Tramp and lady presentation (Dec.19,2012)

TRAMP and LADY Prostate Cancer (PCa)

Models

Speaker: Ahmad Usama Date: Dec.19,2012

ID: 101012422

Outlines

1- Introduction.

2- Models of Prostate Cancer.

3- Mouse Models of PCa.

3- TRAMP Model.

4- LADY Model.

5- Example of LADY Model.

6- Conclusion.

Introduction

What is the Prostate Cancer?

Form of Cancer in the Prostate

(gland in the male reproductive system)

Bones Lymph Nodes

Slowly Growing

Aggressively progression and invasion

Metastasize

Introduction

Prostate Cancer Treatment Strategies

Surgery RadiotherapyAndrogen

Deprivation Therapy

Introduction

Prostate Anatomy at Human & Mouse

Trends in Genetics, Volume 18, Issue 5, S1-S5, 1 May 2002

Introduction

Pathway of human prostate cancer progression


Pathway of human prostate cancer progression

THE JOURNAL OF UROLOGY Vol. 170, 2444–2452, December 2003

Models of Prostate Cancer

Prostate Cancer Models

Prostate CancerMouse Models

Prostate CancerNon- Mouse

Models

Dogs Rats

Lack of metastasesAndrogen independent

Mouse Models of Prostate CancerFive Main Categories of Prostate Cancer Mouse Models:

1- Xenograft Models

The mouse is used as immunodeficient recipient of human tumor , cell lines or primary cell cultures.

Ex : Nude Mice , SCID Mice , NOD Mice , NOD – SCID Mice , …..etc.

2 & 3 – Genetic Engineering Mouse Models (GEMM)

Transgenic models that use prostate specific promoters to express:

A- SV 40 T- antigen (Oncogene)Ex: TRAMP Models and LADY Models

B- and to express another oncogenes. Ex: Mt-PRL and BK5-IGF1 models

4- Traditional Knockout Models.Ex: Pten and Stat5a Models.

5- Conditional Knockout Models.Ex:PSA-Cre and MMTV-Cre Models.

Mouse Models of Prostate Cancer

Transgenic T(Tuomr) Antigen (Tag) Models

Introduction of DNA constructs to induce the expression on Tag

(oncoprotein) under the control of prostate specific promoter

Tumor Antigen(Tag)

Large T antigenSuppressive

interactions with P53 & RB

Small t antigen Interaction with PP2a to induce transformation

Transgenic T Antigen (Tag) Models

TRAMP Model (Transgenic Adenocarcinoma of Mouse Prostate)

SV 40 Large T antigen (Tag)

expression

SV 40 Small t antigen (tag)

expression

Rat Probasin Promoter (rPB)

Androgens

TRAMP Model

* Advantages:1- Transgene expression specifically in the prostate (androgen regulation).

2- The first mouse model to display distant organ metastases.

3- Castration resistance well modeled.

* Disadvantages:- Develop PCa of neuroendocrine origin not from epithelial origin like human.

LADY Model

Generations of rPB promoters

First Generation

Second Generation

Third Generation

PB promoter426 bp

LBP promoter11,500 bp

ARR2PB promoter

More transgene expression rate

* LADY Model differs from TRAMP at some aspects:1- Using second generation of rPB (Large fragmentPB promoter) which causes high transgene expression.

2- Large T antigen is only expressed.

LADY Model

* Advantage:

- LADY models more accurately mimic the majority of human PCa because the cancer slow growing and has mostly epithelial phenotype.

* Disadvantage:

- Castration resistance and metastasis are not modeled well.

[CANCER RESEARCH 61, 2239–2249, March 1, 2001]

Example of LADY Model

LADY Model used called : 12T-10 large probasin prompter – Tag mouse

Pathway of mouse prostate cancer progression

1) Low Grade Intraepithelial

Neoplasia(LGPIN)

2) High Grade Intraepithelial

Neoplasia(HGPIN)

5)Undifferentiated carcinoma

(UC)With NE differentiation

3) Microinvasion(MI)

4) Invasion Carcinoma (IC)(IC , ICad , ICne , IC ad +ne )

Neurodendocrine Cells (NE)

NE

Normal Prostate growthProstate Cancer Progression

Polypeptidegrowth factors

Biogenic Amines VEGF TGF α


Histopathological Progression of Prostatic Neoplasia in the12T-10 LPB-Tag Line


A) LGPIN at the dorsal lobe of a 2-month-old

mouse

B) HGPIN at the dorsal lobe of a 7-month-old

mouse

C) MI at the dorsal lobe of a 8-month-old mouse

D) IC with glandular differentiation at the

dorsolateral lobe of a 11-month-old mouse

E) IC with NE differentiation at the

ventral lobe of a 6-month-old mouse

F) IC with glandular differentiation with

adjacent UC (NE )at the dorsolateral lobe of a 12-

month-old mouse

G & H) UC (NE) at the ventral lobe of a 10-month-

old mouse


CG (Chromogranin) : immunostaining for demonstration

the progression of NE differentiation

Nontransgenic control mice up to 10 months:

– ve or few CG positive NE cells

HGPIN and LGPIN lesions of 2-4 month old at 12T-10 mice:

– ve or few CG positive NE cells

HGPIN,+ve CG , Aged ≥ 5 months

+ve CG staining for Tag and AR at HGPIN lesions

+ve CG staining for IC lesions with NE differentiationat the dorsolateral lobe

-ve CG staining for IC lesions with glandular differentiation

at the dorsolateral lobe


+ve GC staining for NE cells at UC lesions

(fainter than at HGPIN)

+ve GC staining for Tag at UC lesions

(fainter than at HGPIN)

Few +ve GC staining for Cytokeratin at UC lesions

PCNA (Proliferating Cellular Nuclear Antigen ) showing extensive nuclear staining

-ve or few +ve staining for AR at UC lesions


Incidence of Regional and Systemic Metastases in the 12T-10 Line

Correlation of Metastases with Pathology of the Prostate


Protein Profiling by Mass Spectrometry Analysis (MALDI – TOF - MS)For the ventral lobes of 12T – 10 and CD1 mice

From 22,000 to 24,000 m/zSpermine – binding protein

signals


Protein Profiling by Mass Spectrometry Analysis (MALDI – TOF - MS)is similar for all lobes


Protein Profiling by Mass Spectrometry Analysis (MALDI – TOF - MS)after Allografting

Protein profiling at primary NE tumor site

Protein profiling before Allografting

Protein Profiling at Metastasis sit

After Allografting

Conclusion

*The goal of genetically engineered mouse models (GEMM) is to accurately mimic all of stages of human disease in the mouse.

* Age is the main risk factor for prostate cancer.

* PCa is largely dependent on androgens for growth and proliferation; hence, androgen deprivation therapy (chemical castration) is the standard of treatment, and it generally causes prostate tumors to regress.

* AR – negative neuroendocrine (NE) cells is more prominent in PCa than in benign tissues.

Thanks For Your Attention

Welcome for Questions

Tramp and lady presentation (Dec.19,2012)

Health & Medicine

Transcript of Tramp and lady presentation (Dec.19,2012)