Phenomic analysis of chronic granulomatous disease reveals more severe integumentary infections in X-Linked compared with autosomal recessive chronic granulomatous disease

BACKGROUND : Chronic granulomatous disease (CGD) is an inborn error of immunity (IEI), characterised by recurrent bacterial and fungal infections. It is inherited either in an Xlinked (XL) or autosomal recessive (AR) mode. Phenome refers to the entire set of phenotypes expressed, and its study allows us to generate new knowledge of the disease. The objective of the study is to reveal the phenomic differences between XL and AR-CGD by using Human Phenotype Ontology (HPO) terms. METHODS : We collected data on 117 patients with genetically diagnosed CGD from Asia and Africa referred to the Asian Primary Immunodeficiency Network (APID network). Only 90 patients with sufficient clinical information were included for phenomic analysis. We used HPO terms to describe all phenotypes manifested in the patients. RESULTS : XL-CGD patients had a lower age of onset, referral, clinical diagnosis, and genetic diagnosis compared with AR-CGD patients. The integument and central nervous system were more frequently affected in XL-CGD patients. Regarding HPO terms, perianal abscess, cutaneous abscess, and elevated hepatic transaminase were correlated with XL-CGD. A higher percentage of XL-CGD patients presented with BCGitis/BCGosis as their first manifestation. Among our CGD patients, lung was the most frequently infected organ, with gastrointestinal system and skin ranking second and third, respectively. Aspergillus species, Mycobacterium bovis, and Mycobacteirum tuberculosis were the most frequent pathogens to be found. CONCLUSION : Phenomic analysis confirmed that XL-CGD patients have more recurrent and aggressive infections compared with AR-CGD patients. Various phenotypic differences listed out can be used as clinical handles to distinguish XL or AR-CGD based on clinical features.The Society for Relief of Disabled Children and Jeffrey Modell Foundation.https://www.frontiersin.org/journals/immunologydm2022Paediatrics and Child Healt

Effects of simvastatin on PP2A activation in porcine coronary artery.

<p>(A) Effects of simvastatin (10 μM) on the protein expression of p-PP2A/total PP2A in porcine coronary artery. *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0). (B) Effects of AICAR (1 mM) on the protein expression of p-PP2A/total PP2A in porcine coronary artery. *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0). (C) Effect of okadaic acid (O.A., 10 nM) on simvastatin- and AICAR-induced protein expression of p-PP2A/total PP2A in porcine coronary artery. *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0). (D) Effect of compound C (10 μM) on simvastatin- and AICAR protein expression of p-PP2A/total PP2A in porcine coronary artery. *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0).</p

Proposed mechanisms for acute simvastatin-induced closure of K_ATP channels of vascular myocytes.

<p>Simvastatin (lipophilic) crosses the plasma membrane and reaches the sacroplasmic reticulum (SR) of vascular myocytes. Binding of simvastatin to SR leads to the release of ryanodine (Ryr)-sensitive Ca²⁺ into the cytosol. Elevation of Ca²⁺ activates CaMK II which leads to the subsequent activation (phosphorylation) of AMPKα. Phosphorylation of AMPKα-Thr¹⁷² causes [glucose]_o uptake with the participation of SGLT1 and Na⁺/K⁺ ATPase. Increase in cytosolic [glucose] leads to an elevation of ATP levels via oxidative phosphorylation. Elevation of [ATP]_i serves two purposes: (1) closure of vascular K_ATP channels, (2) providing phosphate groups for cellular proteins (e.g. PP2A and AMPK) phosphorylation. Phosphorylation of PP2A occurs downstream of AMPK phosphorylation. PP2A phosphorylation results in PP2A inactivation which “releases” AMPK and thus phosphorylation of AMPKα-Thr¹⁷² resulted. AICAR produces similar effects as simvastatin except the initial step involves LKB1-Ser⁴²⁸ phosphorylation.</p

Effects of simvastatin and AICAR on [Glucose]_o uptake and the role(s) of [glucose]_o and [Na⁺]_o in mediating simvastatin effects on AMPK and PP2A activities.

<p>(A) Effects of simvastatin (10 μM) and AICAR (1 mM) on [³H]-2-deoxy-glucose uptake, with and without compound C (10 μM), of porcine coronary artery myocytes (n = 6 for each treatment). *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls. Summary of the effect of simvastatin and AICAR on the protein expression of p-PP2A/total PP2A in (B) [glucose]_o-free, (C) [Na⁺]_o-free, (D) with phloridzin (1 mM) and (E) with ouabain (10 μM) in porcine coronary artery. *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0).</p

Role(s) of [Ca²⁺]_o and [Ca²⁺]_i in mediating the effects of simvastatin on AMPK and PP2A activities.

<p>(A) Effects of simvastatin (10 μM), with and without ryanodine (100 μM) pre-treatment, on [Ca²⁺]_i changes (F₁/F₀) of porcine coronary artery myocytes, estimated using Fluo-4/AM with confocal laser scanning microscope. (B) Summary of [Ca²⁺]_i changes in response to simvastatin (10 μM) before and after ryanodine (100 μM) challenges. Results are expressed (Area Under Curve, AUC) as mean ± SEM of 13–15 cells (***<i>P</i><0.001). (C) Summary of the effects of ryanodine (100 μM) on simvastatin (10 μM)- or AICAR (1 mM)-induced protein expression of p-AMPK/total AMPK in porcine coronary artery. *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0). (D) Summary of the effects of caffeine (1 mM) on the protein expression of p-AMPK/total AMPK in porcine coronary artery, with and without ryanodine (100 μM). *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0). (E) Effect of ryanodine (100 μM) on simvastatin (10 μM)-induced protein expression of p-PP2A/total PP2A in porcine coronary artery. *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0). (F) Summary of the effect of simvastatin (10 μM), AICAR (1 mM) and caffeine (1 mM) on the protein expression of p-PP2A/total PP2A, with and without KN93 (10 μM) in porcine coronary artery. *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0).</p

Participation of cytochrome P450 3A4.

<p>(A) Biochemical existence of cytochrome 450 (CYP450 3A4) in porcine liver, porcine coronary artery (endothelium denuded) and human left internal mammary artery (endothelium denuded). Beta actin was used as loading control. (B) Effects of simvastatin on the protein expression of p-AMPK/total AMPK, with ketoconazole (Keto, 10 μM, n = 4), in porcine coronary artery (endothelium denuded). *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0). (C) Effects of simvastatin on the protein expression of p-PP2A/total PP2A, with ketoconazole (Keto, 10 μM, n = 4), in porcine coronary artery (endothelium denuded). *<i>P</i><0.05 and **<i>P</i><0.01 compared to controls (i.e. time 0).</p

Effects of simvastatin on K_ATP channel openers-induced vasorelaxation.

<p>(A) Effect of simvastatin (1, 3 and 10 μM) (n = 6 to 8) on cromakalim-induced relaxation of U46619 (10 nM) pre-contracted porcine coronary artery (endothelium-denuded). (B) Effect of simvastatin (1, 3 and 10 μM) (n = 6 to 8) on pinacidil-induced relaxation of U46619 (10 nM) pre-contracted porcine coronary artery (endothelium-denuded). (C) Effect of okadaic acid (10 nM) (n = 6 to 8) on simvastatin-inhibited cromakalim-induced relaxation of U46619 (10 nM) pre-contracted porcine coronary artery (endothelium-denuded). (D) Effect of okadaic acid (10 nM) (n = 6 to 8) on simvastatin-inhibited pinacidil-induced relaxation of U46619 (10 nM) pre-contracted porcine coronary artery (endothelium-denuded). *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001 compared to controls.</p

Biochemical existence of HMG-CoA reductase, and the effects of simvastatin and simvastatin Na⁺ on the protein expression of HMG-CoA reductase and p-HMG-CoA reductase.

<p>(A) Biochemical existence of HMG-CoA reductase in porcine liver, porcine coronary artery (endothelium-denuded) and human left internal mammary artery (endothelium-denuded). Beta actin was used as loading control. (B) Effects of simvastatin (SIM) (10 μM) and simvastatin Na⁺ (SIM Na⁺) (10 μM) (incubation, 2 to 30 min) on the protein expression of p-HMG-CoA reductase-Ser⁸⁷¹ and HMG-CoA reductase of porcine coronary artery.</p

Effects of simvastatin on K_ATP channel openings.

<p>(A) Effects of cromakalim (Crom., 10 μM) on whole-cell K_ATP channel openings of single human internal mammary artery myocytes in the presence of glibenclamide (Glib., 3 μM) (n = 5 to 6). (B) Effects of cromakalim (Crom., 10 μM) on whole-cell K_ATP channel openings of single human internal mammary artery myocytes with and without simvastatin (1, 3 and 10 μM). (C) Effects of simvastatin (10 μM) and glibenclamide (Glib., 3 μM) on cromakalim (Crom., 10 μM)-induced whole-cell K_ATP channel openings of single porcine artery myocytes (in the presence of okadaic acid, 10 nM). Number of cells studied is indicated in parenthesis. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001 compared to controls. (D) Effects of cromakalim (Crom., 10 μM) on whole-cell K_ATP channel openings of single porcine coronary artery myocytes in the presence of AICAR (1 mM). Number of cells studied is indicated in parenthesis. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001 compared to controls.</p