cars and armored cars lymphoma and myeloma 2014 new york ny october 23, 2014 renier brentjens md phd...
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CARs and armored CARs
Lymphoma and Myeloma 2014New York NY
October 23, 2014
Renier Brentjens MD PhDAssociate MemberLeukemia Service
Chief, Cellular Therapeutics CenterDepartment of Medicine
Memorial Sloan Kettering Cancer Center
Renier Brentjens MD PhD
•Stockholder: Juno Therapeutics (scientific co-founder)•Royalties: Juno Therapeutics•Honoraria: none•Reserch Funding: Juno Therapeutics•Consultant fees: Juno Therapeutics•Discussion of off-label drug use: Tocilizumab
Conflict of Interest Disclosure
Generation of a tumor targeted chimeric antigen receptor (CAR)
VH VL5’ LTR
α-tumor scFv
CD8 ζ chain 3’ LTRψ
SD SA
α-TAA mAb
TCR complex
CAR retroviral vector
α-TAA scFv—CD8-ζ
TAA CAR
VH VL5’ LTR
αTAA scFv
CD8 ζ chain 3’ LTRψ
SD SA
Generation of TAA-targetedT cells for treatment of Cancer
αTAA scFv
CD8
CD3 ζ
1. Construct a chimeric antigen receptor (CAR)
SFG-CAR
2. Subclone CAR gene into a retroviral vector (SFG)
3. Transduce and expand patient T cells ex vivo
4. Infuse transduced T cells to eradicate TAA+ tumor cells
TAA
Native TCR
TAA CAR
• HLA-independent antigen recognition, therefore universal application
• Active in both CD4+ and CD8+ T cells• Target antigens include proteins, carbohydrates
and glycolipids • Rapid generation of tumor specific T cells • Minimal risk of autoimmunity or GvHD• A living drug, single infusion
Advantages of CAR T cell therapy
Expression of CD19 and other B cell markers on B lineage cells
Y Y
Y
Y
Y
Y
Y
Y
YY
YY
YY
Y Y
Y Y
preB-ALLB cell lymphomas and
leukemias myelomas
Stem Cell pre B immature B mature B plasma cellpro B
CD19
CD22
CD20
Evolution in CAR design
Clinical trials using CD19 targeted T cells in relapsed or refractory B-ALL
Sci Transl Med. 2013 Mar 20;5(177):177ra38
Patient characteristics and treatment outcomes
Maude et al N Engl J Med. 2014, 371:1507-17
UPenn studies of relapsed B-ALL
• 25 pediatric and 5 adult relapsed or refractory B-ALL patients treated
• 19-4-1BBz CAR design• 90% CR• 6 month EFS 67%• 6 month OSR 78%
Lee et al Lancet. Published online October 13, 2014
NCI studies of relapsed B-ALL
• 20 pediatric and young adult relapsed or refractory B-ALL patients treated.
• 19-28z CAR design• 70% CR (14/20)• 60% MRD- CR• 5 month EFS 78% in MRD- patients
The Problem
Clinical trials using CD19 targeted T cells in low grade B cell malignancies
Kalos et al Sci Trans Med 2011
UPenn clinical trial results
Patient Prior Chemotherapy Conditioning Chemotherapy
Response
1 Fludarabine, Rituximab, Alemtuzumab, R-CVP,
Lenolidomide, PCR
Bendamustine CR (3+ years)
2 Alemtuzumab Bendamustine/Rituximab PR (7 months)
3 Rituximab/Fludarabine, Rituximab/Bendamustine.
Alemtuzumab
Pentostatin/Cyclophosphamide
CR (3+ years)
Updated UPenn Trials in CLL (ASH 2013)
• Abstract 4162– CD19 CAR T cells treating relapsed/refractory CLL
• Utilizing a 4-1BBz CAR construct, 14 CLL patients treated• 3/14 patients obtained CR (21%), 5/14 patients obtained PR (36%), 6/14 patients
with no response (43%)• 6/14 patients with persistent detectable CAR T cells (5-35 months)• No CR patients with reported relapsed disease• No dose response reported
• Abstract 873– Dose randomized dose optimization trial of CLL patients with either high or
low dose CAR T cell infusions• Utilizing a 4-1BBz CAR construct, 27 CLL patients treated• Patients randomized to either low dose (5 x 107 CAR T cells) or high dose (5 x 108
CAR T cells) • No dose response benefit seen in these treated patients• Overall response rate (CR + PR) was 40%• No correlation with CRS and RR was observed
MSKCC clinical trial results: CLL
Cyclophosphamide
No Cyclophosphamide
Brentjens et al Blood. 2011 Nov 3;118(18):4817-28
CAR questions
• What is the etiology of differential responses by CAR T cell therapy to relapsed B-ALL versus CLL?
• What is the role of bulky disease and CAR T cell anti-tumor efficacy?
• The role of the hostile tumor micro-environment and CAR T cell function
• How to build a better T cell?
The hostile tumor microenvironment
The tumor microenvironment contains multiple inhibitory factors designed to potentially suppress effector T cells.
– CD4+ CD25hi FoxP3+ regulatory T cells (Tregs)– MDSCs– TAMs– Expression of inhibitory ligands by tumor (PD-L1)– Tumor secretion of T cell suppressive cytokines (TGF-β and
IL-10)
The solution?Armored CAR T cells
Moving Forward: Armored CARs
IL-12
• A heterodimeric cytokine secreted by activated APCs, neutrophils and macrophages.
• Induces Th1 CD4+ T cell response enhancing IL-2 and IFN-γ secretion
• Enhances T cell clonal expansion and effector function in concert with TCR signaling (signal 1) and CD28 co-stimulation (signal 2), serving as a signal 3.
• Avoids/reverses T cell anergy• May overcome Treg mediated effector T cell inhibition• Recruits and activates NK cells• Clinical trials in cancer using systemic IL-12 therapy has
been limited by severe inflammatory side effects
Syngeneic EL4(hCD19) tumor model
mCD19-/- hCD19+/-
mCD19-/- hCD19+/-
IV injection
EL4(hCD19)
Harvest splenocytes
IV injection
Retroviraltransduction
with chimeric receptor
Assess T cell homing to tumor
Assess T cell eradication of
tumor
Assess T cell proliferation
in vivo
Assess long-term survival
of T cells
Assess the efficacyof suicide vectors
Assess memory T cell response to rechallenge
with tumor
53%
Determine the sideeffects of therapy
Per
cent
Sur
viva
l
Days since tumor cell injection
Lymphodepletion enhances anti-tumor efficacy of 19z1+ T cells
100 101 102 103 104100
101
102
103
104
1Élive cells
FL1-H: FL1-HeightF
L2
-H: h
CD
19
PE
24.3 0.11
0.06375.5
24.3%
hC
D19
100 101 102 103 104
100
101
102
103
104
6Élive cells
FL1-H: mCD19
FL
2-H
: hC
D1
9
2.07 0.02
0.2997.6
2.07%
hC
D19
19z1IRESIL-12 modified T cells secrete biologically active IL-12 and exhibit enhanced targeted cytotoxic function and resistance to Tregs
1:1 2.5:1 5:1 10:1
***
*
A
B
C D
Pegram et al Blood 2012
A B
Syngeneic IL-12 secreting CD19 targeted T cells induce B cell aplasias and tumor eradication
Pegram et al Blood 2012
IL-12 secreting CAR T cells in vivo efficacy
Enhanced CMphenotype, enhanced
cytotoxicity, enhanced persistence
Resistance to Treg and TGFβ inhibition
NK cell Recruitment
and activation
IL-12 secretion
IL-12 secretion
Targeted tumor cytotoxicity
Targeted tumor cytotoxicity
Targeted tumor cytotoxicity
Tumor cell
NK cell
ActivatedTIL
AnergicTIL
IL-12 secretion
Reversal of anergy
CAR-IRES IL-12
IL-12 genetically modified T cells: Armored CAR T cells
Conclusions
• Autologous CD19 targeted CAR modified T cells have demonstrated very promising anti-tumor efficacy in B cell ALL with more modest responses in patients with low grade B cell malignancies.
• Etiologies of CAR T cell resistance may be related to the hostile tumor microenvironment.
• Application of CAR T cell therapy for low grade B cell malignancies as well as moving forward towards application to solid tumor malignancies requires “armored” CAR T cells designed to both overcome the hostile tumor microenvironment and exhibit enhanced anti-tumor efficacy and long term persistence.
• “Armored” CAR T cells appear to have enhanced anti-tumor efficacy based on pre-clinical tumor models.
• Future studies using “armored” CAR T cell technology will focus on translation of these armored CAR T cells to the clinical setting both in the context hematological as well as solid tumor malignancies.
Renier BrentjensHollie PegramMythili KoneruSarwish RafiqSwati PendharkarEric SmithJames LeeYan NikhaminJae ParkKevin CurranPeter Chang
Michel SadelainMarco DavilaMichael GongJean Baptiste Latouche
Leukemia ServiceDavid ScheinbergJae ParkMartin TallmanMark FrattiniPeter MaslakMark HeaneyJoe JurcicNicole LamannaMarco DavilaDan Douer
Cell Therapy and Cell Engineering Facility(Isabelle Riviere, Director)
R&D, ManufacturingXiuyan WangJolanta StefanskiMalgorzata OlszewskaOriana Borquez-OjedaTeresa WasielewskaJinrong Qu
QA/QCShirley BartidoYongzeng Wang(Mark Przybylowski)James HoseyDomenick Pirraglia(Vanessa Capacio)
Clinical ResearchYvette Bernal
FundingCA59350 (MS) ; P30 CA-008748 (CT); 3RO1CA138738-02S1(RJB); Alliance for Cancer Gene Therapy ; Terry Fox Run for Cancer Research; William H. Goodwin and Alice Goodwin, and the Commonwealth Cancer Foundation for Research and the ETC of MSKCC; Damon Runyon Clinical Investigator Award (RJB); William Lawrence & Blanche Hughes Foundation (RJB); CLL-Global Research Foundation (RJB)
Lymphoma ServiceCraig MoskowitzAriela Noy
GYN serviceSamith SandadiRoisin O’Clearbhail
Adult BMT ServiceSergio GeraltCraig Sauter
Department of ClinicalLaboratoriesLillian ReichDavid WuestKathy Smith
BiostatisticsGlenn Heller
Brentjens’ Lab Members