Table 1.
Busulfan animal transplantation and engraftment parameters
| Animal ID |
11021142 |
10U004 |
H84D |
|||
|---|---|---|---|---|---|---|
| Vector | pCDH-EF1α-T2A- no marker gene library 15 (without WPRE) | pCDH-EF1α-T2A- tNGFR library 17 (without WPRE) | pCDH-EF1α-T2A- no marker gene library 15 (without WPRE) | pCDH-EF1α-T2A- tNGFR library 17 (without WPRE) | pCDH-EF1α-T2A- no marker gene library 21 (with WPRE) | pCDH-EF1α-T2A- tNGFR library 11 (with WPRE) |
| Date of transplant | 02/01/2019 | 11/29/2019 | 07/31/2020 | |||
| Transduction MOI | 25 | 25 | 25 | 25 | 20 | 20 |
| Transduction condition | FN+ Cytokines + protamine sulfatea | FN+ Cytokines + protamine sulfatea | FN + Cytokines + p407 + PGE2b | |||
| Transduction efficiencyc | N/A | 31.5% | N/A | 31.2% | N/A | 66.0% |
| Number of cells collected (× 106) | 28 | 27.5 | 28 | |||
| Number of cells infused (× 106) | 36.0 | 36.0 | 45.0 | 41.0 | 25.9 | 25.6 |
| Transplantation dose (CD34+ cells × 106/kg) | 15.0 | 16.4 | 10.5 | |||
| Busulfan conditioning dose (mg/kg) × 4 days | 5.5 | 5.5 | 6.0 | |||
| Day until neutrophil recovery to >500/μL | Day 11 | Never Below 500/μL | Never below 500/μL | |||
| Last follow-up (months post- transplant) | 36 | 26 | 18 | |||
| Total number of barcoded clones detected | 2,118 | 2,250 | 18,803 | |||
| Number of barcoded clones detected at the last follow-up, which include all lineages (T, B, Mo, Gr, CD16+, and CD56+NK) | 13 (24m) | 52 (22.5m) | 1,778 (18m) | |||
| d Estimated Minimum frequency long-term CD34+ engraftment (at last follow-up) | 0.00011% | 0.00037% | 0.01% | |||
a
Fibronectin (FN) coated plate + % HSA + cytokines (Flt-3, SCF, TPO all at 100 ng/mL) + protamine sulfate (4 μg/mL).
b
Fibronectin (FN) coated plate + cytokines (Flt-3, SCF, TPO all at 100 ng/mL) + P407 (100 μg/mL) + PGE2 (10 μM).
c
Transduction efficiency is measure by FACS for NGFR.
d
Estimated Minimum frequency long-term CD34+ engraftment (at last follow-up) is calculated by using the “unique clones number that tracked at the last follow-up” to divide “initial infused transduced CD34+ cells numbers.”