Figure 1.
In vitro effect of apilimod on IL-12p70, IL-10, GM-CSF, and IL-6 in human whole blood cells.
Human whole blood from a normal volunteer was stimulated with 0.1% SAC in the presence of different concentrations of apilimod. Supernatants were tested for IL-12p70 (circles), IL-10 (triangles), GM-CSF (diamonds) and IL-6 (squares). Results are representative of one of three individual experiments with whole blood from different volunteers each time.
Figure 2.
Ex vivo stimulation of whole blood drawn 2h post 70mg dose as compared to the pre-dose in cytokine production.
Whole blood drawn pre-dose and 2h post 70mg dose (n = 10) were stimulated with 0.1% SAC within 24h of the draw, and the supernatants were analyzed for IL-12p70 (a), IL-10 (b), and GM-CSF (c). *, p ≤0.05; statistically significant differences between pre- and 2h post-dose.
Figure 3.
Histological improvement by apilimod treatment.
Histology and immunohistochemistry of one patient (1046) showing improved histology and clinical measures (58%reduction in PASI score) at week 12 in the 70mg QD apilimod treated group. Skin biopsies from non-lesions (left) and lesions (middle) at baseline and lesion at week 12 (right) were stained with H&E, K16, anti-CD3 Ab, anti-CD11c Ab, or anti- IL-12p40 Ab. Cells staining positive for CD3, CD11c and IL-12p40 are indicated (arrows).
Table 1.
Clinical response based on assessment of skin biopsies and PASI and PGA.
Table 2.
Mean skin-infiltrating T cell and dendritic cell numbers.
Figure 4.
Changes in epidermal CD11c+ cells in responders and non-responders in 70mg QD apilimod cohort.
Results shown are the number of epidermal CD11c+ cells in individual patients categorized as either responders (left, n = 7) or non-responders (right, n = 7) in the 70mg QD cohort at baseline (week 0), week 2, 6, and 12.
Figure 5.
Changes in the expression levels of IL-12/IL-23p40, IL-23p19, and IL-10 at week 2 in 70mg QD apilimod cohort.
RNA was prepared from biopsies obtained from the psoriatic skin lesions at baseline (week 0) and week 2, and RT-PCR was performed for IL-12/IL-23p40 (a), IL-23p19 (b), and IL-10 (c). The expression levels were normalized to house keeping gene, hARP. Results shown are the expression levels of individuals in 70mg QD cohort (n = 11, 3 histological responders, 8 histological non-responders). *, p ≤0.05; **, p ≤0.01; statistically significant differences between baseline and week 2.
Figure 6.
Changes in the expression levels of TH1, TH17, and other inflammatory genes at week 12 in responders and non-responders.
RT-PCR was performed with biopsies obtained from non-lesion skin and psoriatic lesions. Results shown are the percentile range (25th–75th%, medians indicated) of histological responders (n = 11) and non-responders (n = 27) for IL-12/IL-23p40, IL-23p19, K16, IL-8, IL-17, IFN-γ, iNOS and TNF-α. There were 26 non-responders for IL-17, and 10 responders and 23 non-responders for IL-10 in the analysis of the change due to 0 value at baseline. *, p < 0.05; **, p <0.01; statistically significant differences between before (week 0) and after (week 12) apilimod treatment in responder and non-responder groups.
Figure 7.
Cellular phenotype in normal controls and psoriasis patients before and after 70mg QD treatment.
Whole blood cells from normal controls and psoriasis patients were analyzed by cytometry for the cellular phenotype. Results shown are the percentile ranges based on cell counts (cells/µL) of peripheral CD4+ T cells, CD8+ T cells, B cells, monocytes, eosinophils and neutrophils from normal controls (n = 16) and psoriasis patients in 70mg QD cohort (n = 12) at baseline (week 0) and week 12. Plot: bottom line, 10th%; bottom box, 25th%; top box, 75th%; top line, 90th%. *, p < 0.05; **, p <0.01; statistically significant differences between before (week 0) and after (week 12) 70mg QD apilimod treatment in psoriasis patients.