TCF3 haploinsufficiency defined by immune, clinical, gene-dosage, and murine studies
Brigette Boast, Shubham Goel, Luis Ignacio González‐Granado, Julie E. Niemela, Jennifer Stoddard, Emily S.J. Edwards, Sandali Seneviratne, Dominik Spensberger, Juan Francisco Quesada‐Espinosa, Luis M. Allende, John McDonnell, Alexandria Haseley, Harry Lesmana, Magdalena Walkiewicz, Emad Muhammad, Julian J. Bosco, Thomas A. Fleisher, Shai Cohen, Steven M. Holland, Menno C. van Zelm, Anselm Enders, Hye Sun Kuehn, Sergio D. Rosenzweig
Abstract
BackgroundTCF3 is a transcription factor contributing to early lymphocyte differentiation. Germline monoallelic dominant negative and biallelic loss-of-function (LOF) null TCF3 mutations cause a fully penetrant severe immunodeficiency. We identified 8 individuals from 7 unrelated families with monoallelic LOF TCF3 variants presenting with immunodeficiency with incomplete clinical penetrance.ObjectiveWe sought to define TCF3 haploinsufficiency (HI) biology and its association with immunodeficiency.MethodsPatient clinical data and blood samples were analyzed. Flow cytometry, Western blot analysis, plasmablast differentiation, immunoglobulin secretion, and transcriptional activity studies were conducted on individuals carrying TCF3 variants. Mice with a heterozygous Tcf3 deletion were analyzed for lymphocyte development and phenotyping.ResultsIndividuals carrying monoallelic LOF TCF3 variants showed B-cell defects (eg, reduced total, class-switched memory, and/or plasmablasts) and reduced serum immunoglobulin levels; most but not all presented with recurrent but nonsevere infections. These TCF3 LOF variants were either not transcribed or translated, resulting in reduced wild-type TCF3 protein expression, strongly suggesting HI pathophysiology for the disease. Targeted RNA sequencing analysis of T-cell blasts from TCF3-null, dominant negative, or HI individuals clustered away from healthy donors, implying that 2 WT copies of TCF3 are needed to sustain a tightly regulated TCF3 gene–dosage effect. Murine TCF3 HI resulted in a reduction of circulating B cells but overall normal humoral immune responses.ConclusionMonoallelic LOF TCF3 mutations cause a gene-dosage–dependent reduction in wild-type protein expression, B-cell defects, and a dysregulated transcriptome, resulting in immunodeficiency. Tcf3+/- mice partially recapitulate the human phenotype, underscoring the differences between TCF3 in humans and mice. TCF3 is a transcription factor contributing to early lymphocyte differentiation. Germline monoallelic dominant negative and biallelic loss-of-function (LOF) null TCF3 mutations cause a fully penetrant severe immunodeficiency. We identified 8 individuals from 7 unrelated families with monoallelic LOF TCF3 variants presenting with immunodeficiency with incomplete clinical penetrance. We sought to define TCF3 haploinsufficiency (HI) biology and its association with immunodeficiency. Patient clinical data and blood samples were analyzed. Flow cytometry, Western blot analysis, plasmablast differentiation, immunoglobulin secretion, and transcriptional activity studies were conducted on individuals carrying TCF3 variants. Mice with a heterozygous Tcf3 deletion were analyzed for lymphocyte development and phenotyping. Individuals carrying monoallelic LOF TCF3 variants showed B-cell defects (eg, reduced total, class-switched memory, and/or plasmablasts) and reduced serum immunoglobulin levels; most but not all presented with recurrent but nonsevere infections. These TCF3 LOF variants were either not transcribed or translated, resulting in reduced wild-type TCF3 protein expression, strongly suggesting HI pathophysiology for the disease. Targeted RNA sequencing analysis of T-cell blasts from TCF3-null, dominant negative, or HI individuals clustered away from healthy donors, implying that 2 WT copies of TCF3 are needed to sustain a tightly regulated TCF3 gene–dosage effect. Murine TCF3 HI resulted in a reduction of circulating B cells but overall normal humoral immune responses. Monoallelic LOF TCF3 mutations cause a gene-dosage–dependent reduction in wild-type protein expression, B-cell defects, and a dysregulated transcriptome, resulting in immunodeficiency. Tcf3+/- mice partially recapitulate the human phenotype, underscoring the differences between TCF3 in humans and mice.