Targeting B cell receptor signaling in cancer: preclinical ......Targeting B cell receptor signaling...
Transcript of Targeting B cell receptor signaling in cancer: preclinical ......Targeting B cell receptor signaling...
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Targeting B cell receptor signaling in cancer: preclinical and clinical advances
Jan Burger
Innovations in Hematology
Dan Tel Aviv Hotel, Tel Aviv, Israel
May 29, 2018
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Targets in the BCR signaling pathway
Burger & O’Brien Nat Rev Clin Onc 2018
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VH D JH C
NN
Somatic mutations
1/51 1/27 1/6
B-Cell Diversity: IGHV
Rearrangement and Mutation
VH in chronic lymphocytic leukemia (CLL)
– U-CLL, with 98% or more sequence homology with the corresponding consensus germline sequence
– M-CLL, with less than 98% sequence homology
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Su
rviv
ing
(%
)
Survival of CLL Patients With Mutated vsUnmutated IGHV
All patients (N=84) Stage-A CLL patients (n=62)
Su
rviv
ing
(%
)
0 50 100 150 200 250 3000
20
40
60
80
100
Months
P=0.0008
Months
P=0.001
0 50 100 150 200 250 3000
20
40
60
80
100
Mutated
UnmutatedUnmutated
Mutated
Hamblin TJ et al. Blood. 1999;94:1848-1854.
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In CLL:
Sites of proliferation =
sites of BCR activation
CLL lymph nodes
From: Soma LA et al,
Human Pathology. 2006;37:152-159
Proliferation
centers
Burger & O’Brien Nat Rev Clin Onc 2018
Fro
m: Y
He
rish
an
ue
t a
l., B
loo
d. 2
01
1
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• Pro-B cells to pre-B cells : generation of μ heavy chain by rearranging VDJ segments
• The pre-B cell receptor (pre-BCR) μ heavy chain paired with the surrogate light chain (λ5 and VpreB)
• Mature B cells express IgM and IgD
• Subsequent B cell development occurs in the secondary lymphoid organs (SLOs)
• In the germinal centre (GC), the enzyme activation- induced cytidine deaminase (AID) introduces mutations in IGHV and IGLV to diversify the Ig repertoire (somatic hypermutation)
• AID also mediates class switch recombinationto generate IgA, IgG or IgE
• Selected GC B cells give rise to memory B cells and long-lived plasma cells
• Plasma cells home to the bone marrow or reside in SLOs, where they secrete antibodies
Burger & Wiestner, NRC 2018
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• Centroblasts: somatic hypermutation in the dark zone
• Centrocytes in the light zone: antigen selection and affinity maturation
• Differentiation into memory B cells or plasma cells
• CLL cells proliferate in proliferation centers, in close contact with T cells and nurselike cells
• Antigen triggers BCR signaling in CLL cells in SLO
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Antigen-dependent and oncogenic BCR signaling in B cell malignancies
a) Chronic infections can lead to clonal B cell expansion. Autoantigen can stimulate polyreactive BCRs. Autonomous BCR signalling,
b) Antigen-dependent chronic active BCR signalling is increased by oncogenic mechanisms. Mutations in CD79A and CD79B contribute to the activation of BCR signalling in ABC-DLBCL.
c) Oncogenic tonic BCR signaling. In Burkitt lymphoma (BL), expression of MYC and transcription factor 3 (TCF3) cooperate to promote tonic BCR signalling; MYC upregulates the microRNAs miR-19a and miR-19b, which in turn decrease expression of the PI3K negative regulator PTEN.
Burger & Wiestner, NRC 2018
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The B cell receptor signaling pathway
Burger & Wiestner, NRC 2018
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IgM and IgD receptors are highly expressed in CLL
subsets
M-C
LL
U-C
LL
CD38
neg
CD38
pos
ZAP70
neg
ZAP70
pos
del13
q,tri1
2,neg
del11
q,del
17p
0
100
200
300
MF
IR
**** ns ** *
IgMIgM IgD
M-C
LL
U-C
LL
CD38
neg
CD38
pos
ZAP70
neg
ZAP70
pos
del13
q,tri1
2,neg
del11
q,del
17p
0
50
100
150
200
MF
IR
** ns *** ns
IgD
Ten Hacken E, J Immunol. 2016
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IgD stimulation induces strong HS1 protein activation
anti-IgM
(minutes)
0 2 5 0 2 5
2 5 2 5100
150
200
250
300
350
HS
1 p
ho
sp
ho
ryla
tio
n
(% u
ntr
eate
d c
on
tro
l)
Minutes of anti-IgM
Minutes of anti-IgD
**
anti-IgM
(minutes)
anti-IgD
(minutes)
anti-IgD
(minutes)
P-HS1
(Y397)
HS1
IgM and IgD: upstream signaling responses
Ten Hacken E, J Immunol. 2016
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IgM induces prolonged pathway activation
P-ERK
ERK
0 2 5 15 30 0 2 5 15 30
anti-IgM
(minutes)
anti-IgD
(minutes)
5 15 302 60
100
200
300
400
BCR stimulation (minutes)
ER
K p
ho
sp
ho
ryla
tio
n
(%u
ntr
ea
ted
co
ntr
ol)
* *
*
IgM
IgD
IgM and IgD: downstream signaling responses
Ten Hacken E, J Immunol. 2016
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IgM signaling exclusively induces CCL3 and CCL4 secretion
an
ti-I
gM
an
ti-I
gD
0 1000 2000 3000 4000
Untreated
0.5 µg/mL
1 µg/mL
5 µg/mL
10 µg/mL
0.5 µg/mL
1 µg/mL
5 µg/mL
10 µg/mL
CCL3 (pg/mL)
0 2000 4000 6000 8000 10000
Untreated
0.5 µg/mL
1 µg/mL
5 µg/mL
10 µg/mL
0.5 µg/mL
1 µg/mL
5 µg/mL
10 µg/mL
CCL4 (pg/mL)an
ti-I
gM
an
ti-I
gD
IgM and IgD: CCL3 and CCL4 chemokine secretion
Ten Hacken E, J Immunol. 2016
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IgM induces CCL3 secretion through BCL6 down-regulation
3 6 9 12 240
500
1000
1500
BCR stimulation (hours)
CC
L3
(pg
/mL
)
anti-IgM
anti-IgD
BCL6
GAPDH
anti-IgM stimulation
(hours)
0 3 6 9 12 24
Untreated
(hours)
3 6 9 12 24
3 6 9 12 24
anti-IgD stimulation
(hours)
0 3 6 9 12 24
Untreated
(hours)
BCL6
GAPDH
BCL
6
IgM and IgD: mechanism of CCL3 production
Ten Hacken E et al., J Immunol. 2016 Sep 15;197(6):2522-31
Ten Hacken E, J Immunol. 2016
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Variable CCL3 protein expression in CLL lymph nodes
• In 42 cases: 24 CCL3 positive (56%) and 19 CCL3 negative (44%) cases. • Membranous staining, impression of secretion of CCL3 into the
environment• Prolymphocytes and paraimmunoblasts particularly in the proliferation
centers were positive
Hartmann EM,… & Rosenwald A. Leukemia& Lymphoma 12:1-9, 10/2015
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• CCL3 positive cases (B) shows more prominent T cell infiltrates (CD3, CD4 and CD8) • CCL3 positive cases (B) have higher number of CD57 positive cells and a higher
proliferation fraction (Ki-67-pos.)
Hartmann EM,… & Rosenwald A. Leukemia& Lymphoma 12:1-9, 10/2015
Association between CCL3 and T cell densityCCL3-neg. CCL3-pos. CCL3-neg. CCL3-pos.
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BCR-RELATED BIOMARKERS:CCL3, CCL4 (MIP-1α,β)
CCL3
CCL4
pg
/mL
time (days)
pre-treatment
S. Ponader et al., Blood 119: 1182-9, 2012
pre-treatment
Ibrutinib trial
Idelalisib trial
Hoellenriegel J et al.; Blood 118(13):3603-12, 09/2011
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Heavy water labeling of CLL cells prior to ibrutinib therapy
MDACC
KineMed, Inc.
Feinstein Institute
Burger J JCI Insight 2017; 2(2):e89904
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Heavy water labeling of CLL cells prior to ibrutinib therapy:Effects of ibrutinib on birth and death rates
• CLL cell proliferation (“birth”) rates: before ibrutinib therapy 0.39% down to 0.05% on ibrutinib
• Death rates increased from 0.18% to 1.5%
• First direct in vivo measurements of ibrutinib’s anti-leukemia activity
• Profound inhibition of CLL cell proliferation
• Promotion of high rates of CLL cell death
Burger J JCI Insight 2017; 2(2):e89904
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Role of tumour-infiltrating B cells in pancreatic ductal adenocarcinoma
Burger & Wiestner, NRC 2018
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Clonal evolution on ibrutinib
Ahn, Underbayev, et al Blood 2017
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Mutations in BTK & PLCG2 confer BTKi resistance
From Wiestner, Haematologica 2015
Chang, ASCO 2013; Woyach, NEJM 2014; Furman, NEJM 2014
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Early clonal shifts during treatment with ibrutinib
Landau DA Nature Comm 2017
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Ibrutinib-resistant patients
MDACC cohortPt # Age
(yrs)/
Gender
/
Rai
stage
Prior therapy Pre-ibrutinib
FISH cytogenetics
IGHV
(M, U)
Treatment Best
response
to
ibrutinib
Time to
PD on
ibrutinib
1 59/M
Rai III
FCR del (17p),
del (13q)
ND Ibrutinib PR 983
2 36/F
Rai IV
FCR,
R+HDMP
del (11q)* U Ibrutinib +
rituximab
PR 176
3 85/F
Rai IV
R, BR, CLB,
R+HDMP
del (17p),
del (13q), trisomy
12*
U Ibrutinib PR 554
4 58/M
Rai IV
FCR, FR,
CHOP, allo-
Tx, BR,
revlimid,
ofatumuma
b
del (17p),
del (11q),
del (13q)*
U Ibrutinib PR 669
5 58/M
Rai II
FCR, F, R, B del (11q),
del (13q)
U Ibrutinib PR 392
*Complex cytogenetics pre-ibrutinib:Pt 2: 46,XX,del(2q37),del(6q21q23), add(7q36),del(11q21q25)Pt 3: 45,X,add(X)(q24), add(6p23), add(12p13), del(17p11.2), -21Pt 4: 45,XY,del(6q13q23), add(8p21), del(11q21q24), -17,-18,+mar[5]
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Clonal evolution in CLL patients during
ibrutinib therapy: Patient #1
Burger, Landau …. & Wu Nature Comm 2017
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Clonal evolution during ibrutinib therapy:
transdifferentiation into histiocytic sarcoma
Burger, Landau …. & Wu Nature Comm 2017
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Droplet-based detection of resistance subclonesat the time of ibrutinib treatment initiation
Conclusion: our analyses support the presence of resistant sub-clones at treatment initiation, in line with theoretical predictions
Burger JA Nature Comm 2017
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SUMMARY• Ibrutinib and other BCR signaling inhibitors including
idelalisib results in major clinical benefit for patients with CLL
• However, this therapy also exerts strong selective pressure, which can promote the outgrowth of resistant subclones
• Long-term toxicity and resistance development are problems with long-term use of ibrutinib (plus: high costs)
• Intermittant therapy could help addressing these issues
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Thank-
you!Collaborators:• Würzburg University: A Rosenwald, E
Hartmann• CLLGRF: F Caligaris-Cappio, N
Chiorazzi, Z Estrov, N Kay• MDACC: M Keating, W Wierda, S
O’Brien, H Kantarjian, V Gandhi, A Ferrajoli, K Balakrishnan
• UCSD: T Kipps, L Rassenti• UC Irvine: D Wodarz, N Komarova• DFCI, Broad I: C Wu, D Landau
My laboratory: Mariela Sivina, Julia Hoellenriegel, Stefan Koehrer, Ekaterina Kim, Elisa ten Hacken, ShubhchintanRandhawaFunding: CPRIT, MD Anderson Moonshot,Leukemia & Lymphoma Society
Dept. of Leukemia, MDACC