Research ArticleAutoimmunity

A CD40L-targeting protein reduces autoantibodies and improves disease activity in patients with autoimmunity

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Science Translational Medicine  24 Apr 2019:
Vol. 11, Issue 489, eaar6584
DOI: 10.1126/scitranslmed.aar6584
  • Fig. 1 Generation and biochemical characterization of VIB4920, a human Tn3 specific for CD40L.

    (A) Inhibition of CD40/CD40L interactions was measured by Proteon. The average of duplicate wells is shown. (B) Human embryonic kidney (HEK) 293 cells expressing CD40 and an NF-κB reporter were stimulated with recombinant CD40L overnight in the presence of anti-CD40L Tn3 proteins. The percent inhibition of luciferase activity is shown. Data represent the mean of duplicate wells. One of two independent studies is shown. (C) Human peripheral blood mononuclear cells (PBMCs) were stimulated with recombinant CD40L and expression of CD86 on CD19+ cells was assessed by flow cytometry. (D) Inhibition of CD40/CD40L interactions by enzyme-linked immunosorbent assay (ELISA). Data represent the mean of duplicate wells. (E) Binding of monovalent Tn3 protein 342 to tumor necrosis factor (TNF) superfamily members was evaluated by ELISA. (F) Proposed structure of VIB4920 based on crystallization of 342 Tn3 [Protein Data Bank (PDB) ID 6BRB] and published crystal structure of HSA (PDB ID 1AO6). The linker is shown in blue. (G) Cartoon presentation of CD40/CD40L and 342/CD40L structures aligned through a common CD40L molecule. CD40L is shown in green, Tn3 is shown in magenta, and CD40 receptor is shown in cyan. OD, optical density.

  • Fig. 2 VIB4920 inhibits CD40 signaling and activation of human B cells and does not induce platelet aggregation in ex vivo studies.

    (A) CD40+ HEK293 cells were stimulated with CD40L, and NF-κB luciferase activity was assessed. Shown is the percent inhibition of the luciferase signal. Data represent the mean of duplicate wells from one of two independent studies. (B) Human PBMCs were stimulated with recombinant MEGACD40L, and the percentage of CD19+/CD86+ cells was measured by flow cytometry. Data represent the mean of duplicate wells from one of three independent studies. (C) Human B cells were stimulated with interleukin-21 (IL-21) and MEGACD40L. The dotted line represents adenosine 5′-triphosphate (ATP) concentrations from unstimulated cells. Data shown are the means and SD of triplicate wells and are representative of two independent experiments. (D and E) Human B cells were left unstimulated (nil) or were stimulated with IL-21, anti-IgM, and MEGACD40L, and plasma cell number was quantified. 5c8 is an anti-CD40L mAb used as a positive control for inhibition of CD40L in this assay. (D) The percentage of IgD CD38hi plasma cells with the indicated molecules at 1 nM. (E) Plasma cell (PC) number at various concentrations as indicated. Data shown are the means and SD of triplicate wells and are representative of two independent experiments. (F and G) Washed human platelets were incubated with preformed immune complexes of anti-CD40L and MEGACD40L, and aggregation was measured. Percent aggregation is shown. (F) Where indicated, platelets were preincubated with anti-CD32a antibody (IV.3) before the addition of immune complex. Adenosine diphosphate (ADP) is a positive control for platelet aggregation. Data are representative of two independent experiments. ***P < 0.001, ****P < 0.0001, by two-tailed unpaired Student’s t test.

  • Fig. 3 VIB4920 demonstrates a favorable PK profile in healthy volunteers and patients with RA.

    (A and B) Circulating concentrations of VIB4920 were determined by ELISA at the indicated time points in (A) the SAD study of healthy volunteers and (B) the MAD study in RA. The dotted line represents the lower limit of sensitivity for the assay. Error bars represent SD of the mean, which was not calculated for groups with n = 2 subjects. (C) sCD40L concentrations were assessed in the SAD study of healthy volunteers at the indicated time points by ELISA. The dotted line represents the lower limit of detection for this assay. LLOQ, lower limit of quantification.

  • Fig. 4 VIB4920 inhibits B cell proliferation and T cell–dependent antibody response in healthy human participants in a dose-dependent manner.

    Healthy volunteers were immunized with KLH 14 days before treatment with placebo or VIB4920 and were reimmunized 15 days after dosing. Three- and 10-mg dose groups only contained n = 2 subjects and were therefore not included in the anti-KLH titer plots. (A) Anti-KLH IgG and (B) anti-KLH IgM titers were assessed by ELISA at various time points. (C) Dose-response model for inhibition of anti-KLH IgG at day 43. (D and E) The frequency of (D) proliferating B cells (Ki67+ CD19+) or (E) class-switched memory B cells (Ki67+ CD19+ CD27+ IgD) in circulation was quantified by flow cytometry at various time points as indicated in volunteers receiving either placebo or the highest dose of VIB4920. (F) Plasma cell signature score in whole blood is shown, as evaluated by TaqMan polymerase chain reaction. Mean and SE expression values for placebo and high-dose VIB4920 groups are shown. *P < 0.05 and **P < 0.01 versus placebo, by Mann-Whitney U test.

  • Fig. 5 VIB4920 demonstrates dose-dependent reductions in ADA.

    (A to C) The presence of ADAs was determined by ELISA. (A) Each subject from the SAD study with healthy volunteers is depicted by an individual line. Subjects with high ADAs (>480 median titer; top dotted line) are indicated with a magenta line; subjects with low ADAs (<480 median titer) are indicated with a dark blue line; subjects with undetectable ADAs are noted with a light blue line. The lower dotted line represents lower titer limit, below which samples are considered negative. (B and C) ADA data from the MAD study in RA. (B) The percentage of subjects with a positive ADA titer at any postdose time point during the study. (C) ADA titer over time in subjects with detectable ADA. LOD, limit of detection.

  • Fig. 6 VIB4920 reduces disease index scores and autoantibodies in patients with RA.

    Disease assessments as indicated were determined from RA donors given placebo or VIB4920 at the indicated visit days. BSL, baseline. (A) DAS28-CRP, (B) CDAI, (C) patient global assessment, (D) physician global assessment, (E) RF autoantibodies, and (F) Vectra disease activity (DA) score. (A to F) Shown is the change in each measure from baseline (means and SE). Data were analyzed using a mixed model for repeated measures analysis with corresponding baseline result included as a covariate.

  • Table 1 RA cohort demographics and clinical characteristics.

    DAS28-CRP, disease activity score 28-joint count–C-reactive protein; CDAI, clinical disease activity index; ESR, erythrocyte sedimentation rate; RF, rheumatoid factor; ACPA, anti–citrullinated peptide antibodies; DMARDs, disease-modifying antirheumatic drugs.

    PlaceboVIB4920 (75 mg)VIB4920 (500 mg)VIB4920 (1000 mg)VIB4920 (1500 mg)VIB4920 (total)
    n15810121242
    Age, mean (SD)58.3 (8.0)57.9 (7.4)53.7 (9.8)52.1 (11.5)50.9 (8.6)53.2 (9.6)
    Sex, n (%) female14 (93.3)6 (75.0)6 (60.0)10 (83.3)10 (83.3)32 (76.2)
    Years since onset of
    symptoms, mean (min,
    max)
    11.5 (1.1, 39.8)13.1 (1.6, 29.5)11.7 (4.0, 20.8)15.0 (2.5, 50.0)10.4 (1.8, 23.2)12.5 (1.6, 50.0)
    Years since diagnosis, mean
    (min, max)
    11.1 (1.1, 39.8)12.8 (1.6, 28.5)9.3 (2.2, 19.9)12.6 (0.5, 44.0)8.5 (1.2, 23.2)10.7 (0.5, 44.0)
    Baseline disease activity
    DAS28-CRP, mean (SD)5.7 (1.18)5.4 (0.85)5.0 (0.98)5.5 (0.56)5.0 (0.58)5.2 (0.76)
    Patient global assessment, mean (SD)64.3 (28.6)47.5 (15.9)52.2 (19.5)61.9 (17.2)59.9 (16.8)56.3 (17.7)
    Physician global assessment, mean (SD)64.1 (21.1)57.9 (17.8)54.6 (12.5)66.0 (12.0)59.8 (11.7)60.0 (13.5)
    CDAI39.8 (13.0)39.3 (11.9)29.3 (9.4)37.7 (7.3)30.1 (5.6)33.8 (9.3)
    Swollen joints10.0 (4.2)9.5 (5.3)7.1 (2.5)8.8 (2.7)6.6 (1.0)7.9 (3.1)
    Tender joints16.9 (7.1)19.3 (7.4)11.5 (4.9)16.2 (5.2)11.5 (5.7)14.3 (6.4)
    CRP (mg/liter)19.1 (34.3)4.7 (5.5)9.2 (10.4)7.7 (6.1)8.5 (12.8)7.7 (9.2)
    ESR (mm/hour)39.0 (25.6)27.0 (15.4)29.1 (19.9)35.3 (17.3)37.8 (25.7)33.0 (20.1)
    RF (U/ml)255 (292.0)505 (670.5)394 (752.9)187 (199.0)242 (207.9)312 (488.2)
    RF positive, n (%)13 (86.7%)7 (87.5%)7 (70.0%)10 (83.3%)11 (91.7%)35 (83.3%)
    ACPA positive, n (%)15 (100%)6 (75%)10 (100%)12 (100%)12 (100%)40 (95.2%)
    DMARDs14 (93.3%)8 (100%)10 (100%)11 (91.7%)11 (91.7%)40 (95.2%)
    Corticosteroids5 (33.3%)3 (37.5%)2 (20.0%)5 (41.7%)3 (25.0%)13 (31.0%)
  • Table 2 VIB4920 demonstrates an acceptable safety profile in patients with RA.

    VIB4920 demonstrates an acceptable safety profile in patients with RA..

    Subjects withPlacebo
    (n = 15)
    VIB4920
    (75 mg; n = 8)
    VIB4920
    (500 mg; n = 10)
    VIB4920
    (1000 mg; n = 12)
    VIB4920
    (1500 mg; n = 12)
    VIB4920
    (total; n = 42)
    ≥1 event13 (86.7%)3 (37.5%)7 (70.0%)6 (50.0%)10 (83.3%)26 (61.9%)
    ≥1 related event5 (33.3%)1 (12.5%)5(50.0%)3 (25.0%)6 (50.0%)15 (35.7%)
    ≥1 event of ≥grade 3 severity*001 (10.0%)1 (8.3%)1 (8.3%)3 (7.1%)
    Death (grade 5 severity)000000
    ≥1 serious event00001 (8.3%)1 (2.4%)
    ≥1 serious and/or ≥grade 3
    severity event
    001 (10.0%)1 (8.3%)1(8.3%)3 (7.1%)
    ≥1 related serious event000000
    ≥1 event leading to discontinuation of
    investigational product
    01 (12.5%)001 (8.3%)2 (4.8%)

    *Subject 20022150007 on 500-mg dose reported three grade 3 adverse events concomitantly (10 to 24 February 2017); PTs: sinusitis, sinus polyp, and nasal turbinate hypertrophy; subject 20024460007 on 1000-mg dose reported a grade 3 event of worsening diabetes mellitus.

    †Subject 20022150009 reported serious adverse event of preferred term encephalitis.

    Supplementary Materials

    • stm.sciencemag.org/cgi/content/full/11/489/eaar6584/DC1

      Materials and Methods

      Fig. S1. Structural characterization of CD40L-Tn3 molecule and interactions.

      Fig. S2. Mouse surrogate Tn3 shows potent neutralizing activity in vivo in response to immunization.

      Fig. S3. Study design for phase 1a study to evaluate safety of VIB4920.

      Fig. S4. Phase 1b study design to evaluate VIB4920 in patients with RA.

      Fig. S5. Dose-dependent reduction in DAS28-CRP and RF autoantibodies in RA in response to VIB4920.

      Fig. S6. Impact of VIB4920 on tender/swollen joint counts and CRP in phase 1b study in patients with RA.

      Table S1. Fusion with serum albumin greatly improves half-life of Tn3 molecule.

      Table S2. VIB4920 demonstrates a good safety profile in healthy volunteers.

      Table S3. VIB4920 demonstrates an acceptable safety profile in patients with RA.

      Table S4. DAS28 categories at day 85.

      Data file S1. Primary data.

    • The PDF file includes:

      • Materials and Methods
      • Fig. S1. Structural characterization of CD40L-Tn3 molecule and interactions.
      • Fig. S2. Mouse surrogate Tn3 shows potent neutralizing activity in vivo in response to immunization.
      • Fig. S3. Study design for phase 1a study to evaluate safety of VIB4920.
      • Fig. S4. Phase 1b study design to evaluate VIB4920 in patients with RA.
      • Fig. S5. Dose-dependent reduction in DAS28-CRP and RF autoantibodies in RA in response to VIB4920.
      • Fig. S6. Impact of VIB4920 on tender/swollen joint counts and CRP in phase 1b study in patients with RA.
      • Table S1. Fusion with serum albumin greatly improves half-life of Tn3 molecule.
      • Table S2. VIB4920 demonstrates a good safety profile in healthy volunteers.
      • Table S3. VIB4920 demonstrates an acceptable safety profile in patients with RA.
      • Table S4. DAS28 categories at day 85.

      [Download PDF]

      Other Supplementary Material for this manuscript includes the following:

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