Din é College / Mayo Clinic: Developing Cancer Researchers

1. Diné College / Mayo Clinic:Developing Cancer Researchers Edward R. Garrison, PhD, MPH (PI) Diné College Christi A. Patten, PhD (PI) Mayo Clinic Rochester Cynthia Claus, MPH (Co-PI) Mayo Clinic Scottsdale P20 Research Training Grant Funded by the National Cancer Institute

3. Mayo Rochester Mayo Scottsdale Mayo Jacksonville

4. Rationale To increase the number of Navajo students entering the health care field by providing opportunities to obtain experience in a scientific field of study. This will ultimately reduce cancer risk behaviors and decrease the mortality rate for the Native American population.

5. Project Aims • Aim 1: Cancer Education - Faculty • Aim 2: Cancer Education - Student • Aim 3: Cancer Research Training- Student Research Training

6. Aim 3: Student Cancer Research Training • Identify, evaluate and follow Diné College students • Identify Mayo mentors and/or labs • The Native CREST summer cancer research training program was piloted at Mayo Rochester in 2008 • Refine research training based on student and mentor feedback

7. Aim 3: Student Cancer Research Training (cont.) • Implement program for Diné College students at Mayo Clinic Rochester again in 2010 (2-3 students for 10 weeks) • Develop the process by which students present their research experiences back to their communities • Develop long-term tracking system

8. GTP GDP PO4 Rab5 Rin2 Regulation of HGF/c-Met Signaling Latoya Dale1,2, Sandra Severson1, Jin Tomshine1 and Bruce Horazdovsky1. 1 Department of Biochemistry and Molecular Biology. Mayo Clinic, Rochester, MN. 2 Native CREST-SURF Program. Diné College. Shiprock, NM. ABSTRACT Rab5 is a critical regulator of early endocytic events. Rab5 is a GTPase that is activated through binding GTP and inactivated through hydrolyzing GTP to GDP. Rab5 is activated by guanine nucleotide exchange factors (GEFs) that promote release of GDP to permit binding of GTP. Proteins containing Vps9 domains serve as the GEFs for Rab5. Most Vps9 domain proteins contain additional signal transduction domains. The Rin proteins (Rin1, Rin2, and Rin3) harbor SH2 domains, proline-rich regions and Ras association domains in addition to the Vps9 domain. These Rin proteins are therefore thought to integrate signaling from receptor tyrosine kinases with endocytosis, and Rin1 has been demonstrated to modulate epidermal growth factor (EGF) signaling and EGF receptor trafficking. The function of Rin2 is less well understood, although Rin2 has been implicated in hepatocyte growth factor (HGF, also known as scatter factor or SF) signaling. HGF is the ligand for the c-MET tyrosine kinase receptor (also referred to as HGFR), and a strong link has been established between hyperactive HGF/c-MET signaling and cancer. To understand the role of Rin2 in HGF signaling and c-MET trafficking, further analyses of HGF-stimulated Erk and Akt activation in human hepatocytes (HuH7) depleted of Rin2. I have also been subcloning Rin2 into a mammalian expression vector for future over-expression experiments. Extension of these studies will help define the role of Rin2 in HGF signaling. Figure 3. RIN2 depletion with shRNA. HuH7 cells were infected with lentivirus encoding RIN2-specific shRNA (H35) or a non-targeting shRNA (NT). RIN2 depletion was determined by Western analysis (N=12). Figure 5. RIN2 Localization. HuH7 cells were transfected with an EGFP-RIN2 expression construct and visualized by fluorescence microscopy after deconvolution using DeltaVision software. Guanine Nucleotide Exchange Factors (GEFs) Figure 4. RIN2 depletion impacts AKT signaling. AKT phosphorylation was monitored by Western analysis following HGF stimulation (10ng/ml) in the presence of RIN2 (H35) or non-targeting (NT) shRNAs. • RESULTS In an effort to understand the role of Rin2 in HGF signaling and c-Met trafficking, we demonstrated the following: • Rin2 was depleted by 43% in HuH7 cells using shRNA (Figure 3). • Rin2 depletion by shRNA impacted AKT signaling in HuH7 cells (Figure 4). • Rin2 was localized to the plasma membrane in HuH7 cells (Figure 5). BACKGROUND Receptors respond to the external environment, transducing signals across the membrane to permit intracellular responses. Appropriate signaling from the receptors allows cell maintenance and/or growth. Inappropriate receptor signaling can lead to either excessive cell proliferation and cancer if too much signaling occurs or loss of cell survival signals and cell death if too little signaling occurs. Receptor endocytosis is one mechanism by which appropriate receptor signaling is maintained. In a general sense, endocytosis serves to maintain an appropriate complement of receptors on the plasma membrane. In addition, endocytosis of activated receptors can directly affect signaling. In receptors systems where signaling can occur at the plasma membrane, endocytosis and subsequent delivery to the lysosome serves to attenuate signaling through degrading the activated receptors. In other receptor systems where delivery to an endosomal compartment is required for signaling, endocytosis promotes signal transduction. Integration of receptor trafficking and receptor signaling is required to maintain appropriate cellular responses. Rab5 Rab5•GDP Rab5•GTP • WORK IN PROGRESS • Coexpress EGFP-Rin2 with Rab5, Rab11, and Rab4 for colocalization studies in HuH7 cells. • Optimize serum starvation conditions for further analyses of HGF-stimulated ERK and AKT activation in HuH7 cells depleted of Rin2. GTPase Activating Proteins (GAPs) Figure 1. Rab5 is Activated by the Vps9-domain proteins Figure 2. Receptor Systems and Vps9 Domain Proteins

9. Diné College SREP: Summer Research Enhancement Program For Cancer and Diabetes Prevention

10. SURF: Summer Undergraduate Research Fellowship

11. Native CRESTCancer Research Experience & Student Training Diné College - Mayo Clinic

12. “Preparing Tomorrow’s Native Researchers Today.” Program Dates: May 29 – August 3, 2012 Application Deadline: February 15, 2012

13. The Native CREST program has been developed to increase the number of students at Diné College preparing to pursue advanced degrees in the health fields by providing opportunities to obtain experience in cancer research and training. Students appointed to this program are interested in the public health and health care fields and are considering a future health-related career through biomedical research, nursing, or pursuing PhD, MD, or MPH degrees.

14. The Program • 10-week summer research training experience at Mayo Clinic in Rochester, Minnesota • Diné College students will work with a Mayo Clinic researcher/faculty member to develop their research interests and experiences • Up to 3 Diné College students for the Summer of 2010 • Students are linked with researcher/faculty who is interested in mentoring a student on a cancer related research project • Program stipend of $4,000 • Students are responsible for arranging their travel, housing and meals (Spirit of EAGLES: ~$1,000)

15. Housing (Rochester, MN): • Grand Kahler Hotel Double:  $750/month  Single:  $1320/month • Residences of Old Town Hall Private Room Apt:  $625/month • Second Street Inn & Suites Single:  $600/month

16. Mayo Clinic Native American Programs Mayo Clinic Cancer Center Rochester, Minnesota Spirit of EAGLES Survivors Education Advocacy Grants Leadership Elders Scholarships Funded by: CA 77410-02

18. Area of Minnesota - 86,943 square miles 12th biggest state in USA

19. Origin of the Name - Minnesota is from a Dakota Sioux Indian word that means "cloudy water" or "sky water" and refers to local rivers. • State Capital - St. Paul (Largest City – Minneapolis) • Major Industries - farming (corn, soybeans, sugar beets, wheat, dairy products), paper pulp, mining (iron ore) • State Nickname - Gopher State. North Star State • State Motto - "L'Etoile du Nord" - The Star of the North • State Bird - Common loon • State Insect – Monarch butterfly • State Fish - Walleye • State Flower - Pink and white lady's slipper

20. Rochester, Minnesota Est. Population: 99,000Median Age:34 yrs One of the Top 3 Most Livable Cities for 5 yrs straight.

21. Mayo Rochester

22. Mayo Rochester

23. Mayo Rochester

27. Initial Steps for Potential Students: • Native CREST APPLICATION FORM • Essay • School Transcript • (2) Native CREST RECOMMENDATION FORM(s) • (2) Letter(s) of Recommendation • Spirit of Eagles Application

28. Are you interested in a pursuing a degree in the health field? Native CREST Cancer Research Experience & Student Training 10-week summer research training experience at Mayo Clinic in Rochester, MN Diné College - Mayo Clinic May 29 – August 3, 2012 For more information contact: Mark C. Bauer, PhD Diné College – Shiprock Campus P.O. Box 580 Shiprock, NM 87420 mcbauer@dinecollege.edu (505) 368-3589 Application Deadline: February 1, 2010

29. Diné College – Shiprock Campus: Mark C. Bauer, PhD Faculty, Math and Public Health mcbauer@dinecollege.edu (505) 368-3589 Mayo Clinic Rochester, MN: Christi A. Patten, PhD Director, Behavioral Health Research patten.christi@mayo.edu (507) 538-7370 Contacts

30. Thank you.