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Somatic ARAF mutations in pediatric Langerhans cell histiocytosis: clinicopathologic, genetic and functional profiling

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Abstract

ARAF mutations have been identified in a limited subset of patients with Langerhans cell histiocytosis (LCH), a rare disorder characterized by abnormal proliferation of Langerhans cells. LCH is primarily instigated by mutations in the mitogen-activated protein kinase (MAPK) signaling pathway, with BRAFV600E and MAP2K1 mutations constituting most cases. ARAF mutations in LCH highlight the heterogeneity of the disease and provide insights into its underlying molecular mechanisms. However, the occurrence of ARAF-positive LCH cases is extremely rare, with only two reported globally. Although they may be linked to a more aggressive form of LCH and a more severe clinical progression, the clinical significance and functional consequences of these mutations remain uncertain. We performed next-generation sequencing (NGS) to explore driver mutations in 148 pediatric LCH patients and recognized a series of mutations, including an identical novel somatic ARAF mutation, c.1046_1051delAGGCTT (p.Q349_F351delinsL), in four pediatric LCH patients. It was considered an ARAF hotspot mutation. All reported ARAF-positive patients worldwide exhibited characteristic pathological features of LCH, albeit with involvement across multiple systems. In vitro functional studies showed that this mutation could trigger the MAPKinase pathway and phosphorylate its downstream effectors MEK1/2 and ERK1/2 (relatively weaker than BRAFV600E). Over-activation of mutant A-Raf kinase could be inhibited by the BRAF inhibitor vemurafenib. LCH is uncommon, and ARAF mutation is even rarer. In our study, we have identified a novel hotspot somatic ARAF mutation, which has been verified through functional analysis to be an activating mutation. LCH patients with ARAF mutation typically have an unfavorable prognosis due to limited treatment experiences, although they do not exhibit a high relapse rate. To aid in the development of personalized treatment approaches and prognostic markers for LCH patients, it is recommended to conduct typical pathological and immunohistochemical examinations, as well as genetic tests utilizing a targeted gene panel or whole exome sequencing (WES), for LCH diagnosis, thereby promoting the use of inhibitor treatment strategies.

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Data availability statement

The datasets used to support this paper's results are available in the article and more data will be available upon request. Additional sequencing data files have been deposited in NCBI GenBank under accession number PRJNA875244.

Abbreviations

DCs:

Dendritic cells

ECD:

Erdheim–Chester disease

ERK:

Extracellular signal-regulated kinase

FFPE:

Formalin-fixed paraffin-embedded

IHC:

Histopathology and immunohistochemistry

LCH:

Langerhans cell histiocytosis

MAPKinase:

Mitogen-activated protein kinase

MEK:

Mitogen-activated protein kinase kinase

MS:

Multiple-system involvement

MS-OR− :

Multiple-system without risk-organ involvement

MS-OR+ :

Multiple-system with risk-organ involvement

NGS:

Next-generation sequencing

SS:

Single system involvement

WES:

Whole exome sequencing

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Acknowledgements

We are grateful to patients and families for their cooperation.

Funding

The present study was supported by the Project from Wujieping Medical Foundation of China (Grant No. 20.6750.2021-04-37).

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Author notes

  1. Statement of equal author contribution: R.L. and Y.G. contributed equally to this study, and should be considered joint first author.

Authors and Affiliations

  1. Department of Hematology, Capital Institute of Pediatrics, Chaoyang District, Beijing, 100020, China

    Rong Liu, Shunqiao Feng & Jing Cao

  2. Clinical Central Laboratory, Capital Institute of Pediatrics, Beijing, 100020, China

    Xiaodai Cui

  3. GrandOmics Inc, Haidian District, Beijing, 100081, China

    Yibing Guo, Lin Han, Yanling Sun & Zhenhua Cao

Author notes

  1. Rong Liu and Yibing Guo are Co-first authors.

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    Contributions

    RL designed research, collected and analyzed clinical data. YG designed research, analyzed data and wrote the manuscript. LH analyzed data and performed laboratory work. SF and JC contributed to the care and management of patients. SF also collected clinical data. YS is the liaison between patients and researchers. ZC and XC conceived and supervised research. All authors have approved the submitted manuscript.

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    Correspondence to Zhenhua Cao or Xiaodai Cui.

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    Conflict of interest

    The authors Y.G., L.H., Y.S. and Z.C. are employed by GrandOmics (Beijing, China). All authors declared no competing financial interests.

    Ethics approval and consent to participate

    This study was performed according to the Declaration of Helsinki, and the collection of tissues or blood samples from patients was obtained with patient-informed consent (from their guardians) under approval by the research ethics committee of the Children’s Hospital of Capital Institute of Pediatrics (Identifier: SHERLLM2020005). All patients provided written informed consent for publication of their cases.

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    Liu, R., Guo, Y., Han, L. et al. Somatic ARAF mutations in pediatric Langerhans cell histiocytosis: clinicopathologic, genetic and functional profiling. Clin Exp Med 23, 5269–5279 (2023). https://doi.org/10.1007/s10238-023-01134-w

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