@article {908, title = {Multiplexed Functional Assessment of Genetic Variants in CARD11.}, journal = {Am J Hum Genet}, volume = {107}, year = {2020}, month = {2020 12 03}, pages = {1029-1043}, abstract = {

Genetic testing has increased the number of variants identified in disease genes, but the diagnostic utility is limited by lack of understanding variant function. CARD11 encodes an adaptor protein that expresses dominant-negative and gain-of-function variants associated with distinct immunodeficiencies. Here, we used a "cloning-free" saturation genome editing approach in a diploid cell line to simultaneously score 2,542 variants for decreased or increased function in the region of CARD11 associated with immunodeficiency. We also described an exon-skipping mechanism for CARD11 dominant-negative activity. The classification of reported clinical variants was sensitive (94.6\%) and specific (88.9\%), which rendered the data immediately useful for interpretation of seven coding and splicing variants implicated in immunodeficiency found in our clinic. This approach is generalizable for variant interpretation in many other clinically actionable genes, in any relevant cell type.

}, keywords = {Adenine, B-Cell CLL-Lymphoma 10 Protein, B-Lymphocytes, CARD Signaling Adaptor Proteins, Cell Line, Diploidy, Exons, Genes, Dominant, Genetic Variation, Guanylate Cyclase, Humans, Immunologic Deficiency Syndromes, Jurkat Cells, Lymphoma, NF-kappa B p50 Subunit, Piperidines, Polymorphism, Single Nucleotide, Primary Immunodeficiency Diseases, Sensitivity and Specificity}, issn = {1537-6605}, doi = {10.1016/j.ajhg.2020.10.015}, author = {Meitlis, Iana and Allenspach, Eric J and Bauman, Bradly M and Phan, Isabelle Q and Dabbah, Gina and Schmitt, Erica G and Camp, Nathan D and Torgerson, Troy R and Nickerson, Deborah A and Bamshad, Michael J and Hagin, David and Luthers, Christopher R and Stinson, Jeffrey R and Gray, Jessica and Lundgren, Ingrid and Church, Joseph A and Butte, Manish J and Jordan, Mike B and Aceves, Seema S and Schwartz, Daniella M and Milner, Joshua D and Schuval, Susan and Skoda-Smith, Suzanne and Cooper, Megan A and Starita, Lea M and Rawlings, David J and Snow, Andrew L and James, Richard G} } @article {607, title = {Hypomorphic caspase activation and recruitment domain 11 (CARD11) mutations associated with diverse immunologic phenotypes with or without atopic disease.}, journal = {J Allergy Clin Immunol}, volume = {143}, year = {2019}, month = {2019 Apr}, pages = {1482-1495}, abstract = {

BACKGROUND: Caspase activation and recruitment domain 11 (CARD11) encodes a scaffold protein in lymphocytes that links antigen receptor engagement with downstream signaling to nuclear factor κB, c-Jun N-terminal kinase, and mechanistic target of rapamycin complex 1. Germline CARD11 mutations cause several distinct primary immune disorders in human subjects, including severe combined immune deficiency (biallelic null mutations), B-cell expansion with nuclear factor κB and T-cell anergy (heterozygous, gain-of-function mutations), and severe atopic disease (loss-of-function, heterozygous, dominant interfering mutations), which has focused attention on CARD11 mutations discovered by using whole-exome sequencing.

OBJECTIVES: We sought to determine the molecular actions of an extended allelic series of CARD11 and to characterize the expanding range of clinical phenotypes associated with heterozygous CARD11 loss-of-function alleles.

METHODS: Cell transfections and primary T-cell assays were used to evaluate signaling and function of CARD11 variants.

RESULTS: Here we report on an expanded cohort of patients harboring novel heterozygous CARD11 mutations that extend beyond atopy to include other immunologic phenotypes not previously associated with CARD11 mutations. In addition to (and sometimes excluding) severe atopy, heterozygous missense and indel mutations in CARD11 presented with immunologic phenotypes similar to those observed in signal transducer and activator of transcription 3 loss of function, dedicator of cytokinesis 8 deficiency, common variable immunodeficiency, neutropenia, and immune dysregulation, polyendocrinopathy, enteropathy, X-linked-like syndrome. Pathogenic variants exhibited dominant negative activity and were largely confined to the CARD or coiled-coil domains of the CARD11 protein.

CONCLUSION: These results illuminate a broader phenotypic spectrum associated with CARD11 mutations in human subjects and underscore the need for functional studies to demonstrate that rare gene variants encountered in expected and unexpected phenotypes must nonetheless be validated for pathogenic activity.

}, issn = {1097-6825}, doi = {10.1016/j.jaci.2018.08.013}, author = {Dorjbal, Batsukh and Stinson, Jeffrey R and Ma, Chi A and Weinreich, Michael A and Miraghazadeh, Bahar and Hartberger, Julia M and Frey-Jakobs, Stefanie and Weidinger, Stephan and Moebus, Lena and Franke, Andre and Sch{\"a}ffer, Alejandro A and Bulashevska, Alla and Fuchs, Sebastian and Ehl, Stephan and Limaye, Sandhya and Arkwright, Peter D and Briggs, Tracy A and Langley, Claire and Bethune, Claire and Whyte, Andrew F and Alachkar, Hana and Nejentsev, Sergey and DiMaggio, Thomas and Nelson, Celeste G and Stone, Kelly D and Nason, Martha and Brittain, Erica H and Oler, Andrew J and Veltri, Daniel P and Leahy, T Ronan and Conlon, Niall and Poli, Maria C and Borzutzky, Arturo and Cohen, Jeffrey I and Davis, Joie and Lambert, Michele P and Romberg, Neil and Sullivan, Kathleen E and Paris, Kenneth and Freeman, Alexandra F and Lucas, Laura and Chandrakasan, Shanmuganathan and Savic, Sinisa and Hambleton, Sophie and Patel, Smita Y and Jordan, Michael B and Theos, Amy and Lebensburger, Jeffrey and Atkinson, T Prescott and Torgerson, Troy R and Chinn, Ivan K and Milner, Joshua D and Grimbacher, Bodo and Cook, Matthew C and Snow, Andrew L} }