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CARD9 Expression Pattern, Gene Dosage, and Immunodeficiency Phenotype Revisited

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Abstract

Background

CARD9 deficiency is an autosomal recessive primary immunodeficiency underlying increased susceptibility to fungal infection primarily presenting as invasive CNS Candida and/or cutaneous/invasive dermatophyte infections. More recently, a rare heterozygous dominant negative CARD9 variant c.1434 + 1G > C was reported to be protective from inflammatory bowel disease.

Objective

We studied two siblings carrying homozygous CARD9 variants (c.1434 + 1G > C) and born to heterozygous asymptomatic parents. One sibling was asymptomatic and the other presented with candida esophagitis, upper respiratory infections, hypogammaglobulinemia, and low class-switched memory B cells.

Methods and Results

The CARD9 c.1434 + 1G > C variant generated two mutant transcripts confirmed by mRNA and protein expression: an out-of-frame c.1358–1434 deletion/ ~ 55 kDa protein (CARD9Δex.11) and an in-frame c.1417–1434 deletion/ ~ 61 kDa protein (CARD9Δ18 nt.). Neither transcript was able to form a complete/functional CBM complex, which includes TRIM62. Based on the index patient’s CVID-like phenotype, CARD9 expression was tested and detected in lymphocytes and monocytes from humans and mice. The functional impact of different CARD9 mutations and gene dosage conditions was evaluated in heterozygous and homozygous c.1434 + 1 G > C members of the index family, and in WT (two WT alleles), haploinsufficiency (one WT, one null allele), and null (two null alleles) individuals. CARD9 gene dosage impacted lymphocyte and monocyte functions including cytokine generation, MAPK activation, T-helper commitment, transcription, plasmablast differentiation, and immunoglobulin production in a differential manner.

Conclusions

CARD9 exon 11 integrity is critical to CBM complex function. CARD9 is expressed and affects particular T and B cell functions in a gene dosage-dependent manner, which in turn may contribute to the phenotype of CARD9 deficiency.

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Abbreviations

BAL:

Bronchoalveolar lavage

BCL10:

B cell lymphoma 10

CARD9:

Caspase recruitment domain-containing protein 9

CBM:

CARD9/BCL10/MALT1

CM:

Central memory

CNS:

Central nervous system

CVID:

Common variable immunodeficiency

GWAS:

Genome-wide association studies

HKC:

Heat-killed Candida

MALT1:

Mucosal-associated lymphoid tissue lymphoma translocation protein 1

IBD:

Inflammatory bowel disease

NCs:

Normal controls

nt:

Nucleotides

PBMCs:

Peripheral blood mononuclear cells

PHA:

Phytohemagglutinin

RNASeq:

RNA sequencing assay

TRIM62:

Tripartite motif containing-62

WBC:

White blood cells

WES:

Whole exome sequencing

WT:

Wild type

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Acknowledgements

We thank the patients and their families for their contributions to the study. These studies were supported by the Intramural Research Program, NIH Clinical Center, US National Institutes of Health (NIH); the French National Research Agency (ANR) under the “Investments for the Future” program (ANR-10-IAHU-01), the ANR-FNS LTh-MSMD-CMCD (ANR-18-CE93-0008-01), The Rockefeller University, and the National Institutes of Health (# R01AI127564).The content of this article does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. government.

Funding

These studies were supported by the Intramural Research Program, NIH Clinical Center, US National Institutes of Health (NIH); the French National Research Agency (ANR) under the “Investments for the Future” program (ANR-10-IAHU-01), the ANR-FNS LTh-MSMD-CMCD (ANR-18-CE93-0008–01), The Rockefeller University, and the National Institutes of Health (# R01AI127564).

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Author notes

  1. Shubham Goel and Hye Sun Kuehn contributed equally.

Authors and Affiliations

  1. Immunology Service, Department of Laboratory Medicine, Clinical Center, NIH, MD, Bethesda, USA

    Shubham Goel, Hye Sun Kuehn, Julie Niemela, Jennifer Stoddard, Mary Garofalo, Sophia Samir, Thomas Fleisher & Sergio D. Rosenzweig

  2. Immunology, Allergy and Rheumatology Division, Department of Pediatrics, Baylor College of Medicine, Texas Children’s Hospital, Houston, TX, USA

    Javier Chinen

  3. Laboratory of Clinical Immunology and Microbiology, Fungal Pathogenesis Section, National Institute of Allergy and Infectious Diseases (NIAID), NIH, Bethesda, MD, USA

    Daisuke Yamanaka, Vasileios Oikonomou & Michail S. Lionakis

  4. Laboratory of Human Genetics of Infectious Diseases, Necker Branch, UMR 1163, INSERM, Necker Hospital for Sick Children, 75015, Paris, France

    Melanie Migaud & Anne Puel

  5. University of Paris, Imagine Institute, 75015, Paris, France

    Melanie Migaud & Anne Puel

  6. St. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, NY, USA

    Anne Puel

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  1. Shubham Goel
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Contributions

SG performed experiments and wrote the first draft of the manuscript. HSK designed, performed, and supervised the experiments. JC diagnosed the index patient, collected biologic and clinical data. MG enrolled the patients and organized sample shipments. JN, JS, DY, MM, SS, and VO performed experiments. TAF, AP, MSL, HSK, and SDR revised and analyzed the data, and collaborated with the manuscript writing. SDR supervised the project.

Corresponding author

Correspondence to Sergio D. Rosenzweig.

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Goel, S., Kuehn, H.S., Chinen, J. et al. CARD9 Expression Pattern, Gene Dosage, and Immunodeficiency Phenotype Revisited. J Clin Immunol 42, 336–349 (2022). https://doi.org/10.1007/s10875-021-01173-6

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