Relevant for In Vivo circvec-DNA i CAR-T
To date, five patients with SLE that was refractory to multiple conventional therapies have been enrolled (Fig. S7 and Table S1). Immunosuppressive drugs were stopped within 1 week before the first infusion of HN2301. The schematic treatment schedule of HN2301 is shown in Figure 1A. At 6 hours after the infusion, CD8+ CD19 CAR T cells were detectable in the peripheral blood. The levels of CD19 CAR T cells and CAR mRNA peaked 6 hours after each HN2301 infusion and decreased to baseline levels within 2 to 3 days (Figure 1B and Figs. S8 and S9). Off-target expression of CAR on non-CD8+ T cells was less than 10% (Fig. S10). Within 6 hours after the first treatment, circulating B cells were substantially decreased in patients who received the 2-mg dose (Patients 1 and 2) and were completely depleted (<1 B cell per microliter) in patients who received the 4-mg dose (Patients 3, 4, and 5), and depletion was maintained until 7 to 10 days after administration (Figure 1C). Up-regulation of CD69 was observed on CD8+ T cells but not on CD4+ T cells, which indicates the activation of CAR T cells (Fig. S11).
Supplementary Figures, NHP og humane data viser at:
- CAR-mRNA og CAR-positive T-celler:
- topper raskt etter infusjon
- faller tilbake mot baseline innen få dager
- Derfor må pasientene få:
- gjentatte infusjoner (dag 0, 2, 4) for å opprettholde effekt
mRNA-payload som gir et kortvarig uttrykk (timer, dager), i motsetning til:
-
circRNA Forlenget, men fortsatt forbigående uttrykk
-
circVec-DNA Potensial for vedvarende uttrykk (uker–måneder)
Doseringsregimet i studien bekrefter kort varighet:
- Repetert dosering med 48 timers intervall
- Tett cytokinovervåkning etter hver dose
- Effekt (B-celle-deplesjon) oppnås, men krever re-eksponering
