Poster presented at Seattle Translational Tumor Research (STTR) 2018 investigator's meeting.

1. Background
Methods and Materials
Acknowledgements
Implementation of Patient-Derived Xenograft and Organoid Colorectal Cancer Models
Conclusions
Results
Figure 2. CRC PDX 114 Tumor
H&E Images
H&E slides from PDX CRC114 in two
separate mice at time of final collection
shown at (A) 4x (B) 20x the mucinous
differentiation and lymphoid aggregation.
Tissue samples:
The CRC PDX samples were obtained from University
of Colorado-Denver and ColoCare study.
Establishment of the CRC PDX lines at FHCRC: All animal
experiments were approved by the IACUC at Fred Hutch.
3D PDX derived organoid Culture: The isolation and
maitainance of tumor organoid culture was adapted from
published protocols (4).
Histology: H/E staining were performed on sections from
FFPE blocks.
CRC Mutation Detection: Variant calls were generated for a
custom panel of CRC-related mutations using Illumina MiSeq
custom TruSeq technology.
Figure 5. Mutation analysis of CRC PDX 125 samples.
Figure 1. Experiment Overview
Figure 4. Tumor organoid derived from
PDX CRC 125 (40x image)
Figure 3. PDX Growth
Curves
PDX tumor CRC125 (A)
and CRC114 (B) was
implanted in several mice at
FHCRC. The growth of
each individual tumor in
each mouse was measured
over the lifetime over of
mouse after the PDX was
implanted.
CRC114CRC125
B
Code
Name Braf KRas NRas PIK3CA
Age of
Consent
Colon
or
Rectal
Primary or
Metastatic
TNM
Stage Chemo?
CRC
125 WT WT WT WT 58
Recta
l M IV Yes
CRC
114 V600E WT WT WT 70
Colon
(right) P III No
A B
• We were able to establish CRC PDX lines and PDX derived 3D
tumor organoids system, enabling accurate preclinical models to
facilitate personalized therapy in CRC.
• The histological and mutation features of PDX tumors and 3D
organoids were overall stable over passages.
Colorectal cancer (CRC) is the third most commonly
diagnosed cancer in the world1
. Approximately 1 in 22 men and
1 in 24 women will be diagnosed with CRC within their lifetime2
.
The mortality rate for men is 17.7% and women is 12.4%3
. This
highlights the need for new advances in treatment and
technology to understand the unique and important biological
features of CRC.
PDX cancer models accurately recapitulate key aspects of
human CRC behavior (i.e. response to therapies) and are
rapidly becoming accepted preclinical models to both query
fundamental biological questions and test novel therapeutics.
With the increasing recognition of the heterogeneity of CRC that
is based on clinical, histopathologic and molecular features and
on the role of the tumor microenvironment on the CRC, PDX
models are believed to be an important preclinical model in
cancer research.
3-D organoids are a cutting edge model system that is the
most physiologically relevant ex vivo system for studying factors
that regulate the formation of CRC. The feasibility to grow
organoids from primary tumor and normal tissue or PDX tumors
has only been demonstrated in the past few years, and remains
a challenging technique.
The goal of this study was to establish CRC PDX lines and
PDX-derived organoids at Fred Hutch, and to assess if the
genetic and histological characteristics were stable over
subsequent passages.
Table 1. Clinical and Mutation information of CRC
patients
Results
Funding is provided from the Seattle Translational Tumor Research
program. Thanks to the Shared Resources at Fred Hutch for
processing and generating the DNA sequencing data. We also wish
to thank staff and patients of the ColoCare Study.
A B
A B
References
1. Bray F et al., Global estimates of cancer prevalence for 27 sites
in the adult population in 2008. Int J Cancer 2013; 132:1133–
1145.
2. Kerr DJ, Domingo E, Kerr R. Is sidedness prognostically
important across all stages of colorectal cancer? Lancet Oncol.
2016;17: 1480-1482
3. Colorectal Cancer Facts & Figures 2017-2019. Centers for
Disease Control and Prevention. Behavioral Risk Factor
Surveillance System, 2014.
4. Sato et al., Long-term expansion of epithelial organoids from
human colon, adenoma, adenocarcinoma and Barrett’s
epithelium. Gastroenterology 141, 1762-1772.
Yu, Ming1
; Ayers, Jessica L.1
; Maden, Sean1
; Carter, Kelly1
; Guo, Yuna1
; Vickers, Kathy2
; Li, Christopher2
; Cromwell, Elizabeth A3
; Uthamanthil, Rajesh K3
; Stella, Nephi4
; Westerhoff, Maria5
; Stachler, Matthew6
; Pitts, Todd7
;
Eckhardt, Gail7
;Grady, William M.1
1. Division of Clinical Research; 2. Division of Public Health Sciences; 3. Comparative Medicine, Fred Hutchinson Cancer Research Center, Seattle, WA. USA; 4. Department of Pharmacology, 5. Department of Pathology, University of Washington, Seattle, WA. USA;
6.Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA; 7.Division of Medical Oncology, University of Colorado Denver, Denver, CO, USA.