Table 1.
Summary of mtDNA sequence variation in single cells from the leukemia patients and healthy controls
| Sample | Leukemiatype | Age, y/sex | Haplogroup | No. of cells | No. ofhaplotypes, (%) | No. ofnonaggregate haploytpes by nucleotide substitutions (%) | Mean no. of cells with nonaggregate haplotype | Mean no. of cells with nonaggregate haplotype by nucleotide substitutions |
|---|---|---|---|---|---|---|---|---|
| CD34+ and blasts | ||||||||
| Patients | ||||||||
| LAA | AML M0 | 61/F | H | 95 | 16 (16.84) | 11 (11.58) | 2.58 | 1.05 |
| AGM | AML M1 | 62/M | B4b | 96 | 11 (11.46) | 7 (7.29) | 5.07 | 1.19 |
| MFSS | AML M1 | 36/F | L2a | 96 | 10 (10.42) | 8 (8.33) | 1.04 | 1.04 |
| JCS | AML M1 | 40/M | L3f | 96 | 15 (15.63) | 12 (12.50) | 3.67 | 1.04 |
| DC | AML M1 | 15/M | L2a | 96 | 18 (18.75) | 12 (12.50) | 3.25 | 3.27 |
| OAM at diagnosis | AML M1 | 33/M | D1 | 92 | 11 (11.96) | 9 (9.78) | 1.51 | 1.56 |
| EMB | AML M1 | 16/M | D1 | 95 | 34 (35.79) | 30 (31.58) | 1.05 | 1.05 |
| ERR | AML M1 | 53/F | A2 | 94 | 6 (6.38) | 5 (5.32) | 1.28 | 1.28 |
| UPN21 | AML | 16/F | D1 | 82 | 7 (8.54) | 3 (3.66) | 1.42 | 1.22 |
| UPN1 | AML M2 | 47/F | T2 | 146 | 10 (6.85) | 5 (3.42) | 1.73 | 0.68 |
| UPN16 | 2nd AML/MDS | 74/F | J1c | 92 | 5 (5.43) | 3 (3.26) | 2.42 | 1.09 |
| UPN22 | Relapsed AML | 11/F | H | 65 | 20 (30.77) | 11 (16.92) | 3.32 | 2.63 |
| OAM at relapse | Relapsed AML | 36/M | D1 | 96 | 6 (6.25) | 4 (4.17) | 5.61 | 6.78 |
| UPN18 | AML, prior NHL | 62/F | A2 | 92 | 8 (8.70) | 4 (4.35) | 2.30 | 1.09 |
| UPN20 | CML | 53/M | A2 | 85 | 3 (3.53) | 1 (1.18) | 6.90 | 1.18 |
| UPN17 | CML | 27/F | X2b | 80 | 17 (21.25) | 10 (12.50) | 3.97 | 4.42 |
| UPN2 | AML from CMML | 58/M | J1c | 96 | 14 (14.58) | 13 (13.54) | 2.31 | 2.31 |
| UPN3 | CLL | 64/F | H | 89 | 9 (10.11) | 4 (4.49) | 2.10 | 1.12 |
| UPN19, blasts | APML | 21/F | H | 94 | 22 (23.40) | 15 (15.96) | 2.60 | 1.06 |
| UPN19, normal CD 34+ cells | APML | 21/F | H | 96 | 10 (10.42) | 4 (4.17) | 4.76 | 1.04 |
| Grand mean ± SD | — | — | — | 94 ± 15 | 13.85 ± 8.56 | 9.33 ± 7.04 | 2.94 ± 1.63 | 1.81 ± 1.49 |
| Donors | ||||||||
| Donor 1 | — | 48/F | U5b | 85 | 14 (16.47) | 8 (9.41) | 2.74 | 1.47 |
| Donor 2 | — | 44/M | L2a | 92 | 16 (17.39) | 11 (11.96) | 2.27 | 1.09 |
| Donor 3 | — | 36/M | L0a1 | 93 | 12 (12.90) | 6 (6.45) | 3.19 | 2.13 |
| Donor 4 | — | 55/M | T2 | 93 | 14 (15.05) | 8 (8.60) | 3.32 | 3.02 |
| Donor 5 | — | 35/M | U5b | 93 | 13 (13.98) | 10 (10.75) | 2.92 | 1.18 |
| Donor 6 | — | 35/M | M7a1 | 95 | 15 (15.79) | 10 (10.53) | 2.91 | 1.05 |
| Donor 7 | — | 32/M | U6 | 93 | 40 (43.01) | 35 (37.63) | 1.45 | 1.08 |
| Donor 8 | — | 25/M | H | 93 | 38 (40.86) | 35 (37.63) | 1.58 | 1.08 |
| Donor 9 | — | 57/M | K1a | 93 | 31 (33.33) | 28 (30.11) | 1.36 | 1.34 |
| Donor 10 | — | 39/M | pre-V | 94 | 17 (18.09) | 12 (12.77) | 2.66 | 1.06 |
| Grand mean ± SD | — | — | — | 92 ± 3 | 22.69 ± 11.65 | 17.58 ± 12.40 | 2.44 ± 0.73 | 1.45 ± 0.64 |
| Granulocytes | ||||||||
| Patients | ||||||||
| UPN21 | AML | 16/F | D1 | 24 | 8 (33.33) | 6 (25.00) | 4.75 | 4.17 |
| UPN22 | Relapsed AML | 11/F | H | 70 | 42 (60.00) | 30 (42.86) | 2.00 | 1.89 |
| UPN20 | CML | 53/M | A2 | 92 | 17 (18.48) | 13 (14.13) | 1.80 | 1.09 |
| UPN19 | APML | 21/F | H | 79 | 23 (29.11) | 15 (18.99) | 2.47 | 1.27 |
| Grand mean ± SD | — | — | — | 66 ± 30 | 35.23 ± 17.66 | 25.25 ± 12.56 | 2.76 ± 1.36 | 2.11 ± 1.42 |
| Donors | ||||||||
| Donor 1 | — | 48/F | U5b | 64 | 28 (43.75) | 23 (35.94) | 2.12 | 1.56 |
| Donor 2, PB | — | 44/M | L2a | 82 | 27 (32.93) | 22 (26.83) | 1.77 | 1.33 |
| Donor 2, bone marrow | — | 45/M | L2a | 96 | 16 (16.67) | 12 (12.50) | 2.36 | 1.04 |
| Donor 3 | — | 36/M | L0a1 | 72 | 26 (36.11) | 23 (31.94) | 1.66 | 1.39 |
| Donor 4 | — | 55/M | T2 | 76 | 27 (35.53) | 19 (25.00) | 1.99 | 1.66 |
| Donor 5 | — | 35/M | U5b | 82 | 31 (37.80) | 26 (31.71) | 1.34 | 1.27 |
| Grand mean ± SD | — | — | — | 79 ± 11 | 33.80 ± 9.14 | 27.32 ± 8.25 | 1.87 ± 0.36 | 1.38 ± 0.22 |
The mtDNA haplogroup classification was based on sequence variation in the consensus sequence of the single cells from respective individuals (Table S2). We counted the number of haplotypes (including the aggregate mtDNA haplotype and the nonaggregate haplotypes that differed from aggregate sequence by nucleotide substitutions or indels or both) and the number of nonaggregate haplotypes that differed from the aggregate type only by nucleotide substitutions, based on the mutations detected in a population of cells from each sample (Table S3). The geometric mean number of cells harboring each nonaggregate haplotype or nonaggregate haplotype by nucleotide substitutions was computed as another parameter to show the difference between the leukemia patients and healthy donors. The data of peripheral blood CD34+ cells and granulocytes for healthy donors 1 to 5 are from Ogasawara et al.21 Single blasts at diagnosis and relapse were analyzed for patient OAM. Both blasts (CD33+ CD34−) and normal CD34+ cells were analyzed for patient UPN19.
Granulocytes from peripheral blood and bone marrow were analyzed for healthy donor 2.
MDS indicates myelodysplastic syndrome; NHL, non-Hodgkin lymphoma; CMML, chronic monomyelocytic leukemia; —, not applicable.