Impaired angiogenesis and extracellular matrix metabolism in autosomal-dominant hyper-IgE syndrome
Natalia I. Dmitrieva, … , Guibin Chen, Manfred Boehm
Natalia I. Dmitrieva, … , Guibin Chen, Manfred Boehm
Published May 5, 2020
Citation Information: J Clin Invest. 2020;130(8):4167-4181. https://doi.org/10.1172/JCI135490.
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Research Article Vascular biology

Impaired angiogenesis and extracellular matrix metabolism in autosomal-dominant hyper-IgE syndrome

Abstract

There are more than 7000 described rare diseases, most lacking specific treatment. Autosomal-dominant hyper-IgE syndrome (AD-HIES, also known as Job’s syndrome) is caused by mutations in STAT3. These patients present with immunodeficiency accompanied by severe nonimmunological features, including skeletal, connective tissue, and vascular abnormalities, poor postinfection lung healing, and subsequent pulmonary failure. No specific therapies are available for these abnormalities. Here, we investigated underlying mechanisms in order to identify therapeutic targets. Histological analysis of skin wounds demonstrated delayed granulation tissue formation and vascularization during skin-wound healing in AD-HIES patients. Global gene expression analysis in AD-HIES patient skin fibroblasts identified deficiencies in a STAT3-controlled transcriptional network regulating extracellular matrix (ECM) remodeling and angiogenesis, with hypoxia-inducible factor 1α (HIF-1α) being a major contributor. Consistent with this, histological analysis of skin wounds and coronary arteries from AD-HIES patients showed decreased HIF-1α expression and revealed abnormal organization of the ECM and altered formation of the coronary vasa vasorum. Disease modeling using cell culture and mouse models of angiogenesis and wound healing confirmed these predicted deficiencies and demonstrated therapeutic benefit of HIF-1α–stabilizing drugs. The study provides mechanistic insights into AD-HIES pathophysiology and suggests potential treatment options for this rare disease.

Authors

Natalia I. Dmitrieva, Avram D. Walts, Dai Phuong Nguyen, Alex Grubb, Xue Zhang, Xujing Wang, Xianfeng Ping, Hui Jin, Zhen Yu, Zu-Xi Yu, Dan Yang, Robin Schwartzbeck, Clifton L. Dalgard, Beth A. Kozel, Mark D. Levin, Russell H. Knutsen, Delong Liu, Joshua D. Milner, Diego B. López, Michael P. O’Connell, Chyi-Chia Richard Lee, Ian A. Myles, Amy P. Hsu, Alexandra F. Freeman, Steven M. Holland, Guibin Chen, Manfred Boehm

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Figure 4

Deficient HIF-1α signaling in AD-HIES SFs at transcriptional and protein stabilization levels.

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Deficient HIF-1α signaling in AD-HIES SFs at transcriptional and protein...
(A) RNA-Seq pathway map showing decreased expression of many HIF-1α transcriptional targets in AD-HIES fibroblasts (P = 5.3E-12; FDR = 4.2E-9; GeneGo Metacore software). See Supplemental Table 2 for expression changes. (B) AD-HIES fibroblasts are deficient in ability to stabilize HIF-1α protein in response to cytokines. Cells were treated with TNF-α and IL-6 for 8 hours. Representative Western blot (upper panel) and quantification by densitometry (lower panel) are shown (n = 8). (C) AD-HIES SFs stabilize HIF-1α under hypoxic conditions (1% O2 for 8 hours). Representative Western blot and quantification (n = 3). (D) TNF-α increases STAT3 binding to promoter of HIF1α gene in control SFs but not in AD-HIES (ChIP; see Supplemental Figure 9 and Supplemental Table 3). Data are plotted as fold increase of STAT3 binding after treatment with TNF-α (see Methods) (n = 3). (E) HIF-1α mRNA levels were decreased in AD-HIES fibroblasts. (n = 6). (F–H) TNF-α decreased degradation rate of HIF-1α in control fibroblasts, but not in AD-HIES, indicating STAT3-dependent stabilization of HIF-1α and that this stabilization is STAT3 dependent. To measure rate of HIF-1α protein degradation, de novo protein synthesis was inhibited with cycloheximide (CHX). (F) Experimental design for estimation of HIF-1α stability. To increase level of HIF-1α, cells were treated with PHD inhibitor dimethyloxalylglycine (DMOG) that was washed out before addition of CHX. (G) Representative Western blot images showing dynamics of HIF-1α decrease after addition of CHX. (H) Quantification of Western blots (n = 3; asterisks, relative to control; pound signs, relative to 0 time). See also Supplemental Figure 8D. Data are represented as mean ± SEM. *,#P < 0.05; **P < 0.01; ***P < 0.001, 2-tailed unpaired t test (B, D, E, and H); 1-way ANOVA followed by Holm-Šidák multiple comparisons test (C). See Supplemental Table 1 for information about patient samples used.