Abstract
Peroxisome biogenesis disorders (PBDs) are a group of autosomal-recessive developmental and progressive metabolic diseases leading to the Zellweger spectrum (ZS) phenotype in most instances. Diagnosis of clinically suspected cases can be difficult because of extensive genetic heterogeneity and large spectrum of disease severity. Furthermore, a second group of peroxisomal diseases caused by deficiencies of single peroxisomal enzymes can show an indistinguishable clinical phenotype. The diagnosis of these peroxisomal disorders relies on the clinical presentation, the biochemical parameters in plasma and erythrocyte membranes, and genetic testing as the final step. Analysis of patients’ cells is frequently required during the diagnostic process, e.g., for complementation analysis to identify the affected gene before sequencing. In the cases with unclear clinical or biochemical presentation, patients’ cells are analyzed to prove PBD or to demonstrate biochemical abnormalities that might be elusive in plasma. Cell lines from skin fibroblast that are usually generated for diagnostic workup are not available in all instances, mainly because the required skin biopsy is invasive and sometimes denied by parents. An alternative cellular system has not been analyzed sufficiently. In this study, we evaluated the alternative use of lymphoblastoid cell lines (LCLs), derived from a peripheral blood sample, in the diagnostic process for PBD. LCLs were suitable for immunofluorescence visualization of peroxisomal enzymes, complementation analysis, and the biochemical analysis to differentiate between control and PBD LCL. LCLs are therefore an easily obtainable alternative cellular system for a detailed PBD diagnostic workup with a reliability of diagnostic results equal to those of skin fibroblasts.
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Acknowledgments
We thank M. Schniewind and J. Kaiser for expert technical assistance, S. Thoms for help with microscopy, and R. Steinfeld, M. Henneke, and C. Brendel for sharing control cell lines. We thank C. Krause and A. Ohlenbusch for mutation analyses, D. Kube for access to the Nucleofector apparatus, and L. Florin (Bonn) for establishing LCL. This work has been supported by the Deutsche Forschungsgemeinschaft (Ga354/7-1) and the Bundesministerium für Bildung und Forschung (BMBF) through the German Leukodystrophy Network.
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Synopsis
Lymphoblastoid cell lines are on par with skin fibroblast cell lines for the diagnosis of peroxisome biogenesis disorders requiring immunofluorescence staining of peroxisomal matrix proteins, measurements of peroxisomal metabolites, and complementation analysis.
Details of Contributions of Individual Authors
Planning of the study: S.G., R.K., J.G.
Conduct of the study: S.G., R.K., H.R.
Reporting of work: S.G., R.K., J.G.
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S. Grønborg
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All authors declare that the answers to all questions on the JIMD competing interest form are No, and therefore they have nothing to declare.
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This work has been supported by the Deutsche Forschungsgemeinschaft (Ga354/7-1) and the Bundesministerium für Bildung und Forschung (BMBF) through the German Leukodystrophy Network. The authors confirm independence from the sponsors; the content of this chapter has not been influenced by the sponsors.
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Grønborg, S., Krätzner, R., Rosewich, H., Gärtner, J. (2011). Lymphoblastoid Cell Lines for Diagnosis of Peroxisome Biogenesis Disorders. In: JIMD Reports - Case and Research Reports, 2011/1. JIMD Reports, vol 1. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8904_2011_12
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DOI: https://doi.org/10.1007/8904_2011_12
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