Abstract
Despite neuroblastoma being the most common extracranial solid cancer in childhood, it is still a rare disease. Consequently, the unavailability of tissue for research limits the statistical power of studies. Pathology archives are possible sources of rare tissue, which, if proven to remain consistent over time, could prove useful to research of rare disease types. We applied immunohistochemistry to investigate whether long term storage caused any changes to antigens used diagnostically for neuroblastoma. We constructed and quantitatively assessed a tissue microarray containing neuroblastoma archival material dating between 1950 and 2007. A total of 119 neuroblastoma tissue cores were included spanning 6 decades. Fourteen antibodies were screened across the tissue microarray (TMA). These included seven positive neuroblastoma diagnosis markers (NB84, Chromogranin A, NSE, Ki-67, INI1, Neurofilament Protein, Synaptophysin), two anticipated to be negative (S100A, CD99), and five research antibodies (IL-7, IL-7R, JAK1, JAK3, STAT5). The staining of these antibodies was evaluated using Aperio ImageScope software along with novel pattern recognition and quantification algorithms. This analysis demonstrated that marker signal intensity did not decrease over time and that storage for 60 years had little effect on antigenicity. The construction and assessment of this neuroblastoma TMA has demonstrated the feasibility of using archival samples for research.
Highlights
Neuroblastoma is the most common extracranial solid tumour in childhood and the most common cancer found in children under the age of five [1,2]
We have demonstrated that despite long term storage of these neuroblastoma specimens, the tissues performed to an equivalent level, and open the potential for a large amount of archival material to be used for research purposes into rare disease
It was important to demonstrate that all tissue cores on the array represented the equivalent proportion of cellular and stromal structures, which is commonly typical for neuroblastoma tumours
Summary
Neuroblastoma is the most common extracranial solid tumour in childhood and the most common cancer found in children under the age of five [1,2]. When compared to adult cancers, neuroblastoma is a very rare disease with current figures in Australia indicating that approximately 35–40 new cases are diagnosed each year [11] With these relatively low numbers, obtaining sufficient samples to conduct research is challenging in countries like Australia with a relatively low population. Archived formalin-fixed paraffin-embedded (FFPE) tissues stored in hospital histopathology departments represent valuable collections of biospecimens that allow modern research questions to be investigated in rare tumours It is queried, whether biospecimens prepared and stored over many decades allow a range of tests for protein and DNA elements to be performed to a similar standard as those prepared using current protocols [12,13,14,15]. We have demonstrated that despite long term storage of these neuroblastoma specimens, the tissues performed to an equivalent level, and open the potential for a large amount of archival material to be used for research purposes into rare disease
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