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Title:Presenters:
Date:
Advancing CAR T Cells for the Treatment of Solid Tumors
Saul Priceman, Ph.D. Assistant Professor
Hematology & Hematopoietic Cell Transplantation
Associate Director, Translational Sciences, TCTRL
Beckman Research Institute
City of Hope
Disclosures
I receive Grant/Research Support from PICI and Mustang.
I am a Consultant for Mustang and Apterna.
Outline
• Immunotherapy and CAR T Cells
• CAR T Cell Program at City of Hope
• Vignettes of Solid Tumor Clinical CAR T Cell Programs at COH
• On the Horizon: Preclinical Development and Combination Therapies
Four Pillars of Cancer Therapy:
Today’s Landscape of Immunotherapy
Yu, et al. Nat Rev Drug Disc 2019
Escape mechanisms in the majority of cancers MHC down-regulation / defective antigen presentation of TAAs Immunosuppressive tumor-infiltrating immune cells
Advantages over other immunotherapy strategies:1. Direct attacking of tumor-associated antigens2. MHC-independent3. Rapid cell product manufacturing
Chimeric Antigen Receptor (CAR) T Cells
Adapted from Roberts et al, Leuk & Lymphoma 2017.
Conventional TCR:MHC Chimeric antigen receptor
1st generation
2nd generation
3rd generation
4th generation
Adoptive Therapy using CAR-engineered T Cells
Priceman, et al. Curr Opin Oncol 2015
2016-2020 COH CAR T TRIALS PATIENTS 286+
Hematologic cancers
Solid tumors
CD19-CAR T cells for Relapsed B-Cell Lymphoma and Leukemia
Case Report:61 yr; maleRelapsed high-grade B cell lymphomaLymphodepletion: Flu/CyCD19-28z CAR T cells (200M; Tn/mem)Grade 2 CRS (1x toci); no neurotoxicity
Lymphodepletion: Flu/Cy
CD19:28ζ CAR T cells (200M; Tn/mem-derived)
Case Report: 61 yr; male
Relapse high-grade B cell lymphoma
Pre-treatment Post-CART 28-days
Grade 2 CRS (1x toci); no neurotoxicity
Lymphodepletion: Flu/Cy
CD19:28ζ CAR T cells (200M; Tn/mem-derived)
Case Report: 61 yr; male
Relapse high-grade B cell lymphoma
Pre-treatment Post-CART 28-days
Grade 2 CRS (1x toci); no neurotoxicity
Pre-treatment Post CAR T cells: 28 days
Clinical Trial
Disease Cell Population Cell Dose(CAR+)
Treated Patients ResponseRate
NCT02051257
NHLw/ auto-transplant
(MRD; low/negantigen)
Tn/mem 200M 6 Pending
NCT02153580
CD19+ B cell Neoplasms
(Active disease)
Tn/mem 200M
600M
5
2
1 of 1 CR (4 Pending)
Pending
NCT02146924
B-ALL(Active disease)
Tcm 200 M 3 30% CR (1 of 3)
Tn/mem 200 M 13 100% CR (13 of 13)
Moving Beyond Blood Cancers
Do CAR T cells offer similar promise for treatment of
solid cancers?
Clinical development of CAR T cells
CD19-CAR T Cells for B-Cell Malignancies
2017: Landmark FDA Approvals
DLBCL (Kite/Gilead)
Pediatric B-ALL + DLBCL (Novartis)
Hartmann, et al. EMBO Mol Med, 2017
T Cell Therapy Program at COH
Brain
- Glioma
- Brain Metastasis
Hematological
- Leukemia – AML, ALL
- Lymphoma
- Multiple Myeloma
Solid Tumors
- Prostate
- Breast
- Pancreatic
- Ovarian
- Liver
Phase I Clinical Trial Evaluating IL13Rα2-targeted CAR T cells
Patient Population Intracranial Delivery Route Investigational Agent T cells DosePlanned
Enrollment Study Objectives
Recurrent IL13Rα2+ High-grade glioima
12 -75 yrs
Tcm-derived IL13BBζ CAR T cell Arm 1: biopsy (ICT-b) Arm 2: resection (ICT-r) Arm 3: intraventricular (ICV) Arm 4: dual ICT/ICVTn/mem-derived IL13BBζ CAR T cell Arm5: dual ICT/ICV
AutologousIL13BBζ/CD19t T cells
2 to 100x106
CAR T cells15 patients/
arm
•Feasibility and safety
•Determine maximum tolerated dose (MTD) and a recommended phase II dosing plan (RP2D)
Clinical PI: Dr. Behnam BadieResearch PI: Dr. Christine Brown
OptionalInfusions 4+
clinicaltrials.gov, NCT02208362
Brown et al., NJEM 2016
MRI+Gd
post-ICT CAR-T
T6
T7
MRI+Gd
post-ICV CAR-T
MRI+Gd T6
T7
MRI+Gd
d108
d85 d289
d254
• CAR Design / Delivery – CARs for brain metastasis
– CARs for mCRPC
– CARs for ovarian cancer
• Tumor Antigen Expression Profile and Heterogeneity– Engineering tumor selectivity of CARs
– Multi-targeted CAR T cells
• Immunosuppression / Physical Barriers– Development of immunocompetent mouse models
– Intrinsic optimization of CAR (4th gen)
Challenges for Solid Tumor CAR T Cell Therapy
Normal
tissue
Tumor
CD
3 T
ce
lls
• CAR Design / Delivery – HER2-CARs for brain metastasis (Priceman et al. Clinical Cancer Res 2018) – Phase I Trial Q4 2018
– PSCA-CARs for mCRPC (Priceman et al. OncoImmunology 2017) – Phase I Trial Q3 2019
– TAG72-CARs for ovarian cancer (Murad et al. Front Immunol 2018)
• Tumor Antigen Expression Profile and Heterogeneity– Engineering tumor selectivity of CARs
– Multi-targeted CAR T cells
• Immunosuppression / Physical Barriers– Development of immunocompetent mouse models
– Intrinsic optimization of CAR (4th gen)
Challenges for Solid Tumor CAR T Cell Therapy
Priceman et al. Clin Cancer Res 2018
T Cell Therapy Program at COH
Brain
- Glioma
- Brain Metastasis
Hematological
- Leukemia – AML, ALL
- Lymphoma
- Multiple Myeloma
Solid Tumors
- Prostate
- Breast
- Pancreatic
- Ovarian
- Liver
FME15-50 0.2x10^6 BBM1-ZsGreen-ffluc i.c.@D0
0 10 20 30 40
tx
50 100 150104
105
106
107
108
109
1010
1011
Days post tumor injection
Flu
x (p
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ton
s/se
c) Mock i.c.
Mock i.c.v.
HER2(EQ)BBz i.c.
HER2(EQ)BBz i.c.v.
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MFI: 117
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MFI: 33
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MFI: 395
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MFI: 5
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MFI: 297
% Chromium Release
Effector:Target LC
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HER2-
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Mock i.c.v.
51
Intraventricular Delivery of HER2-BB𝜁 CAR T Cells for the
Treatment of Breast Cancer Brain Metastases
Priceman, et al. Clin Cancer Res 2018
-ICV therapy provides potent antitumor activity against brain metastases and is superior to IV delivery (at 10-fold lower dose)
Mock i.c.
HER2-BBζ i.c.
HER2-BBζ i.c.v.
Mock i.c.v.
FME16-42 BBM1-eGFP-ffluc
HD007.10 CAR T cells
i.c.v. vs. i.v.
0 50 100 150
tx
104
105
106
107
108
109
1010
1011
Days post tumor injection
Flu
x (p
hoto
ns/
sec)
HER2-BBz i.v.
Mock i.v.Tumor Only
HER2-BBz i.c.v.
0 50 100 150
tx
0
50
100
Days post tumor injection
Perc
ent surv
ival Tumor Only
HER2-BBz i.c.v.
HER2-BBz i.v.
Mock i.v.
edTumor only
HER2-BBζ i.v.
HER2-BBζ i.c.v.
Mock i.v.
Phase I Clinical Trials to Evaluate HER2-BB𝜁 CAR T Cells in Brain Cancers
• HER2+ Brain/Leptomeningeal Metastases
(Clinical PI: Jana Portnow, MD, Research PI: Saul Priceman, PhD) – enrolling
• HER2+ GBM – enrolling
OptionalInfusions 4+HER2
HER2BBz HER2BBz HER2BBz
HER2-CAR in BBM phase 1 trial
Priceman et al.
Oncoimmunology 2018
T Cell Therapy Program at COH
Brain
- Glioma
- Brain Metastasis
Hematological
- Leukemia – AML, ALL
- Lymphoma
- Multiple Myeloma
Solid Tumors
- Prostate
- Breast
- Pancreatic
- Ovarian
- Liver
PSCA-41BBζ CAR T Cells Show Increased Control of Disseminated Disease
-4-1BB co-stimulation demonstrates durable anti-tumor activity in patient-derived PSCA+ PCa bone metastasis xenograft model, compared with CD28 co-stimulation
Priceman, et al. OncoImmunology 2018
0.0
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Day 24: FME16-10Day 76: FME16-41
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p < 0.05
NS
0
1000
2000
3000
4000
PS
A (
pg
/mL)
PSCA(ΔCH2)28ζ
PSCA(ΔCH2)BBζ
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Tu
mo
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Mo
ck
tx
0/8
0%
0/9
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PS
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(ΔC
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3/11
27%
PS
CA
(ΔC
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10/11
91%
Survival
NSG
LAPC-9
(intratibial)
PSCA-CAR human
CD3+ T cells
b
a
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Tu
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lyM
ock
PS
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Phase I Clinical Trials to Evaluate PSCA-BB𝜁 CAR T Cells in Solid Tumors
• PSCA+ metastatic castration resistant prostate cancer
(Clinical PI: Tanya Dorff, MD, Research PI: Saul Priceman, PhD) – enrolling
• PSCA+ metastatic pancreatic cancer – TBD
PSCA-CAR in mCRPC phase 1 trial
! 4
Toxicity and disease response evaluations will be evaluated as described above.
Study Population: Patients eligible for the proposed study should have 1) pathologic diagnosis of prostate cancer, (2) metastatic castration resistant prostate cancer (mCRPC) (Note: castration will be defined by a
testosterone <50 ng/dL achieved by orchiectomy or LHRH agonist/antagonist therapy), 3) disease progression
on the last line of therapy based on: rising PSA with 2 consecutive values 7 days apart or measurable disease
with an increase in 20% or more of longest diameters of measurable lesions or non-measurable disease with 1 or more new lesions in soft tissue, or 2 or more new lesions in bone, and (4) prior abiraterone or enzalutamide,
but not both. Patients may also have had 1) chemotherapy for castration-sensitive prostate cancer, but not for
castration-resistant disease, 2) prior radiotherapy, provided it was rendered > 28 days prior to treatment, or 3) prior use of sipuleucel-T.
Objectives: The primary objectives are 1) to evaluate the safety and tolerability of PSCA(ΔCH2)BBζ-CAR T
cells in patients with mCRPC, and 2) identify the recommended Phase II dose (RP2D). The secondary
objectives are 1) to assess clinical response based on Prostate Cancer Working Group 3 (PCWG3) criteria and 2) to assess whether PSCA-CAR T cells expand and persist. Correlative objectives include 1) enumeration and
phenotypic characterization of circulating tumor cells (CTC) pre- and post-therapy and 2) characterization of
humoral and cell-mediated immunity to PSCA and other known prostate cancer antigens.
Endpoints: The primary endpoints are DLTs and all other toxicities post CAR T cells. The secondary endpoints
include: 1) response based on Prostate Cancer Working Group 3 (PCWG3) criteria and 2) persistence of T
cells to 28 days post infusion (defined as CAR T cells >0.1% of total CD3 cells by flow-cytometry; AUC of log10 copies/µg of genomic DNA). The correlative endpoints are 1) assessment of circulating tumor cell (CTC)
conversion (from ≥5 CTCs/7.5 mL to <5 CTCs/7.5 mL, or vice versa) and 2) detection of AR-V7 splice variants.
Toxicity: will be assessed using the National Cancer Institute’s Common Terminology Criteria for Adverse
Events (CTCAE v5). A DLT is defined as: 1) any Grade 3 or higher toxicity occurring within 28 days of T cell infusion with an attribution of definitely or probably related to T cell infusion, excepting expected adverse
events of specified grade and duration, including cytokine release syndrome (CRS); and 2) any Grade 3 or
greater autoimmune toxicity occurring within 28 days of T cell infusion. A toxicity of any grade that is normally expected with advanced prostate cancer or related prior therapy and/or treatment will not be considered a DLT
with respect to protocol continuation, or dose escalation/de-escalation of T cell dose.
Study Design: This is a Phase I dose escalation trial of adoptive T cell therapy. This trial seeks to determine an RP2D to test in future phase II
trials. RP2D will be based on maximum tolerated dose (MTD),
participant data on disease response, late toxicities and 2nd
infusions.
The toxicity equivalence range (TEQR) design of Blanchard and Longmate
22 will be used to evaluate select doses of PSCA(ΔCH2)BBζ-CAR T cells and determine the MTD.
The dose schedule is shown in Table 1. The starting dose will be dose 0. The TEQR design22 can be viewed
as a minimal elaboration of the 3+3 design to include an explicit toxicity target range, and permit intuitive specification of a too-toxic level for closing a dose level. In this implementation of the TEQR design, we define
the target equivalence range of DLT as 0.20-0.35. Toxicity levels of 0.51 or higher will be considered too toxic
Table 1. CAR+ Cell Dose Schedule
Dose -1 Starting Dose 0
Dose 1 Dose 2
25M 50M 200M 800M
Figure 3: T cell product manufacturing and patient treatment plan. BX = biopsy, PB = peripheral blood for correlative assays, CT =!computed tomography scan, PET = positron emission tomography, LTFU = long-term follow-up. *Cyclophosphamide lymphodepleting regimen, 1 or 2 days at the discretion of PI, based on disease burden and co-morbidities. **T cell infusion may be given within a window of 3-10 days after last dose of lymphodepleting regimen
CT CT
Dose -1
Starting
Dose 0a Dose 0b Dose 1 Dose 2
50M 100M 100M +precond. 300M +precond. 600M + precond.
Table 1. CAR+ Cell Dose Schedule
Murad et al.
Front Immunol 2018
T Cell Therapy Program at COH
Brain
- Glioma
- Brain Metastasis
Hematological
- Leukemia – AML, ALL
- Lymphoma
- Multiple Myeloma
Solid Tumors
- Prostate
- Breast
- Pancreatic
- Ovarian
- Liver
Regional delivery of TAG72-CAR T cells delay the growth of human peritoneal
ovarian tumors in xenograft models
0 50 100 150 2000
25
50
75
100
Days post tumor injection
Perc
ent S
urv
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Mock (i.p.)
Mock (i.v.)
TAG72-BBζ (i.p.)
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6 13 29
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100
Days post T cell infusion
CA
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ells
per
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lood
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0 25 50 75
tx
107
108
109
1010
1011
Days post tumor injection
i.p. OVCAR3 tumor
i.p. T cell
i.v.T cell
a
Figure 3
c
0 25 50 75
tx
107
108
109
1010
1011
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photo
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0 25 50 75
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107
108
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107
108
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0 25 50 75
tx
107
108
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0 25 50 75
tx
107
108
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1011
Days post tumor injection
d
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Day 6 Day 13
Mock
(i.p
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ock
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(i.v
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4%
1%92%
14%
0%86%
13%
0%86%
87%
7%5%
82%
15%1%
17%
0%83%
40%
16%44%
b
Mock i.p.
TAG72-BBζ i.p.
Mock i.v.
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Com
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Com
ment2:
1.0
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0.6
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0.2
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Min = -1.4387e+08
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*** ns ns
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0 50 100 150 2000
25
50
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100
Days post tumor injection
Perc
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Days post T cell infusion
CA
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0 25 50 75
tx
107
108
109
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1011
Days post tumor injection
i.p. OVCAR3 tumor
i.p. T cell
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a
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c
0 25 50 75
tx
107
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0 25 50 75
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0 25 50 75
tx
107
108
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0 25 50 75
tx
107
108
109
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1011
Days post tumor injection
d
e
Mock i.p.
TAG72-BBζ i.p.
Mock i.v. TAG72-BBζ i.v.
f9%
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Day 6 Day 13
Mock
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ock
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1%92%
14%
0%86%
13%
0%86%
87%
7%5%
82%
15%1%
17%
0%83%
40%
16%44%
b
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H20131001122422
Tue,Oct 01, 2013 12:24:22
Emfilter=O
penBin:M
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Cam
era: IVIS
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Experiment: FM
E13-57
Com
ment1:
Com
ment2:
1.0
0.8
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Color B
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Max = 1e+06
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Day 2
6
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6
10
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*** ns ns
***
0 50 100 150 2000
25
50
75
100
Days post tumor injection
Perc
ent S
urv
ival
Mock (i.p.)
Mock (i.v.)
TAG72-BBζ (i.p.)
TAG72-BBζ (i.v.)
6 13 29
0.1
1
10
100
Days post T cell infusion
CA
R+
T c
ells
per
mL b
lood
i.p.
i.v.
p < 0.001 p = 0.6 p = 0.8
0 25 50 75
tx
107
108
109
1010
1011
Days post tumor injection
i.p. OVCAR3 tumor
i.p. T cell
i.v.T cell
a
Figure 3
c
0 25 50 75
tx
107
108
109
1010
1011
Flu
x (
photo
ns/s
ec)
0 25 50 75
tx
107
108
109
1010
1011
Flu
x (
photo
ns/s
ec)
0 25 50 75
tx
107
108
109
1010
1011
0 25 50 75
tx
107
108
109
1010
1011
0 25 50 75
tx
107
108
109
1010
1011
Days post tumor injection
d
e
Mock i.p.
TAG72-BBζ i.p.
Mock i.v. TAG72-BBζ i.v.
f9%
8%82%
Day 6 Day 13
Mock
(i.p
.)M
ock
(i.v
.)TA
G72-
BBζ
(i.p
.)TA
G72-B
Bζ
(i.v
.)
mC
D4
5
hCD45
4%
1%92%
14%
0%86%
13%
0%86%
87%
7%5%
82%
15%1%
17%
0%83%
40%
16%44%
b
Mock i.p.
TAG72-BBζ i.p.
Mock i.v.
TAG72-BBζ i.v.
tx
Click # H
H20131001122422
Tue,Oct 01, 2013 12:24:22
Emfilter=O
penBin:M
(8), FOV25,f1, 3s
Cam
era: IVIS
186, SI620E
EV
User: H
HG
roup:Grp4
Experiment: FM
E13-57
Com
ment1:
Com
ment2:
1.0
0.8
0.6
0.4
0.2
x106
Image
Min = -1.4387e+08
Max = 1.6421e+08p/sec/cm̂2/sr
Color B
arMin = 1e+05
Max = 1e+06
bkgsub
flat-fieldedcosm
ic
WAR
NING
: Saturated Lu
minescent Im
age
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OV
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3
98
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ositiv
e
MF
I: 11
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Ga
M-P
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27
44
Ca
ov-3
A2
78
0
79
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ositiv
e
MF
I: 33
10
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po
sitiv
e
MF
I: 39
5
22
% p
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e
MF
I: 5
99
% p
ositiv
e
MF
I: 29
7
% Chromium Release
Effe
cto
r:Targ
et
LC
L-O
KT
3
MA
DH
27
44
Ca
ov-3
SK
OV
3O
VC
AR
3
A2
78
0
0
20
40
60
80
100
50
:1 2
5:1
5:1
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12
0
20
00
IFN
-g
15
00
10
00
60
0
40
0
20
00
pg/mL
TN
F-a
D
MA
DH
27
44
Ca
ov-3
SK
OV
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VC
AR
3
A2
78
0
PB
SM
ock
CD
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CE
7R
Day 3
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n
Day
11
Day 1
2 =
T c
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tratio
n
Day 1
8
Day 2
6
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05
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04
10
6
10
5
108106
13 66433428242117Days:
*** ns ns
***
ba
Murad, et al. Front Immunol 2018
-Intraperitoneal delivery of TAG72-CAR T cells demonstrate potent anti-tumor activity with improved survival of mice bearing peritoneal ovarian tumors, but not intravenous delivery-Tumor recurrences observed in all mice following therapy
0 25 50 75
tx
107
108
109
1010
1011
Days post tumor injection
0 25 50 75106
107
108
109
1010
1011 tx
a
Figure 4
i.p. OV90 tumor
i.p. T cell
1x vs. repeat
0 25 50 75106
107
108
109
1010
1011 txc
0 25 50 75
tx
107
108
109
1010
1011
Flu
x (
photo
ns/s
ec)
0 25 50 75106
107
108
109
1010
1011 tx
0 25 50 75106
107
108
109
1010
1011 tx
0 25 500
1
210
30
50
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
0 25 500
10
20
30
40
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
d
Mock 1xMock repeat
TAG72-BBζ 1x
TAG72-BBζ repeat
b
Mock 1x
TAG72-BBζ 1x
Mock repeat
TAG72-BBζ repeat
tx
0 25 500
1
210
30
50
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
0 25 500
10
20
30
40
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
Click # HH
20131001122422Tue,O
ct 01, 2013 12:24:22Em
filter=Open
Bin:M (8), FOV
25,f1, 3sC
amera: IV
IS 186, S
I620EE
V
User: HH
Group:G
rp4Experim
ent: FME
13-57C
omment1:
Com
ment2:
1.0
0.8
0.6
0.4
0.2
x106
Image
Min = -1.4387e+08
Max = 1.6421e+08p/sec/cm̂2/sr
Color B
arMin = 1e+05
Max = 1e+06
bkgsub
flat-fieldedcosm
ic
WARNING
: Saturated Lu
minescent Im
age
EAB
L1
CA
M/C
E7
scF
v
CD
28
tm/c
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hu
CD
3z c
yt
hug F4 c
zzz
C
OV
CA
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SK
OV
3
98
% p
ositiv
e
MF
I: 11
7
CE
7 +
Ga
M-P
E
MA
DH
27
44
Ca
ov-3
A2
78
0
79
% p
ositiv
e
MF
I: 33
10
0%
po
sitiv
e
MF
I: 39
5
22
% p
ositiv
e
MF
I: 5
99
% p
ositiv
e
MF
I: 29
7
% Chromium Release
Effe
cto
r:Targ
et
LC
L-O
KT
3
MA
DH
27
44
Ca
ov-3
SK
OV
3O
VC
AR
3
A2
78
0
0
20
40
60
80
100
50
:1 2
5:1
5:1
1:1
12
0
20
00
IFN
-g
15
00
10
00
60
0
40
0
20
00
pg/mL
TN
F-a
D
MA
DH
27
44
Ca
ov-3
SK
OV
3O
VC
AR
3
A2
78
0
PB
SM
ock
CD
19R
CE
7R
Day 3
Day 5
= T
cell a
dm
inis
tratio
n
Day
11
Day 1
2 =
T c
ell a
dm
inis
tratio
n
Day 1
8
Day 2
6
5x1
05
5x1
04
10
6
10
5
108106
7 5641342822191511Days:
tx tx tx
f TAG72-BBζ 1x TAG72-BBζ repeat TAG72-BBζ 1x TAG72-BBζ repeat
Day 42 Day 70
CD
3
0 20 40 60 80 100 1200
25
50
75
100
Days post tumor injection
Perc
ent surv
ival
Mock (1x)
Mock (repeat)
TAG72-BBζ (1x)
TAG72-BBζ (repeat)
Mock repeat
e
***
***
Murad, et al. Front Immunol 2018
-CAR T cell repeat infusions more effective in controlling peritoneal tumors, but once ceased, tumors recurred with similar kinetics
Regional delivery of TAG72-CAR T cells delay the growth of human peritoneal
ovarian tumors in xenograft models
0 25 50 75
tx
107
108
109
1010
1011
Days post tumor injection
0 25 50 75106
107
108
109
1010
1011 tx
a
Figure 4
i.p. OV90 tumor
i.p. T cell
1x vs. repeat
0 25 50 75106
107
108
109
1010
1011 txc
0 25 50 75
tx
107
108
109
1010
1011
Flu
x (
photo
ns/s
ec)
0 25 50 75106
107
108
109
1010
1011 tx
0 25 50 75106
107
108
109
1010
1011 tx
0 25 500
1
210
30
50
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
0 25 500
10
20
30
40
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
d
Mock 1xMock repeat
TAG72-BBζ 1x
TAG72-BBζ repeat
b
Mock 1x
TAG72-BBζ 1x
Mock repeat
TAG72-BBζ repeat
tx
0 25 500
1
210
30
50
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
0 25 500
10
20
30
40
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
Click # HH
20131001122422Tue,O
ct 01, 2013 12:24:22Em
filter=Open
Bin:M (8), FOV
25,f1, 3sC
amera: IV
IS 186, S
I620EE
V
User: HH
Group:G
rp4Experim
ent: FME
13-57C
omment1:
Com
ment2:
1.0
0.8
0.6
0.4
0.2
x106
Image
Min = -1.4387e+08
Max = 1.6421e+08p/sec/cm̂2/sr
Color B
arMin = 1e+05
Max = 1e+06
bkgsub
flat-fieldedcosm
ic
WARNING
: Saturated Lu
minescent Im
age
EAB
L1
CA
M/C
E7
scF
v
CD
28
tm/c
yto
hu
CD
3z c
yt
hug F4 c
zzz
C
OV
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3
98
% p
ositiv
e
MF
I: 11
7
CE
7 +
Ga
M-P
E
MA
DH
27
44
Ca
ov-3
A2
78
0
79
% p
ositiv
e
MF
I: 33
10
0%
po
sitiv
e
MF
I: 39
5
22
% p
ositiv
e
MF
I: 5
99
% p
ositiv
e
MF
I: 29
7
% Chromium Release
Effe
cto
r:Targ
et
LC
L-O
KT
3
MA
DH
27
44
Ca
ov-3
SK
OV
3O
VC
AR
3
A2
78
0
0
20
40
60
80
100
50
:1 2
5:1
5:1
1:1
12
0
20
00
IFN
-g
15
00
10
00
60
0
40
0
20
00
pg/mL
TN
F-a
D
MA
DH
27
44
Ca
ov-3
SK
OV
3O
VC
AR
3
A2
78
0
PB
SM
ock
CD
19R
CE
7R
Day 3
Day 5
= T
cell a
dm
inis
tratio
n
Day
11
Day 1
2 =
T c
ell a
dm
inis
tratio
n
Day 1
8
Day 2
6
5x1
05
5x1
04
10
6
10
5
108106
7 5641342822191511Days:
tx tx tx
f TAG72-BBζ 1x TAG72-BBζ repeat TAG72-BBζ 1x TAG72-BBζ repeat
Day 42 Day 70
CD
3
0 20 40 60 80 100 1200
25
50
75
100
Days post tumor injection
Perc
ent surv
ival
Mock (1x)
Mock (repeat)
TAG72-BBζ (1x)
TAG72-BBζ (repeat)
Mock repeat
e
***
***
0 25 50 75
tx
107
108
109
1010
1011
Days post tumor injection
0 25 50 75106
107
108
109
1010
1011 tx
a
Figure 4
i.p. OV90 tumor
i.p. T cell
1x vs. repeat
0 25 50 75106
107
108
109
1010
1011 txc
0 25 50 75
tx
107
108
109
1010
1011
Flu
x (
photo
ns/s
ec)
0 25 50 75106
107
108
109
1010
1011 tx
0 25 50 75106
107
108
109
1010
1011 tx
0 25 500
1
210
30
50
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
0 25 500
10
20
30
40
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
d
Mock 1xMock repeat
TAG72-BBζ 1x
TAG72-BBζ repeat
b
Mock 1x
TAG72-BBζ 1x
Mock repeat
TAG72-BBζ repeat
tx
0 25 500
1
210
30
50
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
0 25 500
10
20
30
40
Days post tumor injection
Rela
tive T
um
or
Gro
wth
Mock (1x)
TAG72-BBz (1x)
Mock (repeat)
TAG72-BBz (repeat)
Click # H
H20131001122422
Tue,Oct 01, 2013 12:24:22
Emfilter=O
penBin:M
(8), FOV25,f1, 3s
Cam
era: IVIS
186, SI620E
EV
User: H
HG
roup:Grp4
Experiment: FM
E13-57
Com
ment1:C
omment2:
1.0
0.8
0.6
0.4
0.2
x106
Image
Min = -1.4387e+08
Max = 1.6421e+08p/sec/cm̂2/sr
Color B
arMin = 1e+05
Max = 1e+06
bkgsub
flat-fieldedcosm
ic
WARNING
: Saturated Lu
minescent Im
age
EAB
L1
CA
M/C
E7
scF
v
CD
28
tm/c
yto
hu
CD
3z c
yt
hug F4 c
zzz
C
OV
CA
R3
SK
OV
3
98
% p
ositiv
e
MF
I: 11
7
CE
7 +
Ga
M-P
E
MA
DH
27
44
Ca
ov-3
A2
78
0
79
% p
ositiv
e
MF
I: 33
10
0%
po
sitiv
e
MF
I: 39
5
22
% p
ositiv
e
MF
I: 5
99
% p
ositiv
e
MF
I: 29
7
% Chromium Release
Effe
cto
r:Targ
et
LC
L-O
KT
3
MA
DH
27
44
Ca
ov-3
SK
OV
3O
VC
AR
3
A2
78
0
0
20
40
60
80
100
50
:1 2
5:1
5:1
1:1
12
0
20
00
IFN
-g
15
00
10
00
60
0
40
0
20
00
pg/mL
TN
F-a
D
MA
DH
27
44
Ca
ov-3
SK
OV
3O
VC
AR
3
A2
78
0
PB
SM
ock
CD
19R
CE
7R
Day 3
Day 5
= T
cell a
dm
inis
tratio
n
Day
11
Day 1
2 =
T c
ell a
dm
inis
tratio
n
Day 1
8
Day 2
6
5x1
05
5x1
04
10
6
10
5
108106
7 5641342822191511Days:
tx tx tx
f TAG72-BBζ 1x TAG72-BBζ repeat TAG72-BBζ 1x TAG72-BBζ repeat
Day 42 Day 70
CD
3
0 20 40 60 80 100 1200
25
50
75
100
Days post tumor injection
Perc
ent surv
ival
Mock (1x)
Mock (repeat)
TAG72-BBζ (1x)
TAG72-BBζ (repeat)
Mock repeat
e
***
***
a ba
Tumor Heterogeneity in Ovarian Cancer and Antigen Loss following
TAG72-CAR T cell therapy
Murad, et al. Front Immunol 2018
a b
-Early tumor recurrences showed loss of TAG72 expression, but later regained expression (in vivo and in vitro)-Suggestive that repeat infusions of greater T cell persistence may durably control tumor growth
Antigen Heterogeneity in Ovarian Cancer Metastasis
TAG72
2+
MUC16
1+
TnMUC1
2+
HER2
3+
CD3
15 Cases:
TAG72: 80% (12/15),
MUC16: 93% (14/15),
TnMUC1: 100% (15/15),
HER2: 20% (3/15)
9099
OM
907
1 O
MTAG72
3+
MUC16
3+
TnMUC1
3+
HER2
0
CD3
TAG72 HER2
TAG72 MUC16
1x
20x
1x
20x
TAG72
2+
MUC16
1+
TnMUC1
2+
HER2
3+
CD3
15 Cases:
TAG72: 80% (12/15),
MUC16: 93% (14/15),
TnMUC1: 100% (15/15),
HER2: 20% (3/15)
9099
OM
907
1 O
M
TAG72
3+
MUC16
3+
TnMUC1
3+
HER2
0
CD3
TAG72 HER2
TAG72 MUC16
1x
20x
1x
20x
Challenges for Solid Tumor CAR T Cell Therapy
• CAR Design / Delivery – HER2-CARs for brain metastasis (Priceman et al. Clinical Cancer Res 2018) – Phase I Trial Q4 2018
– PSCA-CARs for mCRPC (Priceman et al. OncoImmunology 2017) – Phase I Trial Q3 2019
– TAG72-CARs for ovarian cancer (Murad et al. Front Immunol 2018)
• Tumor Antigen Expression Profile and Heterogeneity– Engineering tumor selectivity of CARs
– Multi-targeted CAR T cells
• Immunosuppression / Physical Barriers– Development of immunocompetent mouse models
– Intrinsic optimization of CAR (4th gen)
Challenges for Solid Tumor CAR T Cell Therapy
Oncolytic Virotherapy
• Oncolytic viruses selectively infect and replicate in tumors, promoting immunogenic cell death (ICD)
• ICD can induce tumor-associated antigen release, recruitment of APCs, and elicit adaptive antitumor immunity
• T-VEC – FDA approved HSV-I expressing GM-CSF for metastatic melanoma
• Enhanced antitumor immunity by introduction of genes (apoptosis-inducing, chemokines, cytokines)
• Benefits of oncolytic viruses + CAR T cells have been demonstrated in preclinical models
– CAR T cells carrying OV
– OV-expressing RANTES/IL-15 + CAR
– OV-expressing co-stimulation/checkpoint blockade + CAR
Fong Lab: How Was Oncolytic Vaccinia Virus CF33 Generated?
29
CF33 Oncolytic Orthopoxvirus for TNBC
Choi et al., Mol Ther Oncolytics, 2018
In vitro activity of CF33 against
triple negative breast cancer cell lines
In vivo anti-tumor efficacy of CF33 against
TNBC tumor xenografts
30
Oncolytic Viruses Deliver CAR Targets to “Targetless” Solid Tumors
Park et al. unpublished
a
b
MOI 0 CD19tMOI 0.025
Vaccinia Virus
CD19
DAPI
MOI 1
J2R
hCD19t
PSE
Vaccinia Oncolytic Virus
CF33-(SE)hCD19t
Park et al. unpublished
Oncolytic Viruses Deliver CAR Targets to “Targetless” Solid Tumors
OV
alo
ne
OV
+ M
ock
OV
+ C
AR
OV
alo
ne
OV
+ M
ock
OV
+ C
AR
0 hrs 24 hrs 48 hrs 72 hrs 0 hrs 24 hrs 48 hrs 72 hrs
MDA-MB-468 U251T
c d
OV19t Drive CD19-CAR T cell Anti-Tumor Responses in Solid Tumors
Park et al. unpublished
0 30 40 50 60 70 800
200
400
600
0 30 40 50 60 70 800
200
400
600
Days post tumor injection
0 30 40 50 60 70 800
200
400
600
0 30 40 50 60 70 800
200
400
600
0 30 40 50 60 70 800
200
400
600
0 30 40 50 60 70 800
200
400
600
0 30 40 50 60 70 800
200
400
600
Days post tumor injection
Tum
or
Volu
me (
mm
3)
No treatment
Mock alone
CAR alone
OV19t alone
OV19t + Mock
OV19t + CAR
OV T cells
p < 0.05
a b
0 30 40 50 60 70 800
200
400
600
Tum
or
Volu
me (
mm
3)
No treatment Mock alone CAR alone
OV19t alone OV19t + Mock OV19t + CAR
-Combination of OV carrying CD19t and CD19-CAR T cells promotes tumor regression in xenograft model of TNBC
Tumor injection OV19t CAR T cells
Monitor tumor growthD0 D36 D46
• CAR Design / Delivery – HER2-CARs for brain metastasis (Priceman et al. Clinical Cancer Res 2018) – Phase I Trial Q4 2018
– PSCA-CARs for mCRPC (Priceman et al. OncoImmunology 2017) – Phase I Trial Q3 2019
– TAG72-CARs for ovarian cancer (Murad et al. Front Immunol 2018)
• Tumor Antigen Expression Profile and Heterogeneity– Engineering tumor selectivity of CARs
– Multi-targeted CAR T cells
• Immunosuppression / Physical Barriers– Development of immunocompetent mouse models
– Intrinsic optimization of CAR (4th gen)
Challenges for Solid Tumor CAR T Cell Therapy
“Hot”
“Cold”
CD8 T cells
Th1
NK cells
DCs
IFNg
TNFa
IL-2
IL-12
CXCL9/10
TAMs
G/M-MDSCs
Tregs
STAT3
TGFb
PD-1/PD-L1
CTLA-4
VEGF
IDO
Tumor mutational burden? Tumor/stroma expression? Low T cells? High myeloid/T suppressive cells?
Immunologically Hot vs. Cold Tumors
Prostate Cancer /
Pancreatic Cancer
Melanoma“Checkpoint pathway inhibitors have overwhelmingly failed as single agents in cold tumors”
“Will CAR T cells work?”
Immunosuppressive Solid Tumor Microenvironment
Patterns of immune activity: (1) Immune-excluded, (2) Immune-infiltrated, (3) Immune-desert
Tumor-associated macrophage
Regulatory T
cell
Cytotoxic T
cellAPC
Myeloid-derived
suppressor cell
Tumor antigen
Dying tumor cell
Stromal cell
NK cell
Tumor
cells
Ongoing studies
• Generate syngeneic models and humanized models to faithfully recapture tumor microenvironment
• Built-in inducible cytokines– Address persistence/survival/function defects of tumor-infiltrating T cells
• Oncolytic viruses– VV (Fong collab) and HSV (Caligiuri/Yu collab)
• Endogenous immunity sparked by CAR, OV, and radiation therapy??
• Clinical trial correlatives to inform on next-generation immunotherapy strategies
Acknowledgements
Current members
John Murad, PhD
Anthony Park, MS (GSR)
Yuki Yamaguchi, PhD
Lauren Adkins, MS
Jackson Gibson, BS
Rudra Bhowmick, PhD
Eric Lee, BS
Jason Yang, BS
Lupita Lopez, BS (GSR)
Past members
Ethan Gerdts (UCSD Med)
Dileshni Tilakawardane, MS (TCTReg)
Brook Jeang, MS (UCI PhD grad)
Kelly Kennewick, BS (UCLA PhD grad)
Anna Kozlowska, PhD
Achini Bandara, PhD
Priceman Lab
Funding
CollaboratorsCOH: Jana Portnow, MD, Tanya Dorff, MD, Mihaela Cristea, MD, John Burnett, PhD, Marcin Kortylewski, PhD, Monty Pal, MD, Vincent Chung, MD, Hua Yu, PhD, LalehMelstrom, MD, Ed Manuel, PhD, Mark LaBarge, PhD, Paul Yazaki, PhD, Jack Shively, PhD, Anna Wu, PhD
UCLA: Owen Witte, MD, Robert Reiter, MD
USC: Peter Kuhn, PhD
FHCC: John Lee, MD
Caltech: Lior Pachter, PhD
Emory/GIT: Hadyn Kissick, PhD, Gabriel Kwong, PhD
Industry PartnersMustang Bio, Inc
Forman Lab (TCTRL)Stephen Forman, MD
Christine Brown, PhD
Xiuli Wang, MD PhD
Elizabeth Budde, MD
Stephen Lin, PhD
Jamie Wagner
Araceli Naranjo
Renate Starr
Wen-Chung Chang, MS
Sanda Thomas, PhD
Vanessa Jonsson, PhD
Brenda Aguilar
Julie Ostberg, PhD
Larry Stern, PhD
Kirsten Rood, PhD
All of the TCTRL
Not shown: Jason Yang
