic acid) for 14 d; total RNA was extracted from epididymal adipose tissue of 70 animals, pooled into 4 groups per treatment, and analyzed on the Agilent Mouse Oligo microarray slide (G4121A; Agilent Technologies, Palo Alto, CA). Hargrave et al. (Hargrave et al., "Effect of Dietary Conjugated Linoleic Acid on Adiposity and the Adipose-transcriptome," J Dairy Sci 88(Suppl. l):280 (2005), which is hereby incorporated by reference in its entirety) fed 12 wk old mice (UNL MC line) diets containing 0 or 2% of a mixed isomer CLA (-50% trans-10, cis-\2) for 16 d; adipose tissue RNA from five littermate pairs was analyzed on the Affymetrix Mouse 430A 2.0 GeneChip (Affymetrix).

Statistical analysis

[00070] Data were analyzed using the fit model procedure of JMP^®

(Version 5.0, SAS Institute). The model to test treatment means included the random effect of cow, and fixed effect of period and treatment. Additionally, the geometric mean of three housekeeping genes (18S (18S ribosomal subunit), ACTB (β-actin), and B2M (B2-microglobulin)) was calculated using GeNorm (Vandesompele et al., "Accurate Normalization of Real-time Quantitative RT-

PCR Data by Geometric Averaging of Multiple Internal Control Genes," Genome Biol 3(7):research0034.1-0034.11 (2002), which is hereby incorporated by reference in its entirety) and used as a covariant in the model. Data points with Studentized Residuals greater than 2.5 were considered outliers and excluded from analysis. Few points were excluded in analysis and rarely more than one per response variable. Preplanned contrasts included the effect of CLA (CON vs. CLA), and the effect of LF/HO diet (CON vs. LF/HO). The relationship between expression of individual genes (SREBPl and S 14) and lipogenic enzymes (fatty acid synthase (FASN) and lipoprotein lipase (LPL)) was tested with the above model by replacing treatment with the predictor gene of interest. Relationships between expression of genes was declared significant at P < 0.01 and a trend at P < 0.05. For statistical analysis of S14 data from House et al. (House et al., "Functional Genomic Characterization of Delipidation Elicited by trans- 10, cis- 12-conjugated Linoleic Acid (?10,cl2-CLA) in a Polygenic Obese Line of Mice," Physiol Genomics 21 :351-61 (2005), which is hereby incorporated by reference in its entirety, the P value and fold change for the S 14 annotated probe was downloaded (GSE1580). For the data of Hargrave et al. (Hargrave et al., "Effect of Dietary Conjugated Linoleic Acid on Adiposity and the Adipose- transcriptome," J Dairy Sci 88(Suppl. l):280 (2005), which is hereby incorporated by reference in its entirety), downloaded fluorescent intensities of the S 14 annotated probe set were log (base 2) transformed and treatment means compared using a t-test.

Example 1 ~ Milk Fat Synthesis and Expression of Lipogenic Enzymes

[00071] The short term infusion of trans-10, cis-\2 CLA decreased milk fat content and yield by 23 and 24%, respectively (Table 3). The LF/HO diet decreased milk fat concentration and yield to an even greater extent, 31 and 38%, respectively. Analysis of the milk fatty acid profile revealed that the reduction in milk fat involved decreased secretion of both de novo and preformed fatty acids (Table 3). However, the decrease was greater for the short and medium chain fatty acids resulting in a shift in milk fat composition to an increased proportion of long chain fatty acids (Table 4). Expression of key regulated enzymes in mammary fatty acid metabolism were analyzed by qRT-PCR to verify down- regulation of lipogenic enzymes. Expression of FASN and LPL were down- regulated for both CLA and LF/HO treatments, whereas expression of stearoyl- CoA desaturase (SCD) was only down-regulated with LF/HO treatment (Figure

¹ Treatments were CON = control, CLA = trans-\0, cis-\2 conjugated linoleic acid, and LF/HO = low forage, high oil diet. Values represent LS means. 2 Probability of a treatment effect (Trt) and that CLA or LF/HO treatment differed from control.

³ Fatty acids <16 carbons originate from mammary de novo synthesis, fatty acids >16 carbons originate from extraction from plasma, and 16 carbon fatty acids originate from both sources.

TABLE 4

Effect of trans- 10, cis- 12 conjugated linoleic acid and a low forage, high oil diet on milk fatty acid profile of dairy cows

Treatment¹ P ¹

Fatty acid CON CLA LF/HO SEM Trt CLA LF/HO g/100 g fatty acids

4:0 1.69 1.63 1.45 0.07 0.04 0.51 0.02

6:0 1.43 1.18 1.05 0.05 <0.001 0.001 <0.001

8:0 0.98 0.79 0.69 0.04 0.004 0.002 <0.001

10:0 2.54 2.07 1.60 0.13 <0.001 0.01 <0.001

12:0 3.26 2.83 2.19 0.14 <0.001 0.03 <0.001

14:0 11.68 11.79 9.97 0.31 <0.001 0.73 <0.001

15:0 1.27 1.11 0.94 0.05 <0.001 0.006 <0.001

16:0 34.08 31.80 26.69 0.66 <0.001 0.004 <0.001

16:1, cis 2.37 2.26 2.94 0.32 0.06 0.64 0.05

17:0 0.53 0.56 0.42 0.02 <0.001 0.006 <0.001

18:0 8.18 9.61 4.55 0.59 <0.001 0.03 <0.001

\%:\, trans-4 0.02 0.02 0.03 0.01 0.55 0.70 0.48

\%:\, trans-5 <0.01 0.01 0.04 0.01 <0.001 0.64 <0.001

18:1, trans- 6-8 0.24 0.30 0.66 0.02 <0.001 0.03 <0.001

18:1, trans-9 0.21 0.25 0.64 0.03 <0.001 0.29 <0.001

18:1, trans- 10 0.41 0.47 5.86 0.95 0.001 0.96 <0.001

18:1, trans- 11 1.21 1.62 9.15 0.93 <0.001 0.73 <0.001

18:1, cis-9 18.51 19.29 12.26 0.80 <0.001 0.31 <0.001

1%:2, cis-9,cis-12 2.11 2.47 2.09 0.11 0.01 0.009 0.85

18:3 0.37 0.43 0.30 0.02 <0.001 0.01 0.003

20:0 0.14 0.16 0.11 0.01 <0.001 0.03 0.004 cis-9, trans-\ \ CLA 0.68 0.77 3.65 0.28 <0.001 0.79 <0.001 trans-lO, cis-l2 CLA <0.01 0.15 0.03 0.00 <0.001 <0.001 0.04

Others 6.57 6.65 10.18 0.31 <0.001 0.84 <0.001

¹ Treatments were CON = control, CLA = trans-lQ, cis-12 conjugated linoleic acid, and LF/HO = low forage, high oil diet. Values represent LS means.

Probability of a treatment effect (Trt) and that CLA or LF/HO treatment differed from control.

Fatty acids <16 carbons originate from mammary de novo synthesis, fatty acids >16 carbons originate from extraction from plasma, and 16 carbon fatty acids originate from both sources. Example 2 ~ Expression of SREBPl and SREBPl Regulatory Proteins

[00072] Expression of SREBPl was down-regulated in mammary tissue during both trans- 10, cis-\2 CLA treatment and diet-induced MFD (Figure 2). Research over the last decade has elaborated elements of the SREBP regulatory system. qRT- PCR was used to examine message abundance of several genes encoding SREBP processing proteins and SREBP transcriptional co-activators. Insulin responsive gene (INSIG) 1 was decreased during trans- 10, cis-Yl CLA treatment and the LF/HO diet, and mammary expression of INSIG2, SREBP cleavage activating protein (SCAP) and peroxisome proliferative activated receptor, gamma, coactivator (PGC) lα were moderately decreased during diet- induced MFD (Figure 3). However, expression of PGCl β was not altered by treatment.

Example 3 - Expression of S14 During Milk Fat Depression

[00073] Expression of S 14 was down-regulated in mammary tissue during treatment with trans- 10, cis-Yl CLA and LF/HO-induced MFD to an extent similar to the reduction in SREBPl expression (Figure 2) and greater than the decrease in milk fat synthesis. Relatively little is known about S 14 in bovine tissues so its tissue expression profile was examined (Figure 4). Results demonstrated that adipose tissue and liver were the predominant sites of S 14 expression. Expression of S14 in lactating mammary tissue was over 3-fold greater than observed for non-lactating mammary tissue, and substantially greater (>75-fold) than the low level observed in the lung. The tissue profile for a gene with sequence homology to S 14 was also examined, midline 1 interacting protein 1 (MIDlIPl), and it was observed that it was expressed at similar levels in all tissues profiled expect skeletal muscle which had approximately 17-fold greater expression than mammary tissue (Figure 4).

Example 4 - Expression of S14 in Mouse Adipose Tissue

[00074] Trans-10, cis-\2 CLA is often referred to as having anti-obesity effects because of its ability to reduce body fat accretion in several species (Wang et al, "Conjugated Linoleic Acid and Obesity Control: Efficacy and Mechanisms," Int J Obes Relat Metab Disord 28 :941 -55 (2004), which is hereby incorporated by reference in its entirety). To see if the down-regulation of S 14 expression was more broadly applicable to CLA treatment, publicly available microarray data was mined from experiments by House et al. (House et al.,

"Functional Genomic Characterization of Delipidation Elicited by trans- 10, cis- 12-conjugated Linoleic Acid (£lO,cl2-CLA) in a Polygenic Obese Line of Mice," Physiol Genomics 21 :351-61 (2005), which is hereby incorporated by reference in its entirety) and Hargrave et al. (Hargrave et al., "Effect of Dietary Conjugated Linoleic Acid on Adiposity and the Adipose-transcriptome," J Dairy Sci 88(Suppl. l):280 (2005), which is hereby incorporated by reference in its entirety). These investigations treated mice with trans-10, cis-\2 CLA and both studies reported that treatment caused a substantial reduction in body fat content. Expression data was specifically extracted for adipose S 14 and it was found to be markedly reduced (Figure 5). Furthermore, the extent of the reduction in S14 expression was similar for the 14 d treatment of 9 wk old NCSU Ml 6 mice (House et al., "Functional Genomic Characterization of Delipidation Elicited by trans-10, cώ-12-conjugated Linoleic Acid (?10,cl2-CLA) in a Polygenic Obese Line of Mice," Physiol Genomics 21 :351-61 (2005), which is hereby incorporated by reference in its entirety) and 16 d treatment of 12 wk old UNL MC mice

(Hargrave et al., "Effect of Dietary Conjugated Linoleic Acid on Adiposity and the Adipose-transcriptome," J Dairy Sci 88(Suppl. l):280 (2005), which is hereby incorporated by reference in its entirety).

[00075] Diet-induced MFD is a naturally occurring situation in dairy production that involves an interrelationship between rumen digestive processes and mammary synthesis of milk fat. MFD is observed for a range of diets and the basis was unclear until recent research observed that diet-induced MFD coincided with a shift in ruminal biohydrogenation and the rumen production of unique fatty acid intermediates. Referred to as the "Biohydrogenation Theory" (Bauman et al., "Nutritional Regulation of Milk Fat Synthesis," Annu Rev Nutr 23 :203-27 (2003), which is hereby incorporated by reference in its entirety), the first specific ruminal biohydrogenation intermediate demonstrated to inhibit milk fat synthesis was trans-10, cis-Yl CLA (Baumgard et al., "Identification of the Conjugated Linoleic Acid Isomer that Inhibits Milk Fat Synthesis," Am J Physiol Regul Integr Comp Physiol 278 :R179-84 (2000), which is hereby incorporated by reference in its entirety). While milk trans- 10, cis -12 CLA concentration correlates with diet- induced MFD, it does not explain the absolute extent of MFD and additional, unidentified inhibitory intermediates have been proposed (Bauman et al., "Nutritional Regulation of Milk Fat Synthesis," Annu Rev Nutr 23:203-27 (2003) and Peterson et al., "Diet-induced Milk Fat Depression in Dairy Cows Results in Increased trans- 10, cis-Yl CLA in Milk Fat and Coordinated Suppression of mRNA Abundance for Mammary Enzymes Involved in Milk Fat Synthesis," J Nutr 133:3098-102 (2003), which are hereby incorporated by reference in their entirety).

[00076] Milk fat depression is characterized by a specific reduction in milk fat synthesis, and milk fat yield decreased 38% with the LF/HO diet was observed and 24% with the 3-d infusion of trans-10, cis-\2 CLA. In dairy cows, about one- half of milk fatty acids originate from de novo synthesis in the mammary gland and the other half are derived by uptake of preformed fatty acids from circulation (Bauman et al., "Nutritional Regulation of Milk Fat Synthesis," Annu Rev Nutr 23:203-27 (2003), which is hereby incorporated by reference in its entirety). Consistent with previous research, a reduction in the secretion of both de novo and preformed fatty acids during MFD was observed, with short and medium chain fatty acids being more markedly reduced. This suggests that many of the different biochemical processes involved in the synthesis of milk fat must be down- regulated, and based on this a regulatory role for SREBP was postulated (Baumgard et al., "Trans-10, cis-12 Conjugated Linoleic Acid Decreases

Lipogenic Rates and Expression of Genes Involved in Milk Lipid Synthesis in Dairy Cows," J Dairy Sci 85:2155-63 (2002), which is hereby incorporated by reference in its entirety).

[00077] The SREBP family of transcription factors function as global regulators of lipid metabolism (Eberle et al., "SREBP Transcription Factors:

Master Regulators of Lipid Homeostasis," Biochimie 86: 839-48 (2004), which is hereby incorporated by reference in its entirety). Of the three isoforms, SREBPIa and Ic are transcribed from the same gene through the use of alternative promoters. SREBPIc predominately regulates enzymes involved in fat synthesis and is expected to be the predominant transcript expressed in mammary tissue. However, it was not possible to design a qRT-PCR assay to distinguish between these two iso forms because the available annotated sequence is limited; thus, they are referred to collectively as SREBPl. The full length SREBP protein is anchored in the endoplasmic reticulum in tight association with SCAP that acts as a chaperone protein (Eberle et al., "SREBP Transcription Factors: Master Regulators of Lipid Homeostasis," Biochimie 86: 839-48 (2004); Horton et al., "SREBPs: Activators of the Complete Program of Cholesterol and Fatty Acid Synthesis in the Liver," J Clin Invest 109: 1125-31 (2002) and Goldstein et al., "Protein Sensors for Membrane Sterols," Cell 124:35-46 (2006), which are hereby incorporated by reference in their entirety). The SREBPl/SCAP complex is held in the ER through association with a third protein, either INSIGl or INSIG2. SREBP is activated by dissociation of INSIG from the SREBP/SCAP complex allowing translocation of SREBP/SCAP complex to the Golgi apparatus where two functionally distinct proteases act in sequence to release a nuclear active fragment (nSREBP). In turn, nSREBP translocates to the nucleus where it binds to sterol-regulatory elements (SRE) in the promoter/enhancer regions of target genes, recruits coactivators and stimulates transcription (Eberle et al., "SREBP Transcription Factors: Master Regulators of Lipid Homeostasis," Biochimie 86: 839-48 (2004), which is hereby incorporated by reference in its entirety).

[00078] SREBPl is proposed as one of the predominant mechanisms of inhibition of fat synthesis by PUFA (Jump et al., "Fatty Acid Regulation of Hepatic Gene Transcription," J Nutr 135:2503-6 (2005), which is hereby incorporated by reference in its entirety). The SREBP-regulatory system was first examined in bovine mammary epithelial cells (MAC-T cell line) and a decreased abundance of nSREBP 1 protein during trans- 10, cis-\2 CLA inhibition of fatty acid synthesis was observed (Peterson et al., "The Inhibitory Effect of trans- 10, cis-\2 CLA on Lipid Synthesis in Bovine Mammary Epithelial Cells Involves Reduced Proteolytic Activation of the Transcription Factor SREBP-I," J Nutr 134:2523-27 (2004), which is hereby incorporated by reference in its entirety). In the current in vivo investigation, decreased expression of SREBPl and INSIGl during both trans- 10, cis-Yl CLA treatment and diet-induced MFD is demonstrated. Expression of SREBPl and INSIGl correlates with the concentration of the nSREBPl protein as both SREBPIc and INSIGl genes contain an SRE in their proximal promoter (Amemiya-Kudo et al., "Promoter Analysis of the Mouse Sterol Regulatory Element-binding Protein- Ic Gene," J Biol Chem 275:31078-85 (2000) and Kast-Woelbern et al., "Rosiglitazone Induction of Insig-1 in White Adipose Tissue Reveals a Novel Interplay of Peroxisome Proliferator-activated Receptor Gamma and Sterol Regulatory Element-binding Protein in the Regulation of Adipogenesis," J Biol Chem 279:23908-15 (2004), which are hereby incorporated by reference in their entirety). INSIGl is normally expressed at higher levels than INSIG2, has a faster turnover and is more dynamic (Goldstein et al., "Protein Sensors for Membrane Sterols," Cell 124:35-46 (2006), which is hereby incorporated by reference in its entirety). SCAP and INSIG typically function in stoichiometric concentrations to each other (Goldstein et al., "Protein Sensors for Membrane Sterols," Cell 124:35- 46 (2006), which is hereby incorporated by reference in its entirety), and it was observed that SCAP and INSIG2 expressions were decreased during diet-induced MFD, but not by CLA treatment. The difference between treatments may relate to the greater decrease in milk fat yield or the longer duration of MFD for the LF/HO diet.

[00079] The PGC-I family of transcription coactivators are environmentally responsive factors regulating tissue metabolism (Lin et al., "Metabolic Control Through the PGC-I Family of Transcription Coactivators," Cell Metab 1 :361-70 (2005), which is hereby incorporated by reference in its entirety). Specifically, PGC- lβ is increased in response to high fat intake and coactivates SREBPl (Lin et al., "Hyperlipidemic Effects of Dietary Saturated Fats Mediated Through PGC-lBeta Coactivation of SREBP," Cell 120:261-73 (2005), which is hereby incorporated by reference in its entirety).

[00080] In lactating mouse mammary tissue, corn oil decreased expression of FASN, SREBPIc, and PGC- lβ but increased expression of PGC-I α. In the current study, expression of PGC-I β was unaffected by treatment and PGC- lα was only slightly decreased by the LF/HO treatment. Thus, the results offer little or no support for altered transcription of the PGC family of coactivators in the regulation of milk fat synthesis.

[00081] The synthesis and secretion of milk fat by the mammary gland involves an integration of biochemical processes including the uptake and transport of circulating fatty acids, cellular fatty acid synthesis, desaturation, and esterifϊcation. Characterization of lipogenic genes during MFD highlights the coordinated down-regulation in the expression of key enzymes associated with these processes (Table 5). SREBPl is highly expressed in the lactating bovine mammary gland and, as illustrated in Table 5, many of these key enzymes are transcriptionally regulated by SREBPl (Liang et al., "Diminished Hepatic Response to Fasting/refeeding and Liver X Receptor Agonists in Mice with Selective Deficiency of Sterol Regulatory Element-binding Protein-lc," J Biol Chem 277:9520-8 (2002) and Horton et al., "Combined Analysis of Oligonucleotide Microarray Data from Transgenic and Knockout Mice Identifies Direct SREBP Target Genes," Proc Natl Acad Sci USA 100:12027-32 (2003), which are hereby incorporated by reference in their entirety). In the mouse, SREBPl is upregulated at the initation of lactation (Rudolph et al., "Metabolic Regulation in the Lactating Mammary Gland: A Lipid Synthesizing Machine," Physiol Genomics 28:323 - 336 (2007), which is hereby incorporated by reference in its entirety) and disruption of the SREBPIc gene results in a 41% decrease in milk fat concentration (Rudolph et al., "SREBP 1-c Plays a Regulatory but Not Essential Role in Mammary Lipogenesis During Lactation," Endocrinology 604 (2005), which is hereby incorporated by reference in its entirety). Interestingly, a maximum reduction of 50% in milk fat synthesis is observed during diet-induced MFD (Bauman et al., "Nutritional Regulation of Milk Fat Synthesis," Annu Rev Nutr 23:203-27 (2003), which is hereby incorporated by reference in its entirety) and during dose response studies with exogenous trans-10, cis-Yl CLA (de Veth et al., "Effect of CLA on Milk Fat Synthesis in Dairy Cows: Comparison of Inhibition by Methyl Esters and Free Fatty Acids, and Relationships Among

Studies," Lipids 39:365-72 (2001), which is hereby incorporated by reference in its entirety). TABLE 5

Summary of SREBPl -regulated lipogenic genes in bovine mammary tissue that demonstrate a coordinated reduction in expression during diet-induced MFD and with CLA treatment

¹ Reference citations are as follows: A = Baumgard et al. (Baumgard et al., "Thms-lO, cis-12 Conjugated Linoleic Acid Decreases Lipogenic Rates and Expression of Genes Involved in Milk Lipid Synthesis in Dairy Cows," J Dairy Sci 85:2155-63 (2002), which is hereby incorporated by reference in its entirety) B = Peterson et al. (Peterson et al., "Diet-induced Milk Fat Depression in Dairy Cows Results in Increased trans-lQ, cis-12 CLA in Milk Fat and Coordinated Suppression of mRNA Abundance for Mammary Enzymes Involved in Milk Fat Synthesis," J Nutr 133:3098-102 (2003), which is hereby incorporated by reference in its entirety) C = Piperova et al. (Piperova et al., "Mammary Lipogenic Enzyme Activity, Trans Fatty Acids and Conjugated Linoleic Acids are Altered in Lactating Dairy Cows Fed a Milk Fat-depressing Diet," J Nutr 130:2568-74 (2000), which is hereby incorporated by reference in its entirety), D = Ahnadi et al. (Ahnadi et al., "Addition of Fish Oil to Diets for Dairy Cows. II. Effects on Milk Fat and Gene Expression of Mammary Lipogenic Enzymes," J Dairy Res 69:521-31 (2002), which is hereby incorporated by reference in its entirety) and E = present invention.

[00082] Mammalian regulation typically includes redundant systems for the amplification of cellular signals and the regulation of biochemical processes. S 14 has been implicated in the transcriptional regulation of lipogenic genes

(Cunningham et al., ""Spot 14" Protein: A Metabolic Integrator in Normal and Neoplastic Cells," Thyroid 8:815-25 (1998); Kinlaw et al., "Direct Evidence For a Role of the "Spot 14" Protein in the Regulation of Lipid Synthesis," J Biol Chem 270:16615-18 (1995) and Martel et al, "S14 Protein in Breast Cancer Cells: Direct Evidence of Regulation by SREBP-Ic, Superinduction with Progestin, and Effects on Cell Growth," Exp Cell Res 312:278-88 (2006), which are hereby incorporated by reference in their entirety), and S 14 was identified as a trans- 10, cis-12 CLA responsive candidate gene in microarray analysis of bovine mammary cell cultures. In rodents, S 14 was highly expressed in lipid-synthesizing tissues, including lactating mammary tissue (Cunningham et al., ""Spot 14" Protein: A Metabolic Integrator in Normal and Neoplastic Cells," Thyroid 8:815-25 (1998), which is hereby incorporated by reference in its entirety). In a survey of bovine tissue, S 14 was highly expressed in liver and adipose tissue and moderately expressed in mammary tissue. In addition, previous studies established that S 14 expression was up-regulated in mammary tissue during lactation in humans (Wells et al., "Expression of "Spot 14" (THRSP) Predicts Disease Free Survival in Invasive Breast Cancer: Immunohistochemical Analysis of a New Molecular Marker," Breast Cancer Res Treat 98(2) :231-240 (2006), which is hereby incorporated by reference in its entirety) and mice (Rudolph et al., "Metabolic Regulation in the Lactating Mammary Gland: A Lipid Synthesizing Machine," Physiol Genomics 28:323 - 336 (2007), which is hereby incorporated by reference in its entirety), and this was also observed for bovine mammary tissue. MIGlIPl, a protein with sequence homology to S 14, has been proposed as a lipogenic factor with possible redundancy to S 14 function (Zhu et al., "The Spot 14 Protein is Required for de novo Lipid Synthesis in the Lactating Mammary Gland," Endocrinology 146:3343-50 (2005), which is hereby incorporated by reference in its entirety). However, in the present invention the lack of treatment effects on mammary expression of MIGlIPl and the tissue profile of MIGlIPl offer little support for a role in regulation of lipogenesis in the lactating bovine mammary gland.

[00083] The exact biochemical function of S 14 has not been established.

Originally identified as a protein acutely responsive to thyroid hormone (Cunningham et al., ""Spot 14" Protein: A Metabolic Integrator in Normal and Neoplastic Cells," Thyroid 8:815-25 (1998), which is hereby incorporated by reference in its entirety), S 14 is primarily a nuclear protein that forms homo- and hetero-dimers (Cunningham et al, ""Spot 14" Protein: A Metabolic Integrator in Normal and Neoplastic Cells," Thyroid 8:815-25 (1998), which is hereby incorporated by reference in its entirety) and interacts with transcription factors (Compe et al., "Spot 14 Protein Interacts and Co-operates with Chicken Ovalbumin Upstream Promoter-transcription Factor 1 in the Transcription of the L-type Pyruvate Kinase Gene Through a Specificity Protein 1 (SpI) Binding Site," Biochem J 358: 175-83 (2001), which is hereby incorporated by reference in its entirety). The S 14 promoter also contains a SRE (Jump et al., "Functional Interaction Between Sterol Regulatory Element-binding Protein- Ic, Nuclear Factor Y, and 3,5,3'-Triiodothyronine Nuclear Receptors," J Biol Chem

276:34419-27 (2001), which is hereby incorporated by reference in its entirety) and expression is highly responsive to nSREBPl (Martel et al., "S14 Protein in Breast Cancer Cells: Direct Evidence of Regulation by SREBP-Ic, Superinduction with Progestin, and Effects on Cell Growth," Exp Cell Res 312:278-88 (2006), which is hereby incorporated by reference in its entirety). Perhaps the strongest evidence of S 14 function comes from studies of rat heptocytes where transfection with S 14 antisense oligonucleotide prevented expression of lipogenic enzymes (Kinlaw et al., "Direct Evidence For a Role of the "Spot 14" Protein in the Regulation of Lipid Synthesis," J Biol Chem 270:16615-18 (1995) and Brown et al., ""Spot 14" Protein Functions at the Pretranslational Level in the Regulation of Hepatic Metabolism by Thyroid Hormone and Glucose," J Biol Chem 272:2163-6 (1997), which are hereby incorporated by reference in their entirety). In addition, mice with a partial S 14 knock-out have decreased milk fat concentration due to decreased de novo fatty acid synthesis, although surprisingly, activities of mammary lipogenic enzymes were unaltered (Zhu et al., "The Spot 14 Protein is Required for de novo Lipid Synthesis in the Lactating Mammary Gland," Endocrinology 146:3343-50 (2005), which is hereby incorporated by reference in its entirety).

[00084] It was found that mammary expression of S 14 was down-regulated during diet-induced MFD and treatment with trans-10, cis-\2 CLA. This is the first study of the regulation of mammary expression and the first to examine the regulatory role of CLA. However, hepatic expression of S 14 has been extensively investigated and shown to be responsive to a range of metabolic hormones and dietary nutrients, including PUFA (Cunningham et al., ""Spot 14" Protein: A Metabolic Integrator in Normal and Neoplastic Cells," Thyroid 8:815-25 (1998), which is hereby incorporated by reference in its entirety). Trans-10, cis-\2 CLA reduces body fat accretion in several species (Wang et al., "Conjugated Linoleic Acid and Obesity Control: Efficacy and Mechanisms," IntJ Obes Relat Metab Disord 28:941-55 (2004), which is hereby incorporated by reference in its entirety), although the CLA dose in studies of its anti-obesity affect is substantially greater than that required to reduce milk fat synthesis (0.5-2.0% of diet in rodents vs. 4.5xlO^"4% of diet in the present invention). Publicly available microarray data from mice was used (House et al., "Functional Genomic Characterization of Delipidation Elicited by trans-10, cώ-12-conjugated Linoleic Acid (?10,cl2-CLA) in a Polygenic Obese Line of Mice," Physiol Genomics 21 :351-61 (2005); Hargrave et al., "Effect of Dietary Conjugated Linoleic Acid on Adiposity and the Adipose-transcriptome," J Dairy Sci 88(Suppl. l):280 (2005), which are hereby incorporated by reference in their entirety) and demonstrated that CLA treatment resulted in a significant reduction in expression of S 14 in adipose tissue. Thus, S 14 may be more broadly implicated in the mechanism by which CLA is able to effect lipid metabolism.

[00085] Multivariate analysis in the present study revealed a significant relationship in bovine mammary tissue between expression of S 14 and expression of FASN and LPL (R² = 0.86 and 0.42, respectively; Table 6). Altered expression of S 14 has also been associated with other unique phenotypes involving regulation of fat synthesis. For example, abnormalities in the regulation of adipose S 14 expression have been reported in obese subjects (Kirschner et al., "Adipose S 14 mRNA is Abnormally Regulated in Obese Subjects," Thyroid 9:143-8 (1999), which is hereby incorporated by reference in its entirety). Likewise, gene expression profiling identified differential expression of S 14 in livers of chickens selected for growth (Cogburn et al., "Systems-wide Chicken DNA Microarrays, Gene Expression Profiling, and Discovery of Functional Genes," Poult Sci 82:939-51 (2003), which is hereby incorporated by reference in its entirety), adipose tissue of chickens selected for adiposity (Carre et al., "Development of 112 Unique Expressed Sequence Tags from Chicken Liver Using an Arbitrarily Primed Reverse Transcriptase-polymerase Chain Reaction and Single Strand Conformation Gel Purification Method," Anim Genet 32:289-97 (2001), which is hereby incorporated by reference in its entirety), and muscle of cattle that differ in marbling (Wang et al., "Transcriptional Profiling of Skeletal Muscle Tissue from Two Breeds of Cattle," Mamm Genome 16:201-10 (2005), which is hereby incorporated by reference in its entirety). Lastly, S 14 is also a component of the lipogenic phenotype observed in aggressive breast cancers (Wells et al., "Expression of "Spot 14" (THRSP) Predicts Disease Free Survival in Invasive Breast Cancer: Immunohistochemical Analysis of a New Molecular Marker," Breast Cancer Res Treat 98(2) :231-240 (2006), which is hereby incorporated by reference in its entirety) and knockdown or overexpression of S 14 result in corresponding effects on breast cancer cell growth (Martel et al., "S14 Protein in Breast Cancer Cells: Direct Evidence of Regulation by SREBP-Ic, Superinduction with Progestin, and Effects on Cell Growth," Exp Cell Res 312:278-88 (2006), which is hereby incorporated by reference in its entirety). The anti-carcinogenic affect of trans-10, cis-Yl CLA has been well characterized for in vitro and in vivo models (Ip et al., "Prevention of Mammary Cancer with Conjugated Linoleic Acid: Role of the Stroma and the Epithelium," J Mammary Gland Biol Neoplasia 8:103-18 (2003), which is hereby incorporated by reference in its entirety). Based on the present invention, the possible role of S 14 in the mechanism merits examination.

TABLE 6

Multivariate analysis of the relationship of the mRNA for regulatory proteins to expression of fatty acid synthase and lipoprotein lipase¹

¹ The relationship between expression of regulatory proteins (thyroid hormone responsive spot 14 = S 14, sterol response element binding protein = SREBP, and insulin induced gene = INSIG) and lipogenic enzymes (fatty acid synthase = FASN and lipoprotein lipase = LPL) was tested by including the mRNA abundance for each gene as a fixed effect in a model that also included the random effect of cow and fixed effect of period. RMSE = root mean square error and EST = estimate.

[00086] In conclusion, decreased expression of SREBPl and proteins associated with SREBPl activation during MFD combined with the presence of SREBP response elements in lipogenic genes down-regulated during MFD, provide strong evidence for SREBPl as a central signaling pathway regulating fatty acid synthesis in the lactating bovine mammary gland. Furthermore, down- regulation of S 14 during diet induced MFD and trans- 10, cis-Yl CLA treatment is consistent with a role for S 14 in mammary fatty acid synthesis, possibly as a SREBPl secondary cellular signal or a lipogenic factor.

[00087] Although the invention has been described in detail, for the purpose of illustration, it is understood that such detail is for that purpose and variations can be made therein by those skilled in the art without departing from the spirit and scope of the invention which is defined by the following claims.

Claims

WHAT IS CLAIMED:

1. A method of screening cancer patients to determine whether the form of cancer they have is suitable for treatment with conjugated linoleic acids, said method comprising: providing a tissue or fluid sample from the patient; evaluating the sample for tumor cells with a disregulation of lipid metabolism; and identifying patients with tumor cells with a disregulation of lipid metabolism as having a form of cancer suitable for treatment with conjugated linoleic acids.

2. The method of claim 1, wherein the disregulation of lipid metabolism is in the form of overexpression of fat synthesis enzymes, high lipogenic activity, or accumulation of lipids in tumor cells.

3. A method of treating a patient for cancer comprising: selecting a patient having tumor cells with a disregulation of lipid metabolism and administering conjugated linoleic acids to the selected patient under conditions effective to treat cancer.

4. The method of claim 3, wherein the patient has breast cancer, prostate cancer, ovarian cancer, colon cancer, endometrium cancer, lung cancer, bladder cancer, stomach cancer, osteophagus cancer, oral tongue cancer, oral cavity cancer, skin cancer, mesotheliomas, retinoblastomas, and/or nephroblastomas .

5. The method of claim 3, wherein the conjugated linoleic acids is selected from the group consisting of the trans-10, cis-12 conjugated linoleic acid, cis-9, trans- 11 conjugated linoleic acid, and mixtures thereof.

6. The method of claim 3, wherein said selecting comprises: providing a tissue or fluid sample from the patient; evaluating the sample for tumor cells with a disregulation of lipid metabolism; and identifying patients with tumor cells with a disregulation of lipid metabolism as having a form of cancer suitable for treatment with conjugated linoleic acids.

7. A method of screening cancer patients to determine whether the form of cancer they have is suitable for treatment with conjugated linoleic acids, said method comprising: providing a tissue or fluid sample from the patient; evaluating the sample for overexpression of the Thyroid Hormone Responsive Spot 14 gene; and identifying patients who overexpress the Thyroid Hormone Responsive Spot 14 gene as having a form of cancer suitable for treatment with conjugated linoleic acids.

8. A method of treating a patient for cancer comprising: selecting a patient overexpressing the Thyroid Hormone Responsive Spot 14 gene and administering conjugated linoleic acids to the selected patient under conditions effective to treat cancer.

9. The method of claim 8, wherein the patient has breast cancer, prostate cancer, ovarian cancer, colon cancer, endometrium cancer, lung cancer, bladder cancer, stomach cancer, osteophagus cancer, oral tongue cancer, oral cavity cancer, skin cancer, mesotheliomas, retinoblastomas, and/or nephroblastomas .

10. The method of claim 8, wherein the conjugated linoleic acids is selected from the group consisting of trans- 10, cis-12 conjugated linoleic acid, cis-9, trans- 11 conjugated linoleic acid, and mixtures thereof.

11. The method of claim 8, wherein said selecting comprises: providing a tissue or fluid sample from the patient; evaluating the sample for overexpression of the Thyroid

Hormone Responsive Spot 14 gene; and identifying patients who overexpress the Thyroid Hormone

Responsive Spot 14 gene as having a form of cancer suitable for treatment with conjugated linoleic acids.

12. A method of screening cancer patients to determine whether the form of cancer they have is suitable for treatment with conjugated linoleic acids, said method comprising: providing a tissue or fluid sample from the patient; evaluating the sample for overexpression of the Sterol Responsive Element Binding Protein gene; and identifying patients who overexpress the Sterol Responsive

Element Binding Protein gene as having a form of cancer suitable for treatment with conjugated linoleic acids.

13. A method of treating a patient for cancer comprising: selecting a patient overexpressing the Sterol Responsive

Element Binding Protein gene and administering conjugated linoleic acids to the selected patient under conditions effective to treat cancer.

14. The method of claim 13, wherein the patient has breast cancer, prostate cancer, ovarian cancer, colon cancer, endometrium cancer, lung cancer, bladder cancer, stomach cancer, osteophagus cancer, oral tongue cancer, oral cavity cancer, skin cancer, mesotheliomas, retinoblastomas, and/or nephroblastomas .

15. The method of claim 13, wherein the conjugated linoleic acids are selected from the group consisting of trans-10, cis-12 conjugated linoleic acid, cis-9, trans- 11 conjugated linoleic acid, and mixtures thereof.

16. The method of claim 13, wherein said selecting comprises: providing a tissue or fluid sample from the patient; evaluating the sample for overexpression of the Sterol Responsive Element Binding Protein gene; and identifying patients who overexpress the Sterol Responsive Element Binding Protein gene as having a form of cancer suitable for treatment with conjugated linoleic acids.

17. A method of screening obesity patients to determine whether the form of obesity they have is suitable for treatment with conjugated linoleic acids, said method comprising: providing a tissue or fluid sample from the patient; evaluating the sample for overexpression of the Thyroid Hormone Responsive Spot 14 gene; and identifying patients who overexpress the Thyroid Hormone Responsive Spot 14 gene as having a form of obesity suitable for treatment with conjugated linoleic acids.

18. A method of treating a patient for obesity comprising: selecting a patient overexpressing the Thyroid Hormone Responsive Spot 14 gene and administering conjugated linoleic acids to the selected patient under conditions effective to treat obesity.

19. The method of claim 18, wherein the conjugated linoleic acids are selected from the group consisting of trans-10, cis-12 conjugated linoleic acid, cis-9, trans- 11 conjugated linoleic acid, and mixtures thereof.

20. The method of claim 18, wherein said selecting comprises: providing a tissue or fluid sample from the patient; evaluating the sample for overexpression of the Thyroid Hormone Responsive Spot 14 gene; and identifying patients who overexpress the Thyroid Hormone Responsive Spot 14 gene as having a form of obesity suitable for treatment with conjugated linoleic acids.

21. A method of screening obesity patients to determine whether the form of obesity they have is suitable for treatment with conjugated linoleic acids, said method comprising: providing a tissue or fluid sample from the patient; evaluating the sample for overexpression of the Sterol Responsive Element Binding Protein gene; and identifying patients who overexpress the Sterol Responsive Element Binding Protein gene as having a form of obesity suitable for treatment with conjugated linoleic acids.

22. A method of treating a patient for obesity comprising: selecting a patient overexpressing the Sterol Responsive Element Binding Protein gene and administering conjugated linoleic acids to the selected patient.

23. The method of claim 22, wherein the conjugated linoleic acids is selected from the group consisting of trans-10, cis-12 conjugated linoleic acid, cis-9, trans- 11 conjugated linoleic acid, and mixtures thereof.

24. The method of claim 22, wherein said selecting comprises: providing a tissue or fluid sample from the patient; evaluating the sample for overexpression of the Sterol Responsive Element Binding Protein gene; and identifying patients who overexpress the Sterol Responsive Element Binding Protein gene as having a form of obesity suitable for treatment with conjugated linoleic acids.