ribosomal dysfunction results in an immunologic susceptibility to lung cancer stephanie chang md,...

Ribosomal Dysfunction Results in an Immunologic

Susceptibility to Lung CancerStephanie Chang MD, Ryuji Higashikubo PhD, Saeed

Arefanian MD, Andrew Gelman PhD, Daniel Kreisel MD, PhD, Alexander Krupnick MD

April 27th,2015

Department of SurgeryDivision of Cardiothoracic Surgery

Department of SurgeryDivision of Cardiothoracic Surgery

• No financial disclosures

Disclosures

Department of SurgeryDivision of Cardiothoracic Surgery

Susceptibility to Lung Cancer

Lung Cancer Resistant

Lung Cancer Susceptible

C3H

Previous work from our lab demonstrates natural killer (NK) cells play a role in this process Cancer Research 2012

Smoker with Bullous Emphysema

Non-Smoker with Lung Cancer

Department of SurgeryDivision of Cardiothoracic Surgery

Sly1380 AA55 kD

SH3 SAM

SH3 SAM

• X chromosome linked protein with unknown function

Expression of Sly 1 Correlates with Immunologic Resistance to Lung

Cancer

Protein protein binding domains

Department of SurgeryDivision of Cardiothoracic Surgery

• The aim of this study was to determine the role of Sly 1 in the susceptibility to lung cancer development

Objective

Department of SurgeryDivision of Cardiothoracic Surgery

Expression of Sly 1 Correlates with Immunologic Resistance to Lung

Cancer

Lung CancerResistant

Lung CancerSusceptible

Sly 1 Level by RT-PCR(normalized to -Actin/standardized patient)

% T

NF

+

NK

Cel

ls

Humans

Department of SurgeryDivision of Cardiothoracic Surgery

Expression of Sly 1 Correlates with Immunologic Resistance to Lung

Cancer

Lung CancerResistant

Lung CancerSusceptible

Sly 1 Level by RT-PCR(normalized to -Actin/standardized patient)

% T

NF

+

NK

Cel

ls

Human Sly 1 Function

Department of SurgeryDivision of Cardiothoracic Surgery

Sly 1 Contributes to NK Cytotoxicity Sly1- (knockout of the gene)

Wild-type littermates

Department of SurgeryDivision of Cardiothoracic Surgery

Activating Receptors Expression

Sly 1 Contributes to NK Cytotoxicity

Cytokine Expression

Sly1- (knockout of the gene)

Wild-type littermates

Department of SurgeryDivision of Cardiothoracic Surgery

Days After Injection

Lewis Lung Carcinoma Tumor Growth

Tu

mor

Volu

me (

mm

3)

Control Group

Days After Injection

Tu

mor

Volu

me (

mm

3)

NK Depletion Group

Sly 1 Expression Influences Lung Cancer Growth Via NK Cells

Department of SurgeryDivision of Cardiothoracic Surgery

Lung Cancer Lysis in vitro

Sly 1 Contributes to NK Cytotoxicity of Lung Cancer

Department of SurgeryDivision of Cardiothoracic Surgery

• Immunoprecipitation of WT and Sly 1 KO NK cell lysates with anti-Sly 1 Ab to evaluate Sly 1 binding partners

• No detection of signaling proteins from activating receptors or cytokine pathways

Sly 1 is a Ribosome Associated Protein

Department of SurgeryDivision of Cardiothoracic Surgery

Sly 1 Affects mRNA levels

Department of SurgeryDivision of Cardiothoracic Surgery

How Could a Ribosomal Protein Impair Gene Expression?

1) mRNA binding - No RNA-binding capacity

ribosome

mRNA40s

60s2) Ribosomal instability - Affects the transcription factor E2F as a protective mechanism

Department of SurgeryDivision of Cardiothoracic Surgery

mRNA

RB P

PP

P

PPPP

P

kinase

E2Fpromoter

ribosome

Normal Cell

How Could a Ribosomal Protein Impair Gene Expression?

Department of SurgeryDivision of Cardiothoracic Surgery

E2FRB

PP

P

PPPP

P

kinase

ribosome

Ribosomal Stress Associated with Malignant Transformation or Cell Stress

X p53

p27p21

Stop cellular proliferation and activation

How Could a Ribosomal Protein Impair Gene Expression?

Department of SurgeryDivision of Cardiothoracic Surgery

E2FRB

PP

P

PPPP

P

kinase

ribosome

X p53

p27p21

phospho-RBRB

Free E2F

Wild-typelittermate Sly1-

Actin

How Could a Ribosomal Protein Impair Gene Expression?

Ribosomal Stress Associated with Malignant Transformation or Cell Stress

Department of SurgeryDivision of Cardiothoracic Surgery

High-dose IL-2 Can Stabilize the Ribosome

Lewis Lung Carcinoma Lysis in vitro

Activating Receptors Expression

At Rest + IL-2 At Rest + IL-2

Department of SurgeryDivision of Cardiothoracic Surgery

• Sly1 is a ribosomal protein that affects NK ribosome stability at resto Ribosomal stress is a sign of malignant transformation which can

destroy the cell or decrease its activation/viability

• Prior to our research, most X linked ribosomopathies were considered severely disabling but we now demonstrate that subtle defects can manifest such as NK dysfunction and immunologic susceptibility to lung cancer

• IL2 can alleviate ribosomal stress and may be useful in lung cancer immunotherapy

Conclusions

Department of SurgeryDivision of Cardiothoracic Surgery

• Thoracic Surgery Foundation for Research and Education• NIH Ruth L Kirschstein NRSA

• Thoracic Immunology Lab• Eberhard-Karls University Tuebingen

• Sandra Beer-Hammer, PhD• Daniel Schall, PhD

Acknowledgements

Department of SurgeryDivision of Cardiothoracic Surgery

Questions?