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A novel human primary immunodeficiency syndrome caused by deficiency of the endosomal adaptor protein p14

Abstract

Lysosome-related organelles have versatile functions, including protein and lipid degradation, signal transduction and protein secretion. The molecular elucidation of rare congenital diseases affecting endosomal-lysosomal biogenesis has given insights into physiological functions of the innate and adaptive immune system. Here, we describe a previously unknown human primary immunodeficiency disorder and provide evidence that the endosomal adaptor protein p14, previously characterized as confining mitogen-activated protein kinase (MAPK) signaling to late endosomes, is crucial for the function of neutrophils, B cells, cytotoxic T cells and melanocytes. Combining genetic linkage studies and transcriptional profiling analysis, we identified a homozygous point mutation in the 3′ untranslated region (UTR) of p14 (also known as MAPBPIP), resulting in decreased protein expression. In p14-deficient cells, the distribution of late endosomes was severely perturbed, suggesting a previously unknown role for p14 in endosomal biogenesis. These findings have implications for understanding endosomal membrane dynamics, compartmentalization of cell signal cascades, and their role in immunity.

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Figure 1: Clinical presentation.
Figure 2: Phenotype of immune cells and melanocytes.
Figure 3: Haplotypes, mutational analysis and determination of RNA instability.
Figure 4: Reconstitution of lysosome-related organelles in neutrophils upon retroviral p14 gene transfer.
Figure 5: Defective signal transduction and late endosomal distribution in p14-deficient fibroblasts.
Figure 6: Aberrant subcellular distribution of late endosomes in p14-deficient fibroblasts and reconstitution upon retroviral gene transfer.

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Acknowledgements

We thank M. Schlesier (University of Freiburg) for the B-cell phenotyping, I. Sandrock, G. Köhne, F. Noyan, K. Boztug and M. Ballmaier for laboratory support, C. Roifman (Hospital for Sick Children) and R. Gatti (University of California at Los Angeles) for providing control samples on Mennonite families, and E. Ungewickell, C. Baum, J. Bohne, C. Kardinal and H. Holtmann for critical discussions. We thank B. Tümmler (Hannover Medical School), C. Baum (Hannover Medical School) and I. Touw (Erasmus University Medical Center) for reagents, M. Zimmermann for help in statistical evaluations and M. Offterdinger for help in determining subcellular distribution of endosomes. This research was supported by Deutsche Forschungsgemeinschaft grants KFO110 and GR1617/3, BMBF, the Elternverein Krebskranke Kinder Hannover, the Austrian Proteomics Platform (APP, GEN-AU), the Special Research Program “Cell Proliferation and Cell Death in Tumors” (SFB021, Austrian Science Fund) and in part by the intramural research program of the US National Institutes of Health, National Library of Medicine (NLM).

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Authors

Contributions

A.A., G. Brandes and J.T. contributed equally to this work. G. Bohn sequenced candidate genes and performed most molecular and cellular functional studies. A.A. characterized the 3′ UTR mutation by assessing RNA metabolism and functional reporter assays. G. Brandes performed electron microscopy and immunofluorescence studies. J.T. performed fine-mapping, screened candidate genes and helped to edit the manuscript. E.G. performed the genome-wide scan. A.A.S. carried out the genetic linkage analysis computations, chose the markers for genetic linkage fine mapping and wrote parts of the manuscript. C.R. performed E. coli lysis assays. N.T. and D.T. did immunofluorescence studies on endosomes. C.Z. cared for patients, and collected and curated data in SCN patient registry. R.A.D. assisted Bohn, A.A. and C.R. R.G. and J.B. performed microarray experiments. L.A.H. gave advice on endosome biology, and educated and supervised N.T. and D.T. K.W. provided laboratory resources, resources for SCN registry and significant help to initiate and carry out the study. B.G. initiated the project together with C.K., educated and supervised J.T. and E.G., assisted A.A.S. in linkage analysis, provided grant and laboratory resources and edited the manuscript. C.K. designed and directed the study, obtained clinical samples, taught and supervised Bohn, A.A., C.R. and R.A.D., provided laboratory and financial resources and wrote the manuscript.

Corresponding author

Correspondence to Christoph Klein.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Assessment of granzyme B and perforin in cytotoxic T-cells, and TEM of neutrophil granulocytes of patients #10, 12 and 13. (PDF 1673 kb)

Supplementary Fig. 2

Statistical analysis of E. coli lysis assay. Data presented in Fig. 4b were transformed by grouping the samples from HD and genetically corrected cells from patient #5 (black lines). (PDF 61 kb)

Supplementary Fig. 3

Analysis of G-CSF-receptor (G-CSFR) transgenic fibroblast cell lines. (PDF 88 kb)

Supplementary Figure 4

Statistical analysis of endosome distribution with respect to the distance from the nucleus. (PDF 2523 kb)

Supplementary Table 1

Immunophenotyping, in vitro T-cell proliferation, serum immunoglobulin levels and vaccination titers. (PDF 116 kb)

Supplementary Table 2

LOD scores and target genes. (PDF 98 kb)

Supplementary Table 3

Transcriptional profiling of EBV-transformed B-cell lines. Expression intensities of transcripts in EBV-transformed B-cell lines derived from parents (#1, 2) werecompared to B-cell lines derived from two patients (#12, 13). (PDF 131 kb)

Supplementary Methods (PDF 164 kb)

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Bohn, G., Allroth, A., Brandes, G. et al. A novel human primary immunodeficiency syndrome caused by deficiency of the endosomal adaptor protein p14. Nat Med 13, 38–45 (2007). https://doi.org/10.1038/nm1528

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