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P53 loss of heterozygosity (LOH) in formalin-fixed paraffin-embedded leiomyosarcoma (LMS): a novel report

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Abstract

Background

The occurrence of p53 loss of heterozygosity (LOH) is a common genetic event in malignancy. LOH occurs when a heterozygous locus loses one of its two parental alleles, becoming homozygous at that locus, by either copy number loss (CNL-LOH) or by becoming copy number neutral (CNN-LOH). A role for CNL-LOH (cnLOH) has been postulated in cancer aetiology. Loss of heterozygosity (LOH) results in irreversible genetic loss.

Aims

LOH was determined in DNA extracted from formalin-fixed paraffin-embedded (FFPE) leiomyosarcoma (LMS) specimens in a retrospective study from 30 patients, to assess the prognostic significance of LOH. The findings were analysed and their validity assessed. LOH was an adverse prognostic factor in LMS. Prospective uniform standardisation of formalin-fixation techniques is required.

Methods

DNA was extracted from 169 formalin-fixed paraffin blocks of 30 patients with LMS, following extensive tissue microdissection. Genomic DNA was amplified using the polymerase chain reaction (PCR) technique. Fluorescence-based microsatellite PCR was used to detect and quantitate heterozygosity loss.

Results

LOH was detected at gene locus 17p13 in 16 LMS (Four grade 2 and 12 grade 3 LMS). LOH was not detected in 14 LMS cases (one grade 1, five grade 2 and eight grade 3 LMS). LOH was associated with shorter patient survival.

Conclusions

The results reported herein endorse the value of utilizing FFPE DNA in identifying LOH as a prognostic factor in LMS. The results have implications for tumour biobanking and precision medicine in patients with sarcomas.

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Data Availability

Determination of growth fractions and use of refined PCR techniques to detect mutations and allelic loss of p53 in paraffin-embedded synovial sarcoma and leiomyosarcoma’ John N McMahon MD Thesis, University College Dublin (UCD) 2022: Chapter 5: PCR-LOH: pp206-209; Chapter 7: Variability of LOH findings in microdissected formalin-fixed paraffin embedded (FFPE) DNA: pp 241-256; pp 269-274. Cox proportional hazards model for Leiomyosarcoma: p365-366.

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Acknowledgements

Dr McMahon wishes to acknowledge the significant theoretical and technical contributions of Professor R McManus (TCD), and supervision by Professor, NA Parfrey (UCD) and Professor D Croke (RCSI). The guidance of Dr. P Kelehan (National Maternity Hospital Dublin), Professor PA Daly (St James’s Hospital Dublin), and Professor PA Dervan (UCD, deceased) is gratefully acknowledged.

Funding

This work was supported by grants from The Irish Cancer Society, The Health Research Board (Ireland) in 1992–1994, and by Trinity College Dublin, by the Histopathology Department in University College, Dublin, and by the Department of Biochemistry, Royal College of Surgeons in Ireland in 1993–1995.

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Authors and Affiliations

  1. Research Laboratory, St Vincent’s University Hospital, University College Dublin, PathologyDublin, Ireland

    John N. McMahon & John F. Stephens

  2. Department of Histopathology, St James’s Hospital and Trinity College Dublin, Dublin, Ireland

    Eoin F. Gaffney

  3. Audio-Visual Centre, University College Dublin, Dublin, Ireland

    William J. Aliaga-Kelly

  4. School of Medicine, University of Limerick, V94 T9PX, Limerick, Ireland

    Amirhossein Jalali

  5. Department of Biochemistry, Royal College of Surgeons in Ireland, Dublin, Ireland

    Bernadette Curran

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Contributions

JNMcM1 performed PCR experiments, analysed the data, wrote and presented the manuscript, EFG2 contributed to data analyses and paper presentation, WJA-K3 performed digital imaging and contributed to paper presentation, JFS1 performed polymerase chain reaction experiments, AJ4 performed the statistical analysis, BC5 performed polymerase chain reaction and heterozygosity analysis. All named authors made significant accountable contributions to the conception, design, accuracy, paper integrity and to the acquisition, analysis and interpretation of data used, resulting in the final version of this work for publication. The authors wish to state that there is no conflict of interest.

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Correspondence to John N. McMahon.

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Ethical statement

All 30 patients gave informed consent to provide clinical treatment and subsequent follow-up details to their clinical consultant and to have their data coordinated centrally in a dedicated database by a research assistant appointed by grants from The Irish Cancer Society. Ethical approval from Irish Research and Ethics Committees for research that posed only a negligible risk to patients, was not required. Selected specific additional patient data, including survival data, was obtained from medical records and then fully anonymised. There was no patient or public involvement in the design phase or logistics of this project or at any stage during the project itself. No additional data is available.

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McMahon, J.N., Gaffney, E.F., Aliaga-Kelly, W.J. et al. P53 loss of heterozygosity (LOH) in formalin-fixed paraffin-embedded leiomyosarcoma (LMS): a novel report. Ir J Med Sci 193, 65–71 (2024). https://doi.org/10.1007/s11845-023-03370-1

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