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. 2023 Mar 1;13(3):580-597.
doi: 10.1158/2159-8290.CD-22-0964.

CD19/CD20 Bispecific Chimeric Antigen Receptor (CAR) in Naive/Memory T Cells for the Treatment of Relapsed or Refractory Non-Hodgkin Lymphoma

Sarah M Larson  1   2 , Christopher M Walthers  3 , Brenda Ji  3 , Sanaz N Ghafouri  1 , Jacob Naparstek  4 , Jacqueline Trent  4 , Jia Ming Chen  4 , Mobina Roshandell  3 , Caitlin Harris  3 , Mobina Khericha  3 , Thomas Schweppe  3 , Beata Berent-Maoz  1   4 , Stanley B Gosliner  5 , Amr Almaktari  3 , Melanie Ayala Ceja  3 , Martin S Allen-Auerbach  6 , Jonathan Said  7 , Karla Nawaly  4 , Monica Mead  1 , Sven de Vos  1   2 , Patricia A Young  1 , Caspian Oliai  1 , Gary J Schiller  1   2 , John M Timmerman  1   2 , Antoni Ribas  1   2   4   8   9 , Yvonne Y Chen  2   3   4   5
Affiliations

CD19/CD20 Bispecific Chimeric Antigen Receptor (CAR) in Naive/Memory T Cells for the Treatment of Relapsed or Refractory Non-Hodgkin Lymphoma

Sarah M Larson et al. Cancer Discov. .

Abstract

To address antigen escape and loss of T-cell functionality, we report a phase I clinical trial (NCT04007029) evaluating autologous naive and memory T (TN/MEM) cells engineered to express a bispecific anti-CD19/CD20 chimeric antigen receptor (CAR; CART19/20) for patients with relapsed/refractory non-Hodgkin lymphoma (NHL), with safety as the primary endpoint. Ten patients were treated with 36 × 106 to 165 × 106 CART19/20 cells. No patient experienced neurotoxicity of any grade or over grade 1 cytokine release syndrome. One case of dose-limiting toxicity (persistent cytopenia) was observed. Nine of 10 patients achieved objective response [90% overall response rate (ORR)], with seven achieving complete remission [70% complete responses (CR) rate]. One patient relapsed after 18 months in CR but returned to CR after receiving a second dose of CART19/20 cells. Median progression-free survival was 18 months and median overall survival was not reached with a 17-month median follow-up. In conclusion, CART19/20 TN/MEM cells are safe and effective in patients with relapsed/refractory NHL, with durable responses achieved at low dosage levels.

Significance: Autologous CD19/CD20 bispecific CAR-T cell therapy generated from TN/MEM cells for patients with NHL is safe (no neurotoxicity, maximum grade 1 cytokine release syndrome) and demonstrates strong efficacy (90% ORR, 70% CR rate) in a first-in-human, phase I dose-escalation trial. This article is highlighted in the In This Issue feature, p. 517.

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Conflict of interest statement

COMPETING INTERESTS

Y.Y.C. is an inventor on a patent application for CART19/20 and holds several patent applications in the area of CAR-T cell therapy. Y.Y.C. is a founder of, holds equity in, and receives consulting fees from ImmPACT Bio. She is a member of the scientific advisory board of and holds equity in Catamaran Bio, Notch Therapeutics, Pluto Immunotherapeutics, Prime Medicine, Sonoma Biotherapeutics, and Waypoint Bio. She has consulted for Novartis and Gritstone Bio. S.M.L. holds equity in 1200 Pharma and TORL BioTherapeutics, and has received research funding from Abbvie, Bioline, Bristol Myers Squibb (BMS), Janssen, Novartis, Pfizer, and Sanofi. C.M.W. is a current employee of and holds equity in Orca Bio. J.T. is a current employee of and holds equity in ImmPACT Bio. M.R. is a current employee of and holds equity in Fate Therapeutics. G.J.S. holds equity in Amgen, BMS, and Johnson & Johnson; has consulted for or received honoraria from Kite, Astellas, AbbVie, Incyte, BMS, Stemline, Karyopharm, Agios, Amgen, AstraZenecca, Novartis, Ono Pharma, Celgene, and Jazz; and has received research funding from Actinium, Actuate, AbbVie, AltruBio, Arog, Astellas, AVM Biopharma, Cellectis, Celgene, Cellerant, Constellation, CTI, Forma, Cyclacel, Daiichi-Sankyo, Deciphera, Ifly, FujiFilm, Gamida, Gilead, Genetech/Roche, Geron, Glycomimetics, Incyte, Janssen, Karyopharm, Kite, Mateon, Medimmune, Millenium, Novartis, Onconova, Pfizer, PreCOG, Regimmune, Samus, Sellas, Sangamo, Semline, Takeda, Tolero, and Trovagene. A.R. has received honoraria from consulting with Cstone, Merck, and Vedanta, is or has been a member of the scientific advisory board and holds stock in Advaxis, Appia, Apricity, Arcus, Compugen, CytomX, Highlight, ImaginAb, ImmPact, ImmuneSensor, Inspirna, Isoplexis, Kite-Gilead, Lutris, MapKure, Merus, PACT, Pluto, RAPT, Synthekine and Tango, has received research funding from Agilent and from Bristol-Myers Squibb through Stand Up to Cancer (SU2C), and patent royalties from Arsenal Bio. The other authors declare no conflicts of interest.

Figures

Fig. 1.
Fig. 1.. Design of phase-1 clinical trial evaluating CD19/CD20 bispecific CAR-T cell therapy (CART19/20) in patients with non-Hodgkin lymphoma, chronic lymphocytic leukemia, and small lymphocytic lymphoma.
(A) Schematic of CD19/CD20 bispecific CAR construct. (B) Schematic of phase-1 dose escalation trial design. (C) Consolidated Standards of Reporting Trials (CONSORT) diagram of CART19/20 trial.
Fig. 2.
Fig. 2.. CART19/20 cells manufactured from naïve/memory T cells are enriched in memory phenotype.
(A) Fold expansion (top) and viability (bottom) of cell product during ex vivo manufacturing. Cell counts were normalized to counts on the day of transduction (day 3). Data are shown with color coding by patient (left), by whether starting cell population underwent CD14/CD25 depletion (middle), and by disease indication (right). (B–E) Flow cytometry performed on cryopreserved cell aliquots post thaw to characterize (B) CD3+ purity and transduction efficiency of final cell product, (C) transduction efficiency grouped by batch of lentivirus used in manufacturing, (D) T-cell subtype distribution among all CD3+ T cells, and (E) T-cell subtype distribution among CAR-expressing T cells. Te/exh: effector/exhausted T cells, CD45RA+/CD45RO−/CD62L−; Tem: effector-memory T cells, CD45RA−/CD45RO+/CD62L−; Tcm: central-memory T cells: CD45RA−/CD45RO+/CD62L+; naïve: CD45RA+/CD45RO−/CD62L+. In panels B, D, and E, red underscoring of patient ID indicates products that did not undergo CD14/CD25 depletion.
Fig. 3.
Fig. 3.. Patients refractory to multiple prior lines of treatment respond to CART19/20 cell therapy.
Timeline of individual patients’ response to prior treatment and to CART19/20 cell therapy. The disease indication and dose of CART19/20 cells received are also indicated for each patient. Light green box for patient 009 post CAR-T cell infusion denotes time between the last CR confirmed by PET scan and the time of patient death. The patient had no detectible lymphoma based on IHC analysis performed on bone-marrow biopsy shortly before death due to grade-5 hypocellular marrow. MCL, mantle-cell lymphoma; FL, follicular lymphoma; PMBCL, primary mediastinal B-cell lymphoma; DLBCL, diffuse large B-cell lymphoma; tFL, transformed follicular lymphoma; HGBCL dh, high-grade B-cell lymphoma double-hit.
Fig. 4.
Fig. 4.. CART19/20 cell therapy is highly effective in treating relapsed/refractory non-Hodgkin lymphomas.
(A) Representative PET scans of patients treated with CART19/20. (B) Overall survival and progression-free survival curves from the time of CART19/20 cell infusion. (C) PET scan obtained at screening for Patient 003, indicating pulmonary involvement of PMBCL. (D) Immunohistochemistry (IHC) analysis of Patient 003 tissue biopsies; original magnification x160. Supraclavicular lymph node biopsy obtained at screening and bone-marrow biopsy obtained 14 days post CART19/20 infusion were analyzed by IHC. Results reveal rapid emergence of a CD19−CD20−BCL6−cMYC− tumor population within 14 days of CART19/20 treatment.
Fig. 5.
Fig. 5.. CART19/20 cells exhibit sustained persistence and efficacy with strong safety.
(A) Flow cytometry analysis on peripheral blood samples collected from Patient 004 at screening as well as 7- and 14-days post CART19/20 infusion. CD19 and CD20 surface staining results indicate the presence of CD20+CD19dim/– cells in Patient 004 prior to CART19/20 cell treatment. (B) PET scans of Patient 004 at the time of relapse and at 60 days post second dose of CART19/20 cells. (C) Presence of CAR transgene as quantified by droplet digital PCR. The psi signal integrated through lentiviral transduction was quantified. Inset shows zoomed-in data from the first 30 days post CART19/20 infusion. (D) Presence of CAR-expressing T cells among peripheral blood mononuclear cells (PBMCs) as quantified by flow cytometry. (E) Presence of CD19+ and/or CD20+ cells among lymphocytes as quantified by flow cytometry. A 3% line is shown to denote threshold for B-cell aplasia. (F) Serum cytokine levels as qualified by Luminex multiplex assay.
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