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

Mayo ScottsdaleMayo Rochester

Mayo Jacksonville

RationaleRationale

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

Project AimsProject Aims

• Aim 1: Cancer Education - FacultyAim 1: Cancer Education - Faculty

• Aim 2: Cancer Education - StudentAim 2: Cancer Education - Student

• Aim 3: Cancer Research Training-Aim 3: Cancer Research Training- Student Research TrainingStudent Research Training

Aim 3: Student Cancer Aim 3: Student Cancer Research TrainingResearch Training

• Identify, evaluate and follow DinIdentify, evaluate and follow Dinéé College students College students

• Identify Mayo mentors and/or labs Identify Mayo mentors and/or labs

• The The Native CREST summer cancer research summer cancer research training program was piloted at Mayo Rochester in training program was piloted at Mayo Rochester in 20082008

• Refine research training based on student and Refine research training based on student and mentor feedbackmentor feedback

Aim 3: Student Cancer Aim 3: Student Cancer Research Training (cont.)Research Training (cont.)

• Implement program for DinImplement program for Dinéé College students College students at Mayo Clinic Rochester again in 2010 at Mayo Clinic Rochester again in 2010

(2-3 students for 10 weeks)(2-3 students for 10 weeks)

• Develop the process by which students Develop the process by which students present their research experiences back present their research experiences back to their communitiesto their communities

• Develop long-term tracking systemDevelop long-term tracking system

Rin2 Regulation of HGF/c-Met SignalingLatoya 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.

BACKGROUNDReceptors 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.

ABSTRACTRab5 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.

RESULTSIn 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).

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.

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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.

Rab5•GDPRab5•GDP Rab5•GTPRab5•GTP

Rab5Rab5

Rab5Rab5

Guanine Nucleotide Exchange Factors (GEFs)

GTPase Activating Proteins (GAPs)

GTP GDP

PO4

Figure 1. Rab5 is Activated by the Vps9-domain proteins

Figure 2. Receptor Systems and Vps9 Domain Proteins

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).

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Figure 5. RIN2 Localization. HuH7 cells were transfected with an EGFP-RIN2 expression construct and visualized by fluorescence microscopy after deconvolution using DeltaVision software.

DinDinéé College College

SREP: Summer Research SREP: Summer Research Enhancement ProgramEnhancement Program

ForFor Cancer and Diabetes Prevention Cancer and Diabetes Prevention

SURF: Summer Undergraduate Research

Fellowship

Native CRESTNative CRESTCCancer ancer RResearch esearch EExperience xperience

& & SStudent tudent TTrainingraining

Diné College - Mayo ClinicDiné College - Mayo Clinic

“Preparing Tomorrow’s Native Researchers Today.”

Program Dates:Program Dates:May 29 – August 3, 2012May 29 – August 3, 2012

Application Deadline:Application Deadline:February 15, 2012February 15, 2012

The The Native CREST program has been developed program has been developed to increase the number of students at Diné to increase the number of students at Diné

College preparing to pursue advanced degrees College preparing to pursue advanced degrees in the health fields by providing opportunities to in the health fields by providing opportunities to

obtain experience in cancer research and obtain experience in cancer research and training. Students appointed to this program are training. Students appointed to this program are

interested in the public health and health care interested in the public health and health care fields and are considering a future health-related fields and are considering a future health-related career through biomedical research, nursing, or career through biomedical research, nursing, or

pursuing PhD, MD, or MPH degrees. pursuing PhD, MD, or MPH degrees.

The ProgramThe Program • 10-week summer research training experience at Mayo 10-week summer research training experience at Mayo

Clinic in Rochester, MinnesotaClinic in Rochester, Minnesota Diné College students will work with a Mayo Clinic Diné College students will work with a Mayo Clinic

researcher/faculty member to develop their research interests and researcher/faculty member to develop their research interests and experiencesexperiences

• Up to 3 Diné College students for the Summer of 2010 Up to 3 Diné College students for the Summer of 2010 Students are linked with researcher/faculty who is interested in Students are linked with researcher/faculty who is interested in

mentoring a student on a cancer related research project mentoring a student on a cancer related research project

• Program stipend of $4,000 Program stipend of $4,000

• Students are responsible for arranging their travel, Students are responsible for arranging their travel, housing and meals (Spirit of EAGLES: ~$1,000)housing and meals (Spirit of EAGLES: ~$1,000)

Housing (Rochester, MN):

• Grand Kahler Hotel Grand Kahler Hotel Double:  $750/month  Single:  $1320/month

• Residences of Old Town HallResidences of Old Town HallPrivate Room Apt:  $625/month

• Second Street Inn & Suites Second Street Inn & Suites Single:  $600/month

Mayo Clinic Native American ProgramsMayo Clinic Cancer Center

Rochester, Minnesota

Spirit of EAGLES SurvivorsEducationAdvocacy

GrantsLeadership

EldersScholarships

Funded by: CA 77410-02

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

• State CapitalState Capital - St. Paul ( - St. Paul (Largest CityLargest City – Minneapolis) – Minneapolis)

• Major IndustriesMajor Industries - farming (corn, soybeans, sugar beets, wheat, dairy products), paper - farming (corn, soybeans, sugar beets, wheat, dairy products), paper pulp, mining (iron ore) pulp, mining (iron ore)

• State NicknameState Nickname - Gopher State. North Star State - Gopher State. North Star State

• State MottoState Motto - "L'Etoile du Nord" - The Star of the North - "L'Etoile du Nord" - The Star of the North

• State BirdState Bird - Common loon - Common loon

• State InsectState Insect – Monarch butterfly – Monarch butterfly

• State FishState Fish - Walleye - Walleye

• State Flower State Flower - Pink and white lady's slipper- Pink and white lady's slipper

Origin of the Name - Minnesota is from a Dakota Sioux Indian word that means "cloudy water" or "sky water" and refers to local rivers.

Rochester, Minnesota

Est. Population: 99,000 Median Age: 34 yrs

One of the Top 3 Most Livable Cities for 5 yrs straight.

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

Native CRESTCancer Research Experience

&

Student Training

10-week summer research training experience at Mayo Clinic in Rochester, MN

For more information contact:Mark C. Bauer, PhD

Diné College – Shiprock CampusP.O. Box 580

Shiprock, NM 87420mcbauer@dinecollege.edu

(505) 368-3589

Application Deadline: February 1, 2010

Are you interested in a pursuing a degree in the health field?

Diné College - Mayo ClinicMay 29 – August 3, 2012

Contacts• 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