Modeling altered T-cell development with induced pluripotent stem cells from patients with RAG1-dependent immune deficiencies
| Autor: | Silvia Giliani, Anne Marie Comeau, Itai M. Pessach, Juan Carlos Zúñiga-Pflücker, David G. Schatz, Carmen Dominguez-Brauer, Luigi D. Notarangelo, Yuhang Zhang, Barry P. Sleckman, Kerstin Felgentreff, Gordon Keller, Marion Kennedy, Frederic D. Bushman, Geneve Awong, Waleed Al-Herz, Francesca A. Ververs, Likun Du, Yu Nee Lee, Andrea L. Bredemeyer, Jared H. Rowe, Patrick M. Brauer, Lisa Ott de Bruin, Erik L. Clarke |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
T cell Genes RAG-1 Receptors Antigen T-Cell alpha-beta T-Lymphocytes Immunology Induced Pluripotent Stem Cells Biology CD38 Biochemistry Recombination-activating gene 03 medical and health sciences 0302 clinical medicine medicine Humans Induced pluripotent stem cell Cells Cultured Homeodomain Proteins Severe combined immunodeficiency V(D)J recombination DNA Breaks Infant hemic and immune systems Cell Biology Hematology medicine.disease Molecular biology Omenn syndrome V(D)J Recombination 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis Mutation Severe Combined Immunodeficiency CD8 |
| Zdroj: | Blood. 128(6) |
| ISSN: | 1528-0020 |
| Popis: | Primary immunodeficiency diseases comprise a group of heterogeneous genetic defects that affect immune system development and/or function. Here we use in vitro differentiation of human induced pluripotent stem cells (iPSCs) generated from patients with different recombination-activating gene 1 (RAG1) mutations to assess T-cell development and T-cell receptor (TCR) V(D)J recombination. RAG1-mutants from severe combined immunodeficient (SCID) patient cells showed a failure to sustain progression beyond the CD3(--)CD4(-)CD8(-)CD7(+)CD5(+)CD38(-)CD31(-/lo)CD45RA(+) stage of T-cell development to reach the CD3(-/+)CD4(+)CD8(+)CD7(+)CD5(+)CD38(+)CD31(+)CD45RA(-) stage. Despite residual mutant RAG1 recombination activity from an Omenn syndrome (OS) patient, similar impaired T-cell differentiation was observed, due to increased single-strand DNA breaks that likely occur due to heterodimers consisting of both an N-terminal truncated and a catalytically dead RAG1. Furthermore, deep-sequencing analysis of TCR-β (TRB) and TCR-α (TRA) rearrangements of CD3(-)CD4(+)CD8(-) immature single-positive and CD3(+)CD4(+)CD8(+) double-positive cells showed severe restriction of repertoire diversity with preferential usage of few Variable, Diversity, and Joining genes, and skewed length distribution of the TRB and TRA complementary determining region 3 sequences from SCID and OS iPSC-derived cells, whereas control iPSCs yielded T-cell progenitors with a broadly diversified repertoire. Finally, no TRA/δ excision circles (TRECs), a marker of TRA/δ locus rearrangements, were detected in SCID and OS-derived T-lineage cells, consistent with a pre-TCR block in T-cell development. This study compares human T-cell development of SCID vs OS patients, and elucidates important differences that help to explain the wide range of immunologic phenotypes that result from different mutations within the same gene of various patients. |
| Databáze: | OpenAIRE |
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