Supplementary for Rare severe Gardner-Diamond syndrome-like presentation with hG5c_2 Neutrophils
Description
Supplementary Table 1. Detailed laboratory and imaging findings. Supplementary Table 2. Markers of scRNA-Seq, Proteomics, and Metabolitics. Supplementary Figure 1. (A) Multiple well-demarcated hemorrhagic lesions, blisters and swelling of the ankle joint were observed in the patient. (B) The ecchymosis evolved from the wrist, elbow, and shoulder to the chest in a linear-to-curvilinear, hook-shaped configuration, and this phenomenon occurred twice before. Supplementary Figure 2. (A) Magnetic Resonance Imaging (MRI) of the brain revealed ventriculomegaly. (B) Pathological examination of skin lesions. 5 min post-onset: sparse lymphocytic infiltration and scattered neutrophils were observed around superficial dermal blood vessels. Small blood vessels exhibited dilation and congestion. 60 min post-onset: Sparse lymphocytic infiltration persisted around superficial dermal blood vessels, accompanied by vascular dilatation and congestion. Erythrocyte extravasation (black arrows) and scattered swelling/degenerative keratinocytes (red circles) were present within the epidermis, whereas the epidermal basal layer remained intact, without evidence of full-thickness coagulative necrosis or subepidermal cleft formation. Scale bar = 500 μm. (C) 200 µL of 80% washed erythrocyte suspension was injected on the left side (yellow arrows) of the patient's back, and 200 µL of normal saline was injected on the right side (blue arrows) of the patient's back. Time points are indicated as follows: 1 (pre-injection), 2 (0 min post-injection), 3 (1.5 h post-injection), 4 (2 h post-injection), 5 (9.3 h post-injection), and 6 (24 h post-injection). Supplementary Figure 3. scRNA-Seq of the PBMC. (A) The ratio of major cell subsets in the PBMC. The percentage of neutrophils was drastically elevated. (B) The ratio of the neutrophils in the PBMC. The percentage of hG5c neutrophils was obviously increased. (C) The ratio of the hG5c neutrophils in the PBMC and highly-expressed genes in the hG5c_2 neutrophils. The percentage of hG5c_2 neutrophils was increased, and the hG5c_2 neutrophils highly expressed genes for neutrophil activation. Supplementary Figure 4. scRNA-Seq and RNA-Seq of the PBMC and proteomics of the serum. (A) Differentially-expressed genes and GSEA of scRNA-Seq data. During the onset of skin ulceration, the chemokine, neutrophil trap formation, and TNFα pathways were highly enriched. (B) GO analysis of RNA-Seq and proteomics of the blood. GO analysis showed a highly-enriched chemokine pathway when compared with that of the normal control (both the parents and three healthy individuals). The Reactome of proteomics showed that the neutrophil degranulation pathway was enriched. Genes in the chemokine pathway were elevated through validation by qPCR. Detail explanation of Supplementary Videos is included in the file named "Explanation of Supplementary Videos".