Changes in prostate gene expression in men undergoing an intensive nutrition and lifestyle intervention Dean Ornish*†‡, Mark Jesus M. Magbanua§, Gerdi Weidner*, Vivian Weinberg¶, Colleen Kemp*, Christopher Green§, Michael D. Mattie§, Ruth Marlin*, Jeff Simkoʈ, Katsuto Shinohara§, Christopher M. Haqq§ and Peter R. Carroll§ §Department of Urology, The Helen Diller Family Comprehensive Cancer Center, and ʈDepartment of Pathology, University of California, 2340 Sutter Street, San Francisco, CA 94115; *Preventive Medicine Research Institute, 900 Bridgeway, Sausalito, CA 94965; †Department of Medicine, School of Medicine, University of California, 505 Parnassus Avenue, San Francisco, CA 94143; and ¶Biostatistics Core, The Helen Diller Family Comprehensive Cancer Center, University of California, 513 Parnassus Avenue, Box 0127, San Francisco, CA 94143 Communicated by J. Craig Venter, The J. Craig Venter Institute, Rockville, MD, April 2, 2008 (received for review February 13, 2008) Epidemiological and prospective studies indicate that comprehensive indolent low-risk prostate cancers, defined by strict clinical and lifestyle changes may modify the progression of prostate cancer. pathologic criteria designed to minimize the risk for metastatic However, the molecular mechanisms by which improvements in diet disease as a result of study participation (9). The 30 men who and lifestyle might affect the prostate microenvironment are poorly enrolled did not undergo surgery or radiation therapy to treat their understood. We conducted a pilot study to examine changes in low-risk tumors; rather, they underwent comprehensive lifestyle prostate gene expression in a unique population of men with low-risk changes (low-fat, whole-foods, plant-based nutrition; stress man- prostate cancer who declined immediate surgery, hormonal therapy, agement techniques; moderate exercise; and participation in a or radiation and participated in an intensive nutrition and lifestyle psychosocial group support). Participants donated serial prostate intervention while undergoing careful surveillance for tumor pro- needle biopsies at baseline and after 3 months of the lifestyle gression. Consistent with previous studies, significant improvements intervention, from which nanogram quantities of mRNA were in weight, abdominal obesity, blood pressure, and lipid profile were purified. At the time this clinical trial began, commercial expression observed (all P < 0.05), and surveillance of low-risk patients was safe. array platforms were not sensitive to nanogram RNA quantities. Gene expression profiles were obtained from 30 participants, pairing Therefore, a reproducible linear RNA amplification and printed RNA samples from control prostate needle biopsy taken before cDNA array platform was used, as in our previous studies of intervention to RNA from the same patient’s 3-month postinterven- melanoma (10), where subsequent studies have confirmed the tion biopsy. Quantitative real-time PCR was used to validate array validity of the gene expression findings (11, 12). Furthermore, observations for selected transcripts. Two-class paired analysis of quantitative real-time PCR (QRT-PCR) was used to provide initial global gene expression using significance analysis of microarrays confirmation of the study results, comparing in pairwise fashion detected 48 up-regulated and 453 down-regulated transcripts after each man’s postintervention to his own preintervention sample. the intervention. Pathway analysis identified significant modulation This article examines the relationship of comprehensive diet and of biological processes that have critical roles in tumorigenesis, lifestyle changes to gene expression in the prostate. including protein metabolism and modification, intracellular protein Results traffic, and protein phosphorylation (all P < 0.05). Intensive nutrition and lifestyle changes may modulate gene expression in the prostate. In the GEMINAL study, 273 men were screened, 96 declined to Understanding the prostate molecular response to comprehensive participate, 146 did not meet inclusion criteria, and 31 were lifestyle changes may strengthen efforts to develop effective preven- enrolled. The participants’ demographics included a mean age of tion and treatment. Larger clinical trials are warranted to confirm the 62.3 years (range 49–80) and a mean PSA level of 4.8 ng/ml (range results of this pilot study. 0.5–21.4) on the day of the initial biopsy. As expected from the trial eligibility criteria, all patients had a Gleason score of 6. Eighty-four exercise ͉ lifestyle changes ͉ prostate cancer ͉ SHOC2 ͉ stress management percent of the men identified their ethnicity as Caucasian, 9% Hispanic, 3% Asian, and 4% African-American. Two-thirds of the men were married, and 72% were currently employed. pidemiological evidence (1, 2) and migrant studies (3) indicate Trial eligibility required PSA Յ10 [or PSA Յ15 if the patient had Ethat the incidence of clinically significant prostate cancer is a history of benign prostatic hyperplasia (BPH)] at the time of much lower in parts of the world where people eat a predominantly screening. One outlier patient with a history of BPH and a prostate low-fat, plant-based diet. We (4, 5) and others (6) have shown volume of 90 cc, as assessed by transrectal ultrasound, had screening MEDICAL SCIENCES previously that diet and lifestyle interventions in men with early- stage prostate cancer decrease prostate-specific antigen (PSA) and decrease the rate of PSA increase. These studies provided some Author contributions: D.O. and M.J.M.M. contributed equally to this work; D.O., G.W., evidence that comprehensive lifestyle changes may have therapeu- R.M., C.M.H., and P.R.C. designed research; D.O., M.J.M.M., C.K., C.G., M.D.M., R.M., J.S., tic potential in early prostate cancers. However, although these K.S., C.M.H., and P.R.C. performed research; M.J.M.M., V.W., C.G., and C.M.H. analyzed interventions are associated with decreased circulating insulin-like data; and D.O., M.J.M.M., G.W., V.W., C.K., C.M.H., and P.R.C. wrote the paper. growth factor 1 (IGF1) (7), and although serum from men after Conflict of interest statement: Although none of the following is a true conflict of interest, in the spirit of full disclosure, D.O. writes general-interest books on preventive medicine, intervention has reduced the ability to stimulate prostate cell-line receives lecture honoraria, and consults with food companies to make more healthful growth in vitro (4), the actual molecular effects of these interven- foods. tions in prostate tissue have not been previously examined. Freely available online through the PNAS open access option. Many men with indolent prostate cancers detected by PSA Data deposition: The data reported in this paper have been deposited in the Gene screening will not exhibit disease progression during their lifetime; Expression Omnibus (GEO) database, www.ncbi.nlm.nih.gov/geo (accession no. GSE10306). their treatment and associated side effects are unnecessary (8). We ‡To whom correspondence should be addressed. E-mail: [email protected]. report here the results of the Gene Expression Modulation by This article contains supporting information online at www.pnas.org/cgi/content/full/ Intervention with Nutrition and Lifestyle (GEMINAL) study, a 0803080105/DCSupplemental. prospective single-arm pilot clinical intervention study in men with © 2008 by The National Academy of Sciences of the USA www.pnas.org͞cgi͞doi͞10.1073͞pnas.0803080105 PNAS ͉ June 17, 2008 ͉ vol. 105 ͉ no. 24 ͉ 8369–8374 Downloaded by guest on September 30, 2021 Table 1. Cardiovascular risk factors and psychological functioning at baseline and 3 months 95% CI of change Baseline, 3 months, Mean change, Variable Mean Ϯ SD Mean Ϯ SD 3-month baseline Lower Upper P* Cardiovascular risk factors BMI 26.5 Ϯ 3.6 23.9 Ϯ 3.0 Ϫ2.6 Ϫ3.0 Ϫ2.2 Ͻ0.001 Systolic blood pressure, mmHg 129.5 Ϯ 15.1 120.3 Ϯ 12.6 Ϫ9.2 Ϫ13.8 Ϫ4.6 Ͻ0.001 Diastolic blood pressure, mmHg 68.2 Ϯ 10.7 62.8 Ϯ 10.3 Ϫ5.4 Ϫ8.4 Ϫ2.3 Ͻ0.002 LDL, mg/dl 121.7 Ϯ 28.2 87.6 Ϯ 25.1 Ϫ34.2 Ϫ41.6 Ϫ26.8 Ͻ0.001 HDL, mg/dl 48.5 Ϯ 12.7 40.2 Ϯ 12.1 Ϫ8.3 Ϫ11.5 Ϫ5.2 Ͻ0.001 Total cholesterol, mg/dl 191.7 Ϯ 33.1 146.5 Ϯ 31.6 Ϫ45.2 Ϫ54.4 Ϫ36.0 Ͻ0.001 LDL/HDL ratio 2.7 Ϯ 1.0 2.3 Ϯ 0.8 Ϫ0.4 Ϫ0.6 Ϫ0.1 Ͻ0.002 Triglycerides, mg/dl 107.0 Ϯ 72.1 93.6 Ϯ 46.3 Ϫ13.4 Ϫ32.3 5.5 0.158 C-reactive protein (ln) 0.12 Ϯ 1.1 Ϫ0.14 Ϯ 1.3 Ϫ0.26 Ϫ0.64 0.12 0.168 Psychological functioning SF-36 Mental component summary 49.7 Ϯ 11.8 56.2 Ϯ 5.5 6.5 2.6 10.3 Ͻ0.003 Physical component summary 54.7 Ϯ 4.9 55.4 Ϯ 5.5 0.8 Ϫ1.5 3.0 0.485 Impact of Event Scale Intrusive thoughts 6.2 Ϯ 5.6 3.7 Ϯ 4.8 Ϫ2.5 Ϫ4.2 Ϫ0.8 Ͻ0.01 Avoidance 10.1 Ϯ 6.8 7.1 Ϯ 5.8 Ϫ3.0 Ϫ5.9 Ϫ0.1 Ͻ0.05 *Paired-samples t test, two-tailed. PSAs of 11.6–12.1 ng/ml, meeting eligibility criteria, but had a PSA score for physical health (Ͼ55.7) and near the lowest-quartile cutoff of 21.4 ng/ml on the day of initial biopsy. After the 3-month score for mental health (Ͻ49.9) (24). The psychological distress intervention, this patient’s PSA decreased to 13.9 ng/ml, and his reported in this sample was comparable with the level of distress prostate volume decreased to 75 cc. All other participants had reported by other men diagnosed with prostate cancer (mean screening and initial biopsy-day PSA values of Ͻ10 ng/ml.