Tissue ACE phenotyping in lung cancer

Autoři: Sergei M. Danilov aff001;  Roman Metzger aff004;  Eckhard Klieser aff005;  Karl Sotlar aff005;  Ilya N. Trakht aff006;  Joe G. N. Garcia aff002
Působiště autorů: Department of Medicine, Division of Pulmonary, Critical Care, Sleep and Allergy, University of Illinois at Chicago, IL, United States of America aff001;  Department of Medicine, University of Arizona Health Sciences, Tucson, AZ, United States of America aff002;  Medical Center, Moscow University, Moscow, Russia aff003;  Department of Pediatric and Adolescent Surgery, Paracelsus Medical University, Salzburg, Austria aff004;  Institute of Pathology, Paracelsus Medical University, University Hospital Salzburg, Salzburg, Austria aff005;  Department of Medicine, Columbia University, New York, NY, United States of America aff006
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0226553



Pulmonary vascular endothelium is the main metabolic site for Angiotensin I-Converting Enzyme (ACE)-mediated degradation of several biologically-active peptides (angiotensin I, bradykinin, hemo-regulatory peptide Ac-SDKP). Primary lung cancer growth and lung cancer metastases decrease lung vascularity reflected by dramatic decreases in both lung and serum ACE activity. We performed precise ACE phenotyping in tissues from subjects with lung cancer.


ACE phenotyping included: 1) ACE immunohistochemistry with specific and well-characterized monoclonal antibodies (mAbs) to ACE; 2) ACE activity measurement with two ACE substrates (HHL, ZPHL); 3) calculation of ACE substrates hydrolysis ratio (ZPHL/HHL ratio); 4) the pattern of mAbs binding to 17 different ACE epitopes to detect changes in ACE conformation induced by tumor growth (conformational ACE fingerprint).


ACE immunostaining was dramatically decreased in lung cancer tissues confirmed by a 3-fold decrease in ACE activity. The conformational fingerprint of ACE from tumor lung tissues differed from normal lung (6/17 mAbs) and reflected primarily higher ACE sialylation. The increase in ZPHL/HHL ratio in lung cancer tissues was consistent with greater conformational changes of ACE. Limited analysis of the conformational ACE fingerprint in normal lung tissue and lung cancer tissue form the same patient suggested a remote effect of tumor tissue on ACE conformation and/or on “field cancerization” in a morphologically-normal lung tissues.


Local conformation of ACE is significantly altered in tumor lung tissues and may be detected by conformational fingerprinting of human ACE.

Klíčová slova:

Adenocarcinoma of the lung – Adenocarcinomas – Blood – Lung and intrathoracic tumors – Non-small cell lung cancer – Secondary lung tumors – Small cell lung cancer – Squamous cell lung carcinoma


1. Bakhle YS and Vane JR. (Eds), 1977 Metabolic Functions of the lung. New York: Marcel Dekker Inc.

2. Junot C, Nicolet L, Ezaqn E, Gonzales M-F, Menard J, Azizi M (1999) Effect of Angiotensin-Converting Enzyme inhibition on plasma, urine, and tissue concentrations of hemoregulatory Peptide Acetyl-Ser-Asp-Lys-Pro in rats. J Pharmacol Exp Ther 291: 982–989. 10565814

3. Franke FE, Metzger R, Bohle R-M, Kerkman L, Alhenc-Gelas F, Danilov SM. Angiotensin I-Converting Enzyme (CD 143) on endothelial cells in normal and in pathological conditions. In: Leucocyte Typing VI: White Cell Differentiation Antigens. ( Kishimoto T et al., Eds.) Garland Publishing Inc. New York, 1997; pp. 749–751.

4. Metzger R, Franke FF, Bohle R-M, Alhenc-Gelas F, Danilov SM (2011) Heterogeneous distribution of Angiotensin I-converting enzyme (CD143) in the human and rat vascular systems: vessels, organs and species specificity. Microvasc Res 82: 206–215.

5. Prohazka J, Krepela E, Sedo A, Viklicky J, Fiala P (1991) Aminopeptidases and angiotensin-I converting enzyme in primary lung tumors and lung parenchyma. Neoplasma. 38: 501–508. 1683470

6. Ashutosh K and Kieghley JF (1976) Diagnostic value of serum angiotensin converting enzyme activity in lung diseases. Thorax 31: 552–557. doi: 10.1136/thx.31.5.552 186911

7. Romer FK. (1981) Angiotensin-converting enzyme and its association with outcome in lung cancer. Br J Cancer 43: 135–142. doi: 10.1038/bjc.1981.21 6258623

8. Mansfield CM, Kimler BF, Henderson SD, Vats TS, Svoboda DJ (1984) Angiotensin-I-converting enzyme in cancer patients. J Clin Oncol 2: 452–456. doi: 10.1200/JCO.1984.2.5.452 6327925

9. Schweisfurth H, Heinrich J, Brugger E, Steinl C, Maiwald L (1985). The value of angiotensin-I-converting enzyme determinations in malignant and other diseases. Clin Physiol Biochem 3: 184–192. 2990799

10. Varela AS and Bosco Lopez Saez JJ (1993) Utility of serum activity of angiotensin-converting enzyme as a tumor marker. Oncology 50: 430–435. doi: 10.1159/000227224 8233282

11. Lever AF, Hole DJ, Gillis CR, McCallum IR, McInnes GT, MacKinnon PL (1998) Do inhibitors of angiotensin I-converting enzyme protect against risk of cancer? Lancet 352: 162–163. doi: 10.1016/S0140-6736(05)77800-4

12. Pinter M, Jain RK. (2017) Targeting the renin-angiotensin system to improve cancer treatment: implication for immunotherapy. Science Transl Med 9: eaan5616.

13. Miao L, Chen W, Zhou L, Wan H, Gao B, Feng Y (2016) Impact of angiotensin I-converting enzyme inhibitors and angiotensin II type-1 receptor blockers on survival of patients with NSCLC. Sci Rep 6: 21359. doi: 10.1038/srep21359 26883083

14. Menter AR, Carroll NM, Sakoda LC, Delate T, Hornbrook MC et al. (2017) Effect of angiotensin system inhibitors on survival in patients receiving chemotherapy for advanced non-small cell lung cancer. Clin Lung Cancer 18: 189–197. doi: 10.1016/j.cllc.2016.07.008 27637408

15. Soubrier F, Alhenc-Gelas F, Hubert C, Allegrini J, John M, et al. (1988) Two putative active centers in human angiotensin I-converting enzyme revealed by molecular cloning. Proc Natl Acad Sci USA 85: 9386–9390. doi: 10.1073/pnas.85.24.9386 2849100

16. Sturrock ED, Anthony S, Danilov SM. Peptidyl-dipeptidase A/Angiotensin I-converting enzyme. In Handbook of Proteolytic Enzymes, Eds. Neil D. Rawlings, Guy Salvesen, 3rd Edition, (Academic Press, Oxford, 2012) Chapter 98, pp.480–494 (3rd Edition,).

17. Bernstein K. Bernstein KE., Ong FS, Blackwell WL Shah KH et al. (2012) A modern understanding of the traditional and nontraditional biological functions of angiotensin-converting enzyme. Pharmacol Rev 65: 1–46. doi: 10.1124/pr.112.006809 23257181

18. Ryan JW, Ryan US, Shultz DR, Whiteker RC, Chung A (1975) Subcellular localization of pulmonary angiotensin-converting enzyme (Kininase II). Biochem J 146: 497–499. doi: 10.1042/bj1460497 168877

19. Caldwell PR, Seegal BC, Hsu KC, Das H, Soffer RL (1976) Angiotensin-converting enzyme: vascular endothelial localization. Science 191: 1050–1051. doi: 10.1126/science.175444 175444

20. Defendini R, Zimmerman EA, Weare JA, Alhenc-Gelas F, Erdos EG (1983) Angiotensin-converting enzyme in epithelial and neuroepithelial cells. Neuroendocrinology 37: 32–40. doi: 10.1159/000123512 6310427

21. Hooper NM, Turner AJ (1987) Isolation of two differentially glycosylated forms of peptidyl-dipeptidase A (angiotensin-converting enzyme) from pig brain: a re-evaluation of their role in neuropeptide metabolism. Biochem J 241: 625–633. doi: 10.1042/bj2410625 2439065

22. Silverstein E, Friedland J, Setton C (1978) Angiotensin-converting enzyme in macrophages and Freund’s adjuvant granuloma. Isr J Med Sci 14: 314–318. 205523

23. Danilov SM, Sadovnikova E, Scharenbourg N, Balysnikova IV, Svinareva DA, et al. (2003) Angiotensin-converting enzyme (CD143) is abundantly expressed by dendritic cells and discriminates human monocytes-derived dendritic cells from acute myeloid leukemia-derived dendritic cells. Exp Hem 31: 1301–1309.

24. Danilov SM, Franke FE, Erdos EG. Angiotensin-Converting Enzyme (CD143). In: Leucocyte Typing VI: White Cell Differentiation Antigens. ( Kishimoto T. et al., Eds.) Garland Publishing Inc.New York. 1997; pp.746–749.

25. Hooper NM, Keen J, Pappin DJ, Turner AJ (1987) Pig kidney angiotensin converting enzyme. Purification and characterization of amphipathic and hydrophilic forms of the enzyme establishes C-terminal anchorage to the plasma membrane. Biochem J. 247: 85–93. doi: 10.1042/bj2470085 2825659

26. Parkin ET, Turner AJ, Hooper NM (2004) Secretase-mediated cell surface shedding of the angiotensin-converting enzyme. Protein Pept Lett 11: 423–432. doi: 10.2174/0929866043406544 15544563

27. Alhenc-Gelas F, Richard J Courbon D, Warnet JM, Corvol P (1991) Distribution of plasma angiotensin I-converting enzyme levels in healthy men: Relationship to environmental and hormonal parameters. J Lab Clin Med 117:33–39. 1846167

28. Romer FK (1984) Clinical and biochemical aspects of sarcoidosis. With special reference to angiotensin-converting enzyme (ACE). Acta Med Scand Suppl 690: 3–96. 6097101

29. Beneteau-Burnat B, Baudin B (1991) Angiotensin-converting enzyme: clinical applications and laboratory investigation in serum and other biological fluids. Crit Rev Clin Lab Sci 28: 337–356. doi: 10.3109/10408369109106868 1663362

30. Danilov SM, Balyasnikova IB, Danilova AS, Naperova IA, Arablinskaya E, et al. (2010) Conformational fingerprinting of the angiotensin-converting enzyme (ACE): Application in sarcoidosis. J Proteome Res 9: 5782–5793. doi: 10.1021/pr100564r 20873814

31. Petrov MN, Shilo VY, Tarasov AV Schwartz DE, Garcia JGN et al. (2012) Conformational changes of blood ACE in chronic uremia. PLoS One 7: e49290 doi: 10.1371/journal.pone.0049290 23166630

32. Kryukova OV, Tikhomirova VE, Golukhova EZ, Evdokimov VV, Kalantarov GF, et al. (2015) Tissue Specificity of human angiotensin I-converting enzyme. PLoS One 10: e0143455. doi: 10.1371/journal.pone.0143455 26600189

33. Danilov SM, Lünsdorf H, Akinbi HT, Nesterovitch AB, Epshtein Y, et al. (2016) Lysozyme and bilirubin bind to ACE and regulate its conformation and shedding. Sci Rep. 6: 34913. doi: 10.1038/srep34913 27734897

34. Tikhomirova VE, Kost OA, Kryukova OV, Bulaeva NI, Zholbaeva AZ et al. (2017) ACE phenotyping in human heart. PLoS One 12: e0181976. doi: 10.1371/journal.pone.0181976 28771512

35. Danilov SM, Tikhomirova VE, Metzger R, Naperova IA, Bukina TM et al. (2018) ACE phenotyping in Gaucher disease. Mol Genet Metab 123: 501–510. doi: 10.1016/j.ymgme.2018.02.007 29478818

36. Metzger R, Bohle RM, Pauls K, Eichner G, Alhenc-Gelas F et al. (1999) Angiotensin-converting enzyme in non-neoplastic kidney diseases. Kidney Int 56: 1442–1454. doi: 10.1046/j.1523-1755.1999.00660.x 10504496

37. Danilov S, Jaspard E, Churakova T, Towbin H, Savoie F, et al. (1994) Structure-function analysis of angiotensin I-converting enzyme using monoclonal antibodies. J Biol Chem 269: 26806–26814. 7523412

38. Wei L, Alhenc-Gelas F, Corvol P, Clauser E (1991) The two homologous domains of human angiotensin I-converting enzyme are both catalytically active. J Biol Chem. 266: 9002–9008. 1851160

39. Jaspard E, Wei L, Alhenc-Gelas F. (1993) Differences in properties and enzymatic specificities between the two active sites of human angiotensin I-converting enzyme: studies with bradykinin and other natural peptides. J Biol Chem 268: 9496–9503. 7683654

40. Georgiadis D, Beau F, Czarny B, Cotton J, Yiotakis A, Dive V (2003) Roles of the two active sites of somatic angiotensin-converting enzyme in the cleavage of angiotensin I and bradykinin: insights from selective inhibitors. Circ Res 93: 148–154. doi: 10.1161/01.RES.0000081593.33848.FC 12805239

41. Skirgello OE, Binevski PV, Pozdnev VF, Kost OA (2005) Kinetic probes for inter-domain cooperation in human somatic angiotensin-converting enzyme. Biochem J 391: 641–647. doi: 10.1042/BJ20050702 16033330

42. Danilov SM, Balyasnikova IV, Albrecht RFII, Kost OA (2008) Simultaneous determination of ACE activity with two substrates provides information on the status of somatic ACE and allows detection of inhibitors in human blood. J Cardiovasc Pharmacol 52: 90–103. doi: 10.1097/FJC.0b013e31817fd3bc 18645413

43. Danilov S, Savoie F, Lenoir B, Jeunemaitre X, Azizi M, et al. (1996) Development of enzyme-linked immunoassays for human angiotensin I converting enzyme suitable for large-scale studies. J Hypertens 14: 719–727. doi: 10.1097/00004872-199606000-00007 8793694

44. Ching SF, Hayes LW, Slakey LL (1983) Angiotensin-converting enzyme in cultured endothelial cells. Synthesis, degradation and transfer to culture medium. Arteriosclerosis 3: 581–588. doi: 10.1161/01.atv.3.6.581 6316884

45. Fishman A. Dynamics of pulmonary circulation. In: Hamilton WF, Dow P., eds/ Handbook of physiology, Washington DC, Am Physiol Sco 1963: 2: 1667.

46. Danilov SM, Tovsky SI, Schwartz DE, Dull RO (2017) ACE phenotyping as a guide toward personalized therapy with ACE inhibitors. J Cardiovasc Pharm Ther 22: 374–386.

47. Danilov SM, Kadrev AV, Kurilova OV, Tikhomirova VE, Kryukova OV et al. (2019) Tissue ACE phenotyping in prostate cancer. Oncotarget 10: 6349–6361. doi: 10.18632/oncotarget.27276 31695843

48. Danilov SM, Wade MS, Schwager SL, Douglas RG, Nesterovitch AB et al. (2014). A novel angiotensin I-converting enzyme mutation (S333W) impairs N-domain enzymatic cleavage of the anti-fibrotic peptide, AcSDKP. PLoS One 9: e88001. doi: 10.1371/journal.pone.0088001 24505347

49. Danilov SM, Deinum J, Balyasnikova IV, Sun Z-L, Kramers C, et al. (2005) Detection of mutated angiotensin I-converting enzyme by serum/plasma analysis using a pair of monoclonal antibodies. Clin Chem 51: 1040–1043. doi: 10.1373/clinchem.2004.045633 15914791

50. Danilov S.M. Gordon K, Nesterovitch AB, Lünsdorf H, Chen Z et al. (2012) An angiotensin I-converting enzyme mutation (Y465D) causes a dramatic increase in blood ACE via accelerated ACE shedding. PLoS One 6: e25952.

51. Danilov SM, Watermeyer JM, Balyasnikova IB, Gordon K, Kugaevskaya EV, et al. (2007) Fine epitope mapping of mAb 5F1 reveals anticatalytic activity toward the N domain of human angiotensin-converting enzyme. Biochemistry 46: 9019–9031. doi: 10.1021/bi700489v 17630779

52. Skirgello OE, Balyasnikova IV, Binevski PV, Sun ZL, Baskin II et al. (2006) Inhibitory antibodies to human angiotensin-converting enzyme: fine epitope mapping and mechanism of action. Biochemistry 45: 4831–4847. doi: 10.1021/bi052591h 16605251

53. Naperova IA, Balyasnikova IV, Schwartz DE, Watermeyer J, Sturrock DE, et al. (2008) Mapping of conformational mAb epitopes to the C domain of human angiotensin I-converting enzyme (ACE). J Proteome Res 7: 3396–3411. doi: 10.1021/pr800142w 18576678

54. Kost OA, Tikhomirova VE, Kryukova OV, Gusakov AV, Bulaeva NI et al. (2018) A conformational fingerprint of angiotensin-converting enzyme. Russian J Bioorg Chem 44: 52–63.

55. Danilov SM, Tikhomirova VE, Kryukova OV, Balatsky AV, Bulaeva NI, et al. (2018) Conformational fingerprint of blood and tissue ACEs: personalized approach. PLoS ONE.13: e0209861. doi: 10.1371/journal.pone.0209861 30589901

56. Ashwell G and Harford J (1982) Carbohydrate-specific receptors of the liver. Annu Rev Biochem, 51: 531–554. doi: 10.1146/annurev.bi.51.070182.002531 6287920

57. Stowell SR, Ju T, Cummings RD (2015) Protein glycosylation in cancer. Annu Rev Pathol 10: 473–510. doi: 10.1146/annurev-pathol-012414-040438 25621663

58. Bull C, Stoel MA, den Brok MH, Adema GJ (2014) Sialic acids sweeten a tumor’s life. Cancer Res 74: 3199–3204. doi: 10.1158/0008-5472.CAN-14-0728 24830719

59. Takashima S and Tsuji S (2011) Functional diversity of mammalian sialyl transferases. Trends Glycosci Glycotechnol 23:178–193.

60. Cohen M and Varki A (2010) The sialome–far more than the sum of its parts. OMICS 14: 455–464. doi: 10.1089/omi.2009.0148 20726801

61. Nelson MA, Wymer J, Clements N (1996) Detection of K-ras gene mutations in non-neoplastic lung tissue and lung cancers. Cancer Lett 103: 115–121. doi: 10.1016/0304-3835(96)04202-4 8616804

62. Tang X, Shigematsu H, Bekele BN, Roth JA, Minna JDet al. (2005) EGFR tyrosine kinase domain mutations are detected in histologically normal respiratory epithelium in lung cancer patients. Cancer Res 65: 7568–7572. doi: 10.1158/0008-5472.CAN-05-1705 16140919

63. Cooper CS, Eeles R, Wedge DC, Van Loo P, Gundem G et al. (2015) Analysis of the genetic phylogeny of multifocal prostate cancer identifies multiple independent clonal expansions in neoplastic and morphologically normal prostate tissue. Nature Genet 47: 367–372. doi: 10.1038/ng.3221 25730763

64. Bader M (2010) Tissue renin-angiotensin-aldosterone systems: targets for pharmacological therapy. Ann Rev Pharmacol Toxicol 50: 439–465.

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