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Proteomic profile of pre - B2 lymphoblasts from children with acute lymphoblastic leukemia (ALL) in relation with the translocation (12; 21).

Costa O, Schneider P, Coquet L, Chan P, Penther D, Legrand E, Jouenne T, Vasse M, Vannier JP - Clin Proteomics (2014)

Bottom Line: The tandem mass spectrometry peak lists extracted using the DataAnalysis program, were compared with the protein database Mascot Daemon.We focused on twelve spots corresponding to sixteen identified candidate proteins among the 26 found differentially expressed (p ≤ 0.05) regarding the presence of t(12;21).By drawing up the protein map of leukemic cells, these new data identified marker candidates of leukemic aggressiveness and new t(12;21) patients subgroups.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire MERCI, Faculté de Médecine et de Pharmacie de Rouen, 123 boulevard Gambetta, Rouen, Cedex 76183, France.

ABSTRACT

Background: Until now, the major prognostic factors for pediatric acute lymphoblastic leukemia (ALL), age, white blood cell count and chromosomal alterations are initially taken into account for the risk stratification of patients. In the light of protein marker studies to classify subtypes of Acute Myeloblastic Leukemia efficiently, we have compared the lymphoblastes proteome in Childhood ALL in accordance with the presence of t(12;21), indicator of good prognosis, usually.

Methods: Protein expression in pre-B2 lymphoblastic cells, collected from residual bone marrow cells after diagnostic procedures, was analyzed using two dimensional gel electrophoresis protocol. Protein spots whose average normalized volumes were statistically different in the two patients groups (n = 13; student t test p < 0.01), were excised. Tryptic peptides were then analyzed using a nano-LC1200 system coupled to a 6340 Ion Trap mass spectrometer equipped with a HPLC-chip cube interface. The tandem mass spectrometry peak lists extracted using the DataAnalysis program, were compared with the protein database Mascot Daemon.

Results: We focused on twelve spots corresponding to sixteen identified candidate proteins among the 26 found differentially expressed (p ≤ 0.05) regarding the presence of t(12;21). Among over expressed proteins, two proteins were implicated in cellular growth arrest (i.e. calponine 2, p ≤ 0.001 and phosphatidylinositol transfer protein beta, p ≤ 0.001) in accordance with good prognosis, while two other proteins favored cell cycle proliferation (i.e. methionine adenosyl transferase 2β, p ≤ 0.005 and heterogeneous nuclear ribonucleo-proteins A2 p ≤ 0.01) and could therefore be good marker candidates of aggressiveness. Level of expression of proteasome subunit beta type-2 (p ≤ 0.01) and protein casein kinase 2α (p ≤ 0.01) which both favored apoptosis, deubiquitinating enzyme OTUB1 (p ≤ 0.05) and MLL septin-like fusion protein MSF-B, septin 9 i4 (p ≤ 0.01) were in accord with a good prognosis related to t(12;21) lymphoblasts.

Conclusion: By drawing up the protein map of leukemic cells, these new data identified marker candidates of leukemic aggressiveness and new t(12;21) patients subgroups. These preliminary results will be in the near future confirmed by using a larger sample of pre-B2 childhood ALLs from national lymphoblastic cell collections.

No MeSH data available.


Related in: MedlinePlus

Pie chart representing the principal protein’s functions with differential expression in lymphoblasts from pre-B2 ALL children, depending on the presence or absence of the translocation t(12; 21). Proteins are characterized by LC-MSMS and Mascot score from 2-DE spots with: A- Differential expression with p ≤ 0.05 (n = 28); B- Differential expression with p ≤ 0.01 (n = 15). ND = none defined.
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Figure 2: Pie chart representing the principal protein’s functions with differential expression in lymphoblasts from pre-B2 ALL children, depending on the presence or absence of the translocation t(12; 21). Proteins are characterized by LC-MSMS and Mascot score from 2-DE spots with: A- Differential expression with p ≤ 0.05 (n = 28); B- Differential expression with p ≤ 0.01 (n = 15). ND = none defined.

Mentions: A total of 4000 spots were detected on 2-DE maps (Figure 1). Protein maps were gathered with regard to the presence (n = 6) or the absence (n = 7) of t(12;21) (Table 1). Twenty six spots were found differently expressed in the two groups of patients (Table 2 for the top 12 spots; the whole spot informations can be seen in Online Additional file 1: Table S1) (student-t test, p ≤ 0.05). Repeatability assay was performed as indicated in Patients, material and methods, giving reliable results (see Online Additional file 1: Figure S1). Most of these proteins are involved in the cell cycle regulation (25%), metabolism (32%) and apoptosis (14%) (Figure 2A). The last ones (18%) are implicated in cytoskeleton organization, protein elongation or posttranslational events while 11% are not yet identified. However, among the 26 differentially expressed spots, only eleven spots (Figure 1A, see also Online Additional file 1: Figure S2 which located the last 15 spots), representing fifteen candidate proteins, were expressed with a sufficient reliable statistic power (student-t test, p ≤ 0.01, with a Power higher than 80%) in the two groups of patients (Figure 3). The average normalized volumes values and statistic ranks are listed Table 2. Five of them were expressed mostly in t(12;21) ALLs. The last seven spots were expressed mostly in the absence of this translocation. Corresponding proteins (Table 2, and Online Additional file 1: Table S1), identified by Mass Spectrometry with relevant Mascot score (more than 3 ions scores > 48) (see Online Additional file 1: Figure S3) were of great interest in terms of leukemogenesis since involved in mechanisms related to either the cell cycle regulation, or apoptosis, or energetic metabolism (i.e., 43%, 29% or 28% of them, respectively) (Figure 2B). Principal components analysis (PCA) (see Patients, materials and methods) indicated that the amount of all the 26 spots of interest, when gathered together, could discriminate 2 groups of patient, correlating with the presence or absence of t(12;21) (Figure 4A,B).


Proteomic profile of pre - B2 lymphoblasts from children with acute lymphoblastic leukemia (ALL) in relation with the translocation (12; 21).

Costa O, Schneider P, Coquet L, Chan P, Penther D, Legrand E, Jouenne T, Vasse M, Vannier JP - Clin Proteomics (2014)

Pie chart representing the principal protein’s functions with differential expression in lymphoblasts from pre-B2 ALL children, depending on the presence or absence of the translocation t(12; 21). Proteins are characterized by LC-MSMS and Mascot score from 2-DE spots with: A- Differential expression with p ≤ 0.05 (n = 28); B- Differential expression with p ≤ 0.01 (n = 15). ND = none defined.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4128613&req=5

Figure 2: Pie chart representing the principal protein’s functions with differential expression in lymphoblasts from pre-B2 ALL children, depending on the presence or absence of the translocation t(12; 21). Proteins are characterized by LC-MSMS and Mascot score from 2-DE spots with: A- Differential expression with p ≤ 0.05 (n = 28); B- Differential expression with p ≤ 0.01 (n = 15). ND = none defined.
Mentions: A total of 4000 spots were detected on 2-DE maps (Figure 1). Protein maps were gathered with regard to the presence (n = 6) or the absence (n = 7) of t(12;21) (Table 1). Twenty six spots were found differently expressed in the two groups of patients (Table 2 for the top 12 spots; the whole spot informations can be seen in Online Additional file 1: Table S1) (student-t test, p ≤ 0.05). Repeatability assay was performed as indicated in Patients, material and methods, giving reliable results (see Online Additional file 1: Figure S1). Most of these proteins are involved in the cell cycle regulation (25%), metabolism (32%) and apoptosis (14%) (Figure 2A). The last ones (18%) are implicated in cytoskeleton organization, protein elongation or posttranslational events while 11% are not yet identified. However, among the 26 differentially expressed spots, only eleven spots (Figure 1A, see also Online Additional file 1: Figure S2 which located the last 15 spots), representing fifteen candidate proteins, were expressed with a sufficient reliable statistic power (student-t test, p ≤ 0.01, with a Power higher than 80%) in the two groups of patients (Figure 3). The average normalized volumes values and statistic ranks are listed Table 2. Five of them were expressed mostly in t(12;21) ALLs. The last seven spots were expressed mostly in the absence of this translocation. Corresponding proteins (Table 2, and Online Additional file 1: Table S1), identified by Mass Spectrometry with relevant Mascot score (more than 3 ions scores > 48) (see Online Additional file 1: Figure S3) were of great interest in terms of leukemogenesis since involved in mechanisms related to either the cell cycle regulation, or apoptosis, or energetic metabolism (i.e., 43%, 29% or 28% of them, respectively) (Figure 2B). Principal components analysis (PCA) (see Patients, materials and methods) indicated that the amount of all the 26 spots of interest, when gathered together, could discriminate 2 groups of patient, correlating with the presence or absence of t(12;21) (Figure 4A,B).

Bottom Line: The tandem mass spectrometry peak lists extracted using the DataAnalysis program, were compared with the protein database Mascot Daemon.We focused on twelve spots corresponding to sixteen identified candidate proteins among the 26 found differentially expressed (p ≤ 0.05) regarding the presence of t(12;21).By drawing up the protein map of leukemic cells, these new data identified marker candidates of leukemic aggressiveness and new t(12;21) patients subgroups.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire MERCI, Faculté de Médecine et de Pharmacie de Rouen, 123 boulevard Gambetta, Rouen, Cedex 76183, France.

ABSTRACT

Background: Until now, the major prognostic factors for pediatric acute lymphoblastic leukemia (ALL), age, white blood cell count and chromosomal alterations are initially taken into account for the risk stratification of patients. In the light of protein marker studies to classify subtypes of Acute Myeloblastic Leukemia efficiently, we have compared the lymphoblastes proteome in Childhood ALL in accordance with the presence of t(12;21), indicator of good prognosis, usually.

Methods: Protein expression in pre-B2 lymphoblastic cells, collected from residual bone marrow cells after diagnostic procedures, was analyzed using two dimensional gel electrophoresis protocol. Protein spots whose average normalized volumes were statistically different in the two patients groups (n = 13; student t test p < 0.01), were excised. Tryptic peptides were then analyzed using a nano-LC1200 system coupled to a 6340 Ion Trap mass spectrometer equipped with a HPLC-chip cube interface. The tandem mass spectrometry peak lists extracted using the DataAnalysis program, were compared with the protein database Mascot Daemon.

Results: We focused on twelve spots corresponding to sixteen identified candidate proteins among the 26 found differentially expressed (p ≤ 0.05) regarding the presence of t(12;21). Among over expressed proteins, two proteins were implicated in cellular growth arrest (i.e. calponine 2, p ≤ 0.001 and phosphatidylinositol transfer protein beta, p ≤ 0.001) in accordance with good prognosis, while two other proteins favored cell cycle proliferation (i.e. methionine adenosyl transferase 2β, p ≤ 0.005 and heterogeneous nuclear ribonucleo-proteins A2 p ≤ 0.01) and could therefore be good marker candidates of aggressiveness. Level of expression of proteasome subunit beta type-2 (p ≤ 0.01) and protein casein kinase 2α (p ≤ 0.01) which both favored apoptosis, deubiquitinating enzyme OTUB1 (p ≤ 0.05) and MLL septin-like fusion protein MSF-B, septin 9 i4 (p ≤ 0.01) were in accord with a good prognosis related to t(12;21) lymphoblasts.

Conclusion: By drawing up the protein map of leukemic cells, these new data identified marker candidates of leukemic aggressiveness and new t(12;21) patients subgroups. These preliminary results will be in the near future confirmed by using a larger sample of pre-B2 childhood ALLs from national lymphoblastic cell collections.

No MeSH data available.


Related in: MedlinePlus