scholarly journals In Vitro Drug Sensitivity Testing Can Predict Induction Failure and Early Relapse of Childhood Acute Lymphoblastic Leukemia

Blood ◽  
1997 ◽  
Vol 89 (8) ◽  
pp. 2959-2965 ◽  
Author(s):  
Teruaki Hongo ◽  
Shuhei Yajima ◽  
Minoru Sakurai ◽  
Yasuo Horikoshi ◽  
Ryoji Hanada
Keyword(s):  
Drug Resistance ◽  
Acute Lymphoblastic Leukemia ◽  
Drug Sensitivity ◽  
Lymphoblastic Leukemia ◽  
Sensitivity Testing ◽  
Childhood All ◽  
Drug Sensitivity Testing ◽  
Induction Failure ◽  
Sensitive Group

Abstract It is vital to develop effective therapy for children with acute lymphoblastic leukemia (ALL), in whom no remission occurs or who suffer relapse with current protocols. Cellular drug resistance is thought to be an important cause of induction failure and relapse. We performed in vitro tests of bone marrow samples in 196 children with newly diagnosed ALL with a 4-day culture and a methyl-thiazol-tetrazolium assay. We tested 16 drugs and calculated the 70% lethal dose (LD70) for 14 drugs and the leukemic cell survival (LCS) rate for dexamethasone and prednisolone. For each single drug, patients were classified into two groups, sensitive or resistant, by median concentration of LD70 or LCS. When patients were classified into three groups by sensitivity to four drugs of DPAV (dexamethasone, prednisolone, L-asparaginase, and vincristine), 3-year event-free survival (EFS; 95% confidence intervals) of the super sensitive group (SS; sensitive to all 4 drugs) was 0.833 (0.690 to 0.976), that of the intermediate sensitive group (IS; sensitive to 2 or 3 drugs) was 0.735 (0.609 to 0.863), and that of the relatively resistant group (RR; sensitive to no drugs or to 1 drug) was 0.541 (0.411 to 0.670; P = .0008). We then investigated the relationship between the above four-drug sensitivity and the time of relapse. The SS and IS patients tended to maintain continuous complete remission, and RR patients tended to undergo induction failure and early and late relapse (P = .004). Initial white blood cell count, immunologic classification, and age were also predictive factors, but the patient numbers showed no statistical correlation between these factors and the four-drug sensitivity groups (SS, IS, and RR). When we took three groups SS/IS/RR and investigated the EFS for various clinical groups, DPAV sensitivity strongly influenced EFS in the standard-risk ALL (P = .016). In vitro drug sensitivity testing provides additional prognostic information about childhood ALL, and early detection of drug resistance at the time chemotherapy commences may provide a successful strategy for individualizing treatment, as the results indicate de novo resistance to front-line drugs and suggest alternative, second-line drugs.

scholarly journals In vitro cellular drug resistance in children with relapsed/refractory acute lymphoblastic leukemia

Blood ◽  
1995 ◽  
Vol 86 (10) ◽  
pp. 3861-3868 ◽  
Author(s):  
E Klumper ◽  
R Pieters ◽  
AJ Veerman ◽  
DR Huismans ◽  
AH Loonen ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Acute Lymphoblastic Leukemia ◽  
Poor Prognosis ◽  
De Novo ◽  
Lymphoblastic Leukemia ◽  
Childhood All ◽  
Acquired Drug Resistance ◽  
The Poor ◽  
Response To Chemotherapy

Cellular drug resistance is thought to be an important cause of the poor prognosis for children with relapsed or refractory acute lymphoblastic leukemia (ALL), but it is unknown when, to which drugs, and to what extent resistance is present. We determined in vitro resistance to 13 drugs with the MTT assay. Compared with 141 children with initial ALL, cells from 137 children with relapsed ALL were significantly more resistant to glucocorticoids, L-asparaginase, anthracyclines, and thiopurines, but not to vinca-alkaloids, cytarabine, ifosfamide, and epipodophyllotoxins. Relapsed ALL cells expressed the highest level of resistance to glucocorticoids, with a median level 357- and >24-fold more resistant to prednisolone and dexamethasone, respectively, than initial ALL cells, whereas the resistance ratios for the other drugs differed from 0.8- to 1.9-fold, intraindividual comparisons between initial and relapsed samples from 16 children with ALL showed that both de novo and acquired drug resistance were involved. Specific in vitro drug-resistance profiles were associated with high-risk relapsed ALL groups. In vitro drug resistance was also related to the clinical response to chemotherapy in relapsed/refractory childhood ALL. We conclude that drug resistance may explain the poor prognosis for children with relapsed/refractory ALL. These day may be helpful to design alternative treatment regimens for relapsed childhood ALL.

scholarly journals In vitro drug-resistance profile in infant acute lymphoblastic leukemia in relation to age, MLL rearrangements and immunophenotype

Leukemia ◽  
2004 ◽  
Vol 18 (3) ◽  
pp. 521-529 ◽  
Author(s):  
N L Ramakers-van Woerden ◽  
H B Beverloo ◽  
A J P Veerman ◽  
B M Camitta ◽  
A H Loonen ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Acute Lymphoblastic Leukemia ◽  
Lymphoblastic Leukemia ◽  
Resistance Profile ◽  
Drug Resistance Profile ◽  
Infant Acute Lymphoblastic Leukemia

scholarly journals In vitro drug sensitivity of cells from children with leukemia using the MTT assay with improved culture conditions

Blood ◽  
1990 ◽  
Vol 76 (11) ◽  
pp. 2327-2336 ◽  
Author(s):  
R Pieters ◽  
AH Loonen ◽  
DR Huismans ◽  
GJ Broekema ◽  
MW Dirven ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Cell Survival ◽  
Mtt Assay ◽  
Large Scale ◽  
Drug Sensitivity ◽  
Lymphoblastic Leukemia ◽  
Leukemia Cells ◽  
Response Curves ◽  
Mtt Reduction

Abstract The knowledge about drug resistance in childhood leukemias and acute lymphoblastic leukemia (ALL) in general is limited. This is because of the lack of a suitable in vitro drug sensitivity assay, which is in part due to low in vitro ALL cell survival. We recently adapted the highly efficient 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay to test cells from ALL patients and showed that its results were comparable with those of the DiSC assay, up to now the most valid but laborious assay. In this study, in vitro drug sensitivity was assessed in cells from 82 children with leukemia, 79 of whom had ALL, with the MTT assay. Dose response curves were obtained for 6-mercaptopurine, 6-thioguanine (6-TG), prednisolone (Pred), daunorubicin (DNR), vincristine (VCR), cytosine arabinoside (Ara-C), L- asparaginase (L-Asp), mafosfamide, and mustine. A cytotoxic effect of methotrexate could be detected in only a few cases. Large interindividual differences in drug sensitivity were detected. Compared with leukemia cells from newly diagnosed patients, leukemia cells from relapsed patients were significantly more in vitro resistant to 6-TG, Pred, Ara-C, mafosfamide and mustine but not to DNR, VCR, and L-Asp. Improvements of culture medium and methods to increase MTT reduction were studied. From 10 components tested, addition of insulin and bovine serum albumin to serum-containing medium improved ALL cell survival. Addition of succinate did not increase the amount of MTT reduction. We conclude that the in vitro MTT assay highly facilitates large-scale studies on drug resistance of ALL patients that can lead to rational improvements in existing treatment protocols.

scholarly journals TEL/AML-1 dimerizes and is associated with a favorable outcome in childhood acute lymphoblastic leukemia

Blood ◽  
1996 ◽  
Vol 88 (11) ◽  
pp. 4252-4258 ◽  
Author(s):  
TW McLean ◽  
S Ringold ◽  
D Neuberg ◽  
K Stegmaier ◽  
R Tantravahi ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Lymphoblastic Leukemia ◽  
Chromosomal Translocation ◽  
Fusion Transcript ◽  
Childhood Acute Lymphoblastic Leukemia ◽  
Childhood All ◽  
Polymerase Chain ◽  
B Lineage

Abstract Polymerase chain reaction-based screening of childhood acute lymphoblastic leukemia (ALL) samples showed that a TEL/AML1 fusion transcript was detected in 27% of all cases, representing the most common known gene rearrangement in childhood cancer. The TEL/AML1 fusion results from a t(12;21)(p13;q22) chromosomal translocation, but was undetectable at the routine cytogenetic level. TEL/AML1-positive patients had exclusively B-lineage ALL, and most patients were between the ages of 2 and 9 years at diagnosis. Only 3/89 (3.4%) adult ALL patients were TEL/AML1-positive. Most importantly, TEL/AML1-positive children had a significantly lower rate of relapse compared with TEL/AML1-negative patients (0/22 v 16/54, P = .004). Co- immunoprecipitation experiments demonstrated that TEL/AML-1 formed homodimers in vitro, and heterodimerized with the normal TEL protein when the two proteins were expressed together. The elucidation of the precise mechanism of transformation by TEL/AML1 and the role of TEL/AML1 testing in the treatment of childhood ALL will require additional studies.

Sensitivity of Childhood Acute Lymphoblastic Leukemia (ALL) to Idarubicin Is Significantly Higher Than to Daunorubicin Treatment Based on Ex Vivo Apoptosis Induction.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4495-4495
Author(s):  
Aram Prokop ◽  
Banu Bagci ◽  
Guenaelle Lingfeld ◽  
Lucia Badiali ◽  
Karin Garbrecht ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Response Rate ◽  
De Novo ◽  
Ex Vivo ◽  
Lymphoblastic Leukemia ◽  
Cell Populations ◽  
Apoptosis Induction ◽  
Good Tolerability ◽  
Childhood All

Abstract Anthracyclines, especially daunorubicin, play a very important role in the treatment of acute lymphoblastic leukemia (ALL) and the relapsed ALL in childhood. In the present study, primary lymphoblasts isolated from 65 children with de novo ALL (median: 5.8 years; range: 1.9 – 16.9 years) and relapsed ALL (median: 12.7 years; range: 1.3 – 17.9 years) were treated with daunorubicin (10 mmol/l) or idarubicin (2 mmol/l) in vitro. We could show that both anthracylines induce apoptosis, as evidenced by measurement of genomic DNA fragmentation. Interestingly, daunorubicin only induced modest apoptosis, whereas idarubicin displayed a significantly stronger apoptosis inducing effect. Furthermore the treatment of daunorubicin-resistant lymphoblasts with idarubicin resulted in good response in most of the resistant cell populations. Out of the 65 patients analysed in this study 23 were female (13 de novo ALL, 10 relapsed ALL) and 42 were male (29 de novo ALL, 13 relapsed ALL). Primary lymphoblasts were obtained by bone marrow aspiration and separated by centrifugation over Ficoll. Within these cell populations following immunologic subgroups were found: 35 c-ALL, 10 pre-B-ALL, 7 pro-B-ALL, 10 T-ALL and 3 pre-T-ALL. Daunorubicin induced apoptosis in 33 out of 65 lymphoblast populations (response rate 50.8 %). Nevertheless, a far higher response rate was observed for idarubicin with 59/65 (90,8 %) (p < 0.008), if response is defined as apoptosis induction higher than 1 %. Daunorubicin-resistance was found in 32/65 (49,2 %), resistance to both was observed in 6/65 (9,2 %). Treatment of daunorubicin-resistant lymphoblasts with idarubicin resulted in significant apoptosis induction in 26 out of 32 cell populations (81,3 %). We clearly demonstrated here that the in vitro treatment of lymphoblasts from children with de novo or relapsed ALL with idarubicin induces significantly higher response rates than daunorubicin treatment. The ex vivo sensitivity of daunorubicin-resistant lymphoblasts of childhood ALL to idarubicin treatment reflects the better potency of idarubicin to induce apoptosis and to overcome daunorubicin resistance. These data prompted us to study the clinical relevance of idarubicin in ongoing clinical trials to improve existing therapeutic regiments. First clinical data point to a good tolerability of idarubicin in the treatment of relapsed ALL in childhood.

In Vitro Drug Sensitivity as a Predicitive Tool of Early Clinical Response in Childhood Acute Lymphoblastic Leukemia.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4232-4232
Author(s):  
Faith C. Galderisi ◽  
Linda C. Stork ◽  
Ju Li ◽  
Motomi Mori ◽  
Solange Mongoue-Tchokote ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Clinical Response ◽  
Induction Therapy ◽  
Drug Sensitivity ◽  
Lymphoblastic Leukemia ◽  
Mixed Effects ◽  
Mixed Effects Model ◽  
Drug Sensitivity Assay ◽  
Pediatric All

Abstract Background: Residual disease or rapidity of response to induction therapy is among the most powerful predictors of outcome in pediatric Acute Lymphoblastic Leukemia (ALL). Various methods to determine response during induction have been in use in clinical investigation. We hypothesize that drug sensitivity at the cellular level predicts rapidity of response to induction therapy in ALL. We recently developed a high resolution flow cytometry based cytotoxicity assay for in vitro cellular drug response profiling for pediatric ALL. This method has a turnaround time of 48 hours and the ability to measure drug effect specific to leukemic cells regardless of number of admixed normal cells. We report preliminary data that correlate results of this drug sensitivity assay with rapidity of response to induction therapy among patients with ALL. Methods: We performed in vitro tests, applying a multiparameter flow cytometric drug cytotoxicity assay, on bone marrow (BM) samples of 23 patients with newly diagnosed standard (n = 10), high (n = 11), and very high (n = 2) risk ALL, as defined by the Children’s Oncology Group (COG) and NCI risk classification. Fourteen patients were rapid early responders (RER) and 9 were slow early responders (SER) by COG criteria at day 15 and 29. Cryopreserved cells from BM samples were thawed and determined to have adequate viability by trypan blue dye exclusion. Drugs were tested at three different concentrations, each in triplicate. Concentrations tested were based on an empirically derived cut-off concentration (EDCC) adopted from published in vitro studies, chosen to produce a large scatter of survival index values among samples. Leukemic blasts were specifically identified by surface markers, CD 45, CD 19 and CD 10 or CD 3, while cytotoxicity was measured with Annexin V based apoptosis. Leukemic cell survival index (LCSI = Average Replicate /Average Control x 100) was determined at 48 hours after in vitro exposure to individual standard induction agents for pediatric ALL: vincristine, asparaginase, dexamethasone, prednisone and daunomycin. LCSI differences between RER and SER were compared using Wilcoxon rank sum test for each drug and concentration. The mixed effects model was used to evaluate the overall difference of LCSI between RER and SER over the three concentrations (referred to as “averaged concentrations”). Results: For dexamethasone, a significantly lower LCSI was seen in the RER compared with the SER cohort at individual and averaged concentrations: RER mean LCSI = 40.2%, SER mean LCSI = 70.1% (p = 0.01, mixed effects model). A trend toward a lower mean LCSI in the RER compared with the SER group was noted for asparaginase and vincristine at individual and averaged concentrations (p < 0.1). Mean LCSI was not different between the RER and SER groups for daunomycin and prednisone at individual or averaged concentrations. Conclusions: This in vitro drug sensitivity assay provides a response profile for dexamethasone that correlates with in vivo response to induction therapy. Research is ongoing with more patient samples in order to achieve a greater statistical power to detect a smaller difference for all drugs tested. Further research will also correlate clinical response with LCSI using drug combinations in vitro. Results of these studies will determine the potential value of this assay for early risk stratification and modification of therapy in de novo or relapsed ALL.

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