Manish Butte

Front Pediatr. 2022 Jul 27;10:883173. doi: 10.3389/fped.2022.883173. eCollection 2022.

ABSTRACT

BACKGROUND: During the process of generating diverse T and B cell receptor (TCR and BCR, respectively) repertoires, double-strand DNA breaks are produced. Subsequently, these breaks are corrected by a complex system led by the non-homologous end-joining (NHEJ). Pathogenic variants in genes involved in this process, such as the NHEJ1 gene, cause severe combined immunodeficiency syndrome (SCID) along with neurodevelopmental disease and sensitivity to ionizing radiation.

OBJECTIVE: To provide new clinical and immunological insights on NHEJ1 deficiency arising from a newly diagnosed patient with severe immunodeficiency.

MATERIALS AND METHODS: A male infant, born to consanguineous parents, suspected of having primary immunodeficiency underwent immunological and genetic workup. This included a thorough assessment of T cell phenotyping and lymphocyte activation by mitogen stimulation tests, whole-exome sequencing (WES), TCR repertoire Vβ repertoire via flow cytometry analysis, and TCR and BCR repertoire analysis via next-generation sequencing (NGS).

RESULTS: Clinical findings included microcephaly, recurrent pneumonia, and failure to thrive. An immune workup revealed lymphopenia, reduced T cell function, and hypogammaglobulinemia. Skewed TCR Vβ repertoire, TCR gamma (TRG) repertoire, and BCR repertoire were determined in the patient. Genetic analysis identified a novel homozygous missense pathogenic variant in XLF/Cernunnos: c.A580Ins.T; p.M194fs. The patient underwent a successful hematopoietic stem cell transplantation (HSCT).

CONCLUSION: A novel NHEJ1 pathogenic variant is reported in a patient who presented with SCID phenotype that displayed clonally expanded T and B cells. An adjusted HSCT was safe to ensure full T cell immune reconstitution.

PMID:35967585 | PMC:PMC9363661 | DOI:10.3389/fped.2022.883173

Front Immunol. 2022 Jul 29;13:919237. doi: 10.3389/fimmu.2022.919237. eCollection 2022.

ABSTRACT

Actin-related protein 2/3 complex subunit 1B (ARPC1B) deficiency is a recently described inborn error of immunity (IEI) presenting with combined immunodeficiency and characterized by recurrent infections and thrombocytopenia. Manifestations of immune dysregulation, including colitis, vasculitis, and severe dermatitis, associated with eosinophilia, hyper-IgA, and hyper-IgE are also described in ARPC1B-deficient patients. To date, hematopoietic stem cell transplantation seems to be the only curative option for patients. ARPC1B is part of the actin-related protein 2/3 complex (Arp2/3) and cooperates with the Wiskott-Aldrich syndrome protein (WASp) in the regulation of the actin cytoskeleton remodeling and in driving double-strand break clustering for homology-directed repair. In this study, we aimed to investigate radiosensitivity (RS) in ARPC1B-deficient patients to assess whether it can be considered an additional disease trait. First, we performed trio-based next-generation-sequencing studies to obtain the ARPC1B molecular diagnosis in our index case characterized by increased RS, and then we confirmed, using three different methods, an increment of radiosensitivity in all enrolled ARPC1B-deficient patients. In particular, higher levels of chromatid-type aberrations and γH2AX foci, with an increased number of cells arrested in the G2/M-phase of the cell cycle, were found in patients’ cells after ionizing radiation exposition and radiomimetic bleomycin treatment. Overall, our data suggest increased radiosensitivity as an additional trait in ARPC1B deficiency and support the necessity to investigate this feature in ARPC1B patients as well as in other IEI with cytoskeleton defects to address specific clinical follow-up and optimize therapeutic interventions.

PMID:35967303 | PMC:PMC9372879 | DOI:10.3389/fimmu.2022.919237

Cytokine. 2022 Aug 11;158:155975. doi: 10.1016/j.cyto.2022.155975. Online ahead of print.

ABSTRACT

OBJECTIVE: Our study aimed to reveal the roles of long noncoding RNA (lncRNA) and immune-relevant genes in systemic lupus erythematosus (SLE).

METHODS: Expression profiling dataset GSE65391 consisted of 72 healthy blood samples and 924 SLE blood samples was downloaded from the Gene Expression Omnibus. Differentially expressed RNAs (DERs) in SLE samples were identified using the Limma package. Immune-relevant DERs were uncovered after assessing the immune cell infiltration types of different samples. Modules significantly related to the disease were extracted via weighted gene co-expression network analysis (WGCNA), followed by module-trait analysis. LncRNA-mRNA co-expression network was constructed for DERs in immune-relevant modules. Genes related to SLE were further revealed via the Comparative Toxicogenomics Database (CTD). Validation assay was conducted by another independent expression profiling dataset, GSE46907 (consisted of 5 healthy blood samples and 5 SLE blood samples) and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis of 15 healthy blood samples and 15 SLE blood samples.

RESULTS: A total of 606 DERs, including 19 lncRNAs and 587 mRNAs, were identified. Obvious differences were observed in the proportions of 20 immune cell types, and 378 immune-relevant DERs were revealed. Genes in the lncRNA-mRNA co-expression network were significantly enriched in primary immunodeficiency, hematopoietic cell lineage, B cell receptor signaling pathway, SLE (up-regulated FCGR3A, down-regulated CD40LG, up-regulated HIST1H2BE, down-regulated human leukocyte antigen [HLA]-DOA, down-regulated HLA-DOB, up-regulated HIST1H3D, and up-regulated HIST1H2BC), and intestinal immune network for IgA production. SLE-related gene, CD40LG, retrieved from CTD has co-expression interactions with seven differentially expressed lncRNAs (HCG27, LINC02555, LINC02210, DHRS4-AS1, MIR600HG, DANCR, and LINC01278). CD40LG and HLA-DOA, were observed down-regulated, and FCGR3A, and HIST1H2BE were up-regulated in validation dataset GES46907. Moreover, CD40LG was validated to be down-regulated using qRT-PCR.

CONCLUSIONS: Our results provide new insight in understanding the pathogenesis of SLE and might be helpful for developing new therapeutic approaches of SLE by modulating immune response.

PMID:35964416 | DOI:10.1016/j.cyto.2022.155975

J Neurol. 2022 Aug 12. doi: 10.1007/s00415-022-11325-2. Online ahead of print.

ABSTRACT

BACKGROUND: There is a gap in the literature regarding genetic underpinnings of pediatric autoimmune CNS diseases. This study explored rare gene variants implicated in immune dysregulation within these disorders.

METHODS: This was a single-center observational study of children with inflammatory CNS disorder who had genetic testing through next generation focused exome sequencing targeting 155 genes associated with innate or adaptive immunity. For in silico prediction of functional effects of single-nucleotide variants, Polymorphism Phenotyping v2, and Sorting Intolerant from Tolerant were used, and Combined Annotation Dependent Depletion (CADD) scores were calculated. Identified genes were analyzed using Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis.

RESULTS: Of 54 patients, 42 (77.8%) carried variant(s), among which 12 (22.2%) had 3-8 variants. Eighty-eight unique single-nucleotide variants of 55 genes were identified. The most variants were detected in UNC13D, LRBA, LYST, NOD2, DOCK8, RNASEH2A, STAT5B, and AIRE. The majority of variants (62, 70.4%) had CADD > 10. KEGG pathway analysis revealed seven genes associated with primary immunodeficiency (Benjamini 1.40E – 06), six genes with NOD-like receptor signaling (Benjamini 4.10E – 04), five genes with Inflammatory Bowel Disease (Benjamini 9.80E – 03), and five genes with NF-kappa B signaling pathway (Benjamini 1.90E – 02).

DISCUSSION: We observed a high rate of identification of rare and low-frequency variants in immune regulatory genes in pediatric neuroinflammatory CNS disorders. We identified 88 unique single-nucleotide variants of 55 genes with pathway analysis revealing an enrichment of NOD2-receptor signaling, consistent with involvement of the pathway within other autoinflammatory conditions and warranting further investigation.

PMID:35960392 | DOI:10.1007/s00415-022-11325-2

Respir Res. 2022 Aug 9;23(1):202. doi: 10.1186/s12931-022-02126-2.

ABSTRACT

BACKGROUND: The efficacy and safety of complement inhibition in COVID-19 patients is unclear.

METHODS: A multicenter randomized controlled, open-label trial. Hospitalized COVID-19 patients with signs of systemic inflammation and hypoxemia (PaO₂/FiO₂ below 350 mmHg) were randomized (2:1 ratio) to receive standard of care with or without the C5 inhibitor zilucoplan daily for 14 days, under antibiotic prophylaxis. The primary outcome was improvement in oxygenation at day 6 and 15.

RESULTS: 81 patients were randomly assigned to zilucoplan (n = 55) or the control group (n = 26). 78 patients were included in the safety and primary analysis. Most were men (87%) and the median age was 63 years. The mean improvement in PaO₂/FiO₂ from baseline to day 6 was 56.4 mmHg in the zilucoplan group and 20.6 mmHg in the control group (mean difference + 35.8; 95% confidence interval (CI) – 9.4 to 80.9; p = 0.12), an effect also observed at day 15. Day 28 mortality was 9% in the zilucoplan and 21% in the control group (odds ratio 0.4; 95% CI 0.1 to 1.5). At long-term follow up, the distance walked in a 6-min test was 539.7 m in zilucoplan and 490.6 m in the control group (p = 0.18). Zilucoplan lowered serum C5b-9 (p < 0.001) and interleukin-8 (p = 0.03) concentration compared with control. No relevant safety differences between the zilucoplan and control group were identified.

CONCLUSION: Administration of zilucoplan to COVID-19 patients in this proof-of-concept randomized trial was well tolerated under antibiotic prophylaxis. While not reaching statistical significance, indicators of respiratory function (PaO₂/FiO₂) and clinical outcome (mortality and 6-min walk test) suggest that C5 inhibition might be beneficial, although this requires further research in larger randomized studies.

PMID:35945604 | DOI:10.1186/s12931-022-02126-2

J Clin Immunol. 2022 Aug 10. doi: 10.1007/s10875-022-01338-x. Online ahead of print.

ABSTRACT

X-linked chronic granulomatous disease is a rare disease caused by mutations in the CYBB gene. While more extensive knowledge is available on genetics, pathogenesis, and possible therapeutic options, mitochondrial activity and its implications on patient monitoring are still not well-characterized. We have developed a novel protocol to study mitochondrial activity on whole blood of XCGD patients before and after transplantation, as well as on XCGD carriers. Here we present results of these analyses and of the restoration of mitochondrial activity in hyperinflamed X-linked Chronic Granulomatous Disease after hematopoietic stem cell transplantation. Moreover, we show a strong direct correlation between mitochondrial activity, chimerism, and DHR monitored before and after transplantation and in XCGD carriers. In conclusion, based on these findings, we suggest testing this new ready-to-use marker to better characterize patients before and after treatment and to investigate disease expression in carriers.

PMID:35945378 | DOI:10.1007/s10875-022-01338-x

Clin Gastroenterol Hepatol. 2022 Aug 6:S1542-3565(22)00730-3. doi: 10.1016/j.cgh.2022.07.030. Online ahead of print.

NO ABSTRACT

PMID:35944833 | DOI:10.1016/j.cgh.2022.07.030

Vox Sang. 2022 Aug 9. doi: 10.1111/vox.13337. Online ahead of print.

ABSTRACT

BACKGROUND AND OBJECTIVES: To evaluate the efficacy, safety and pharmacokinetics of a new, highly purified 10% IVIg (BT595, Yimmugo®) administered in children and adults with Primary immunodeficiency diseases (PID).

MATERIALS AND METHODS: Prospective, uncontrolled, multicentre Phase III trial. Patients aged 2 to <76 years with PID were switched from their pre-trial IVIg replacement therapy to BT595. In all, 67 patients (49 adults, 18 children) received doses between 0.2 and 0.8 g/kg body weight for approximately 12 months at intervals of 3 or 4 weeks. Dosing and dosing intervals were based on each patient’s pre-trial infusion schedule. The primary end point was the rate of acute serious bacterial infections (SBIs); secondary efficacy, safety and pharmacokinetic outcomes were also evaluated.

RESULTS: The primary efficacy end point was met, and the unadjusted SBI rate was 0.01 per subject-year (adjusted SBI rate 0.015 per subject-year, with an upper limit of the one-sided 99% confidence interval of 0.151). A single adult patient experienced one event classified as an SBI. All secondary end points, including those related to infections, supported the efficacy. Infusion rates were increased up to 8 ml/kg/h. Overall, 8% of infusions were associated with ≥1 infusional adverse event (AE) (start during or within 72 h post-infusion), comprising mainly headache (2.4%), fatigue (0.9%) and nausea (0.5%). There were no infusional AEs at infusion rates of >4.0 ml/kg/h, and only one patient required a single premedication. The observed patterns, severity and frequency of treatment-emergent adverse events are consistent with the established safety profile for IVIgs and did not show clinically relevant differences between all age groups.

CONCLUSION: BT595 is effective, safe and well tolerated for treating patients with PID.

PMID:35944615 | DOI:10.1111/vox.13337

Genes Immun. 2022 Aug 8. doi: 10.1038/s41435-022-00178-4. Online ahead of print.

ABSTRACT

RALY is a multifunctional RNA-binding protein involved in cancer metastasis, prognosis, and chemotherapy resistance in various cancers. However, the molecular mechanism of which is still unclear. We have established RALY overexpression cell lines and studied the effect of RALY on proliferation and apoptosis in HeLa cells. Then we used RNA-seq to analyze the transcriptomes data. Lastly, RT-qPCR experiments had performed to confirm the RNA-seq results. We found that the overexpression of RALY in HeLa cells inhibited proliferation. Moreover, the overexpression of RALY changed the gene expression profile, and the significant upregulation of genes involved immune/inflammatory response related biological process by NOD-like receptor signaling pathway cytokine-cytokine receptor interaction. The significant downregulation genes involved innate immune response by the Primary immunodeficiency pathway. Notably, IFIT1, IFIT2, IFTI3, IFI44, HERC4, and OASL expression had inhibited by the overexpression of RALY. Furthermore, RALY negatively regulates the expression of transcription factors FOS and FOSB. Notably, we found that 645 alternative splicing events had regulated by overexpression of RALY, which is highly enriched in transcription regulation, RNA splicing, and cell proliferation biological process by the metabolic pathway. We show that RALY regulates the expression of immune/inflammatory response-related genes via alternative splicing of FOS in HeLa cells. The novel role of RALY in regulating immune/inflammatory gene expression may explain its function in regulating chemotherapy resistance and provides novel insights into further exploring the molecular mechanism of RALY in regulating cancer immunity and chemo/immune therapies.

PMID:35941292 | DOI:10.1038/s41435-022-00178-4

Front Immunol. 2022 Jul 22;13:894648. doi: 10.3389/fimmu.2022.894648. eCollection 2022.

ABSTRACT

Primary immunodeficiency may present with treatment-refractory enteropathy. We present two patients with celiac/celiac-like disease diagnosed in early childhood and refractory to the gluten-free diet. One patient had features of multi-system autoimmunity, whereas the other had celiac-like disease as an isolated clinical finding. Both patients underwent genetic testing given disease refractoriness and were ultimately diagnosed with cytotoxic T lymphocyte antigen 4 (CTLA4) haploinsufficiency. They are both now in complete clinical and endoscopic remission on abatacept. CTLA4 haploinsufficiency has incomplete penetrance and significant phenotypic heterogeneity but should be considered in the differential diagnosis of refractory celiac/celiac-like disease, as treatment implications are significant.

PMID:35935971 | PMC:PMC9352891 | DOI:10.3389/fimmu.2022.894648