Paneth Cell Phenotype Correlates with Genetics, Transcriptome Profile, Pathologic Hallmark and Predicts Clinical Outcome in
Patients with Crohn's Disease
Ta-Chiang Liu, M.D., Ph.D.
Assistant Professor
Department of Pathology and Immunology
Washington University in St. Louis
Disclosures
Nothing to disclose
Goal: Identification of a cellular readout that synthesizes genetic and environmental
factors and subclassifies Crohn’s Disease
EnvironmentGenetic Susceptibility
Cellular readout
Disease subclassification
Risk Loci Predicted to Affect Paneth Cell FunctionATG16L1NOD2XBP1ITLN1
Paneth Cells: Critical Roles in Innate Immunity
AntimicrobialsLysozymeα-Defensins
Secretion defect in Atg16L1-deficient Paneth cells
WT Atg16l1 deficient
Cadwell et al., Nature 2008.
NOD2 Variants and Disease Susceptibility
Common CD-associated variants
Rare CD-associated variants (6 known)
Allele carriage rate: <5%
Allele carriage rate: <1%
Relative risk for development of CD:NOD2 Heterozygous = 2-3NOD2 Homozygous = ~20
HypothesisNOD2 risk variants may be associated with an abnormal granule
phenotype in the Paneth cells of CD patients
Materials and Methods De-identified ileocolectomy tissue from CD patients Inclusion criteria:
Access to proximal margin of ileocolic resection Minimum of 100 well-oriented crypts No active or chronic inflammatory disease in the ileal
sections used for lysozyme stain
Scale bars: 10 µMImmunofluorescenceLysozymeHoescht
Gastroenterology, in press
NOD2 Risk Variants are Associated with Abnormal Paneth cell Phenotype
diminished
NOD2 Risk Variants are Associated with Abnormal Paneth cell Phenotype
*p<0.05 compared to 0 risk variants
Gastroenterology, in press
NOD2 and ATG16L1 Risk Alleles are Additive for Abnormal Paneth Cell Phenotypes
Activated Immune Response Profile is Associated with Abnormal Paneth Cells
Phenotype
GO term analysis performed using DAVID Gastroenterology, in press
Paneth cell phenotype is associated with clinical outcome
Gastroenterology, in press
CD patients from WU and CSMC (n=143) underwent resection and received post-op prophylactic therapy
End point: endoscopic evidence of disease recurrence after surgery
Paneth cell phenotype as predictive cellular biomarker for
CD – practical issues
% Abormal Paneth cellsin uninvolved area
% A
bnor
mal
Pan
eth
cells
in in
volv
ed a
rea
0 10 20 30 40 500
20
40
60
80
Paneth cell phenotypes in involved areas correlate with that of the uninvolved areas
P<0.0001
% Abormal Paneth cellsin first resection
% A
bnor
mal
Pan
eth
cells
in s
econ
d re
sect
ion
0 10 20 30 40 500
10
20
30
40
Paneth cell phenotype remains stable over the years
(n=30)P<0.0001
Paneth cell defects may be patchy: how many crypts are needed to generate readout equivalent
to resection specimens?
“Virtual biopsy”
At least 40-50 crypts are needed to generate equivalent results as with resection specimens.
% Abormal Paneth cellsin resection
% A
bnor
mal
Pan
eth
cells
in b
iops
y
0 20 40 60 80 1000
10
20
30
40
50
Paneth cell phenotypes in matched biopsy and resection specimens are consistent
(n=20)P=0.0004
Normal
Disordere
d
Diminish
ed
Diffuse
Excluded
Enlarged
0
20
40
60
80
100
Tota
l Pan
eth
cells
(%)
Normal
Disordere
d
Diminish
ed
Diffuse
Excluded
Enlarged
0
20
40
60
80
100
Tota
l Pan
eth
cells
(%)
Bad Paneth cell phenotype is more prevalent in pediatric CD
Adult (n=124) Pediatric (n=77)
P< 0.0001
(Collaboration with Nita Salzman)
15%47%
Significant increase in bad Paneth cell phenotype over the past 30 years
1980-1982 (n=52)
Normal
Disord
ered
Diminish
ed
Diffuse
Excluded
Enlarged
0
20
40
60
80
100
Paneth Cell Phenotype
Tota
l Pan
eth
cells
(%)
P=0.0328
Normal
Disordere
d
Diminish
ed
Diffuse
Excluded
Enlarged
0
20
40
60
80
100
Tota
l Pan
eth
cells
(%)
2011-2013 (n=124)
15%3%
1980-1982 (n=52)
Summary
• Bad Paneth cell phenotype is associated with CD-associated risk alleles, distinct gene expression profile, pathology hallmark, and clinical outcome.
• Paneth cell phenotype is spatially and temporally stable.
• Paneth cell phenotype analysis can be performed using routine biopsy material.
• Bad Paneth cell phenotype is commonly seen in pediatric patients, and appears to be a new form of disease that have emerged over the last 30 years.
AcknowledgementCedars-Sinai
Dalin LiFadi TowficNir ModianoRachel WinterTalin HaritunainsDeepti DhallStephan TarganDermot P. B. McGovern
Washington University Kelli VanDussen Robi D. Mitra Rich Head Rodney D. Newberry Feng Gao Thaddeus S. Stappenbeck
Harvard/MGH
Ramnik Xavier
Medical College of Wisconsin
Nita Salzman