Clinical, Immunological, and Molecular Variability of RAG Deficiency: A Retrospective Analysis of 22 RAG Patients

Abstract Purpose We described clinical, immunological, and molecular characterization within a cohort of 22 RAG patients focused on the possible correlation between clinical and genetic data. Methods Immunological and genetic features were investigated by multiparametric flow cytometry and by Sanger or next generation sequencing (NGS) as appropriate. Results Patients represented a broad spectrum of RAG deficiencies: SCID, OS, LS/AS, and CID. Three novel mutations in RAG1 gene and one in RAG2 were reported. The primary symptom at presentation was infections (81.8%). Infections and autoimmunity occurred together in the majority of cases (63.6%). Fifteen out of 22 (68.2%) patients presented autoimmune or inflammatory manifestations. Five patients experienced severe autoimmune cytopenia refractory to different lines of therapy. Total lymphocytes count was reduced or almost lacking in SCID group and higher in OS patients. B lymphocytes were variably detected in LS/AS and CID groups. Eighteen patients underwent HSCT permitting definitive control of autoimmune/hyperinflammatory manifestations in twelve of them (80%). Conclusion We reinforce the notion that different clinical phenotype can be found in patients with identical mutations even within the same family. Infections may influence genotype–phenotype correlation and function as trigger for immune dysregulation or autoimmune manifestations. Severe and early autoimmune refractory cytopenia is frequent and could be the first symptom of onset. Prompt recognition of RAG deficiency in patients with early onset of autoimmune/hyperinflammatory manifestations could contribute to the choice of a timely and specific treatment preventing the onset of other complications.

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Clinical, Immunological and Molecular Variability of RAG Deficiency: a Retrospective Analysis of 22 RAG Patients

10.21203/rs.3.rs-584134/v1 ◽

2021 ◽

Author(s):

Cristina Cifaldi ◽

Beatrice Rivalta ◽

Donato Amodio ◽

Algeri Mattia ◽

Lucia Pacillo ◽

...

Keyword(s):

Novel Mutation ◽

Specific Treatment ◽

Cd4 Cells ◽

Multiparametric Flow Cytometry ◽

Genetic Features ◽

Rag Deficiency ◽

Next Generation Sequencing Ngs ◽

Autoimmune Cytopenia ◽

Generation Sequencing

Abstract Purpose RAG deficiency is associated with a variety of clinical phenotypes. We described clinical, immunological and molecular characterization within a cohort of 22 RAG patients focused on the possible correlation between clinical and genetic data. Methods Immunological and genetic features were investigated by Multiparametric Flow Cytometry and by Sanger or Next generation sequencing (NGS) respectively. Results Patients represented a broad spectrum of RAG deficiencies: SCID n=8, OS n=6, LS/AS n=4 and CID n=4. Four novel mutation in RAG1 gene and one in RAG2 were reported. The primary symptom at presentation were infections (81.8%). Infections and autoimmunity occurred together in the majority of cases (63.6%). Fifteen out of 22 (68.2%) patients presented autoimmune/hyperinflammatory manifestations. Four patients experienced severe autoimmune cytopenia refractory to different lines of therapy. Total lymphocytes count was reduced or almost lacking in SCID group. CD4 cells count was higher in OS patients. B lymphocytes were variably detected in AS and CID groups. Eighteen patients underwent HSCT permitting definitive control of autoimmune/hyperinflammatory manifestations in twelve of them (80%). Conclusion RAG deficiency still represents a challenge in the tracing of effective management and follow-up, notably considering the inability to predict the disease course in atypical cases. Immune dysregulation manifestations are common features often refractory to conventional medical management. Severe and early autoimmune refractory cytopenia is frequent and could be the first symptom of onset. Prompt recognition of RAG deficiency in patients with early onset of autoimmune/hyperinflammatory manifestations could contribute to the choice of a timely and specific treatment preventing the onset of other complications.

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New mutations in KCNT2 gene causing early infantile epileptic encephalopathy type 57: Case study and literature review

Acta Biochimica Polonica ◽

10.18388/abp.2020_5364 ◽

2020 ◽

Author(s):

Meryem Alagoz ◽

Nasim Kherad ◽

Sureyya Bozkurt ◽

Adnan Yuksel

Keyword(s):

Novel Mutation ◽

Bioinformatic Analysis ◽

Epileptic Encephalopathy ◽

Phenotype Correlation ◽

Whole Exome ◽

And Function ◽

Next Generation Sequencing Ngs ◽

Generation Sequencing ◽

New Mutations

Purpose. Early infantile epileptic encephalopathy (EIEE) 57 belongs to a group of encephalopathies with early-onset and characterised by severe electroencephalogram abnormalities, seizures, developmental delay and intellectual disability. Method. We carried out Whole Exome analysis using Next Generation Sequencing (NGS) and bioinformatic analysis performed to find mutation associated with the patient phenotypes. The effect of the mutation on protein structure analysed by PolyPhen2 and Swissmodel ExPASy. Results. In this study, we evaluated two unrelated Turkish males diagnosed with EIEE type 57 to investigate the genetic cause of this disease. Whole exome sequencing revealed mutations in KCN2 gene, which is a member of Potassium channels (KCN) gene family associated with epileptic encephalopathies. Two mutations, c.545A>T (p.Asn182Ile and c.2638C>A (p.Leu880Met) were reported here as a novel mutation. Conclusions. Our findings implicate the genotype-phenotype correlation of these mutations. Furthermore, the computational analysis showed their effect on protein binding site and function suggesting their role in the development of early infantile epileptic encephalopathy 57.

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Upregulation of miR-941 in Circulating CD14+ Monocytes Enhances Osteoclast Activation via WNT16 Inhibition in Patients with Psoriatic Arthritis

International Journal of Molecular Sciences ◽

10.3390/ijms21124301 ◽

2020 ◽

Vol 21 (12) ◽

pp. 4301

Author(s):

Shang-Hung Lin ◽

Ji-Chen Ho ◽

Sung-Chou Li ◽

Yu-Wen Cheng ◽

Yi-Chien Yang ◽

...

Keyword(s):

Psoriatic Arthritis ◽

Osteoclast Differentiation ◽

Joint Disease ◽

Healthy Controls ◽

Treatment Target ◽

Potential Biomarker ◽

Osteoclast Activation ◽

And Function ◽

Next Generation Sequencing Ngs ◽

Generation Sequencing

Psoriatic arthritis (PsA) is a destructive joint disease mediated by osteoclasts. MicroRNAs (miRNAs) regulate several important pathways in osteoclastogenesis. We profiled the expression of miRNAs in CD14+ monocytes from PsA patients and investigated how candidate microRNAs regulate the pathophysiology in osteoclastogenesis. The RNA from circulatory CD14+ monocytes was isolated from PsA patients, psoriasis patients without arthritis (PsO), and healthy controls (HCs). The miRNAs were initially profiled by next-generation sequencing (NGS). The candidate miRNAs revealed by NGS were validated by PCR in 40 PsA patients, 40 PsO patients, and 40 HCs. The osteoclast differentiation and its functional resorption activity were measured with or without RNA interference against the candidate miRNA. The microRNA-941 was selectively upregulated in CD14+ monocytes from PsA patients. Osteoclast development and resorption ability were increased in CD14+ monocytes from PsA patients. Inhibition of miR-941 abrogated the osteoclast development and function while increased the expression of WNT16. After successful treatment, the increased miR-941 expression in CD14+ monocytes from PsA patients was revoked. The expression of miR-941 in CD14+ monocytes is associated with PsA disease activity. MiR-941 enhances osteoclastogenesis in PsA via WNT16 repression. The miR-941 could be a potential biomarker and treatment target for PsA.

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Ribosomal profiling adds new coding sequences to the proteome

Biochemical Society Transactions ◽

10.1042/bst20150170 ◽

2015 ◽

Vol 43 (6) ◽

pp. 1271-1276 ◽

Cited By ~ 21

Author(s):

Muhammad Ali S. Mumtaz ◽

Juan Pablo Couso

Keyword(s):

Single Nucleotide ◽

Protein Coding ◽

Large Numbers ◽

New Findings ◽

Genomic Scale ◽

Bioinformatic Approaches ◽

And Function ◽

Next Generation Sequencing Ngs ◽

Small Orfs ◽

Generation Sequencing

Next generation sequencing (NGS) has enabled an in-depth look into genes, transcripts and their translation at the genomic scale. The application of NGS sequencing of ribosome footprints (Ribo-Seq) reveals translation with single nucleotide (nt) resolution, through the deep sequencing of ribosome-bound fragments (RBFs). Some results of Ribo-Seq challenge our understanding of the protein-coding potential of the genome. Earlier bioinformatic approaches had shown the presence of hundreds of thousands of putative small ORFs (smORFs) in eukaryotic genomes, but they had been largely ignored due to their large numbers and difficulty in determining their translation and function. Ribo-Seq has revealed that hundreds of putative smORFs within previously assumed long non-coding RNAs (lncRNAs) and UTRs of canonical mRNAs are associated with ribosomes, appearing to be translated. Here we review some of the approaches used to define translation within Ribo-Seq experiments and the challenges in defining translation of these novel smORFs in lncRNAs and UTRs. We also look at some of the bioinformatic and biochemical approaches used to independently corroborate these exciting new findings and elucidate real translation events.

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