Genetic regulation of surfactant deficiency has been suggested by heritability of neonatal respiratory distress syndrome, persistence of gender and racial disparities in disease risk, and the frequency of surfactant replacement treatment failures. Individually rare, collectively common (2-4/100 infants) mutations in the ATP- binding cassette sub-family A member 3 gene (ABCA3) disrupt surfactant function through diverse mechanisms including reduced lipid transport, ABCA3 misfolding or altered trafficking, or induction of increased endoplasmic reticulum stress and increase risk for neonatal respiratory distress syndrome in term and late preterm infants. Our goal is to develop and implement a human model system that uses static and dynamic lipidomic signatures for functional screening of ABCA3 mutations and that could be used for testing small molecules to correct mutation-encoded, functional defects in any gene expressed in the human alveolar type 2 cell. In the R21 Phase of this Award, using adenoviral vectors with high transduction efficiency and cargo capacity and highly sensitive and specific mass spectrometry based lipidomic profiling, we will test the hypothesis that ABCA3 gene silencing and rescue with wild-type ABCA3 cDNA reconstitute surfactant phospholipid signatures in human, primary alveolar type 2 cells. In the R33 Phase of this Award, using mass spectrometry-based lipidomic profiling, electron microscopy, and surface activity measurements, we will examine disruption of surfactant lipidomic signatures, lipid turnover rates, and lipid secretion kinetics, lamellar body phenotype, and surfactant function by previously characterized and uncharacterized ABCA3 mutations associated with increased risk for neonatal respiratory distress syndrome. The overall impact of this Award will provide a human model system for functional, lipidomics-based screening of genomic hits associated with surfactant deficiency and for development of small- molecule based strategies to correct mutation-encoded, functional surfactant defects.

Public Health Relevance

Neonatal respiratory distress syndrome due to surfactant deficiency is the most common respiratory cause of death and morbidity in infants <1 year of age in the United States. Genetic causes of surfactant deficiency contribute substantially to disease burden but are poorly understood. Using innovative, state of the art methods, we will develop and implement a human model tissue culture system that can provide information to improve diagnostic and treatment strategies for infants with genetic surfactant deficiency. (End of Abstract)

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21HL120760-01
Application #
8606976
Study Section
Special Emphasis Panel (ZHL1-CSR-H (S1))
Program Officer
Gan, Weiniu
Project Start
2014-03-01
Project End
2016-02-29
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
1
Fiscal Year
2014
Total Cost
$417,420
Indirect Cost
$112,210
Name
Washington University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Towe, Christopher T; White, Frances V; Grady, R Mark et al. (2018) Infants with Atypical Presentations of Alveolar Capillary Dysplasia with Misalignment of the Pulmonary Veins Who Underwent Bilateral Lung Transplantation. J Pediatr 194:158-164.e1
Wambach, Jennifer A; Wegner, Daniel J; Yang, Ping et al. (2018) Functional characterization of biallelic RTTN variants identified in an infant with microcephaly, simplified gyral pattern, pontocerebellar hypoplasia, and seizures. Pediatr Res 84:435-441
Frankfater, Cheryl; Jiang, Xuntian; Hsu, Fong-Fu (2018) Characterization of Long-Chain Fatty Acid as N-(4-Aminomethylphenyl) Pyridinium Derivative by MALDI LIFT-TOF/TOF Mass Spectrometry. J Am Soc Mass Spectrom 29:1688-1699
Wambach, Jennifer A; Stettner, Georg M; Haack, Tobias B et al. (2017) Survival among children with ""Lethal"" congenital contracture syndrome 11 caused by novel mutations in the gliomedin gene (GLDN). Hum Mutat 38:1477-1484
Lin, Meei-Hua; Miner, Jeffrey H; Turk, John et al. (2017) Linear ion-trap MSn with high-resolution MS reveals structural diversity of 1-O-acylceramide family in mouse epidermis. J Lipid Res 58:772-782
Wright, Catherine C; Hsu, Fong Fu; Arnett, Eusondia et al. (2017) The Mycobacterium tuberculosis MmpL11 Cell Wall Lipid Transporter Is Important for Biofilm Formation, Intracellular Growth, and Nonreplicating Persistence. Infect Immun 85:
Wambach, Jennifer A; Yang, Ping; Wegner, Daniel J et al. (2016) Functional Characterization of ATP-Binding Cassette Transporter A3 Mutations from Infants with Respiratory Distress Syndrome. Am J Respir Cell Mol Biol 55:716-721
Hsu, Fong-Fu (2016) Characterization of Hydroxyphthioceranoic and Phthioceranoic Acids by Charge-Switch Derivatization and CID Tandem Mass Spectrometry. J Am Soc Mass Spectrom 27:622-32
Flentie, Kelly N; Stallings, Christina L; Turk, John et al. (2016) Characterization of phthiocerol and phthiodiolone dimycocerosate esters of M. tuberculosis by multiple-stage linear ion-trap MS. J Lipid Res 57:142-55
Hsu, Fong-Fu (2016) Complete structural characterization of ceramides as [M-H](-) ions by multiple-stage linear ion trap mass spectrometry. Biochimie 130:63-75

Showing the most recent 10 out of 11 publications