# Presentation: 516820

## RFA 03-001 Collaborations on Nutritional Modulation of Genetic Pathways Leading to Cancer

**John Milner, Ph.D. Chief**
**Nutrition Science Research Group**
**Division of Cancer Prevention**
**m**_[ilnerj](mailto:Milnerj@mail.nih.gov)__[@mail.nih.gov](mailto:Milnerj@mail.nih.gov)_
**301-496-0118**

## Aim of U54 RFA

**interdisciplinary**** ****collaborations**** **
**to expand and facilitate research to define the precise role of diet in cancer prevention by elucidating the influence of bioactive food components on genetic and epigenetic events involved with cancer.****  **

**Aim of U54 RFA**

## Background for Concept

**Inconsistencies**** suggest complexity in nutrition cancer relationship, i.e. dependency on intrinsic and extrinsic factors**
**Need ****to move beyond observations**

**Background for Concept**

## Numerous Dietary Components Can Potentially Alter Genetic and Epigenetic Events

- Essential Nutrients
- Phytochemicals or allelochemicals (bioactive secondary compounds) in plants. Carotenoids, Flavonoids, Indoles, Isothiocyanates, Allyl  Sulfur, etc.
- Zoochemicals- Conjugated linoleic acid, n-3 fatty acids

## Processes Likely Influenced by Nutrients

**Carcinogen Metabolism**

**Cell Signaling**

** ****Apoptosis**

**Nutrients**

**Differentiation**

**Cell Cycle**

## Cancer Risk

**Dietary Constituent**

**Absorbed**
**Dose **

**Biologically Effective Dose**

** **

** **

**Inactive Metabolite**

**Altered **

**Altered **

**Structure **

**Structure **

**Function**

**Function**

**Cancer Risk**

**Genetic**
**Pathways**

**Early Biologic**
**Effect**

## Effect of DHA (2 g/ml) on Tumor Growth

- Effect of DHA (2 g/ml) on Tumor Growth

- MDA 231 cell lack mrg gene (n3 transport protein gene)
- Transfection of MRG-231 with mrg gene increases sensitivity to n-3

- Shi et al Cancer Res. 60:6482, 2000

## < 300 mg 

- < 300 mg

- Caffeine

- > 300 mg

- 6

- -12

- 0

- -6

- Vitamin D Receptor Genotype

- TT        Tt       tt

- TT       Tt        tt

- Spine Bone Mass Density (%)

**Influence of Caffeine on Bone Mass May Depend on Genes**

**Rapuri  et al. Am J Clin Nutr 2001 Nov;74(5):694-700**

**Notes:**
BACKGROUND: The role of caffeine as a risk factor for bone loss is controversial. OBJECTIVE: Our goals were 1) to compare in both a cross-sectional study and a 3-y longitudinal study the bone mineral density (BMD) of postmenopausal women consuming high or low amounts of caffeine and 2) to study the interaction between caffeine intake, vitamin D receptor (VDR)polymorphism, and BMD in the longitudinal study. DESIGN: The results are derived from cross-sectional measurements of BMD in 489 elderly women (aged 65-77 y) and from longitudinal measurements made in 96 of these women who were treatedwith a placebo for 3 y. Changes in BMD were adjusted for confounding factors and were compared between groups with eitherlow (< or =300 mg/d) or high (>300 mg/d) caffeine intakes and between the VDR genotype subgroups of the low- andhigh-caffeine groups. RESULTS: Women with high caffeine intakes had significantly higher rates of bone loss at the spine than did those with low intakes (-1.90 +/- 0.97% compared with 1.19 +/- 1.08%; P = 0.038). When the data were analyzed according to VDR genotype and caffeine intake, women with the tt genotype had significantly (P = 0.054) higher rates of boneloss at the spine (-8.14 +/- 2.62%) than did women with the TT genotype (-0.34 +/- 1.42%) when their caffeine intake was>300 mg/d. CONCLUSIONS: Intakes of caffeine in amounts >300 mg/d ( approximately 514 g, or 18 oz, brewed coffee) accelerate bone loss at the spine in elderly postmenopausal women. Furthermore, women with the tt genetic variant of VDRappear to be at a greater risk for this deleterious effect of caffeine on bone.

## Effect of Nrf2 genotype on induction of GST and NQO1 activities by oltipraz in mouse liver

**Effect of Nrf2 genotype on induction of GST and NQO1 activities by oltipraz in mouse liver**
**Ramos-Gomez *****et al.****** *****Proc Natl Acad Sci U S A 2001;98:3410-5**

## Molecular Targets for Selenium

- Se

- Glutathione Peroxidase

- NFB

- gadd

- Cyclin  E

- Glutathione S-transferase

- Cdk2 Kinase

- Lipoxygenase

- Protein Kinase C

**Credentialing of nutrients and targets- the future?**
**Credentialing is defined as genetic and epigenetic changes  that bring about a phenotypic change**

## Genetic and Epigenetic Effects of Folate Depletion

**Altered Tissue **
**Folate Distribution**

**Altered**
** ****DNA/RNA **
**Methylation**

**Changes in Tissue **
**Folate Status**

**dNTP Pool **
**Imbalance**

**Disruption of**
**DNA Integrity**

**Inadequate**
**Folate Intake**

**Disruption in**
**DNA Repair**

**Increased**
**DNA Damage**

**Altered Gene **
**Expression**

**Increased**
**Uracil**
**Misincorporation**

## Concept Builds On

**Recommendations by:**
**The 1999 Nutrition Implementation Group**** report on “New Directions for Nutritional Research at the National Cancer Institute.”  **
**Small group of external scientists**** convened by the Division of Cancer Prevention, National Cancer Institute (NCI) in 2000**** **
**NCI 2001 Bypass Budget**** **

## Nutrition

**Molecular **
**Targets**

**Defining the Signatures of **
**Cancer Cells**

- Cancer  Imaging

**Genes and the**
**Environment**

- Cancer
- Communications

- Research on
- Tobacco &
- Tobacco
- Related
- Cancers

**Nutrition**

## Basic

- Molecular Biology

- Genetics

- Biostatistics

- Physiology

- Clinical

- Population

- Nutrition

- Chemistry

## Merit of An Interdisciplinary Approach to Resolving Nutrition and Cancer Prevention Issues 

**Embody new and innovative preclinical and clinical approaches to nutrition and genetics**
**Provide a mechanism for collaborative and integrative research programs to delineate how specific dietary components influence genetic pathways**
**Build on resources and linkages within NCI**

## Support Mechanism for Phase I and II

**I.  P20 Planning Grant--**** ****Supports planning for new programs.  Such awards have been particularly useful in the development of Cancer Centers and SPORES.**** **
**II. U54 Cooperative Specialized Center Award Mechanism--****  ****Used to enhance the conduct of already funded research projects through collaborative activities, supports full range of research development from very basic to clinical**

## Phase II:

** ****Large-Scale Collaborative Project**

**5.0 year program**
**Variable Components**
**Essential**
**	****	****Administrative**
**	****	****Collaborative Projects**
**	****Possible**
**	****	****Pilot Projects**
**	****	****Core resources**

## Incomplete List of Areas of Interest

- Characterization of molecular events that govern the ability of specific nutrients to alter cell cycle checkpoints;
- Credentialing of target receptors for cancer prevention that are modified by dietary constituents;
- Methylation patterns that are influenced by dietary manipulations that influence gene expressions and cellular phenotypes;

## Some Other Areas of Interest

- DNA repair mechanisms influenced by dietary constituents;
- Intercellular communications that are influenced by bioactive food components
- Signaling pathways that regulate cancer growth, development, differentiation and apoptosis as regulated by dietary components;

## Possible Structure of Team (Decision Pathway)

**Possible Structure of Team ****(Decision Pathway)**

**Administrative Committee**
**PI  and 2-3 Active Investigators**

**Collaborative Project**

**3-4 components**

**Core Resources**

**Pilot Project**

**1-2 projects**

**ie. Data management, analytical,**
**transgenic**

## The required components are as follows:

- Selecting one or more cancer-relevant genetic pathways for which evidence exists that it may be modified by nutrients.
-  Establishing effective and synergistic collaborations that enhance the capabilities of individuals and leverage available resources for the Team.
-  Evaluating and validating assays, probes, and other techniques in preclinical models, for assessing genetic effects, changes in biochemical processes downstream from the main target; pharmacokinetics or pharmacodynamics of bioactive food components and for the establishment of quantity-temporal relationships.

## o  Identifying how nutrients directed at high-priority genetic pathways/targets affect endpoints/biomarkers that may be used as indicators in a clinical setting.  A Team may choose to achieve translational goal of their research by including 

- acquisition of relevant cells (e.g., from cancers, preinvasive neoplastic lesions, or fields at risk for cancer) by biopsy, aspiration or plasmapheresis,
-  Techniques for imaging molecular and cellular processes such as techniques based on radiotracers or ultrasound or optical imaging, or
-  other indicators as appropriate.

## Exploring new scientific insights into mechanisms and determinants of response

- Exploring new approaches to early clinical trials methodology; for example, the use of alternate trial designs and endpoints, or exploring designs based on molecular classifications of tumor rather than histology per se.
- Responsiveness to share information via publications, meetings and support of training activities and to disseminate new reagents and techniques discovered under this initiative to other investigators.

## Clinical Trails

- Each application must propose an institutional review process for selecting Pilot Projects for funding that generate feasibility data and have the most promising translational research potential.

**Clinical Trails**

## may not 

- may not
- exceed $125,000 or 10 percent (whichever is larger) of the direct cost budget proposed for any 1 year.

## Reason for NCI Involvement 

**To support, stimulate and expedite the awardee’s activities by working cooperatively in a partner role, but not assuming direction, prime responsibility, or a dominant role.  **
**Help by promoting linkages within the Project’s Collaborative team, by fostering linkages with other groups associated with NCI and by providing information about other resources available at NCI**

## Planning Committee

- Planning Committee is expected to meet twice yearly
- Provide oversight of projects and identify areas of collaboration across awards

**Planning Committee**

## Budget ($ Million)

**	****  ****2001      2002    2003     2004     2005      ****Total**
**Phase I      ****	****1.20 ****	****	****	****	****	**** 1.2****	**
**Phase II     ****	****0.0****	****10.5 ****	****10.8****	****11.1****	****11.5****	****  43.9****	**
**	**
**Total****	**** 1.20         10.5 ****	****10.8****	**** 11.1****	****  11.5****	****  45.1**

**12 Awards for Phase I**
**6 Awards for Phase II**
**Review by a Division of Extramural Activities (DEA)/NCI ad hoc review group**