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. 1995 May 9;92(10):4357–4361. doi: 10.1073/pnas.92.10.4357

Bombesin and [Leu8]phyllolitorin promote fetal mouse lung branching morphogenesis via a receptor-mediated mechanism.

K A King 1, J S Torday 1, M E Sunday 1
PMCID: PMC41943  PMID: 7753811

Abstract

Pulmonary neuroendocrine cells are localized predominantly at airway branchpoints. Previous work showed that gastrin-releasing peptide (GRP), a major pulmonary bombesin-like peptide, occurred in neuroendocrine cells exclusively in branching human fetal airways. We now demonstrate that GRP and GRP receptor genes are expressed in fetal mouse lung as early as embryonic day 12 (E12), when lung buds are beginning to branch. By in situ hybridization, GRP receptor transcripts were at highest levels in mesenchymal cells at cleft regions of branching airways and blood vessels. To explore the possibility that bombesin-like peptides might play a role in branching morphogenesis, E12 lung buds were cultured for 48 hr in serum-free medium. In the presence of 0.10-10 microM bombesin, branching was significantly augmented as compared with control cultures, with a peak of 94% above control values at 1 microM (P < 0.005). The bombesin receptor antagonist [Leu13- psi(CH2NH)Leu14]bombesin alone (100 nM) had no effect on baseline branching but completely abolished bombesin-induced branching. A bombesin-related peptide, [Leu8]phyllolitorin also increased branching (65% above control values at 10 nM, P < 0.005). [Leu8]Phyllolitorin also significantly augmented thymidine incorporation in cultured lung buds. Fibronectin, which is abundant at branchpoints, induces GRP gene expression in undifferentiated cell lines. These observations suggest that BLPs secreted by pulmonary neuroendocrine cells may contribute to lung branching morphogenesis. Furthermore, components of branchpoints may induce pulmonary neuroendocrine cell differentiation as part of a positive feedback loop, which could account in part for the high prevalence of these cells at branchpoints.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aguayo S. M., Kane M. A., King T. E., Jr, Schwarz M. I., Grauer L., Miller Y. E. Increased levels of bombesin-like peptides in the lower respiratory tract of asymptomatic cigarette smokers. J Clin Invest. 1989 Oct;84(4):1105–1113. doi: 10.1172/JCI114273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aguayo S. M., King T. E., Jr, Waldron J. A., Jr, Sherritt K. M., Kane M. A., Miller Y. E. Increased pulmonary neuroendocrine cells with bombesin-like immunoreactivity in adult patients with eosinophilic granuloma. J Clin Invest. 1990 Sep;86(3):838–844. doi: 10.1172/JCI114782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Battey J. F., Way J. M., Corjay M. H., Shapira H., Kusano K., Harkins R., Wu J. M., Slattery T., Mann E., Feldman R. I. Molecular cloning of the bombesin/gastrin-releasing peptide receptor from Swiss 3T3 cells. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):395–399. doi: 10.1073/pnas.88.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cardiff R. D., Leder A., Kuo A., Pattengale P. K., Leder P. Multiple tumor types appear in a transgenic mouse with the ras oncogene. Am J Pathol. 1993 Apr;142(4):1199–1207. [PMC free article] [PubMed] [Google Scholar]
  5. Carney D. N., Gazdar A. F., Bepler G., Guccion J. G., Marangos P. J., Moody T. W., Zweig M. H., Minna J. D. Establishment and identification of small cell lung cancer cell lines having classic and variant features. Cancer Res. 1985 Jun;45(6):2913–2923. [PubMed] [Google Scholar]
  6. Cho T., Chan W., Cutz E. Distribution and frequency of neuro-epithelial bodies in post-natal rabbit lung: quantitative study with monoclonal antibody against serotonin. Cell Tissue Res. 1989 Feb;255(2):353–362. doi: 10.1007/BF00224118. [DOI] [PubMed] [Google Scholar]
  7. Coy D. H., Heinz-Erian P., Jiang N. Y., Sasaki Y., Taylor J., Moreau J. P., Wolfrey W. T., Gardner J. D., Jensen R. T. Probing peptide backbone function in bombesin. A reduced peptide bond analogue with potent and specific receptor antagonist activity. J Biol Chem. 1988 Apr 15;263(11):5056–5060. [PubMed] [Google Scholar]
  8. Cuttitta F., Carney D. N., Mulshine J., Moody T. W., Fedorko J., Fischler A., Minna J. D. Bombesin-like peptides can function as autocrine growth factors in human small-cell lung cancer. 1985 Aug 29-Sep 4Nature. 316(6031):823–826. doi: 10.1038/316823a0. [DOI] [PubMed] [Google Scholar]
  9. Desanti A. M., McDowell E. M., Strum J. M. Airway branching patterns and cytodifferentiation in cultured fetal hamster lung. Tissue Cell. 1992;24(6):853–868. doi: 10.1016/0040-8166(92)90021-x. [DOI] [PubMed] [Google Scholar]
  10. Doyle L. A., Giangiulo D., Hussain A., Park H. J., Yen R. W., Borges M. Differentiation of human variant small cell lung cancer cell lines to a classic morphology by retinoic acid. Cancer Res. 1989 Dec 1;49(23):6745–6751. [PubMed] [Google Scholar]
  11. Falconieri Erspamer G., Mazzanti G., Farruggia G., Nakajima T., Yanaihara N. Parallel bioassay of litorin and phyllolitorins on smooth muscle preparations. Peptides. 1984 Jul-Aug;5(4):765–768. doi: 10.1016/0196-9781(84)90019-6. [DOI] [PubMed] [Google Scholar]
  12. Fathi Z., Corjay M. H., Shapira H., Wada E., Benya R., Jensen R., Viallet J., Sausville E. A., Battey J. F. BRS-3: a novel bombesin receptor subtype selectively expressed in testis and lung carcinoma cells. J Biol Chem. 1993 Mar 15;268(8):5979–5984. [PubMed] [Google Scholar]
  13. Impicciatore M., Bertaccini G. The bronchoconstrictor action of the tetradecapeptide bombesin in the guinea-pig. J Pharm Pharmacol. 1973 Nov;25(11):872–875. doi: 10.1111/j.2042-7158.1973.tb09965.x. [DOI] [PubMed] [Google Scholar]
  14. Infeld M. D., Brennan J. A., Davis P. B. Human fetal lung fibroblasts promote invasion of extracellular matrix by normal human tracheobronchial epithelial cells in vitro: a model of early airway gland development. Am J Respir Cell Mol Biol. 1993 Jan;8(1):69–76. doi: 10.1165/ajrcmb/8.1.69. [DOI] [PubMed] [Google Scholar]
  15. Kane M. A., Aguayo S. M., Portanova L. B., Ross S. E., Holley M., Kelley K., Miller Y. E. Isolation of the bombesin/gastrin-releasing peptide receptor from human small cell lung carcinoma NCI-H345 cells. J Biol Chem. 1991 May 25;266(15):9486–9493. [PubMed] [Google Scholar]
  16. King K. A., Hua J., Torday J. S., Drazen J. M., Graham S. A., Shipp M. A., Sunday M. E. CD10/neutral endopeptidase 24.11 regulates fetal lung growth and maturation in utero by potentiating endogenous bombesin-like peptides. J Clin Invest. 1993 May;91(5):1969–1973. doi: 10.1172/JCI116417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klagsbrun M., D'Amore P. A. Regulators of angiogenesis. Annu Rev Physiol. 1991;53:217–239. doi: 10.1146/annurev.ph.53.030191.001245. [DOI] [PubMed] [Google Scholar]
  18. Lauweryns J. M., Peuskens J. C. Argyrophil (kinin and amine producing?) cells in human infant airway epithelium. Life Sci. 1969 Jun 1;8(11):577–585. doi: 10.1016/0024-3205(69)90019-8. [DOI] [PubMed] [Google Scholar]
  19. Lazarus L. H., Wilson W. E., Gaudino G., Irons B. J., Guglietta A. Evolutionary relationship between nonmammalian and mammalian peptides. Peptides. 1985;6 (Suppl 3):295–307. doi: 10.1016/0196-9781(85)90389-4. [DOI] [PubMed] [Google Scholar]
  20. Lebacq-Verheyden A. M., Krystal G., Sartor O., Way J., Battey J. F. The rat prepro gastrin releasing peptide gene is transcribed from two initiation sites in the brain. Mol Endocrinol. 1988 Jun;2(6):556–563. doi: 10.1210/mend-2-6-556. [DOI] [PubMed] [Google Scholar]
  21. McGowan S. E. Extracellular matrix and the regulation of lung development and repair. FASEB J. 1992 Aug;6(11):2895–2904. [PubMed] [Google Scholar]
  22. Montesano R., Matsumoto K., Nakamura T., Orci L. Identification of a fibroblast-derived epithelial morphogen as hepatocyte growth factor. Cell. 1991 Nov 29;67(5):901–908. doi: 10.1016/0092-8674(91)90363-4. [DOI] [PubMed] [Google Scholar]
  23. Nagalla S. R., Gibson B. W., Tang D., Reeve J. R., Jr, Spindel E. R. Gastrin-releasing peptide (GRP) is not mammalian bombesin. Identification and molecular cloning of a true amphibian GRP distinct from amphibian bombesin in Bombina orientalis. J Biol Chem. 1992 Apr 5;267(10):6916–6922. [PubMed] [Google Scholar]
  24. Ng S. Y., Gunning P., Eddy R., Ponte P., Leavitt J., Shows T., Kedes L. Evolution of the functional human beta-actin gene and its multi-pseudogene family: conservation of noncoding regions and chromosomal dispersion of pseudogenes. Mol Cell Biol. 1985 Oct;5(10):2720–2732. doi: 10.1128/mcb.5.10.2720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Roman J., Little C. W., McDonald J. A. Potential role of RGD-binding integrins in mammalian lung branching morphogenesis. Development. 1991 Jun;112(2):551–558. doi: 10.1242/dev.112.2.551. [DOI] [PubMed] [Google Scholar]
  26. Rozengurt E., Sinnett-Smith J. Bombesin stimulation of DNA synthesis and cell division in cultures of Swiss 3T3 cells. Proc Natl Acad Sci U S A. 1983 May;80(10):2936–2940. doi: 10.1073/pnas.80.10.2936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ruff M., Schiffmann E., Terranova V., Pert C. B. Neuropeptides are chemoattractants for human tumor cells and monocytes: a possible mechanism for metastasis. Clin Immunol Immunopathol. 1985 Dec;37(3):387–396. doi: 10.1016/0090-1229(85)90108-4. [DOI] [PubMed] [Google Scholar]
  28. Schuger L., O'Shea S., Rheinheimer J., Varani J. Laminin in lung development: effects of anti-laminin antibody in murine lung morphogenesis. Dev Biol. 1990 Jan;137(1):26–32. doi: 10.1016/0012-1606(90)90004-3. [DOI] [PubMed] [Google Scholar]
  29. Siegfried J. M., Guentert P. J., Gaither A. L. Effects of bombesin and gastrin-releasing peptide on human bronchial epithelial cells from a series of donors: individual variation and modulation by bombesin analogs. Anat Rec. 1993 May;236(1):241–247. doi: 10.1002/ar.1092360129. [DOI] [PubMed] [Google Scholar]
  30. Speirs V., Bienkowski E., Wong V., Cutz E. Paracrine effects of bombesin/gastrin-releasing peptide and other growth factors on pulmonary neuroendocrine cells in vitro. Anat Rec. 1993 May;236(1):53–67. doi: 10.1002/ar.1092360110. [DOI] [PubMed] [Google Scholar]
  31. Spindel E. R., Sunday M. E., Hofler H., Wolfe H. J., Habener J. F., Chin W. W. Transient elevation of messenger RNA encoding gastrin-releasing peptide, a putative pulmonary growth factor in human fetal lung. J Clin Invest. 1987 Oct;80(4):1172–1179. doi: 10.1172/JCI113176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sunday M. E., Hua J., Dai H. B., Nusrat A., Torday J. S. Bombesin increases fetal lung growth and maturation in utero and in organ culture. Am J Respir Cell Mol Biol. 1990 Sep;3(3):199–205. doi: 10.1165/ajrcmb/3.3.199. [DOI] [PubMed] [Google Scholar]
  33. Sunday M. E., Hua J., Reyes B., Masui H., Torday J. S. Anti-bombesin monoclonal antibodies modulate fetal mouse lung growth and maturation in utero and in organ cultures. Anat Rec. 1993 May;236(1):25–34. doi: 10.1002/ar.1092360107. [DOI] [PubMed] [Google Scholar]
  34. Sunday M. E., Hua J., Torday J. S., Reyes B., Shipp M. A. CD10/neutral endopeptidase 24.11 in developing human fetal lung. Patterns of expression and modulation of peptide-mediated proliferation. J Clin Invest. 1992 Dec;90(6):2517–2525. doi: 10.1172/JCI116145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sunday M. E., Kaplan L. M., Motoyama E., Chin W. W., Spindel E. R. Gastrin-releasing peptide (mammalian bombesin) gene expression in health and disease. Lab Invest. 1988 Jul;59(1):5–24. [PubMed] [Google Scholar]
  36. Sunday M. E., Willett C. G. Induction and spontaneous regression of intense pulmonary neuroendocrine cell differentiation in a model of preneoplastic lung injury. Cancer Res. 1992 May 1;52(9 Suppl):2677s–2686s. [PubMed] [Google Scholar]
  37. Wada E., Way J., Shapira H., Kusano K., Lebacq-Verheyden A. M., Coy D., Jensen R., Battery J. cDNA cloning, characterization, and brain region-specific expression of a neuromedin-B-preferring bombesin receptor. Neuron. 1991 Mar;6(3):421–430. doi: 10.1016/0896-6273(91)90250-4. [DOI] [PubMed] [Google Scholar]
  38. Warburton D., Seth R., Shum L., Horcher P. G., Hall F. L., Werb Z., Slavkin H. C. Epigenetic role of epidermal growth factor expression and signalling in embryonic mouse lung morphogenesis. Dev Biol. 1992 Jan;149(1):123–133. doi: 10.1016/0012-1606(92)90269-m. [DOI] [PubMed] [Google Scholar]
  39. Weber S., Zuckerman J. E., Bostwick D. G., Bensch K. G., Sikic B. I., Raffin T. A. Gastrin releasing peptide is a selective mitogen for small cell lung carcinoma in vitro. J Clin Invest. 1985 Jan;75(1):306–309. doi: 10.1172/JCI111690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wharton J., Polak J. M., Bloom S. R., Ghatei M. A., Solcia E., Brown M. R., Pearse A. G. Bombesin-like immunoreactivity in the lung. Nature. 1978 Jun 29;273(5665):769–770. doi: 10.1038/273769a0. [DOI] [PubMed] [Google Scholar]

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