DIAPH1-Deficiency is Associated with Major T, NK and ILC Defects in Humans

Loss of function mutations in Diaphanous related formin 1 (DIAPH1) are associated with seizures, cortical blindness, and microcephaly syndrome (SCBMS) and are recently linked to combined immunodeficiency. However, the extent of defects in T and innate lymphoid cells (ILCs) remain unexplored. Herein, we characterized the primary T, natural killer (NK) and helper ILCs of six patients carrying two novel loss of function mutation in DIAPH1 and Jurkat cells after DIAPH1 knockdown. Mutations were identified by whole exome sequencing. T-cell immunophenotyping, proliferation, migration, cytokine signaling, survival, and NK cell cytotoxicity were studied via flow cytometry-based assays, confocal microscopy, and real-time qPCR. CD4<sup>+</sup> T cell proteome was analyzed by mass spectrometry. p.R351* and p.R322*variants led to a significant reduction in the DIAPH1 mRNA and protein levels. DIAPH1-deficient T cells showed proliferation, activation, as well as TCR-mediated signaling defects. DIAPH1-deficient PBMCs also displayed impaired transwell migration, defective STAT5 phosphorylation in response to IL-2, IL-7 and IL-15. In vitro generation/expansion of Treg cells from naïve T cells was significantly reduced. shRNA-mediated silencing of DIAPH1 in Jurkat cells reduced DIAPH1 protein level and inhibited T cell proliferation and IL-2/STAT5 axis. Additionally, NK cells from patients had diminished cytotoxic activity, function and IL-2/STAT5 axis. Lastly, DIAPH1-deficient patients' peripheral blood contained dramatically reduced numbers of all helper ILC subsets. DIAPH1 deficiency results in major functional defects in T, NK cells and helper ILCs underlining the critical role of formin DIAPH1 in the biology of those cell subsets.

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Publication Details

Journal
Journal of Clinical Immunology
Published
2024-08-09
DOI
https://doi.org/10.1007/s10875-024-01777-8
Citations
8
Primary Topic
Immunodeficiency and Autoimmune Disorders
Type
article
Field-Weighted Citation Impact
1.92

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article

DIAPH1-Deficiency is Associated with Major T, NK and ILC Defects in Humans

Kemal Erdem Başaran, Servet Özcan, Aydan İkincioğulları, Safa Barış et al.
8 citations
Journal of Clinical Immunology
Immunodeficiency and Autoimmune Disorders
1.92
article

DIAPH1-Deficiency is Associated with Major T, NK and ILC Defects in Humans

Kemal Erdem Başaran, Servet Özcan, Aydan İkincioğulları, Safa Barış, Can Berk Leblebici, Zehra Büşra Azizoğlu, Fatma Zehra Okuş, Ahmet Eken, Elif Karakoç-Aydıner, Royala Babayeva, Özlem Canöz, Figen Doğu, Hasret Erkmen, Mehmed Fatih Canatan, Ekrem Ünal, Alper Özcan, Zehra Şule Haskoloğlu, Serdar Ceylaner, Mohammad Bilal Alsavaf, Salim Can, Hüseyin Per, Hakan Gümüş, Gülsüm Gümüş, Mehmet Köse, Musa Karakükçü, Mustafa Burak Acar, Halit Canatan, Şerife Ayaz‐Güner, Ahmet Özen, Ebru Yılmaz
article en
8 citations

Abstract

Loss of function mutations in Diaphanous related formin 1 (DIAPH1) are associated with seizures, cortical blindness, and microcephaly syndrome (SCBMS) and are recently linked to combined immunodeficiency. However, the extent of defects in T and innate lymphoid cells (ILCs) remain unexplored. Herein, we characterized the primary T, natural killer (NK) and helper ILCs of six patients carrying two novel loss of function mutation in DIAPH1 and Jurkat cells after DIAPH1 knockdown. Mutations were identified by whole exome sequencing. T-cell immunophenotyping, proliferation, migration, cytokine signaling, survival, and NK cell cytotoxicity were studied via flow cytometry-based assays, confocal microscopy, and real-time qPCR. CD4<sup>+</sup> T cell proteome was analyzed by mass spectrometry. p.R351* and p.R322*variants led to a significant reduction in the DIAPH1 mRNA and protein levels. DIAPH1-deficient T cells showed proliferation, activation, as well as TCR-mediated signaling defects. DIAPH1-deficient PBMCs also displayed impaired transwell migration, defective STAT5 phosphorylation in response to IL-2, IL-7 and IL-15. In vitro generation/expansion of Treg cells from naïve T cells was significantly reduced. shRNA-mediated silencing of DIAPH1 in Jurkat cells reduced DIAPH1 protein level and inhibited T cell proliferation and IL-2/STAT5 axis. Additionally, NK cells from patients had diminished cytotoxic activity, function and IL-2/STAT5 axis. Lastly, DIAPH1-deficient patients' peripheral blood contained dramatically reduced numbers of all helper ILC subsets. DIAPH1 deficiency results in major functional defects in T, NK cells and helper ILCs underlining the critical role of formin DIAPH1 in the biology of those cell subsets.

Journal of Clinical ImmunologyVol. 44(8)
Izmir Institute of Technology (TR), Ankara University (TR), Cell Technology (China) (HK), Medical Park Gaziantep Hospital (TR), Kayseri Eğitim ve Araştırma Hastanesi (TR), Hasan Kalyoncu University (TR), Intergen (Turkey) (TR), Marmara University (TR), Erciyes University (TR)
Erciyes Üniversitesi, İzmir Yüksek Teknoloji Enstitüsü, Türkiye Bilimsel ve Teknolojik Araştırma Kurumu, Marmara Üniversitesi
Openalex Percentile: Top 14%
Immunodeficiency and Autoimmune Disorders
1.92
DOIOpen Access PDFOpenAlex
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