Energy balance journal club
Download
1 / 58

ENERGY BALANCE JOURNAL CLUB - PowerPoint PPT Presentation


  • 88 Views
  • Uploaded on

A High-Fat Diet Activates Oncogenic Kras and COX2 to Induce Development of Pancreatic Ductal Adenocarcinoma in Mice. ENERGY BALANCE JOURNAL CLUB. Bincy Philip, Christina L. Roland, Jaroslaw Daniluk, Yan Liu, Deyali Chatterjee,

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about ' ENERGY BALANCE JOURNAL CLUB' - luke


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
Energy balance journal club

A High-Fat Diet Activates Oncogenic Kras and COX2 to Induce Development of

Pancreatic Ductal Adenocarcinoma in Mice

ENERGY BALANCE JOURNAL CLUB

Bincy Philip, Christina L. Roland, Jaroslaw Daniluk, Yan Liu, Deyali Chatterjee,

Sobeyda B. Gomez, Baoan Ji, Haojie Huang, Huamin Wang, Jason B. Fleming,

Craig D. Logsdon, Zobeida Cruz-Monserrate

Zobeida Cruz-Monserrate Ph.D.

Instructor

Cancer Biology Department


Pancreatic cancer a deadly disease 2013 estimated us cancer deaths
Pancreatic Cancer: A Deadly Disease Development of2013 Estimated US Cancer Deaths

Men306,920

Women273,430

Lung & bronchus 28%

Prostate 10%

Colon & rectum 9%

Pancreas 6%

Liver & intrahepatic 5%bile duct

Leukemia 4%

Esophagus 4%

Urinary bladder 4%

Non-Hodgkin 3% lymphoma

Kidney 3%

26% Lung & bronchus

14% Breast

9% Colon & rectum

7% Pancreas

5% Ovary

4% Leukemia

3% Non-Hodgkin lymphoma

3% Uterine corpus

2% Liver & intrahepatic bile duct

2% Brain and other nervous system

Source: American Cancer Society, 2013


Pancreatic ductal adenocarcinoma pdac is deadly and difficult to diagnose early
Pancreatic Ductal Adenocarcinoma (PDAC) is Deadly and Difficult to Diagnose Early

  • Highest death to incidence ratio (0.99) of all cancers

  • 5-year survival rate below 6%

    • Median survival ~ 6 months

  • Surgical resection is the only effective treatment

    • < 20% of the patients are eligible

  • Rarely detected at an early stage (small lesions)

    • High rate of dissemination

  • Conventional cancer treatments fail

    • Resistance to chemotherapy

    • Treatment options limited

Prevention and Early Detection


Pancreas microanatomy
Pancreas Microanatomy Difficult to Diagnose Early

Endocrine

Exocrine

Most pancreatic cancers (around 75%)

Develop in the exocrine pancreas

Omary, M.B., et. al., 2007. 117: p. 50-9


Multistep Progression Model of PDAC Difficult to Diagnose Early

Pancreatic Intraepithelial Neoplasias (PanINs)

Normal duct

  • single cell layer

  • low cuboidal

PanIN-1A/1B

  • elongated cells

  • mucin

  • papillary growth

PanIN-2

  • early nuclear abnormalities

PanIN-3

  • luminal budding

  • nuclear atypia

  • mitosis

Carcinoma

  • invasion

  • desmoplasia

PDAC >90% Mutant K-Ras

Hruban, R.H., et al., 2001. 25(5): p. 579-86


Non modifiable pdac risk factors
Non-Modifiable Difficult to Diagnose Early PDAC Risk Factors


Modifiable pdac risk factors
Modifiable Difficult to Diagnose Early PDAC Risk Factors

Obesity


Obesity is a risk factor for many cancers including pancreatic
Obesity is a Risk Factor for Many Cancers Including Pancreatic

Eugenia E. Calle* and Rudolf Kaaks 2004:4:579-591


Mouse models allow conditional specific knock in of oncogenic k ras
Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Cell Type Specific Promoter

Duct

Islet

Acini


Mouse models allow conditional specific knock in of oncogenic k ras1
Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Cell Type Specific Promoter

Duct

Islet

Acini

Elastase-Cre-Er

Tamoxifen Regulated


Mouse models allow conditional specific knock in of oncogenic k ras2
Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Cell Type Specific Promoter

Duct

Islet

Acini

Acinar Cell Specific Cre

Elastase-Cre-Er

Tamoxifen Regulated


Mouse models allow conditional specific knock in of oncogenic k ras3
Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Oncogenic K-Ras

“flox-stopped“

X

Cell Type Specific Promoter

Endogenous Promoter “Knock In”

Stop

K-RasG12D

Poly A

p-K-Ras

Duct

loxP

loxP

loxP = locus of recombination

Islet

Acini

Acinar Cell Specific Cre

Elastase-Cre-Er

Tamoxifen Regulated


Mouse models allow conditional specific knock in of oncogenic k ras4

K-Ras Oncogenic K-RasG12D

Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Oncogenic K-Ras

“flox-stopped“

X

Cell Type Specific Promoter

Endogenous Promoter “Knock In”

Elastase-Cre-Er

Stop

K-RasG12D

Poly A

p-K-Ras

loxP

loxP

Cre mediated deletion via Tamoxifen after birth

loxP = locus of recombination

LSL/BAC

Acinar

mK-Ras

Endogenous

mutant K-Ras expression in acinar cells


Mouse models allow conditional specific knock in of oncogenic k ras5

K-Ras Oncogenic K-RasG12D

Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Oncogenic K-Ras

“flox-stopped“

X

Cell Type Specific Promoter

Endogenous Promoter “Knock In”

Elastase-Cre-Er

Stop

K-RasG12D

Poly A

p-K-Ras

loxP

loxP

Cre mediated deletion via Tamoxifen after birth

loxP = locus of recombination

LSL/BAC

Acinar

mK-Ras

Endogenous

mutant K-Ras expression in acinar cells

Normal Pancreas at early age


Mouse models allow conditional specific knock in of oncogenic k ras6

K-Ras Oncogenic K-RasG12D

Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Oncogenic K-Ras

“flox-stopped“

X

Could this mouse model be used to test risk factors like obesity?

Cell Type Specific Promoter

Endogenous Promoter “Knock In”

Elastase-Cre-Er

Stop

K-RasG12D

Poly A

p-K-Ras

loxP

loxP

Cre mediated deletion via Tamoxifen after birth

loxP = locus of recombination

LSL/BAC

Acinar

mK-Ras

Endogenous

mutant K-Ras expression in acinar cells

Normal Pancreas at early age


High fat induced pdac mouse model

K-Ras Oncogenic K-RasG12D

High Fat-Induced PDAC Mouse Model

LSL/BAC

Acinar

mK-Ras

Endogenous

mutant K-Ras expression in acinar cells


High fat induced pdac mouse model1

K-Ras Oncogenic K-RasG12D

High Fat-Induced PDAC Mouse Model

LSL/BAC

Acinar

mK-Ras

Endogenous

mutant K-Ras expression in acinar cells

Isocaloric


High fat induced pdac mouse model2

K-Ras Oncogenic K-RasG12D

High Fat-Induced PDAC Mouse Model

LSL/BAC

Acinar

mK-Ras

Endogenous

mutant K-Ras expression in acinar cells

Isocaloric



High fat diet increased inflammation fibrosis and panin lesions on mice with k ras g12d mutation
High Fat Diet Increased Weight Compared to Control DietInflammation, Fibrosis, and PanIN Lesions on Mice with K-RasG12D Mutation


High fat diet increased inflammation fibrosis and panin lesions on mice with k ras g12d mutation1
High Fat Diet Increased Weight Compared to Control DietInflammation, Fibrosis, and PanIN Lesions on Mice with K-RasG12D Mutation


High Fat Diet Increased Areas of Weight Compared to Control DietCollagen Deposition on Mice with K-RasG12D Mutation


High Fat Diet Increased Areas of Weight Compared to Control DietActivated Stellate Cells on Mice with K-RasG12D Mutation

Stellate Cells


Endogenous Mutant K-Ras Weight Compared to Control Dietis not Sufficient to Transform Most Cells

Developmental promoter EMBRYONIC expression in all cell types

Multiple PanIN lesions; PDAC- 2/29 (7%) 1 year

Endogenous K-Ras mutations generates cancer with low efficiency

Acinar cell promoter

ADULT expression in acinar cells

No PanIns No tumors – 0/11 (0%) 1 year

(Unless inflammation was induced)


Mutant k ras at endogenous levels requires an inflammatory insult to transform acinar cells
Mutant K-Ras at Endogenous Levels Requires an Inflammatory Insult to Transform Acinar Cells

Bac-Elastase CreERT

100% efficient

100% specific for adult acinar cells

If oncogenic mutant Ras is always “on” then there should be effects on cell function.


Mutant k ras at endogenous levels requires an inflammatory insult to transform acinar cells1
Mutant K- Insult to Transform Ras at Endogenous Levels Requires an Inflammatory Insult to Transform Acinar Cells

mutant Ras NO Stimulant

mutant Ras

+ LPS Stimulant


High Fat Diet Insult to Transform

Link Between Ras and Obesity?


High fat diet increased ras activity in mice with k ras g12d mutation
High Fat Diet Increased Insult to Transform Ras Activity in Mice with K-RasG12D Mutation


High fat diet activates of k ras downstream pathways in mice with k ras g12d mutation
High Fat Diet Activates of Insult to Transform K-Ras Downstream Pathways in Mice with K-RasG12D Mutation


Ras must be active to initiate downstream signaling resulting in progression of pdac
Ras Must Be Insult to Transform Active to Initiate Downstream Signaling Resulting in Progression of PDAC

Cox2

PGE2

Receptors (GF, hormones, etc)

PI3K

Ras-GTP

GEFs

Raf

Ras-GDP

GAPs

RalGDS

INACTIVE

ACTIVE

MAPK


High fat diet increases cox 2 in pancreas on mice with k ras g12d mutation
High Fat Diet Increases Insult to Transform Cox-2 in Pancreas on Mice with K-RasG12D Mutation


High fat diet promotes recruitment of macrophages on mice with k ras g12d mutation
High Fat Diet Promotes Recruitment of Insult to Transform Macrophages on Mice with K-RasG12D Mutation


High fat diet increases cox 2 in pancreas on mice with k ras g12d mutation1
High Fat Diet Increases Insult to Transform Cox-2 in Pancreas on Mice with K-RasG12D Mutation


High fat diet promotes pdac

K-Ras Insult to Transform G12D

High Fat Diet Promotes PDAC

LSL/BAC

Acinar

mK-Ras

Endogenous

mutant K-Ras expression in acinar cells

Normal Pancreas at early age


High fat diet promotes pdac1

K-Ras Insult to Transform G12D

High Fat Diet Promotes PDAC

LSL/BAC

Acinar

mK-Ras

High Fat Diet

Endogenous

mutant K-Ras expression in acinar cells

Normal Pancreas at early age

Cox2

PanINs

Cancer


Cox 2 deletion in acinar cells with knock in of oncogenic k ras

K-Ras Insult to Transform G12D

Cox-2 Deletion in Acinar Cells with “Knock-in” of Oncogenic K-Ras

LSL/BAC

Cox-2 KO

“flox-stopped“

Acinar

mK-Ras

X

Endogenous

mutant K-Ras expression in acinar cells


Cox 2 deletion in acinar cells with knock in of oncogenic k ras1

K-Ras Insult to Transform G12D

K-RasG12D

Cox-2 Deletion in Acinar Cells with “Knock-in” of Oncogenic K-Ras

LSL/BAC

Cox-2 KO

“flox-stopped“

Acinar

mK-Ras

X

Endogenous

mutant K-Ras expression in acinar cells

COXKO/LSL/BAC

Acinar

mK-Ras

Endogenous

mutant K-Ras expression in acinar cells but NO Cox-2 Expression


Cox 2 is required in high fat induced pdac mouse model

K-Ras Insult to Transform G12D

COX-2 is Required in High Fat-Induced PDAC Mouse Model

COXKO/LSL/BAC

Acinar

mK-Ras

Endogenous

mutant K-Ras expression in acinar cells but NO Cox-2 Expression

Isocaloric


Conditional knockout of cox 2 in the acinar cells blocked the effects of high fat diet
Conditional Knockout of Insult to Transform Cox-2 in the Acinar Cells Blocked the Effects of High Fat Diet


Conditional knockout of cox 2 in the acinar cells blocked the effects of high fat diet1
Conditional Knockout of Insult to Transform Cox-2 in the Acinar Cells Blocked the Effects of High Fat Diet


Conditional knockout of cox 2 in the acinar cells blocked the effects of high fat diet2
Conditional Knockout of Insult to Transform Cox-2 in the Acinar Cells Blocked the Effects of High Fat Diet


Systemic cox 2 inhibition decreases the effects of high fat diet
Systemic Insult to Transform Cox-2 inhibition Decreases the Effects of High Fat Diet


Systemic cox 2 inhibition decreases the effects of high fat diet1
Systemic Insult to Transform Cox-2 inhibition Decreases the Effects of High Fat Diet



High fat diet decreases survival of mice susceptible to pdac1
High Fat Diet Decreases Survival of Mice Susceptible to PDAC Insult to Transform

Control Diet

30 days

160

Days

High Fat Diet


High fat diet decreases survival of mice susceptible to pdac2
High Fat Diet Decreases Survival of Mice Susceptible to PDAC Insult to Transform

Control Diet

High Fat Diet

30 days

205

Days

Pancreas

Pancreas

Pancreas


High fat diet decreases survival of mice susceptible to pdac3
High Fat Diet Decreases Survival of Mice Susceptible to PDAC Insult to Transform

Control Diet

High Fat Diet

30 days

205

Days

Pancreas

Pancreas

High Fat Diet

Pancreas

Pancreas


Mouse models allow conditional specific knock in of oncogenic k ras7
Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Cell Type Specific Promoter

Duct

Islet

Acini

Elastase-Cre-Er

Tamoxifen Regulated


Mouse models allow conditional specific knock in of oncogenic k ras8
Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Cell Type Specific Promoter

PDX-1 Cre

Activates Cre during development


Mouse models allow conditional specific knock in of oncogenic k ras9
Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Oncogenic K-Ras

“flox-stopped“

X

Cell Type Specific Promoter

Endogenous Promoter “Knock In”

PDX-1 Cre

Stop

K-RasG12D

Poly A

p-K-Ras

Activates Cre during development

loxP

loxP

loxP = locus of recombination


Mouse models allow conditional specific knock in of oncogenic k ras10

K-Ras Oncogenic K-RasG12D

Mouse Models Allow Conditional Specific “Knock-in” of Oncogenic K-Ras

re

Oncogenic K-Ras

“flox-stopped“

X

Cell Type Specific Promoter

Endogenous Promoter “Knock In”

PDX-1 Cre

Stop

K-RasG12D

Poly A

p-K-Ras

Activates Cre during embryogenesis

loxP

loxP

loxP = locus of recombination

Cre mediated deletion during development

LSL/PDX-1

mK-Ras

All cells

Endogenous

mutant K-Ras expression in every cell of the pancreas


High fat diet decreases survival of mice susceptible to pdac4
High Fat Diet Decreases Survival of Mice Susceptible to PDAC Oncogenic K-Ras

Some

Mice in control diet are still alive

Cre mediated deletion via Tamoxifen after birth

Cre mediated deletion during development


High Fat Diet Accelerated PanIN-2 and PanIN-3 Development Compared to Control Diet Using and Embryonic Promoter

Control Diet

137

Days

High Fat Diet

Cre mediated deletion during development

90

Days


High Fat Diet Accelerated PanIN-2 and PanIN-3 Development Compared to Control Diet Using and Embryonic Promoter

Control Diet

137

Days

High Fat Diet

Cre mediated deletion during development

90

Days


Summary
Summary Compared to Control Diet Using and Embryonic Promoter:

Obesity is a Risk Factor for PDAC in Humans

High Fat Diet Accelerates PanIN formation and

Cancer Development in PDAC Mouse Models

Ras Activity and Cox-2 are essential

for High Fat Diet induced PDAC

HOW DOES FAT LEAD TO RAS ACTIVATION?

Mechanisms?


Future directions with high fat induced pdac model
Future Directions with High Fat Induced PDAC Model Compared to Control Diet Using and Embryonic Promoter

Test Prevention Agents

LSL/BAC


Acknowledgments
Acknowledgments Compared to Control Diet Using and Embryonic Promoter

Dr. Logsdon’s Laboratory

Collaborators

MDACC

Huamin Wang, Pathology

Jason Fleming, Surgical Oncology

Joya Chandra, Pediatrics

David McConkey, Urology

Kathleen M. Schmeler MD, Ob/GYN

Ralph B. Arlinghaus, Translational Molecular Pathology

Adel El-Naggar, Pathology

The Methodist Hospital Research Institute

Ching-Hsuan Tung Ph.D.

Wael R. Abd-Elgaliel Ph.D.

Rita Serta Ph.D.

City of Hope National Medical Center

Ann David Ph.D.

University of Rhode Island

Oleg Andreev

Yana Reshetnyak

Bincy Philips, MS

Funding Sources

NIDDK Minority Supplement

Phi Beta Psi Sorority


ad