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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?