Table 1.
Experimental groups of mice by genotype and treatment*
| Genotype | No. of mice examined (M+F) | Injected with | Dose per mouse (IFU) | Frequency of injections | Age at time of injection | Duration | Age at time of analysis |
|---|---|---|---|---|---|---|---|
| Set 1 (Time course) | |||||||
| C57BL/6J | 6 (3+3) | NA | NA | NA | NA | NA | 10w |
| Abcc6−/−Rag1−/− | 4 (2+2) | NA | NA | NA | NA | NA | 10w |
| Abcc6−/−Rag1−/− | 6 (2+4) | Ad5-CMV-hABCC6 | 4×108 | 1 | 6w | 1w | 7w |
| Abcc6−/−Rag1−/− | 6 (1+5) | Ad5-CMV-hABCC6 | 4×108 | 1 | 6w | 2w | 8w |
| Abcc6−/−Rag1−/− | 4 (2+2) | Ad5-CMV-hABCC6 | 4×108 | 1 | 6w | 4w | 10w |
| Abcc6−/−Rag1−/− | 5 (1+4) | Ad5-CMV-hABCC6 | 4×108 | 1 | 6w | 6w | 12w |
| Abcc6−/−Rag1−/− | 4 (2+2) | Ad5-CMV-hABCC6 | 4×108 | 1 | 6w | 8w | 14w |
| Set 2 (Dose response) | |||||||
| C57BL/6J | 6 (3+3) | NA | NA | NA | NA | NA | 10w |
| Abcc6−/−Rag1−/− | 4 (2+2) | NA | NA | NA | NA | NA | 10w |
| Abcc6−/−Rag1−/− | 8 (4+4) | Ad5-PL-hABCC6 | 1×109 | 1 | 6w | 4w | 10w |
| Abcc6−/−Rag1−/− | 8 (5+3) | Ad5-CMV-hABCC6 | 1×109 | 1 | 6w | 4w | 10w |
| Abcc6−/−Rag1−/− | 8 (5+3) | Ad5-CMV-hABCC6 | 4×108 | 1 | 6w | 4w | 10w |
| Set 3 (Prevention study) | |||||||
| C57BL/6J | 7 (2+5) | NA | NA | NA | NA | NA | 14w |
| C57BL/6J | 9 (4+5) | NA | NA | NA | NA | NA | 18w |
| Abcc6−/−Rag1−/− | 14 (9+5) | NA | NA | NA | NA | NA | 14w |
| Abcc6−/−Rag1−/− | 13 (9+4) | NA | NA | NA | NA | NA | 18w |
| Abcc6−/−Rag1−/− | 10 (6+4) | Ad5-CMV-hABCC6 | 4×108 | 1 | 6w | 8w | 14w |
| Abcc6−/−Rag1−/− | 9 (5+4) | Ad5-CMV-hABCC6 | 4×108 | 2 | 6w, 10w | 4w, 4w | 14w |
| Abcc6−/−Rag1−/− | 13 (9+4) | Ad5-CMV-hABCC6 | 4×108 | 3 | 6w, 10w, 14w | 4w, 4w, 4w | 18w |
| Set 4 (Reversal study) | |||||||
| C57BL/6J | 6 (3+3) | NA | NA | NA | NA | NA | 47w |
| Abcc6−/−Rag1−/− | 8 (3+5) | Ad5-PL-hABCC6 | 4×108 | 1 | 43w | 4w | 47w |
| Abcc6−/−Rag1−/− | 8 (3+5) | Ad5-CMV-hABCC6 | 4×108 | 1 | 43w | 4w | 47w |
*
Mice with desired genotypes were injected with either Ad5-PL-hABCC6 or Ad5-CMV-hABCC6 adenovirus at designated frequency and duration as indicated in the table. The mice were sacrificed for determination of human ABCC6 expression in mouse liver, PPi levels in plasma and the degree of ectopic mineralization in the muzzle skin. M, male; F, female; w, weeks.