Construction and validation of a bone marrow tissue microarray
Institute of Pathology, University Hospital of Basel, Schönbeinstrasse 40, Basel CH‐4031, Switzerland; sdirnhofer@uhbs.ch
Institute of Pathology, University Hospital of Basel, Schönbeinstrasse 40, Basel CH‐4031, Switzerland; sdirnhofer@uhbs.ch
Abstract
Background
The use of tissue microarrays (TMAs) is now a generally accepted method for the investigation of solid tumours. However, little is known about the applicability of the TMA technique for analysis of patients with acute leukaemia. A bone marrow (BM)‐TMA analysis with 15 different immunohistochemical markers was performed. The TMA was validated by comparison with the corresponding full tissue sections.
Materials and methods
A BM‐TMA comprising 148 cases of acute leukaemia, including 115 acute myeloid leukaemia (AML) and 33 acute lymphoblastic leukaemia (ALL) cases, was constructed. Expression of CD3, CD10, CD15, CD20, CD34, CD61, CD68, CD79a, CD99, CD117, CD138, myeloperoxidase, haemoglobin A1, glycophorin and terminal deoxynucleotidyl transferase was immunohistochemically analysed. 50 cases of the TMA were directly compared with the corresponding full tissue section to validate the results.
Results
Morphologically and immunohistochemically, 6 (4%) of 148 cases and 765 (11%) cores of 6912 individual analyses were not evaluable. A direct comparison of TMA cases with conventional full sections showed a concordance of the results of 100%.
Conclusions
The small size of bone‐marrow biopsies and the presence of bony trabeculae do not preclude construction and analysis of acute leukaemia TMAs. Acute leukaemia cases on TMA displayed the characteristic phenotypic profiles expected in different AML and ALL subtypes. Therefore, the TMA technique is also a promising method for high‐throughput analysis of combined marker expression and clinicopathological correlations in patients with leukaemia.
The World Health Organization classification of acute leukaemias includes morphological, immunohistochemical, cytogenetic and molecular features.1 The spectrum of classical molecular diagnostic methods has recently been extended by the use of gene expression profiling (GEP). GEP of patients with acute myeloid leukaemia (AML) has shown differentially expressed genes in various leukaemia cases, thereby allowing delineation of new subtypes with particular biological features and clinical outcome.23 Similarly, in patients with acute lymphoblastic leukaemia (ALL), it has been possible to identify clinically relevant subtypes by the use of GEP.24 On the one hand, these data corroborate the existing acute leukaemia classification system; on the other hand, they prompt to search for new markers at the protein level that might be of diagnostic and/or prognostic relevance for the individual patient.
The tissue microarray (TMA) technology allows the simultaneous analysis of a large number of tumours to investigate the genetic changes and protein expression in a single experiment under highly standardised conditions.56 Advances in genomics and proteomics are increasing the need to evaluate large numbers of molecular targets with regard to their prognostic and predictive value in clinical oncology.7 Originally, TMAs were used in the research of solid tumours. For these tumours, especially carcinomas, this technique is now a recognised and established method. Recently, the TMA approach was also applied successfully to fluorescent in‐situ hybridisation and immunohistochemical studies of lymphomas.8910 The combination of GEP using cDNA arrays with TMA technique in lymphoma research is promising because new possible surrogate markers or combinations thereof can rapidly be evaluated for their use in daily routine diagnostics. This has been shown in various studies for the immunohistochemical evaluation of diffuse large B cell lymphomas confirming the molecular classification.11121314
Up to now, very little information is available about the applicability of the TMA technique for the analysis of bone‐marrow biopsies, because of the specific difficulties in constructing arrays generated from small tissue cylinders, including bone structures. Only two very recent studies describe the use of bone marrow (BM)‐TMA, albeit with small case numbers.1516 Therefore, the aim of the present study was to assess the feasibility of BM‐TMA construction. A TMA consisting of 148 acute leukaemia cases was constructed, and the quality of morphology and immunohistochemistry of 15 markers was investigated. Additionally, we validated the methodology by directly comparing a panel of immunohistochemical marker expression on the tissue cores of the TMA with the corresponding conventional bone‐marrow biopsies.
Acknowledgements
We thank H Weisskopf, T Schürch and J Schwegler for their excellent technical assistance and A Tzankov for editorial help.
Abbreviations
ALL - acute lymphoblastic leukaemia
AML - acute myeloid leukaemia
BM‐TMA - bone marrow tissue microarray
cHL - classical Hodgkin's lymphoma
FAB - French–American–British
GEP - gene expression profiling
H&E - haematoxylin and eosin
MPO - myeloperoxidase
TdT - terminal deoxynucleotidyl transferase
TMA - tissue microarray
Footnotes
Funding: This study was partly supported by a grant from AIRC (Milan).
Competing interests: None.
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