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Vol. 95, No. 2, 2009
Issue release date: February 2009
Free Access
Neonatology 2009;95:117–124
(DOI:10.1159/000153095)

Developmental and Genetic Regulation of Human Surfactant Protein B in vivo

Hamvas A.a · Heins H.B.a · Guttentag S.H.d · Wegner D.J.a · Trusgnich M.A.a · Bennet K.W.a · Yang P.a · Carlson C.S.c · An P.b · Cole F.S.a
aDivision of Newborn Medicine, the Edward Mallinckrodt Department of Pediatrics and St. Louis Children’s Hospital, bDivision of Statistical Genomics, Department of Genetics, Washington University School of Medicine, St. Louis, Mo., cDivision of Public Health Sciences, the Fred Hutchinson Cancer Research Center, University of Washington, Seattle, Wash., and dDivision of Neonatology, Department of Pediatrics, The Children’s Hospital of Philadelphia and the University of Pennsylvania School of Medicine, Philadelphia, Pa., USA
email Corresponding Author

Abstract

Background: Genetic and developmental disruption of surfactant protein B (SP-B) expression causes neonatal respiratory distress syndrome (RDS). Objectives: To assess developmental and genetic regulation of SP-B expression in vivo. Methods: To evaluate in vivo developmental regulation of SP-B, we used immunoblotting to compare frequency of detection of mature and pro-SP-B peptides in developmentally distinct cohorts: 24 amniotic fluid samples, unfractionated tracheal aspirates from 101 infants ≥34 weeks’ gestation with (75) and without (26) neonatal RDS, and 6 nonsmoking adults. To examine genetic regulation, we used univariate and logistic regression analyses to detect associations between common SP-B (SFTPB) genotypes and SP-B peptides in the neonatal RDS cohort. Results: We found pro-SP-B peptides in 24/24 amniotic fluid samples and in 100/101 tracheal aspirates from newborn infants but none in bronchoalveolar lavage from normal adults (0/6) (p < 0.001). We detected an association (p = 0.0011) between pro-SP-B peptides (Mr 40 and 42 kDa) and genotype of a nonsynonymous single nucleotide polymorphism at genomic position 1580 that regulates amino-terminus glycosylation. Conclusions: Pro-SP-B peptides are more common in developmentally less mature humans. Association of genotype at genomic position 1580 with pro-SP-B peptides (Mr 40 and 42 kDa) suggests genetic regulation of amino terminus glycosylation in vivo.


 goto top of outline Key Words

  • Neonatal respiratory diseases
  • Respiratory distress syndrome
  • Surfactant proteins

 goto top of outline Abstract

Background: Genetic and developmental disruption of surfactant protein B (SP-B) expression causes neonatal respiratory distress syndrome (RDS). Objectives: To assess developmental and genetic regulation of SP-B expression in vivo. Methods: To evaluate in vivo developmental regulation of SP-B, we used immunoblotting to compare frequency of detection of mature and pro-SP-B peptides in developmentally distinct cohorts: 24 amniotic fluid samples, unfractionated tracheal aspirates from 101 infants ≥34 weeks’ gestation with (75) and without (26) neonatal RDS, and 6 nonsmoking adults. To examine genetic regulation, we used univariate and logistic regression analyses to detect associations between common SP-B (SFTPB) genotypes and SP-B peptides in the neonatal RDS cohort. Results: We found pro-SP-B peptides in 24/24 amniotic fluid samples and in 100/101 tracheal aspirates from newborn infants but none in bronchoalveolar lavage from normal adults (0/6) (p < 0.001). We detected an association (p = 0.0011) between pro-SP-B peptides (Mr 40 and 42 kDa) and genotype of a nonsynonymous single nucleotide polymorphism at genomic position 1580 that regulates amino-terminus glycosylation. Conclusions: Pro-SP-B peptides are more common in developmentally less mature humans. Association of genotype at genomic position 1580 with pro-SP-B peptides (Mr 40 and 42 kDa) suggests genetic regulation of amino terminus glycosylation in vivo.

Copyright © 2008 S. Karger AG, Basel


 goto top of outline References
  1. Hills B: Surface-active phospholipid: a Pandora’s box of clinical applications. 1. The lung and air spaces. Intern Med J 2002;32:170–178.
  2. Mazela J, Merritt TA, Gadzinowski J, Sinha S: Evolution of pulmonary surfactants for the treatment of neonatal respiratory distress syndrome and paediatric lung diseases. Acta Paediatr 2006;95:1036–1048.
  3. Kramer B: The respiratory distress syndrome (RDS) in preterm infants: physiology, prophylaxis, and new therapeutic approaches. Intensivmed 2007;44:403–408.

    External Resources

  4. Angus D, Linde-Zwirble WT, Clermont G, Griffin MF, Clark RH: Epidemiology of neonatal respiratory failure in the united states: projections from California and New York. Am J Respir Crit Care Med 2001;164:1154–1160.
  5. Hamvas A, Devine T, Cole FS: Surfactant therapy failure identifies infants at risk for pulmonary mortality. Am J Dis Child 1993;147:665–668.
  6. Richardson D, Torday JS: Racial differences in predictive value of the lecithin/sphingomyelin ratio. Am J Obstet Gynecol 1994;170:1273–1278.
  7. Nogee LM de Mello DE, Dehner LP, Colten HR: Brief report: deficiency of pulmonary surfactant protein B in congenital alveolar proteinosis. N Engl J Med 1993;328:406–410.
  8. Clark R: The epidemiology of respiratory failure in neonates born at an estimated gestational age of 34 weeks or more. J Perinatol 2005;25:251–257.
  9. Weaver T, Conkright JJ: Function of surfactant proteins B and C. Annu Rev Physiol 2001;63:555–578.
  10. Guttentag S, Beers MF, Bieler BM, Ballard PL: Surfactant protein B processing in human fetal lung. Am J Physiol 1998;275:L559–L566.
  11. Brasch F, Johnen G, Winn-Brasch A, Guttentag SH, Schmiedl A, Kapp N, Suzuki Y, Muller KM, Richter J, Hawgood S, Ochs M: Surfactant protein B in type II pneumocytes and intra-alveolar surfactant forms of human lungs. Am J Respir Cell Mol Biol 2004;30:449–458.
  12. Ueno T, Linder S, Na CL, Rice WR, Johansson J, Weaver TE: Processing of pulmonary surfactant protein B by napsin and cathepsin H. J Biol Chem 2004;279:16178–16184.
  13. Dilger I, Schwedler G, Dudenhausen JW: Determination of the pulmonary surfactant-associated protein SP-B in amniotic fluid with a competition ELISA. Gynecol Obstet Invest 1994;38:24–27.
  14. Mori K, Ikeda K, Tanaka M: Different expression of surfactant protein B mature peptide and proprotein at 21 weeks’ gestation in human fetal pulmonary epithelial cells. Pediatr Int 2002;44:500–504.
  15. Stahlman M, Gray ME, Whitsett JA: The ontogeny and distribution of surfactant protein B in human fetuses and newborns. J Histochem Cytochem 1992;40:1471–1480.
  16. Griese M, Schumacher S, Tredano M, Steinecker M, Braun A, Guttentag S, Beers MF, Bahuau M: Expression profiles of hydrophobic surfactant proteins in children with diffuse chronic lung disease. Respir Res 2005;6:80.
  17. Lin S, Akinbi HT, Breslin JS, Weaver TE: Structural requirements for targeting of surfactant protein B (SP-B) to secretory granules in vitro and in vivo. J Biol Chem 1996;271:19689–19695.
  18. Akinbi H, Breslin JS, Ikegami M, Iwamoto HS, Clark JC, Whitsett JA, Jobe AH, Weaver TE: Rescue of SP-B knockout mice with a truncated SP-B proprotein. Function of the c-terminal propeptide. J Biol Chem 1997;272:9640–9647.
  19. Korimilli A, Gonzales LW, Guttentag SH: Intracellular localization of processing events in human surfactant protein B biosynthesis. J Biol Chem 2000;275:8672–8679.
  20. Nogee L, Garnier G, Dietz HC, Singer L, Murphy AM, deMello DE, Colten HR: A mutation in the surfactant protein B gene responsible for fatal neonatal respiratory disease in multiple kindreds. J Clin Invest 1994;93:1860–1863.
  21. Melton K, Nesslein LL, Ikegami M, Tichelaar JW, Clark JC, Whitsett JA, Weaver TE: Sp-B deficiency causes respiratory failure in adult mice. Am J Physiol Lung Cell Mol Physiol 2003;285:L543–L549.
  22. Wang G, Christensen ND, Wigdahl B, Guttentag SH, Floros J: Differences in n-linked glycosylation between human surfactant protein-B variants of the C or T allele at the single-nucleotide polymorphism at position 1580: implications for disease. Biochem J 2003;369:179–184.
  23. Ballard P, Merrill JD, Godinez RI, Godinez MH, Truog WE, Ballard RA: Surfactant protein profile of pulmonary surfactant in premature infants. Am J Respir Crit Care Med 2003;168:1123–1128.
  24. Merrill J, Ballard RA, Cnaan A, Hibbs AM, Godinez RI, Godinez MH, Truog WE, Ballard PL: Dysfunction of pulmonary surfactant in chronically ventilated premature infants. Pediatr Res 2004;56:918–926.
  25. Hamvas A, Wegner DJ, Carlson CS, Bergmann KR, Trusgnich MA, Fulton L, Kasai Y, An P, Mardis ER, Wilson RK, Cole FS: Comprehensive genetic variant discovery in the surfactant protein B gene. Pediatr Res 2007;62:170–175.
  26. Hamvas A, Wegner DJ, Trusgnich MA, Madden K, Heins H, Liu Y, Rice T, An P, Watkins-Torry J, Cole FS: Genetic variant characterization in intron 4 of the surfactant protein B gene. Hum Mutat 2005;26:494–495.
  27. Bachurski C, Ross GF, Ikegami M, Kramer BW, Jobe AH: Intra-amniotic endotoxin increases pulmonary surfactant proteins and induces SP-B processing in fetal sheep. Am J Physiol Lung Cell Mol Physiol 2001;280:L279–L285.
  28. Kramer B, Kramer S, Ikegami M, Jobe AH: Injury, inflammation, and remodeling in fetal sheep lung after intra-amniotic endotoxin. Am J Physiol Lung Cell Mol Physiol 2002;283:L452–L459.
  29. Brasch F, Ochs M, Kahne T, Guttentag S, Schauer-Vukasinovic V, Derrick M, Johnen G, Kapp N, Muller KM, Richter J, Giller T, Hawgood S, Buhling F: Involvement of napsin A in the C- and N-terminal processing of surfactant protein B in type-II pneumocytes of the human lung. J Biol Chem 2003;278:49006–49014.
  30. Foster C, Aktar A, Kopf D, Zhang P, Guttentag S: Pepsinogen C: a type 2 cell-specific protease. Am J Physiol Lung Cell Mol Physiol 2004;286:L382–L387.
  31. Guttentag S, Robinson L, Zhang P, Brasch F, Buhling F, Beers M: Cysteine protease activity is required for surfactant protein B processing and lamellar body genesis. Am J Respir Cell Mol Biol 2003;28:69–79.

 goto top of outline Author Contacts

Aaron Hamvas, MD
Division of Newborn Medicine, St. Louis Children’s Hospital
One Children’s Place
St. Louis, MO 63110 (USA)
Tel. +1 314 454 6148, Fax +1 314 454 4633, E-Mail hamvas@kids.wustl.edu


 goto top of outline Article Information

These data were presented at the Pediatric Academic Societies’ Meetings in San Francisco, May 2004 and 2006, and in Washington, D.C., May 2005 (E-PAS 2006:59:2610.4; Pediatr Res 2005;57:2502, and Pediatr Res 2004;55:2664).

Received: December 6, 2007
Accepted: January 21, 2008
Published online: September 6, 2008
Number of Print Pages : 8
Number of Figures : 3, Number of Tables : 5, Number of References : 31


 goto top of outline Publication Details

Neonatology (Fetal and Neonatal Research)

Vol. 95, No. 2, Year 2009 (Cover Date: February 2009)

Journal Editor: Halliday H.L. (Belfast), Speer C.P. (Würzburg)
ISSN: 1661–7800 (Print), eISSN: 1661–7819 (Online)

For additional information: http://www.karger.com/NEO


Copyright / Drug Dosage / Disclaimer

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

Abstract

Background: Genetic and developmental disruption of surfactant protein B (SP-B) expression causes neonatal respiratory distress syndrome (RDS). Objectives: To assess developmental and genetic regulation of SP-B expression in vivo. Methods: To evaluate in vivo developmental regulation of SP-B, we used immunoblotting to compare frequency of detection of mature and pro-SP-B peptides in developmentally distinct cohorts: 24 amniotic fluid samples, unfractionated tracheal aspirates from 101 infants ≥34 weeks’ gestation with (75) and without (26) neonatal RDS, and 6 nonsmoking adults. To examine genetic regulation, we used univariate and logistic regression analyses to detect associations between common SP-B (SFTPB) genotypes and SP-B peptides in the neonatal RDS cohort. Results: We found pro-SP-B peptides in 24/24 amniotic fluid samples and in 100/101 tracheal aspirates from newborn infants but none in bronchoalveolar lavage from normal adults (0/6) (p < 0.001). We detected an association (p = 0.0011) between pro-SP-B peptides (Mr 40 and 42 kDa) and genotype of a nonsynonymous single nucleotide polymorphism at genomic position 1580 that regulates amino-terminus glycosylation. Conclusions: Pro-SP-B peptides are more common in developmentally less mature humans. Association of genotype at genomic position 1580 with pro-SP-B peptides (Mr 40 and 42 kDa) suggests genetic regulation of amino terminus glycosylation in vivo.



 goto top of outline Author Contacts

Aaron Hamvas, MD
Division of Newborn Medicine, St. Louis Children’s Hospital
One Children’s Place
St. Louis, MO 63110 (USA)
Tel. +1 314 454 6148, Fax +1 314 454 4633, E-Mail hamvas@kids.wustl.edu


 goto top of outline Article Information

These data were presented at the Pediatric Academic Societies’ Meetings in San Francisco, May 2004 and 2006, and in Washington, D.C., May 2005 (E-PAS 2006:59:2610.4; Pediatr Res 2005;57:2502, and Pediatr Res 2004;55:2664).

Received: December 6, 2007
Accepted: January 21, 2008
Published online: September 6, 2008
Number of Print Pages : 8
Number of Figures : 3, Number of Tables : 5, Number of References : 31


 goto top of outline Publication Details

Neonatology (Fetal and Neonatal Research)

Vol. 95, No. 2, Year 2009 (Cover Date: February 2009)

Journal Editor: Halliday H.L. (Belfast), Speer C.P. (Würzburg)
ISSN: 1661–7800 (Print), eISSN: 1661–7819 (Online)

For additional information: http://www.karger.com/NEO


Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

References

  1. Hills B: Surface-active phospholipid: a Pandora’s box of clinical applications. 1. The lung and air spaces. Intern Med J 2002;32:170–178.
  2. Mazela J, Merritt TA, Gadzinowski J, Sinha S: Evolution of pulmonary surfactants for the treatment of neonatal respiratory distress syndrome and paediatric lung diseases. Acta Paediatr 2006;95:1036–1048.
  3. Kramer B: The respiratory distress syndrome (RDS) in preterm infants: physiology, prophylaxis, and new therapeutic approaches. Intensivmed 2007;44:403–408.

    External Resources

  4. Angus D, Linde-Zwirble WT, Clermont G, Griffin MF, Clark RH: Epidemiology of neonatal respiratory failure in the united states: projections from California and New York. Am J Respir Crit Care Med 2001;164:1154–1160.
  5. Hamvas A, Devine T, Cole FS: Surfactant therapy failure identifies infants at risk for pulmonary mortality. Am J Dis Child 1993;147:665–668.
  6. Richardson D, Torday JS: Racial differences in predictive value of the lecithin/sphingomyelin ratio. Am J Obstet Gynecol 1994;170:1273–1278.
  7. Nogee LM de Mello DE, Dehner LP, Colten HR: Brief report: deficiency of pulmonary surfactant protein B in congenital alveolar proteinosis. N Engl J Med 1993;328:406–410.
  8. Clark R: The epidemiology of respiratory failure in neonates born at an estimated gestational age of 34 weeks or more. J Perinatol 2005;25:251–257.
  9. Weaver T, Conkright JJ: Function of surfactant proteins B and C. Annu Rev Physiol 2001;63:555–578.
  10. Guttentag S, Beers MF, Bieler BM, Ballard PL: Surfactant protein B processing in human fetal lung. Am J Physiol 1998;275:L559–L566.
  11. Brasch F, Johnen G, Winn-Brasch A, Guttentag SH, Schmiedl A, Kapp N, Suzuki Y, Muller KM, Richter J, Hawgood S, Ochs M: Surfactant protein B in type II pneumocytes and intra-alveolar surfactant forms of human lungs. Am J Respir Cell Mol Biol 2004;30:449–458.
  12. Ueno T, Linder S, Na CL, Rice WR, Johansson J, Weaver TE: Processing of pulmonary surfactant protein B by napsin and cathepsin H. J Biol Chem 2004;279:16178–16184.
  13. Dilger I, Schwedler G, Dudenhausen JW: Determination of the pulmonary surfactant-associated protein SP-B in amniotic fluid with a competition ELISA. Gynecol Obstet Invest 1994;38:24–27.
  14. Mori K, Ikeda K, Tanaka M: Different expression of surfactant protein B mature peptide and proprotein at 21 weeks’ gestation in human fetal pulmonary epithelial cells. Pediatr Int 2002;44:500–504.
  15. Stahlman M, Gray ME, Whitsett JA: The ontogeny and distribution of surfactant protein B in human fetuses and newborns. J Histochem Cytochem 1992;40:1471–1480.
  16. Griese M, Schumacher S, Tredano M, Steinecker M, Braun A, Guttentag S, Beers MF, Bahuau M: Expression profiles of hydrophobic surfactant proteins in children with diffuse chronic lung disease. Respir Res 2005;6:80.
  17. Lin S, Akinbi HT, Breslin JS, Weaver TE: Structural requirements for targeting of surfactant protein B (SP-B) to secretory granules in vitro and in vivo. J Biol Chem 1996;271:19689–19695.
  18. Akinbi H, Breslin JS, Ikegami M, Iwamoto HS, Clark JC, Whitsett JA, Jobe AH, Weaver TE: Rescue of SP-B knockout mice with a truncated SP-B proprotein. Function of the c-terminal propeptide. J Biol Chem 1997;272:9640–9647.
  19. Korimilli A, Gonzales LW, Guttentag SH: Intracellular localization of processing events in human surfactant protein B biosynthesis. J Biol Chem 2000;275:8672–8679.
  20. Nogee L, Garnier G, Dietz HC, Singer L, Murphy AM, deMello DE, Colten HR: A mutation in the surfactant protein B gene responsible for fatal neonatal respiratory disease in multiple kindreds. J Clin Invest 1994;93:1860–1863.
  21. Melton K, Nesslein LL, Ikegami M, Tichelaar JW, Clark JC, Whitsett JA, Weaver TE: Sp-B deficiency causes respiratory failure in adult mice. Am J Physiol Lung Cell Mol Physiol 2003;285:L543–L549.
  22. Wang G, Christensen ND, Wigdahl B, Guttentag SH, Floros J: Differences in n-linked glycosylation between human surfactant protein-B variants of the C or T allele at the single-nucleotide polymorphism at position 1580: implications for disease. Biochem J 2003;369:179–184.
  23. Ballard P, Merrill JD, Godinez RI, Godinez MH, Truog WE, Ballard RA: Surfactant protein profile of pulmonary surfactant in premature infants. Am J Respir Crit Care Med 2003;168:1123–1128.
  24. Merrill J, Ballard RA, Cnaan A, Hibbs AM, Godinez RI, Godinez MH, Truog WE, Ballard PL: Dysfunction of pulmonary surfactant in chronically ventilated premature infants. Pediatr Res 2004;56:918–926.
  25. Hamvas A, Wegner DJ, Carlson CS, Bergmann KR, Trusgnich MA, Fulton L, Kasai Y, An P, Mardis ER, Wilson RK, Cole FS: Comprehensive genetic variant discovery in the surfactant protein B gene. Pediatr Res 2007;62:170–175.
  26. Hamvas A, Wegner DJ, Trusgnich MA, Madden K, Heins H, Liu Y, Rice T, An P, Watkins-Torry J, Cole FS: Genetic variant characterization in intron 4 of the surfactant protein B gene. Hum Mutat 2005;26:494–495.
  27. Bachurski C, Ross GF, Ikegami M, Kramer BW, Jobe AH: Intra-amniotic endotoxin increases pulmonary surfactant proteins and induces SP-B processing in fetal sheep. Am J Physiol Lung Cell Mol Physiol 2001;280:L279–L285.
  28. Kramer B, Kramer S, Ikegami M, Jobe AH: Injury, inflammation, and remodeling in fetal sheep lung after intra-amniotic endotoxin. Am J Physiol Lung Cell Mol Physiol 2002;283:L452–L459.
  29. Brasch F, Ochs M, Kahne T, Guttentag S, Schauer-Vukasinovic V, Derrick M, Johnen G, Kapp N, Muller KM, Richter J, Giller T, Hawgood S, Buhling F: Involvement of napsin A in the C- and N-terminal processing of surfactant protein B in type-II pneumocytes of the human lung. J Biol Chem 2003;278:49006–49014.
  30. Foster C, Aktar A, Kopf D, Zhang P, Guttentag S: Pepsinogen C: a type 2 cell-specific protease. Am J Physiol Lung Cell Mol Physiol 2004;286:L382–L387.
  31. Guttentag S, Robinson L, Zhang P, Brasch F, Buhling F, Beers M: Cysteine protease activity is required for surfactant protein B processing and lamellar body genesis. Am J Respir Cell Mol Biol 2003;28:69–79.