ABCMdb

ABCA3 p.Asp253His

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Publications

PMID: 22068586 [PubMed] Flamein F et al: "Molecular and cellular characteristics of ABCA3 mutations associated with diffuse parenchymal lung diseases in children."

No. Sentence Comment

7 Functional studies of p.D253H and p.T1173R mutations revealed that p.D253H and p.T1173R induced abnormal lamellar bodies. Login to comment
41 Analysis of genomic DNA from the parents and kindred showed that the compound heterozygous p.R1583W/ p.S128Rfs (Fig. 1A) and p.R208W/p.R1521W (Fig. 1B) mutations were inherited, as well as the homozygous mutations p.T1173R (Fig. 1C) and p.D253H (Fig. 1D). Login to comment
54 Genetic analysis results in the 10 children harboring homozygous and compound heterozygous (shaded) or heterozygous ABCA3 mutations Patient NRD Clinical outcome ABCA3 mutation ABCA3 SNPs ABCA3 variants cDNA level Protein level dbSNPs rs# cluster id Missense variants in conserved amino acid 1 Yes ILD c.[3518C.G] + [3518C.G] p.[T1173R] + [T1173R] rs149532, rs13332514 2 Yes ILD c.[757G.C] + [757G.C] p.[D253H] + [D253H] 3 Yes Death c.[1385T.G] + [2890G.A] p.[L462R] + [G964S] rs149532 4 Yes Death c.[4747C.T] + c.[384delC] p.[R1583W] + p.[S128Rfs] rs149532 c.[450G.A] (het) 5 No Death c.[629G.T] + [3079G.C] p.[G210V] + [A1027P] rs149532 6 Yes ILD c.[622C.T] + [4561C.T] p.[R208W] + [R1521W] rs149532, rs323043 7 Yes Death c.[604G.C] + [907C.G] p.[G202R] + [L303V] rs149532, rs323043 (het), rs13332514 8 Yes Death c.[2888A.G] + [?] Login to comment
68 Pedigree of the families with the ABCA3 mutations p.R1583W/p.S128Rfs (A), p.R1521W/R208W (B), p.T1173R/p.T1173R (C) and p.D253H/ p.T1173R (D). Login to comment
78 Electron microscopy was performed on a lung biopsy from the patient harboring the p.D253H mutation and showed abnormal lamellar bodies with dense inclusions (Fig. 3C). Login to comment
79 BALF analysis Western blot analysis of surfactant proteins (Fig. 4) was performed in seven patients, who had the following ABCA3 mutations: p.D253H (patient 2), p.T1173R (patient 1), p.L462R/ p.G964S (patient 3), p.G202R/p.L303V (patient 7), p.Y963C (patient 8), p.R1583W/p.S128Rfs (patient 4) and p.S872G (patient 10), respectively. Login to comment
80 SP-C (Fig. 4A) and SP-B (Fig. 4B) were detected at a size of 3.7 and 8 kDa in variable amounts, the smallest amount being found in the patient with the p.G202R/p.L303V ABCA3 mutation (patient 7) in whom SP-C and SP-B are only faintly visible. Login to comment
84 Characterization of ABCA3 mutations The two mutations p.T1173R and p.D253H were deliberately chosen for subsequent functional studies because they were homozygous. Login to comment
89 (A) HRCT scan, (B) hematoxylin and eosin (HE) staining of lung tissue and (C) electron microscopy of type-II cells from the patient (patient 2) harboring the ABCA3 homozygous mutation p.D253H. Login to comment
97 Thus, p.D253H and p.T1173R mutants were not associated with a localization defect. Login to comment
99 In WT, p.T1173R and p.D253H cells, anti-GFP antibody revealed two bands of 180 kDa (150 kDa ABCA3 + 30 kDa GFP) and 220 kDa (190 + 30 kDa GFP), respectively (Fig. 6). Login to comment
101 Lamellar bodies in ABCA3 WT, p.D253H and p.T1173R cells. Login to comment
104 The p.D253H mutation induced abnormal lamellar bodies with electron-dense inclusions (Fig. 7, dashed arrows), consistent with the results of the patient`s lung biopsy (Fig. 3C). Login to comment
106 Cytokine production by ABCA3 WT, p.D253H and p.T1173R cells. Login to comment
110 A significant, faint increase in IL-8 mRNA expression was also observed between WT and p.D253H cells. Login to comment
112 At the protein level, ELISA results confirmed that A549 cells expressing the p.T1173R mutant produced more IL-8 than did WT cells (Fig. 8A). Login to comment
113 In contrast, IL-8 production by p.D253H cells was similar to that of WT cells (Fig. 8B). Login to comment
123 Human Molecular Genetics, 2012, Vol. 21, No. 4 769 ERK1/2 inhibitor treatment, IL-8 production by p.T1173R cells remained increased compared with WT and p.D253H cells. Login to comment
125 Western blot and relative quantification of ERK1/2 phosphorylation confirmed that the observed IL-8 overproduction in p.T1173R cells was independent of ERK1/2 signaling. Login to comment
137 Intracellular localization of wild-type ABCA3 and of the p.D253H and p.T1173R mutants. Login to comment
138 A549 cells either non-transfected or transfected with mock vector, wild-type protein ABCA3-WT (A) or mutated proteins p.D253H (B) and p.T1173R (C) were analyzed using confocal microscopy. Login to comment
141 Western blot analysis of ABCA3 in cells transiently transfected with ABCA3-WT or the p.D253H or p.T1173R mutation. Login to comment
155 To confirm the hypothesis, it would have been interesting to compare the patterns of siblings harboring the p.D253H mutation. Login to comment
161 The presence of abnormal lamellar bodies in the patient harboring the p.D253H mutation supports a pathogenic effect of this mutation. Login to comment
162 These abnormalities were also observed in vitro in p.D253H- and p.T1173R-transfected cells, suggesting that ABCA3 abnormalities may consistently induce abnormal lamellar bodies. Login to comment
163 However, since we do not have the corresponding biopsy from the patient harboring the p.T1173R, we cannot draw a firm conclusion on this point. Login to comment
166 A549 cells transfected with mock vector (pEGFP-N1), ABCA3-WT (A), or mutated ABCA3-D253H (B) or ABCA3-T1173R-GFP were analyzed using electron microscopy. Login to comment
167 Human Molecular Genetics, 2012, Vol. 21, No. 4 771 We performed in vitro experiments to elucidate the pathophysiological effects of two mutations associated with progression towards ILD, p.D253H and T1173R. Login to comment
172 The p.D253H and p.T1173R mutations induced abnormal lamellar bodies. Login to comment
192 (B) Role for MAPK and NF-kB-dependent signaling in IL-8 secretion by ABCA3-WT, D253H and T1173R cells. Login to comment
227 Mutagenesis primers (Sigma) were as follows: D253H-For-5' -ACCCGCCGTTCATCGCACACCCCTTCC-3' , D253H-Rev-5' -GGAAGGGGTGTGCGATGAACGGCGGGT- 3' ; T1173R-For-5' -ACGTGCGTGCCTTCAGGCGGGACG-3' , and T1173R-Rev-5' -CGTCCCGCCTGAAGGCACGCACGT- 3' . Login to comment
230 Cells (1 × 106 ) were transfected with 1 mg of ABCA3-WT, ABCA3-D253H or ABCA3-T1173R plasmid using a nucleofector device (Lonza, Cologne, Germany) as recommended by the manufacturer. Login to comment
43 Analysis of genomic DNA from the parents and kindred showed that the compound heterozygous p.R1583W/ p.S128Rfs (Fig. 1A) and p.R208W/p.R1521W (Fig. 1B) mutations were inherited, as well as the homozygous mutations p.T1173R (Fig. 1C) and p.D253H (Fig. 1D). Login to comment
56 Genetic analysis results in the 10 children harboring homozygous and compound heterozygous (shaded) or heterozygous ABCA3 mutations Patient NRD Clinical outcome ABCA3 mutation ABCA3 SNPs ABCA3 variants cDNA level Protein level dbSNPs rs# cluster id Missense variants in conserved amino acid 1 Yes ILD c.[3518C.G] + [3518C.G] p.[T1173R] + [T1173R] rs149532, rs13332514 2 Yes ILD c.[757G.C] + [757G.C] p.[D253H] + [D253H] 3 Yes Death c.[1385T.G] + [2890G.A] p.[L462R] + [G964S] rs149532 4 Yes Death c.[4747C.T] + c.[384delC] p.[R1583W] + p.[S128Rfs] rs149532 c.[450G.A] (het) 5 No Death c.[629G.T] + [3079G.C] p.[G210V] + [A1027P] rs149532 6 Yes ILD c.[622C.T] + [4561C.T] p.[R208W] + [R1521W] rs149532, rs323043 7 Yes Death c.[604G.C] + [907C.G] p.[G202R] + [L303V] rs149532, rs323043 (het), rs13332514 8 Yes Death c.[2888A.G] + [?] Login to comment
70 Pedigree of the families with the ABCA3 mutations p.R1583W/p.S128Rfs (A), p.R1521W/R208W (B), p.T1173R/p.T1173R (C) and p.D253H/ p.T1173R (D). Login to comment
81 BALF analysis Western blot analysis of surfactant proteins (Fig. 4) was performed in seven patients, who had the following ABCA3 mutations: p.D253H (patient 2), p.T1173R (patient 1), p.L462R/ p.G964S (patient 3), p.G202R/p.L303V (patient 7), p.Y963C (patient 8), p.R1583W/p.S128Rfs (patient 4) and p.S872G (patient 10), respectively. Login to comment
86 Characterization of ABCA3 mutations The two mutations p.T1173R and p.D253H were deliberately chosen for subsequent functional studies because they were homozygous. Login to comment
91 (A) HRCT scan, (B) hematoxylin and eosin (HE) staining of lung tissue and (C) electron microscopy of type-II cells from the patient (patient 2) harboring the ABCA3 homozygous mutation p.D253H. Login to comment
103 Lamellar bodies in ABCA3 WT, p.D253H and p.T1173R cells. Login to comment
108 Cytokine production by ABCA3 WT, p.D253H and p.T1173R cells. Login to comment
115 In contrast, IL-8 production by p.D253H cells was similar to that of WT cells (Fig. 8B). Login to comment
139 Intracellular localization of wild-type ABCA3 and of the p.D253H and p.T1173R mutants. Login to comment
140 A549 cells either non-transfected or transfected with mock vector, wild-type protein ABCA3-WT (A) or mutated proteins p.D253H (B) and p.T1173R (C) were analyzed using confocal microscopy. Login to comment
143 Western blot analysis of ABCA3 in cells transiently transfected with ABCA3-WT or the p.D253H or p.T1173R mutation. Login to comment
157 To confirm the hypothesis, it would have been interesting to compare the patterns of siblings harboring the p.D253H mutation. Login to comment
164 These abnormalities were also observed in vitro in p.D253H- and p.T1173R-transfected cells, suggesting that ABCA3 abnormalities may consistently induce abnormal lamellar bodies. Login to comment
168 A549 cells transfected with mock vector (pEGFP-N1), ABCA3-WT (A), or mutated ABCA3-D253H (B) or ABCA3-T1173R-GFP were analyzed using electron microscopy. Login to comment
169 We performed in vitro experiments to elucidate the pathophysiological effects of two mutations associated with progression towards ILD, p.D253H and T1173R. Login to comment
174 The p.D253H and p.T1173R mutations induced abnormal lamellar bodies. Login to comment
194 (B) Role for MAPK and NF-kB-dependent signaling in IL-8 secretion by ABCA3-WT, D253H and T1173R cells. Login to comment
228 Mutagenesis primers (Sigma) were as follows: D253H-For-5' -ACCCGCCGTTCATCGCACACCCCTTCC-3' , D253H-Rev-5' -GGAAGGGGTGTGCGATGAACGGCGGGT- 3' ; T1173R-For-5' -ACGTGCGTGCCTTCAGGCGGGACG-3' , and T1173R-Rev-5' -CGTCCCGCCTGAAGGCACGCACGT- 3' . Login to comment
231 Cells (1 &#d7; 106 ) were transfected with 1 mg of ABCA3-WT, ABCA3-D253H or ABCA3-T1173R plasmid using a nucleofector device (Lonza, Cologne, Germany) as recommended by the manufacturer. Login to comment

PMID: 24871971 [PubMed] Wambach JA et al: "Genotype-phenotype correlations for infants and children with ABCA3 deficiency."

No. Sentence Comment

134 Alleles with ABCA3 Variants in Cis Allele Number of Subjects with Allele R43C-P1653L 1 D115E-D253H 1 (2 alleles, 1 subject homozygous) V129M-V1495M 1 W179C-P770L 3 (3 subjects heterozygous) E195K-R1271Q 1 R280C-Q1589X 2 (3 alleles, 1 subject homozygous, 1 subject heterozygous) R288K-S693L 2 (2 subjects heterozygous) c.1474_1475insT-D953N 4 (3 siblings homozygous, 1 subject heterozygous) P766S-L960F 4 (4 subjects heterozygous) H778R-L1252P 1 A54T-R1482W-IVS25-98 C . Login to comment

PMID: 26186947 [PubMed] Mulugeta S et al: "Lost after translation: insights from pulmonary surfactant for understanding the role of alveolar epithelial dysfunction and cellular quality control in fibrotic lung disease."

No. Sentence Comment

267 Summary of reported phenotypic features for surfactant component mutations Mutation (Domain) Clinical Diagnosis Lung Phenotype (in vivo) Subcellular Localization Trafficking Cellular Responses (in vitro) References SFTPA2 F198S (CRD) G231V (CRD) Familial pulmonary fibrosis Total BAL [SP-A] Normal ER retention Intracellular aggregation Not secreted (af9;) ER stress, cleared by ERAD (af9;) TGFbeta1 elaboration 99, 100, 175 SFTPC Group A1 èc;Exon4 (BRICHOS) L188Q (BRICHOS) G100S (BRICHOS) NSIP (Children) IPF/UIP (Adult) Absence of mature SP-C (humans) Arrested lung development (mice) ER stress (humans; mice) 1Sensitivity to bleomycin (mice) Epithelial cytotoxicity ER retention&#a1; aggresomes Intracellular aggregates ERAD requires Erdj 4/5 MG132 blocks degradation 4-PBA improves aggregates (af9;) ER stress (af9;) Apoptosis (af9;) Incomplete or absent proSP-C processing (af9;) IL-8/TGFbeta1 expression (af9;) Polyubiquitinated isoforms 21, 39, 97, 98, 100, 111, 112, 116, 117, 120, 153, 159, 160, 173, 193 Group A2 L110R (BRICHOS) P115L (BRICHOS) A116D (BRICHOS) Unspecified ILD Unspecified ILD Unspecified chILD Phenotype not reported EEA-1 (af9;); Syntaxin2 (afa;) Intracellular aggregation 2 PC secretion (af9;) Aberrant processing, 2 cell viability 1 HSP response (af9;) Congo red aggregates 160, 193 Group B1 E66K (Linker) I73T (Linker) NSIP/PAP (Child) IPF/UIP (Adult) 1 Phospholipid; 1SP-A, PAS positive staining Biopsy: PM and EE localization Misprocessed SP-C (BAL) Misprocessed SP-B (BAL) Plasma membrane&#a1;EE&#a1;LE/MVB (af9;) Aberrantly processed protein (af9;) Late autophagy block 2 Mitophagy 1 Mysfunctional mitochondria 1, 19, 24, 26, 49, 116, 118, 128, 152 Group B2 èc;91-93 (Non-BRICHOS) NSIP/PAP 2 BAL SP-B 1 BAL SP-A 2 Surfactant surface tension (af9;) Intracellular aggregates (af9;) Congo red staining Plasma membraneߥ EEA1 (af9;) compartmentsߥ Not reported 55, 181 Group C P30L (NH2-terminal) Unspecified ILD Phenotype not reported (af9;) ER retention 1 Bip expression (af9;) Polyubiquitinated isoforms 13, 116, 160 ABCA3 Group I (Trafficking Defective) L101P (1st luminal loop) R280C (1st cytosolic loop) L982P (3rd luminal loop) G1221S (11th TM domain) L1553P (COOH-terminal) Q1591P (COOH-terminal) Surfactant deficiency* RDS* chILDߤ Phenotype not reported Phenotype not reported Phenotype not reported (af9;) ER retention Non-LRO cytosolic vesicles (af9;) ER stress 30, 31, 103, 147, 172, 177 Group II (Functionally Defective) R43L (1st luminal loop) D253H (1st luminal loop) E292V (1st cytosolic loop) N568D (ABC1) E690K (ABC1) T1114M (8thTM domain) T1173R (1st luminal loop) L1580P (COOH-terminal) Surfactant deficiency* RDS* chILD (CPI)ߤ Reduced SP-B and SP-C (afa;) ER retention Lysosomes or LROs (normal) Impaired lipid transport Impaired ATP hydrolysis Impaired ATP binding Abnormal LBs 1 IL8 secretion 20, 25, 103, 104, 147, 148, 177 *Seen with homozygous or compound heterozygous ABCA3 expression; ߤfound with heterozugous ABCA3 expression. Login to comment