Heterogeneous patterns of engraftment of CD34− cells from NPM1-mutated AML in immunocompromised mice
| Patient code . | % CD34+ in original sample . | Purity of CD34− cells . | No./type mice evaluated . | No. of cells/mice, × 106 . | No. of mice engrafted/no. evaluated . | Time of evaluation, w . | % hCD45+ in mice BM* . | % hCD45+/CD33+ (gated on hCD45)* . | Pattern of engraftment† . | WB (NPMm) . | NPM1 gene status‡ . |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 8R | 1.9 | 98.5 | 2/NOD | 2 | 2/2 | 16 | 36/73‖ | 97.5/99 | 2 | + | ND |
| 23 | 1.7 | 99 | 1/NOG | 0.2 | 1/1 | 15 | 21 | 0.57 | 2 | + | Mutated |
| 21 | 0.02 | 99.99 | 2/NOG | 2 | 2/2 | 6 | 29/7.5¶ | 13.8/95 | 4 | ND | ND |
| 22§ | 4.5 | 99.9 | 6/NOG | 10 | 6/6 | 4/5/6/7/8/9 | 75/72/86‖/82/87#/94‖ | 82/85/93/90/89/97 | 3 | + | Mutated |
| 27 | 0.02 | 99.98 | 3/NOG | 10 | 3/3 | 4/6/8 | 33/64‖/42 | 73/86/56 | 3 | + | Mutated |
| 28§ | 0.07 | 99.98 | 1/NOG | 10 | 1/1 | 5 | 42 | 46 | 3 | ND | Mutated |
| 3/NOG | 2 | 3/3 | 6/9/13 | 4.3/12/5¶ | 30/35/82 | 3 | ND | Mutated | |||
| 29§ | 0.5 | 99.9 | 1/NOG | 10 | 1/1 | 6 | 14¶ | 0.7 | 4 | NA | NA |
| 4/NOG | 4 | 4/4 | 5/6/7/14 | 16/ND/2.7/10.7¶ | 2.2/–/1.7/1 | 4 | NA | NA | |||
| 2/NOG | 2 | 2/2 | 7/14 | 1.6/16 | 4/2.2 | 4 | NA | NA | |||
| 34§ | 5 | 96 | 2/NOG | 10 | 2/2 | 4 | 0.61/4.9 | –/0.2 | 4 | NA | NA |
| 36 | 0.26 | 99.98 | 3/NOG | 10 | 3/3 | 6 | 4.7/2.3/5.2¶ | 0.3/5.9/9.9 | 4 | NA | NA |
| 37§ | 0.2 | 99.8 | 4/NOG | 10 | 3/4 | 5/7/8/11 | 1.8/3.2/0/1.65 | 95.8/98.7/–/96 | 4 | NA | NA |
| 38§ | 0.16 | 99.9 | 3/NOG | 10 | 3/3 | 7 | 0.68/1.2/20¶ | 1.8/2.5/1 | 4 | NA | NA |
| Patient code . | % CD34+ in original sample . | Purity of CD34− cells . | No./type mice evaluated . | No. of cells/mice, × 106 . | No. of mice engrafted/no. evaluated . | Time of evaluation, w . | % hCD45+ in mice BM* . | % hCD45+/CD33+ (gated on hCD45)* . | Pattern of engraftment† . | WB (NPMm) . | NPM1 gene status‡ . |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 8R | 1.9 | 98.5 | 2/NOD | 2 | 2/2 | 16 | 36/73‖ | 97.5/99 | 2 | + | ND |
| 23 | 1.7 | 99 | 1/NOG | 0.2 | 1/1 | 15 | 21 | 0.57 | 2 | + | Mutated |
| 21 | 0.02 | 99.99 | 2/NOG | 2 | 2/2 | 6 | 29/7.5¶ | 13.8/95 | 4 | ND | ND |
| 22§ | 4.5 | 99.9 | 6/NOG | 10 | 6/6 | 4/5/6/7/8/9 | 75/72/86‖/82/87#/94‖ | 82/85/93/90/89/97 | 3 | + | Mutated |
| 27 | 0.02 | 99.98 | 3/NOG | 10 | 3/3 | 4/6/8 | 33/64‖/42 | 73/86/56 | 3 | + | Mutated |
| 28§ | 0.07 | 99.98 | 1/NOG | 10 | 1/1 | 5 | 42 | 46 | 3 | ND | Mutated |
| 3/NOG | 2 | 3/3 | 6/9/13 | 4.3/12/5¶ | 30/35/82 | 3 | ND | Mutated | |||
| 29§ | 0.5 | 99.9 | 1/NOG | 10 | 1/1 | 6 | 14¶ | 0.7 | 4 | NA | NA |
| 4/NOG | 4 | 4/4 | 5/6/7/14 | 16/ND/2.7/10.7¶ | 2.2/–/1.7/1 | 4 | NA | NA | |||
| 2/NOG | 2 | 2/2 | 7/14 | 1.6/16 | 4/2.2 | 4 | NA | NA | |||
| 34§ | 5 | 96 | 2/NOG | 10 | 2/2 | 4 | 0.61/4.9 | –/0.2 | 4 | NA | NA |
| 36 | 0.26 | 99.98 | 3/NOG | 10 | 3/3 | 6 | 4.7/2.3/5.2¶ | 0.3/5.9/9.9 | 4 | NA | NA |
| 37§ | 0.2 | 99.8 | 4/NOG | 10 | 3/4 | 5/7/8/11 | 1.8/3.2/0/1.65 | 95.8/98.7/–/96 | 4 | NA | NA |
| 38§ | 0.16 | 99.9 | 3/NOG | 10 | 3/3 | 7 | 0.68/1.2/20¶ | 1.8/2.5/1 | 4 | NA | NA |
BM indicates bone marrow; NOD, NOD/SCID; NOG, NOD/SCID/IL2Rγnull; WB (NPMm), Western blot analysis with anti-NPM mutant specific antibody; R, relapse; ND, not done; and NA, not applicable
Evaluated by flow cytometry on cells recovered from femurs or tibias.
See “Patterns of engraftment of CD34− cells from NPM1-mutated AML in immunocompromised mice” for description of patterns of engraftment.
Evaluated by genomic DNA fragment analysis.
Lower cell doses did not engraft (supplemental Table 5).
Used for serial transplantation experiments.
Human cell infiltration of mice BM was much higher when evaluated at immunohistochemistry.
FLT3-ITD-negative.