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From a 2DE-Gel Spot to Protein Function: Lesson Learned From HS1 in Chronic Lymphocytic Leukemia
Published on: October 19, 2014
Proteomic profile of pre - B2 lymphoblasts from children with acute lymphoblastic leukemia (ALL) in relation with the
Odile Costa1, Pascale Schneider2, Laurent Coquet3
1Laboratoire MERCI, Faculté de Médecine et de Pharmacie de Rouen, 123 boulevard Gambetta, Rouen, Cedex 76183, France.
Insights
This study identified novel protein markers in childhood acute lymphoblastic leukemia (ALL) associated with the t(12;21) chromosomal alteration. These markers can help predict prognosis and identify aggressive subtypes of pediatric ALL.
Area of Science:
- Proteomics
- Pediatric Oncology
- Molecular Biology
Background:
- Traditional prognostic factors for pediatric acute lymphoblastic leukemia (ALL) include age, white blood cell count, and chromosomal alterations.
- Protein marker studies are increasingly used for classifying leukemia subtypes, similar to Acute Myeloblastic Leukemia.
- This study investigates the lymphoblast proteome in childhood ALL, focusing on the presence of the t(12;21) translocation, a known indicator of good prognosis.
Purpose of the Study:
- To compare the proteome of lymphoblasts in childhood ALL based on the presence or absence of the t(12;21) translocation.
- To identify novel protein markers associated with good prognosis and leukemic aggressiveness in pediatric ALL.
- To explore potential new subgroups within t(12;21) positive ALL patients.
Main Methods:
- Two-dimensional gel electrophoresis was used to analyze protein expression in pre-B2 lymphoblastic cells from pediatric ALL patients.
- Differentially expressed protein spots (p < 0.01) between patient groups were excised and analyzed by nano-liquid chromatography-mass spectrometry.
- Tandem mass spectrometry data was processed and compared against the Mascot protein database.
Main Results:
- Sixteen candidate proteins were identified from twelve differentially expressed spots (p ≤ 0.05) in relation to t(12;21).
- Proteins linked to growth arrest (calponine 2, phosphatidylinositol transfer protein beta) and apoptosis promotion (proteasome subunit beta type-2, casein kinase 2α, OTUB1, MSF-B, septin 9 i4) were associated with good prognosis.
- Proteins favoring cell cycle proliferation (methionine adenosyl transferase 2β, heterogeneous nuclear ribonucleo-proteins A2) were identified as potential markers of aggressiveness.
Conclusions:
- The proteomic map of leukemic cells revealed potential marker candidates for assessing leukemic aggressiveness.
- New patient subgroups within t(12;21) positive childhood ALL may be identifiable through proteomic profiling.
- These preliminary findings require confirmation in larger cohorts of pediatric ALL patients.
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.
