Research ArticleImmunotherapy

Chimeric antigen receptor T cells persist and induce sustained remissions in relapsed refractory chronic lymphocytic leukemia

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Science Translational Medicine  02 Sep 2015:
Vol. 7, Issue 303, pp. 303ra139
DOI: 10.1126/scitranslmed.aac5415
  • Fig. 1. Overall survival and PFS.

    (A and B) Kaplan-Meier curves for overall survival (A) and PFS (B) were estimated for the 14 subjects who received therapy. Shaded region indicates 95% CI. Number of subjects at risk is presented in a 3-month interval. Vertical tick marks indicate time points with censored data. (Source data, table S6).

  • Fig. 2. CTL019 expansion by qPCR in the first 12 months.

    Peripheral blood CTL019 expansion was measured as copies per microgram of genomic DNA by qPCR in 14 subjects. Values below the quantitative limit of detection (<25 copies/μg DNA) are shown with open circles. (Source data, table S7).

  • Fig. 3. Association between peak CTL019 expansion and response.

    (A and B) Peak CTL019 expansion in the first 3 months measured by flow cytometry (A) in 11 subjects and qPCR (B) in 14 subjects. Flow cytometry detection was not available for subjects 01, 02, and 03 because an antibody to detect CTL019 cells was not available at the time of their treatment. Wilcoxon rank-sum P values are provided. (Source data, table S8).

  • Fig. 4. Long-term persistence of CTL019 cells and polyfunctionality in patients achieving CR.

    (A) Long-term persistence of CTL019 cells by flow cytometry (solid circle) and qPCR (solid triangle) beyond 12 months from infusion. All observed values were above the limit of detection by flow cytometry (0.1%) and above the limit of quantification by qPCR (<25 copies/μg DNA). (B and C) Purified PBMCs from patient UPCC04409-02 from the indicated postinfusion time points were stimulated for 6 hours with APC-expressing CD19 or control antigen (mesothelin) and examined for degranulation (CD107a) and cytokine protein expression. (B) Contour plots displaying CD107a versus MIP-1β (top two rows) or IL-2 versus IFN-γ expression (bottom rows) in CD8+ CAR+ T cells stimulated with K562 cells expressing control antigen (mesothelin) or CD19. (C) Summary graph of functional response of CAR19+ CD8 T cells upon stimulation with CD19+ target cells over time. Data are plotted from (B) and are depicted as the background-subtracted frequency of this subset. (Source data, tables S9 and S10).

  • Fig. 5. Peak cytokines values over the first month after infusion.

    (A to D) Peak values for IL-6 (A), IFN-γ (B), IL-2 receptor antagonist (IL-2RA) (C), and IL-2 (D) by CRS grades are displayed. Gray symbols represent nonresponders; black symbols are those who achieved a complete or partial remission. Individual values are plotted with a symbol that reflects their CRS grade (1 to 4). Wilcoxon rank-sum P values are also provided. Cytokine values were missing for one grade 4 subject who achieved CR (n = 13). (Source data, table S11).

  • Table 1. Summary of patient baseline characteristics (N = 14).
    CharacteristicsStatistics, n (%)
    N14
    Age at infusion (years)
      Mean (SD)66.9 (8.1)
      Median (range)66 (51–78)
    Gender
      Male12 (85)
      Female2 (14)
    No. of previous therapies
      Mean (SD)5.3 (2.8)
      Median (range)5 (1–11)
    P53 or 17p deletion
      No8 (57)
      Yes6 (43)
    IGHV mutation
      No9 (64)
      Yes4 (29)
      Unknown1 (7)
    Lymphocyte-depleting chemotherapy
      Bendamustine6 (43)
      Fludarabine/cyclophosphamide3 (21)
      Pentostatin/cyclophosphamide5 (36)
    Lymphocytes in bone marrow at enrollment (%)*
      Mean (SD)79.5 (17.9)
      Median (range)87.5 (40–95)
    Rai stage
      15 (36)
      49 (64)
    Binet stage
      A1 (7)
      B4 (29)
      C9 (64)

    *Data not available for four subjects.

    • Table 2. Treatment and clinical characteristics of subjects (N = 14).

      DLBCL, diffuse large B cell lymphoma; NED, no evidence of disease; NR, no response. MRD tested by deep sequencing analysis as described in Materials and Methods.

      IDTotal T
      cells
      infused
      (× 108)
      Total CTL019
      cells infused
      (× 108)
      Peak CTL019
      expansion
      (% of CD3+ cells)
      Best overall
      response
      Last follow-up or
      progression
      (months)
      Comments, current status
      015011.3N/ACR53MRD-negative; progression-free
      023.00.142N/ACR52MRD-negative; progression-free
      0325.55.86N/APR5Progression, 5 months; died of disease, 27 months
      0510.03.9214.1PR13Progression, 13 months; alive with disease, 36 months
      063.00.6460.2NR1Died of disease, 8 months
      071.70.1720.3NR1Died of complications from bone marrow transplant, 9 months
      095.01.7081.9CR21MRD-negative; progression-free, 21 months; died of infection
      1030.05.6134.3CR28Bulky adenopathy (11 cm); MRD-negative;
      progression-free
      125.01.1818.3PR6Bulky adenopathy (9 cm); died of
      pulmonary embolus
      1418.01.56<0.1NR7Alive with disease, 26 months
      174.21.031.6NR10Alive with disease, 18 months
      1850.02.770.2NR4Alive with disease, 17 months
      225.00.86434.9PR10Bulky adenopathy (9 cm); progressed 10 months with
      transformed CD19-dim DLBCL; died of disease at 10 months
      2520.02.712.6NR3Alive with disease, 16 months

      *CTL019 refers to transduced, CAR-modified T cells.

      †Subjects 01, 02, and 03 were not evaluated for early expansion by flow cytometry because an antibody to detect CTL019 cells was not available at the time of their treatment.

      • Table 3. IGH deep sequencing analysis of blood and bone marrow shows eradication of CLL and B cells for subjects 01 and 02.

        BM, bone marrow; PB, peripheral blood; Mo, month; Yr, year.

        Patient
        UPCC04409 no.
        Sample
        type
        Time
        point
        Cell equivalents
        sequenced
        Total reads
        of IGH
        Total unique
        IGH reads
        Tumor clone
        reads
        CLL clone
        (% of total)
        01PBBaseline408,57948407,59299.76
        Mo 6285,305736200.00
        Yr 1411200.00
        Yr 3298,667174600.00
        Yr 3.5350,171123800.00
        BMMo 1408,838179300.00
        Mo 6202,535445100.00
        Mo 1218,50623100.00
        Mo 2488200.00
        02PBBaseline1,385,34045441,285,86292.82
        Mo 625,0413800.00
        Mo 32317,71488800.00
        Yr 3346,057160800.00
        Yr 4308,4192121000.00
        BMYr 15200.00
        Yr 26012500.00

      Supplementary Materials

      • www.sciencetranslationalmedicine.org/cgi/content/full/7/303/303ra139/DC1

        Fig. S1. Consort diagram.

        Fig. S2. CTL019 cells are polyfunctional.

        Fig. S3. Example of serum ferritin response to tocilizumab in a patient with CRS.

        Fig. S4. Association of peak CTL019 expansion with CRS and the response observed within the first 3 months after infusion.

        Table S1. Cell dose and product characteristics (N = 14).

        Table S2. Tumor burden assessed by IgH deep sequencing analysis of blood and bone marrow of all study subjects.

        Table S3. Association of pretreatment characteristics with response.

        Table S4A. Penn Grading System for CTL019-associated CRS.

        Table S4B. Definition of high-dose vasopressors.

        Table S5. Association of pretreatment characteristics with CRS (N = 14).

        Table S6. Source data for Fig. 1 (Excel).

        Table S7. Source data for Fig. 2 (Excel).

        Table S8. Source data for Fig. 3 (Excel).

        Table S9. Source data for Fig. 4A (Excel).

        Table S10. Source data for Fig. 4C (Excel).

        Table S11. Source data for Fig. 5 (Excel).

      • Supplementary Material for:

        Chimeric antigen receptor T cells persist and induce sustained remissions in relapsed refractory chronic lymphocytic leukemia

        David L. Porter,* Wei-Ting Hwang, Noelle V. Frey, Simon F. Lacey, Pamela A. Shaw, Alison W. Loren, Adam Bagg, Katherine T. Marcucci, Angela Shen, Vanessa Gonzalez, David Ambrose, Stephan A. Grupp, Anne Chew, Zhaohui Zheng, Michael C. Milone, Bruce L. Levine, Jan J. Melenhorst, Carl H. June*

        *Corresponding author. E-mail: david.porter{at}uphs.upenn.edu (D.L.P.); cjune{at}exchange.upenn.edu (C.H.J.)

        Published 2 September 2015, Sci. Transl. Med. 7, 303ra139 (2015)
        DOI: 10.1126/scitranslmed.aac5415

        This PDF file includes:

        • Fig. S1. Consort diagram.
        • Fig. S2. CTL019 cells are polyfunctional.
        • Fig. S3. Example of serum ferritin response to tocilizumab in a patient with CRS.
        • Fig. S4. Association of peak CTL019 expansion with CRS and the response observed within the first 3 months after infusion.
        • Table S1. Cell dose and product characteristics (N = 14).
        • Table S2. Tumor burden assessed by IgH deep sequencing analysis of blood and bone marrow of all study subjects.
        • Table S3. Association of pretreatment characteristics with response.
        • Table S4A. Penn Grading System for CTL019-associated CRS.
        • Table S4B. Definition of high-dose vasopressors.
        • Table S5. Association of pretreatment characteristics with CRS (N = 14).

        [Download PDF]

        Other Supplementary Material for this manuscript includes the following:

        • Table S6. Source data for Fig. 1 (Excel).
        • Table S7. Source data for Fig. 2 (Excel).
        • Table S8. Source data for Fig. 3 (Excel).
        • Table S9. Source data for Fig. 4A (Excel).
        • Table S10. Source data for Fig. 4C (Excel).
        • Table S11. Source data for Fig. 5 (Excel).

        [Download Tables S6 to S11]

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