Quantitation of HPLC analysis in controls and corrected iPSCs
| . | . | α0 Thal iPS . | β1* . | β2* . | α1† . | α2† . | H1 . | FL . | ||
|---|---|---|---|---|---|---|---|---|---|---|
| Het . | Hom . | Het . | Hom . | Hom . | Hom . | |||||
| Baso-E, 14-day liquid culture | ||||||||||
| α/all globins | % α globin | 0.0 | 4.7 | 8.2 | 5.4 | 12.9 | 14.5 | 13.6 | 9.8 | |
| α/α+ζ | αζ ratio | 0.0 | 12.1 | 18.8 | 11.8 | 24.6 | 31.8 | 28.8 | 19.0 | |
| α+ζ/γ+ϵ+β | Chain imbalance | 73.2 | 63.6 | 76.6 | 83.9 | 110.9 | 83.8 | 89.3 | 106.2 | |
| Ortho-E, 24-day liquid culture | ||||||||||
| α/all globins | % α globin | 0.0 | 13.3 | 25.8 | ND | 26.0 | 42.6 | 37.8 | 46 | 45.4 |
| α/α+ζ | αζ ratio | 0.0 | 79.5 | 75.4 | ND | 94.5 | 98.4 | 81.9 | 91.5 | 95.0 |
| α+ζ/γ+ϵ+β | Chain imbalance | 2.3 | 20.1 | 52.0 | ND | 38.0 | 76.5 | 85.8 | 101.2 | 91.4 |
| . | . | α0 Thal iPS . | β1* . | β2* . | α1† . | α2† . | H1 . | FL . | ||
|---|---|---|---|---|---|---|---|---|---|---|
| Het . | Hom . | Het . | Hom . | Hom . | Hom . | |||||
| Baso-E, 14-day liquid culture | ||||||||||
| α/all globins | % α globin | 0.0 | 4.7 | 8.2 | 5.4 | 12.9 | 14.5 | 13.6 | 9.8 | |
| α/α+ζ | αζ ratio | 0.0 | 12.1 | 18.8 | 11.8 | 24.6 | 31.8 | 28.8 | 19.0 | |
| α+ζ/γ+ϵ+β | Chain imbalance | 73.2 | 63.6 | 76.6 | 83.9 | 110.9 | 83.8 | 89.3 | 106.2 | |
| Ortho-E, 24-day liquid culture | ||||||||||
| α/all globins | % α globin | 0.0 | 13.3 | 25.8 | ND | 26.0 | 42.6 | 37.8 | 46 | 45.4 |
| α/α+ζ | αζ ratio | 0.0 | 79.5 | 75.4 | ND | 94.5 | 98.4 | 81.9 | 91.5 | 95.0 |
| α+ζ/γ+ϵ+β | Chain imbalance | 2.3 | 20.1 | 52.0 | ND | 38.0 | 76.5 | 85.8 | 101.2 | 91.4 |
Summary of HPLC analyses of globin chain expression performed after 14 days or 24 days of liquid culture.
iPSC indicates induced pluripotent stem cell; Thal, thalassemia; FL, fetal liver; Het, heterozygous insertion; Hom, homozygous insertion; and ND, not determined.
Columns β1 and β2 summarize the results for the iPSCs in which the α-globin therapeutic gene is driven by the β-globin promoter. In iPSCs, the α1 and β1 transgenes are in the same orientation as the PPP1R12C genes. In iPSCs, the α2 and β2 transgenes are in opposite orientation (n = 3).
Columns α1 and α2 summarize the results for the iPSCs in which the α-globin therapeutic gene is driven by the α-globin promoter.