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Bone Marrow Failure and Immunodeficiency Associated with Human RAD50 Variants

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Abstract

Purpose

The MRE11-RAD50-NBN (MRN) complex plays a key role in recognizing and signaling DNA double-strand breaks. Pathogenic variants in NBN and MRE11 give rise to the autosomal-recessive diseases, Nijmegen breakage syndrome (NBS) and ataxia telangiectasia-like disorder, respectively. The clinical consequences of pathogenic variants in RAD50 are incompletely understood. We aimed to characterize a newly identified RAD50 deficiency/NBS-like disorder (NBSLD) patient with bone marrow failure and immunodeficiency.

Methods

We report on a girl with microcephaly, mental retardation, bird-like face, short stature, bone marrow failure and B-cell immunodeficiency. We searched for candidate gene by whole-exome sequencing and analyzed the cellular phenotype of patient-derived fibroblasts using immunoblotting, radiation sensitivity assays and lentiviral complementation experiments.

Results

Compound heterozygosity for two variants in the RAD50 gene (p.Arg83His and p.Glu485Ter) was identified in this patient. The expression of RAD50 protein and MRN complex formation was maintained in the cells derived from this patient. DNA damage-induced activation of the ATM kinase was markedly decreased, which was restored by the expression of wild-type (WT) RAD50. Radiosensitivity appeared inconspicuous in the patient-derived cell line as assessed by colony formation assay. The RAD50R83H missense substitution did not rescue the mitotic defect in complementation experiments using RAD50-deficient fibroblasts, whereas RAD50WT did. The RAD50E485X nonsense variant was associated with in-frame skipping of exon 10 (p.Glu485_545del).

Conclusion

These findings indicate important roles of RAD50 in human bone marrow and immune cells. RAD50 deficiency/NBSLD can manifest as a distinct inborn error of immunity characterized by bone marrow failure and B-cell immunodeficiency.

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Data Availability

The datasets for this article are not publicly available due to concerns regarding participant/patient anonymity. Requests to access the datasets should be directed to the corresponding author.

Abbreviations

AT:

Ataxia telangiectasia

ATLD:

Ataxia telangiectasia-like disorder

DSB:

DNA double-strand break

IEI:

Inborn error of immunity

MRN:

MRE11-RAD50-NBN

NBS:

Nijmegen breakage syndrome

NBSLD:

NBS-like disorder

WES:

Whole-exome sequencing

WT:

Wild-type

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Acknowledgements

We thank Mika Nagase and Girmay Asgedom for their technical assistance, Louisa Weinhold and Katja Bezjak for scientific assistance, Detlev Schindler for support with large T immortalized cell lines and Axel Schambach for support in setting up the lentiviral complementation system.

Funding

This study was supported by the Research on Measures for Intractable Disease Project to SK, the Claudia von Schilling Foundation for Breast Cancer Research to TD, and MEXT/JSPS KAKENHI (Grant Number: 22K07887) to HK.

Author information

Author notes

  1. Masatoshi Takagi, Akihiro Hoshino, and Kristine Bousset equally contributed to this work.

Authors and Affiliations

  1. Department of Pediatrics and Developmental Biology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo, Japan

    Masatoshi Takagi, Akihiro Hoshino, Dan Tomomasa & Tomohiro Morio

  2. Department of Community Pediatrics, Perinatal and Maternal Medicine, Tokyo Medical and Dental University (TMDU), Tokyo, Japan

    Masatoshi Takagi

  3. Department of Pediatrics, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan

    Akihiro Hoshino & Xi Yang

  4. Gynaecology Research Unit, Hannover Medical School, 30625, Hannover, Germany

    Kristine Bousset, Jule Röddecke, Hanna Luisa Martin, Iulia Folcut & Thilo Dörk

  5. Division of Immunology, Children’s Hospital of Chongqing Medical University, Chongqing, China

    Xi Yang

  6. Department of Genome Repair Dynamics, Radiation Biology Center, Kyoto University, Kyoto, Japan

    Junya Kobayashi

  7. Department of Pediatrics, Kindai University Faculty of Medicine, Osaka-Sayama, Japan

    Naoki Sakata

  8. Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan

    Kenichi Yoshida & Seishi Ogawa

  9. Laboratory of DNA Information Analysis, Human Genome Center, Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Satoru Miyano

  10. Laboratory of Sequence Analysis, Human Genome Center, Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Satoru Miyano

  11. Department of Pediatrics, Nagoya University Graduate School of Medicine, Nagoya, Japan

    Seiji Kojima

  12. Department of Child Health and Development, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), 1-5-45 Yushima, Bunkyo-Ku, Tokyo, 113-8519, Japan

    Hirokazu Kanegane

Authors
  1. Masatoshi Takagi
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  2. Akihiro Hoshino
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Contributions

MT designed and performed experiments and analyzed the data. AH, KB, JR, HLM, IF, DT, XY and JK performed experiments and analyzed the data. NS provided the patient data. KY, SM and SO performed genetic analysis. MT, AH, TD and HK wrote the manuscript. SK and TM provided critical discussion. TD and HK conceptualized the study. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Thilo Dörk or Hirokazu Kanegane.

Ethics declarations

Ethics Approval

This study was performed in line with the principles of the Helsinki Declaration. Approval was granted by the Tokyo Medical and Dental University Ethics Committee (protocol no. 103).

Consent to Participate

Informed consent was obtained from the patient and her parents.

Consent for Publication

Informed consent was obtained from the patient and her parents.

Conflict of Interest

The authors declare no competing interests.

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Thilo Dörk and Hirokazu Kanegane share senior authorship.

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Takagi, M., Hoshino, A., Bousset, K. et al. Bone Marrow Failure and Immunodeficiency Associated with Human RAD50 Variants. J Clin Immunol 43, 2136–2145 (2023). https://doi.org/10.1007/s10875-023-01591-8

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