Download slides

Edison Liu, M.D.
Integrated
Translational Science
Center
Dave Tuveson M.D.,
Ph.D.
and
Larry Baker D.O.
CSHL-JAX-SWOG-HOPE
September 15, 2016
Our ITSC: Education and Research
•
SWOG Meetings
•
•
•
•
Translational Summer Workshop
•
•
•
Introductory sessions and office hours with ITSC Leadership
Eagle II Reception - Thursday September 15, 2016 (for all workshop
participants and pilot project awardees).
Translational dinner, talks and networking session is being planned for
Spring 2017 SWOG group meeting
Alternating years at CSHL or JAX
12 SWOG Clinical Investigators, 12 basic scientists from JAX, 12 basic
scientists from CSHL
Pilot Projects
•
•
~ 4 awarded per year (1 year award, $100,000 in direct costs)
Must have one clinical and one basic scientist as co-PIs.
Translational Workshop
SWOG/Integrated Translational Science Center
The forum provides an opportunity for:
• CSHL & JAX scientists to present the technology platforms
• SWOG clinicians to discuss clinical questions that need to be addressed
Goal is to pair SWOG clinicians with CSHL/JAX scientists to propose pilot
projects using technologies to tackle clinical questions
ITSC Translational Workshops:
• Banbury Conference Center, CSHL (June 18-20, 2015)
• Highseas Conference Center, JAX (Aug 31-Sept 2, 2016)
• Banbury Conference Center, CSHL (2017, date tba)
• Highseas Conference Center, JAX (2018, date tba)
Pilot Projects
SWOG/Integrated Translational Science Center
Goals:
• Support collaborative projects between SWOG investigators with coPI from Cold Spring Harbor Laboratory or The Jackson Laboratory
• Use technology platforms – THE DISCOVERY ENGINE - available
at CSHL and JAX to obtain preliminary data that can then be used
as the basis for grant funding or development of clinical protocols
2014 (Year 1): 4 Letters of Intent and 4 full applications requested; 2
funded
2015 (Year 2): 16 Letters of Intent and 10 full applications requested (7
from workshop participants); 6 Funded (4 from SWOG clinicians who attended
the workshop)
2016 (Year 3):
7 Letters of Intent Received.
Plan to fund 4 pilots
SWOG Discovery Engine: CSHL and JAX
Clinical
Cancer Genomics:
Computational
Modeling
RNA,DNA,Epigenome
Single Cell
Genomics
Gene Editing
Genome Engineering
Cell-based Screens
Patient Derived
Xenografts
Humanized NSG
Xenografts
Conditional Reprogrammed
Progenitor Cells
Cancer Organoids
6
SWOG Discovery Engine: CSHL and JAX
US
SWOG-CSHL-JAX Discovery
Genomics Cell Engine
Models PDX Models
YOU
Important Clinical
Question
SWOG
Impactful Clinical Trial
EVERYONE
Improved Patient
Outcome
ITSC Pilots with SWOG-JAX PIs
SWOG PI
JAX PI
Title
Gandara, Mack
Bult, Keck
Testing novel therapies for EGFR mutant
lung cancer in PDX models
Algazi
Keck
Testing immune modulators that target
human squamous cell carcinoma of the
head & neck (HNSCC) in hu-NSG mice
Burack, Casulo
Hasham
The AID-RAD51 axis as a better
biomarker in follicular lymphoma
Hassan
Hasham, Bult
Determining the role of AICDA in
development of T790M mutation in lung
cancer PDX models
Hu-Lieskovan
Robson
Tumor microenvironment profiling to
identify biomarkers of anti-PD-1 therapy
Pusztai
Chuang
Genomic analysis of inflammatory breast
cancer (IBC)
8
ITSC Pilot Project 2015:
Lajos Pusztai (Yale) and Jeff Chuang (JAX)
Landscape of somatic mutations in inflammatory
breast cancer whole-genome sequences
Paired-end whole genome sequencing (WGS) of
20 IBC vs. 23 non-IBC
Age, race and ER and HER2 matched,
from TCGA for comparison.
9
10
ITSC Pilot Project 2015:
Lajos Pusztai (Yale) and Jeff Chuang (JAX)
Landscape of somatic mutations in inflammatory
breast cancer whole-genome sequences
Recurrent mutations were detected in the coding region of
GRIN2A gene in 15% IBC samples, vs. 1.7% previously
reported in breast cancers). Other significant mutations in
coding regions included GRHL1, PIK3R2, ESR1, and
FLG2.
Contributions of mutational signature 9, that is associated
with polymerase , were significantly higher in IBC cohort
than non-IBC cohort (p-value=0.056).
To be presented at the San Antonio Breast Cancer Con
11
Pilot Projects - Awarded
2014 Pilots
Induced Cytidine Deaminase as a Biomarker in Follicular Lymphoma
(Owen O’Conner, MD/PhD; & Kevin Mills, PhD, JAX)
Neutrophil Extracellular DNA Traps and Breast Cancer Metastasis (Anne
Schott, MD, Univ Michigan; Mikala Egeblad, PhD, CSHL)
2015 Pilots
The AID-RAD51 Axis as a Better Biomarker in Follicular Lymphoma
(Kevin Mills, PhD, JAX; Richard Burack, MD/PhD and Caral Casulo, MD,
Univ Rochester)
Genomic Analysis of Inflammatory Breast Cancer (IBC) in SWOG S0800
Clinical Trial Tissues to Identify Molecular Alterations that Underlie the
Aggressive Biology of IBC and Could Lead to the Development of New
Diagnostic Tests and Therapeutic Interventions (Lajos Pusztai, MD/PhD
and Mark B. Gerstein, PhD, Yale; Naoto Ueno, MD/PhD, MD Anderson; &
Jeffrey Chuang, PhD, JAX)
SWOG/CSHL Highlight
SWOG/Integrated Translational Science Center
2014 Pilot Project
Park J., Wysocki, R.W., Amoozgar, Z., Maiorino, L., Fein, M.R., Jorns, J., , Schott, A.F., Kinugasa‐Katayama, Y., Lee, Y., Won, N.H., Nakasone, E.S., Hearn, S.A., Küttner,V., Qiu, J., Almeida, A.S., Perurena, N., Kessenbrock, K., Goldberg M.S., and Egeblad.M. Cancer cells trigger neutrophils to induce metastasis‐supporting extracellular DNA traps. Sci. Transl. Med. (Accepted)
Cancer cells induce
metastasis-supporting
neutrophil extracellular DNA traps
Mikala Egeblad and Anne Schott
5 m
Neutrophils and DNA “clouds” surround
metastatic 4T1 breast cancer cells in lungs
4T1 cancer cells
Neutrophils
DNA (DAPI)
Lung imaging, 2 hrs after i.v. injection of cancer cells
Park et al., Science Translational Medicine, In press
Neutrophil Extracellular Traps (NETs) capture
and kill pathogens in the extracellular space
Peptidylarginine deiminase 4
(PAD4)-mediated
histone modification
Bacteria/LPS
Bacterial
trapping
NADPH
oxidase
NADPH + 2O2 =>
NADP+ + H+ + 2O2-
Cathepsin G
Myeloperoxidase
Elastase
Chromosome
decondensation
Protease association
Expelled DNA w.
Cathepsin G
Myeloperoxidase
Elastase
J Clin Invest. 2013 Jul 1. pii: 67484
Are NETs found in human
breast cancer?
ITSC pilot project w. Anne Schott, Julie Jorns et al.
NETs are present in human breast cancer
Primary human breast tumor
Neutrophil
NET forming neutrophil
20μm
Myeloperoxidase Citrullinated histone H3 DNA (DAPI)
Julie Jorns
Anne Schott
Youngseok Lee
Nam H. Won
Anne Schott
Julie Schor
Bobby Wysocki
Laura Maiorino
Presence of NETs in metastatic human breast
tumors is associated with the triple negative
breast cancer subtype
Park et al., Sci Transl, accepted
A transwell co-culture model to study
cancer cell / neutrophil interactions
Cancer cells
Cancer cell invasion?
22h
Matrigel
(basement
membrane)
Serum free media
Neutrophils
Neutrophil activation?
50 m
+DNase I
Invasive capacity (% of serum-free)
The DNA scaffold of NETs is required for
neutrophil-induced cancer cell invasion
p=0.03
p=0.02
200
150
100
50
0
DAPI
Histone
Neutrophil Elastase
Park et al., Sci Transl, accepted
Conditioned medium (CM) from neutrophils
induced to form NETs promotes invasion
Cancer cells
Cancer cells
CM
Neutrophils
Juwon Park
Yumi Kunigasa
How do cancer cells
induce NETs?
How do cancer cells induce NETs?
G-CSF secreted by cancer cells
Induces NET formation
Inhibition of G-CSF
p=0.03
NET formation (A.U.)
NET formation (A.U.)
p=0.03
Recombinant G-CSF
4T1 cancer cells
Park et al., Sci Transl, accepted
Can NETs be targeted in vivo?
Recombinant DNase I is used
to treat cystic fibrosis
Plasma DNase I (ng/ml)
DNase I-coated nanoparticles have higher
activity in blood than free DNase I
75U DNase I (free or coated on nanoparticles) injected i.p. per mouse. Plasma DNase I
concentration was determined at indicated time after injection in radial diffusion assay.
Park et al., Sci Transl, accepted
Nets per tissue area
(NETs per mm2)
DNase I-coated nanoparticles reduce NETs
after cancer cell injection
Park et al., Sci Transl, accepted
DNase I-mediated digestion of NETs
reduces breast cancer metastasis
Ctrl. nanoparticles
DNase I-coated
nanoparticles
4 mm
Metastatic burden 4 weeks after cancer cells inj.
Nanoparticles inj. i.p. daily for 2 weeks
after cancer cell inj.
Park et al., Sci Transl, accepted
Conclusions and Perspectives
Neutrophil Extracellular Traps promote metastasis
Neutropenia, induced by some types of chemotherapy, is
treated with G-CSF - and NETs are induced by G-CSF…
Is this a problem?
Cancer patients with post-surgical infection have up to 3fold increased risk of metastatic recurrence – could NETs
be targets to reduce this risk?
How do NETs work? Are they scaffolds for concentrated
enzymatic activities?
Acknowledgements
Egeblad Lab
Juwon Park
Robert Wysocki
Laura Maiorino
Miriam Fein
Jean Albrengues
Collaborators
Michael Goldberg & Zohreh Amoozgar (DFCI)
Julie Jorns & Anne Schott (U. Michigan)
Youngseok Lee & Nam H. Won (Korea U.)
Mark Looney & Max Krummel (UCSF)
George Peeters (Solamere Technologies)
Emilis Bruzas
Victoria Kuettner
Mario Shields
Jacqueline Cappellani
Contributing alumni:
Yumi Kunigasa
Elizabeth Nakasone
Ana Almeida
Jing Qiu
Funding
SWOG/JAX/CSHL ITSC
DOD BRCP Era of Hope Scholar
Integrated Translational Science Program:
SWOG, JAX and CSHL
We Acknowledge the GENEROUS support of