Eradication of B-ALL using chimeric antigen receptor–expressing T cells targeting the TSLPR oncoprotein

Key Points Adoptive transfer of T cells genetically modified to express anti-TSLPR chimeric antigen receptors can cure B-ALL in xenograft models. Anti-TSLPR CAR constructs containing a CH2CH3 spacer domain were inactive against TSLPR-overexpressing B-ALL.

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103 Inclusion of a Dap10 costimulatory domain enhances anti-tumor efficacy of chimeric PD1-expressing T cells in multiple types of solid tumors

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0103 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A114-A114

Author(s):

Amorette Barber

Keyword(s):

T Cells ◽

T Cell ◽

Cell Lines ◽

Inflammatory Cytokines ◽

Chimeric Antigen Receptor ◽

Adoptive Transfer ◽

Antigen Receptor ◽

Antigen Receptors ◽

Chimeric Antigen Receptors

BackgroundAdoptive transfer of T cells is a promising anti-tumor therapy for many cancers. To enhance tumor recognition by T cells, chimeric antigen receptors (CAR) consisting of signaling domains fused to receptors that recognize tumor antigens can be expressed in T cells. One receptor that is a prospective target for a new chimeric antigen receptor is PD1 because the ligands for the PD1 receptor are expressed on many cancer types. Therefore, we developed a murine chimeric PD1 receptor (chPD1) consisting of the PD1 receptor extracellular domain and the activation domain of CD3 zeta. In addition, current chimeric antigen receptor therapies utilize various costimulatory domains to enhance anti-tumor efficacy. Therefore, we also compared the inclusion of CD28, Dap10, 4-1BB, GITR, ICOS, or OX40 costimulatory domains in the chPD1 receptor to determine which costimulatory domain induced optimal anti-tumor immunity.MethodsTo determine if this novel CAR could potentially target a wide variety of tumors, the anti-tumor efficacy of chPD1 T cells against murine lymphoma, melanoma, kidney, pancreatic, liver, colon, breast, ovarian, prostate, and bladder cancer cell lines was measured.ResultsOf the eighteen cell lines tested, all expressed PD1 ligands on their cell surface, making them potential targets for chPD1 T cells. Regardless of the costimulatory domain in the CAR, all of the chPD1 T cells induced similar levels of T cell proliferation and tumor cell lysis. However, differences were observed in the cytokine secretion profiles depending on which costimulatory receptor was included in the CAR. While most of the chPD1 T cell receptor combinations secreted both pro-inflammatory (IFNγ, TNFα, IL-2, GM-CSF, IL-17, and IL-21) and anti-inflammatory cytokines (IL-10), chPD1 T cells containing a Dap10 costimulatory domain secreted high levels of proinflammatory cytokines but did not secrete a significant amount of anti-inflammatory cytokines. Furthermore, T cells expressing chPD1 receptors with a Dap10 domain also had the strongest anti-tumor efficacy in vivo. ChPD1 T cells did not survive for longer than 14 days in vivo, however treatment with chPD1 T cells induced long-lived protective host-anti-tumor immune responses in tumor-bearing mice.ConclusionsTherefore, adoptive transfer of chPD1 T cells could be a novel therapeutic strategy to treat multiple types of cancer and inclusion of the Dap10 costimulatory domain in chimeric antigen receptors may induce a preferential cytokine profile for anti-tumor therapies.Ethics ApprovalThe study was approved by Longwood University’s IACUC.

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A novel method to generate T-cell receptor–deficient chimeric antigen receptor T cells

Blood Advances ◽

10.1182/bloodadvances.2017012823 ◽

2018 ◽

Vol 2 (5) ◽

pp. 517-528 ◽

Cited By ~ 14

Author(s):

Takahiro Kamiya ◽

Desmond Wong ◽

Yi Tian Png ◽

Dario Campana

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Chimeric Antigen Receptor ◽

Cell Receptor ◽

Surface Expression ◽

Antigen Receptors ◽

Chimeric Antigen Receptors ◽

Key Points ◽

Novel Method

Key Points Newly designed PEBLs prevent surface expression of T-cell receptor in T cells without affecting their function. Combined with chimeric antigen receptors, PEBLs can rapidly generate powerful antileukemic T cells without alloreactivity.

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Novel Antigen-Specific Expansion of T Cells Transduced with a CD19 Chimeric Antigen Receptor

Blood ◽

10.1182/blood.v116.21.3262.3262 ◽

2010 ◽

Vol 116 (21) ◽

pp. 3262-3262

Author(s):

Daniel W. Lee ◽

James N Kochenderfer ◽

Christoph Rader ◽

Rimas J Orentas ◽

Crystal L Mackall

Keyword(s):

T Cells ◽

Chimeric Antigen Receptor ◽

Antigen Receptor ◽

K562 Cells ◽

Antigen Receptors ◽

Chimeric Antigen Receptors ◽

Car T Cells ◽

Car T ◽

Expansion Protocol ◽

Specific Expansion

Abstract Abstract 3262 Relapsed acute lymphoblastic leukemia (ALL) remains a difficult challenge for both pediatric and adult patients. Chimeric antigen receptors (CARs) are genetically engineered molecules expressed in transduced T lymphocytes. CARs express both a target binding motif and TCRzeta signals needed for T cell activation, with or without other costimulatory domains. CARs that target CD19 (expressed on most ALL blasts) are being studied in several clinical trials in adults and are planned for pediatric relapsed/refractory ALL. To date, CARs have been expanded ex vivo using antigen-independent techniques. This project sought to develop antigen specific expansion of cells transduced with a CD19-specific CAR by employing artificial APCs (aAPCs) and to explore whether the method of expansion impacted functionality of CD19-CAR T cells. aAPCs used in these studies express the high affinity Fc receptor (CD64) and the costimulatory molecule CD137L (aAPC-41BBL). We created an Fc-CD19 fusion protein that when loaded onto the aAPCs, engages the chimeric antigen receptor and induces antigen specific activation. Antigen-specific vs. non-specific approaches to activation and expansion of CAR expressing T cells were compared using three different expansion protocols (EPs). CD19-CAR transduced T cells were expanded with A) irradiated aAPC-41BBL loaded with anti-OKT3, B) irradiated aAPC-41BBL loaded with rFc-CD19 or C) irradiated allogeneic PBMC feeder cells with anti-OKT3. Briefly, OKT3 and IL-2 activated T cells were transduced with CD19CAR, control CAR (Her2-specific), or non-transduced (Mock). All cultures were maintained in IL-2. Functionality was assessed in a 4-hour 51Cr release assay against 4 distinct CD19+ ALL cell lines, K562 cells, and K562 stably transfected with CD19 (K562-CD19) or NGFR (K562-NGFR). Results demonstrate similar significant levels of CD19-specific cytotoxicity in the 4h assay at E:T ratios as low as 2.5:1, regardless of the expansion protocol used (20-40% lysis of all CD19+ targets when expanded by OKT3/aAPC, 40–60% lysis with rFc-CD19/aAPC, and 35–45% lysis with allogeneic feeders and OKT3). To evaluate cytotoxicity in long term culture, CD19-CAR T cells expanded using the three protocols were co-cultured with ALL cell targets for 4 days, then flow cytometry was performed to enumerate surviving ALL cells as determined by CD22+ staining. NALM6 and K562-CD19 cells were entirely eliminated even at an E:T ratio of 2.5:1 by CD19-CAR T cells, regardless of expansion protocol, and not by any of the 3 Mock EPs. Therefore, these results demonstrate that the cytolytic potential of CAR transduced T cells is similar, regardless of whether expansion occurs via CD3 signaling or via the chimeric receptor itself. Interestingly, we did observe substantial NK mediated killing in these assays, which correlated with CD56+ cell content and was eliminated by cold target inhibition using K562 cells. Studies are underway to determine whether differences in NK killing varies with expansion protocol. In summary, Fc-bearing artificial antigen presenting cells combined with CAR specific Fc fusion proteins provide a potential off-the-shelf reagent for antigen specific expansion of T cells with chimeric antigen receptors. This could overcome variable transduction efficiencies and allow administration of a more homogenous population of CAR specific T cells. Disclosures: No relevant conflicts of interest to declare.

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DIPG-36. ANTI-GD2 CHIMERIC ANTIGEN RECEPTOR T CELLS AS A POTENT IMMUNOTHERAPY REGIMEN IN XENOGRAFT MODELS OF HISTONE 3 K27M MUTANT DIFFUSE MIDLINE GLIOMA

Neuro-Oncology ◽

10.1093/neuonc/noy059.129 ◽

2018 ◽

Vol 20 (suppl_2) ◽

pp. i56-i56

Author(s):

Christopher Mount ◽

Robbie Majzner ◽

Shree Sundaresh ◽

Evan Arnold ◽

Meena Kadapakkam ◽

...

Keyword(s):

T Cells ◽

Chimeric Antigen Receptor ◽

Antigen Receptor ◽

Histone 3 ◽

Xenograft Models ◽

Diffuse Midline Glioma

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T cells expressing CD123-specific chimeric antigen receptors exhibit specific cytolytic effector functions and antitumor effects against human acute myeloid leukemia

Blood ◽

10.1182/blood-2012-12-474056 ◽

2013 ◽

Vol 122 (18) ◽

pp. 3138-3148 ◽

Cited By ~ 194

Author(s):

Armen Mardiros ◽

Cedric Dos Santos ◽

Tinisha McDonald ◽

Christine E. Brown ◽

Xiuli Wang ◽

...

Keyword(s):

T Cells ◽

Myeloid Leukemia ◽

Autologous Tumor ◽

Antigen Receptors ◽

Chimeric Antigen Receptors ◽

Antitumor Effects ◽

Effector Functions ◽

Key Points ◽

Car T ◽

Human Acute Myeloid Leukemia

Key Points CD123 CAR T cells specifically target CD123+ AML cells. AML patient-derived T cells can be genetically modified to lyse autologous tumor cells.

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ICOS-based chimeric antigen receptors program bipolar TH17/TH1 cells

Blood ◽

10.1182/blood-2013-10-535245 ◽

2014 ◽

Vol 124 (7) ◽

pp. 1070-1080 ◽

Cited By ~ 166

Author(s):

Sonia Guedan ◽

Xi Chen ◽

Aviv Madar ◽

Carmine Carpenito ◽

Shannon E. McGettigan ◽

...

Keyword(s):

T Cells ◽

Effector Function ◽

Th1 Cells ◽

Antigen Receptors ◽

Chimeric Antigen Receptors ◽

Effector Functions ◽

Key Points

Key Points ICOS-based CARs program bipolar TH17/TH1 cells with augmented effector function and in vivo persistence. The expression of selected CAR endodomains can program T cells for their subsequent differentiation fates and effector functions.

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