Pulmonary fibrosis is a serious medical problem affecting more than 200,000 patients in the U.S. At present, there are few treatments available to counteract pulmonary fibrosis. Modulation of the renln-anglotensin system has been suggested, but the efficacy of such strategies remains uncertain and somewhat contradictory. My project is to evaluate whether other substrates of ACE beside angiotensin II may play a significant role. My approach was to develop two new strains of mice by homologous recombination in order to mutate the specific amino acids responsible for zinc binding (and thus catalysis) in each domain of ACE. In mice termed N-KO, the N-termlnal domain of ACE is Inactivated. In C-KO mice, It is the C-terminal domain that is no longer catalytic. The cardiovascular and renal phenotypes of N-KO and C-KO mice are indistinguishable from wild-type mice. These mouse models are ideal to study specific functions of ACE independent of blood pressure changes, a typical bias In studying the RAS with pharmacologic manipulations. I have now shown that the tetrapeptide AcSDKP, a specific substrate of the N-terminal catalytic site of ACE protects mice from bleomycin-induced lung Injury. The next step of my investigation is to analyze the molecular mechanisms responsible for AcSDKP protection. My hypothesis is that it is mediated by an inhibition ofthe TFG-beta/Smad pathway. This will be evaluated by determining the phosphorylation of Smad proteins and their translocation to the nucleus. The last part of my project consists of determining if these observations in N-KO mice could be transposed to other animal models. I will investigate if administration of AcSDKP to wild-type mice protects them from bleomycin-induced lung injury. Preliminary observations suggest that AcSDKP administration alone limits the inflammation without preventing the development of fibrosis. This may be due to the high expression of ACE in the lung and the rapid degradation of AcSDKP in that organ. I will therefore test the co-administration of AcSDKP and ACE inhibitors.

Public Health Relevance

This proposal will yield fundamental knowledge about the importance of ACE and its multiple substrates in tissue injury. My studies are relevant to a variety of human diseases, including diseases ofthe lung, heart and kidney. Another outcome of this investigation is to demonstrate a new aproach to limiting secondary Injury of bleomycin administration and therefore extend the clinical reachm of this antineoplastic drug.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Transition Award (R00)
Project #
5R00HL088000-06
Application #
8010966
Study Section
Special Emphasis Panel (NSS)
Program Officer
Colombini-Hatch, Sandra
Project Start
2006-12-01
Project End
2011-12-31
Budget Start
2011-01-01
Budget End
2011-12-31
Support Year
6
Fiscal Year
2011
Total Cost
$249,000
Indirect Cost
Name
Cedars-Sinai Medical Center
Department
Type
DUNS #
075307785
City
Los Angeles
State
CA
Country
United States
Zip Code
90048
Bernstein, Kenneth E; Koronyo, Yosef; Salumbides, Brenda C et al. (2014) Angiotensin-converting enzyme overexpression in myelomonocytes prevents Alzheimer's-like cognitive decline. J Clin Invest 124:1000-12
Bernstein, Kenneth E; Gonzalez-Villalobos, Romer A; Giani, Jorge F et al. (2014) Angiotensin-converting enzyme overexpression in myelocytes enhances the immune response. Biol Chem 395:1173-8
Koronyo-Hamaoui, Maya; Shah, Kandarp; Koronyo, Yosef et al. (2014) ACE overexpression in myelomonocytic cells: effect on a mouse model of Alzheimer's disease. Curr Hypertens Rep 16:444
Shen, Xiao Z; Ong, Frank S; Bernstein, Ellen A et al. (2012) Nontraditional roles of angiotensin-converting enzyme. Hypertension 59:763-8
Gonzalez-Villalobos, Romer A; Billet, Sandrine; Kim, Catherine et al. (2011) Intrarenal angiotensin-converting enzyme induces hypertension in response to angiotensin I infusion. J Am Soc Nephrol 22:449-59
Bernstein, Kenneth E; Shen, Xiao Z; Gonzalez-Villalobos, Romer A et al. (2011) Different in vivo functions of the two catalytic domains of angiotensin-converting enzyme (ACE). Curr Opin Pharmacol 11:105-11
Okwan-Duodu, Derick; Datta, Vivekanand; Shen, Xiao Z et al. (2010) Angiotensin-converting enzyme overexpression in mouse myelomonocytic cells augments resistance to Listeria and methicillin-resistant Staphylococcus aureus. J Biol Chem 285:39051-60
Weiss, Daiana; Bernstein, Kenneth E; Fuchs, Sebastian et al. (2010) Vascular wall ACE is not required for atherogenesis in ApoE(-/-) mice. Atherosclerosis 209:352-8
Li, Ping; Xiao, Hong D; Xu, Jianguo et al. (2010) Angiotensin-converting enzyme N-terminal inactivation alleviates bleomycin-induced lung injury. Am J Pathol 177:1113-21
Bernstein, Kenneth E; Fuchs, Sebastien (2010) Angiotensin II and JAK2 put on the pressure. Nat Med 16:165-6